Biomedical subjects

L Y Wang

Publications and source records attributed to L Y Wang.

At least 19 recordsLinked to original sources

PubMed1997

3-Phosphohistidine cannot replace phosphotyrosine in high-affinity binding to phosphotyrosine binding or Src homology 2 domains.

Posttranslational phosphorylation of proteins is an important event in many cellular processes. Phosphorylated tyrosine residues can serve as association sites for other proteins in signal transduction cascades of tyrosine kinase receptors. Formation of phosphohistidine residues in proteins has been found in eukaryotic and prokaryotic organisms. Furthermore, it has been suggested that phosphohistidine might substitute for phosphotyrosine in conferring high-affinity binding to proteins involved in signal transduction. We have analyzed the ability of 3-phosphohistidine to associate with the known phosphotyrosine-specific phosphotyrosine binding and src homology 2 protein domains. From our binding studies using synthetic peptides, we conclude that 3-phosphohistidine cannot replace phosphotyrosine in conferring high-affinity binding to the phosphotyrosine binding domain of shc or the src homology 2 domain of phospholipase C-gamma1.

Amino Acid Sequence↗

PubMed1997

p53 mutations, chronic hepatitis B virus infection, and aflatoxin exposure in hepatocellular carcinoma in Taiwan.

Recent studies have implicated aflatoxin B1 (AFB1) exposure as an etiological agent in hepatocellular carcinoma (HCC) and suggested an interaction with chronic hepatitis B virus (HBV) infection. Worldwide AFB1 exposure correlates with a specific mutation at codon 249 in the p53 tumor suppressor gene in liver tumors. This study investigated the roles of HBV and AFB1 in the HCC carcinogenic pathway involving p53 mutations. In cases and controls, chronic HBV infection was assessed by serum hepatitis B surface antigen (HBsAg) and AFB1 exposure by immunohistochemical detection of AFB1-DNA adduct in liver tissue. p53 protein mutations in tumor tissues of HCC cases were identified by immunohistochemistry and DNA mutations by single-stranded conformational polymorphism and sequencing analysis. Both chronic HBsAg carrier status and liver AFB1-DNA adducts were significantly higher in cases than in controls with odds ratios (OR) of 8.4 and 3.9, respectively (P < 0.01). Moreover, HCC risk was greatest in individuals with both AFB1-DNA adducts and HBsAg, suggesting a viral-chemical interaction. Mutant p53 protein, mutations in the p53 gene, and specific codon 249 mutations were detected in 37, 29, and 13%, respectively, of the HCC cases. Most of the DNA mutations were transversions, and the only major clustering site for mutations was codon 249. AFB1-DNA adducts were associated with p53 protein (OR = 2.9, P = 0.054) and DNA mutations (OR = 2.9, P = 0.082) but with borderline significance. All of the codon 249 mutations (n = 12) occurred in HBsAg-seropositive carriers, resulting in an OR of 10.0 (P < 0.05), suggesting that HBV may be involved in the selection of these mutations. The ORs between HBsAg and p53 DNA and protein mutations were 2.6 (P = 0.077) and 1.8 (P > 0.05), respectively. Both p53 DNA and protein mutations were related to tumor stage, suggesting that they are late events. These studies provided further support for the role of aflatoxin exposure in HCC in Taiwan and insight into viral-chemical interactions and molecular pathogenesis.

Aflatoxin B1↗

PubMed1997

Enhancement of EAE and induction of autoantibodies to T-cell epitopes in mice infected with a recombinant vaccinia virus encoding myelin proteolipid protein.

SJL/J mice were infected with a recombinant vaccinia virus encoding myelin proteolipid protein (PLP) (VVplp). Antibody responses to whole PLP and to encephalitogenic peptides, p139-151, p178-191 or p104-117 were measured after vaccination and following challenge with these three PLP peptides. Competitive ELISAs showed that antibodies to p139-151 and p178-191 represented the majority of antibodies in the anti-PLP antibody response following VVplp vaccination, since the antibodies to intact PLP could be inhibited 56, 35 and 1%, respectively, by p139-151, p178-191 and p104-117. After peptide challenge, epitope specific anti-peptide antibodies were enhanced. These anti-peptide antibodies also reacted with the intact PLP molecule. Interestingly, the mean titer of anti-p139-151 antibody in p139-151 challenged mice was significantly higher than that observed for anti-p178-191 in p178-191 and for anti-p104-117 in p104-117 challenged mice. Following peptide challenge, the anti-PLP IgG response shifted from an IgG1 to an IgG2a and 2b phenotype. In these mice, both the clinical disease and histological pattern of experimental allergic encephalomyelitis (EAE) were enhanced. The enhancement was most pronounced in the pathologic scores in the p139-151 challenged group followed by p104-117 challenged mice. Thus, humoral immune responses to PLP encephalitogenic peptides can be generated with virus encoding a self central nervous system (CNS) protein.

Animals↗

PubMed1997

Regulation of heme biosynthesis in Salmonella typhimurium: activity of glutamyl-tRNA reductase (HemA) is greatly elevated during heme limitation by a mechanism which increases abundance of the protein.

In Salmonella typhimurium and Escherichia coli, the hemA gene encodes the enzyme glutamyl-tRNA reductase, which catalyzes the first committed step in heme biosynthesis. We report that when heme limitation is imposed on cultures of S. typhimurium, glutamyl-tRNA reductase (HemA) enzyme activity is increased 10- to 25-fold. Heme limitation was achieved by a complete starvation for heme in hemB, hemE, and hemH mutants or during exponential growth of a hemL mutant in the absence of heme supplementation. Equivalent results were obtained by both methods. To determine the basis for this induction, we developed a panel of monoclonal antibodies reactive with HemA, which can detect the small amount of protein present in a wild-type strain. Western blot (immunoblot) analysis with these antibodies reveals that the increase in HemA enzyme activity during heme limitation is mediated by an increase in the abundance of the HemA protein. Increased HemA protein levels were also observed in heme-limited cells of a hemL mutant in two different E. coli backgrounds, suggesting that the observed regulation is conserved between E. coli and S. typhimurium. In S. typhimurium, the increase in HemA enzyme and protein levels was accompanied by a minimal (less than twofold) increase in the expression of hemA-lac operon fusions; thus HemA regulation is mediated either at a posttranscriptional step or through modulation of protein stability.

Acclimatization↗

PubMed1997

Clinical utility of color Doppler sonography in the differentiation of hepatocellular carcinoma from metastases and hemangioma.

The clinical utility of color Doppler sonography in the differentiation of hepatocellular carcinoma from metastases and hemangioma was investigated in 72 hepatocellular carcinomas (80 lesions), 30 metastases (82 lesions), and 39 hemangiomas (54 lesions). Overlaps of color patterns were found among hepatocellular carcinoma, metastases and hemangioma. Pulsatile waves from lesions with the basket, vessels within tumor, or spot patterns, or lesions measuring less than 3 cm with detectable signals, did not favor the diagnosis of hemangioma. In conclusion, color Doppler sonography can aid in the differentiation of hepatocellular carcinoma from hemangioma but may be unreliable in the differentiation of hepatocellular carcinoma from hypervascular metastases.

Adolescent↗

PubMed1996

Aflatoxin exposure and risk of hepatocellular carcinoma in Taiwan.

To investigate the carcinogenic effect of environmental aflatoxin exposure, 56 cases of hepatocellular carcinoma (HCC) diagnosed between 1991 and 1995 were identified and individually matched by age, sex, residence and date of recruitment to 220 healthy controls from the same large cohort in Taiwan. Blood samples were analyzed for hepatitis B and C viral markers and for aflatoxin-albumin adducts; urine was tested for aflatoxin metabolites. We obtained information about sociodemographic characteristics, habitual alcohol drinking, cigarette smoking and diet in a structured interview. Hepatitis B virus surface antigen (HBsAg) carriers had a significantly increased risk for HCC. After adjustment for HBsAg serostatus, the matched odds ratio (ORm) was significantly elevated for subjects with high levels of urinary aflatoxin metabolites. When stratified into tertiles, a dose-response relationship with HCC was observed. The ORm for detectable aflatoxin-albumin adducts was not significant after adjustment for HBsAg serostatus. HBsAg-seropositive subjects with high aflatoxin exposure had a higher risk than subjects with high aflatoxin exposure only or HBsAg seropositivity only. In male HBsAg-seropositive subjects, adjusted ORs were 2.8 (95% confidence interval [CI] = 0.9-9.1) for detectable compared with non-detectable aflatoxin-albumin adducts and 5.5 (CI = 1.3-23.4) for high compared with low urinary aflatoxin metabolite levels. Our results suggest that environmental aflatoxin exposure may enhance the hepatic carcinogenic potential of hepatitis B virus. A large-scale study will be needed to evaluate the effect of aflatoxin exposure on HBsAg non-carriers.

Aflatoxins↗

PubMed1996

A natural kinase-deficient variant of fibroblast growth factor receptor 1.

A fibroblast growth factor receptor 1 variant missing 37 amino acids from the carboxy-terminal tyrosine kinase catalytic domain was discovered in human lung fibroblasts and several other human cell lines. The receptor variant binds specifically to acidic fibroblast growth factor but has no tyrosine kinase activity. It was found that cellular transfectants expressing the fibroblast growth factor receptor 1 variant are mitogenically inactive and ligand binding to the receptor causes neither receptor autophosphorylation nor phospholipase C-gamma transphosphorylation. The fibroblast growth factor receptor 1 variant therefore represents an inactive receptor for acidic fibroblast growth factor. Since both kinase and kinase-deficient receptor forms are expressed in cells, it is conceivable that the kinase-deficient receptor plays an important role in regulating cellular responses elicited by acidic fibroblast growth factor stimulation.

Base Sequence↗

PubMed1996

p53 mutations in gastric cancers from Taiwan.

Mutations of the p53 gene were investigated in 80 surgical specimens of primary gastric cancer by polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) analysis. Mutations were detected in 18 tumors (22.5%) and localized to exons 5, 7 and 8. Mutations did not follow a random distribution among different subtypes, but instead clustered in the group of papillary adenocarcinomas, in which 7/12 (58.3%) cases were mutated. Positivity for p53 mutation was significantly higher in intestinal-type (37.5%) than in diffuse-type carcinomas (12.5%). These results suggest that gene alterations of p53 are not rare and may participate in the carcinogenesis of intestinal-type carcinomas of the stomach. Twenty of 21 p53 mutations were represented by single nucleotide changes, mostly missense mutations (19 events) and one nonsense mutation. Transversional mutations constitute the majority of p53 mutations (65%) and only 20% of mutations show G:C to A:T transitions. It is possible that the etiologies of gastric cancer in different geographical areas are different.

Adult↗

PubMed1996

The 30 kDa protein co-purified with chick liver glutathione S-transferases is a carbonyl reductase.

An unidentified 30 kDa protein was co-purified with chick liver glutathione S-transferases from S-hexylglutathione affinity column. The protein was isolated to apparent homogeneity with chromatofocusing. The molecular mass of the protein was determined to be 30 277 +/- 3 dalton by mass spectrometry. The protein was digested with Achromobacter proteinase I. Amino-acid sequence analyses of the resulting peptides show a high degree of identity with those of human carbonyl reductase. The protein is active with menadione as substrate. Thus, it is identified as chick liver carbonyl reductase.

Alcohol Oxidoreductases↗

PubMed1996

Protection against oxidative injury and permeability alteration in cultured alveolar epithelium by transferrin-catalase conjugate.

The successful prevention of hydrogen peroxide-induced alveolar permeability alterations and cell injury by transferrin-catalase conjugate is described in this study. Permeability alterations and cell injury were induced in cultured alveolar epithelial monolayers by hydrogen peroxide. Transepithelial transport of a permeability marker, [14C] mannitol, and cellular nuclear fluorescence of a membrane integrity indicator, propidium iodide, were used to quantitate epithelial permeability and damage respectively. Hydrogen peroxide (0.1 - 10 mM) induced a dose-dependent increase in both alveolar permeability and cellular damage; however, the oxidant effect on monolayer permeability did not require prior cell damage. Electron spin resonance measurements using the spin trap 5,5-dimethyl-l-pyrroline-N-oxide indicated the formation of hydroxyl radicals in hydrogen peroxide-treated cells. Chelation of the cellular pool of iron by deferoxamine inhibited radical formation and helped protect the cells from oxidative changes. Prior treatment of the cells with catalase (0.1 U-10 U/ml) had minimal protective effects on cell injury and permeability alterations. In contrast, transferrin-catalase conjugate, at the same concentration range, exhibited much improved protective effects on the cells in response to oxidant stress. This enhanced protection was found to correlate well with an increase in cellular uptake of the enzyme conjugate via the transferrin receptor endocytosis pathway. Effective protection by the enzyme conjugate was shown to require both the antioxidant enzyme moiety and the cognate moiety for the cell surface receptor. These findings indicate the potential therapeutic merit of transferrin-catalase conjugate for the treatment of pathological processes in the lung, whenever oxidative stress is involved.

Animals↗

PubMed1996

Elevated aflatoxin exposure and increased risk of hepatocellular carcinoma.

To elucidate the importance of aflatoxin in the etiology of hepatocellular carcinoma (HCC), a community-based cohort study combined with molecular dosimetry of aflatoxin exposure was performed in the Penghu Islets where the HCC mortality rate is highest in Taiwan. A total of 6,487 residents aged 30 to 65 years were recruited in the two-stage screening survey and underwent regular follow-up examination. Among 33 newly diagnosed HCC cases, 31 (94%) were chronic hepatitis B surface antigen (HBsAg) carriers and 3 (9%) were positive for antibodies against hepatitis C virus (HCV). Among 20 HCC patients and 86 matched healthy controls whose serum samples were tested for aflatoxin B1 (AFB1)-albumin adducts by competitive enzyme-linked immunosorbant assay (ELISA), 13 (65%) HCC patients and 32 (37%) matched controls were seropositive, showing a statistically significant multivariate-adjusted odds ratio of 5.5 with a 95% confidence interval of 1.2 to 24.5. The results imply the elevated risk of HCC among Penghu residents may be attributable to their heavy exposure to aflatoxins and high HBsAg carrier rate.

Adult↗

PubMed1996

Inhibition by nystatin of Kv1.3 channels expressed in Chinese hamster ovary cells.

The patch-clamp technique was used to study the effects of nystatin on a cloned delayed rectifier potassium channel (Kv1.3) expressed in Chinese hamster ovary (CHO) cells. Kv1.3 currents recorded in the whole-cell configuration, using an intracellular solution containing nystatin, were subjected to a time- and concentration-dependent reduction in their amplitude and in the time constants of apparent inactivation. Direct application of nystatin to the cytoplasmic side of excised inside-out patches inhibited Kv1.3 currents and this inhibition was immediately reversible upon washout of the drug. In contrast, currents mediated by another delayed rectifier (Kv3.1) were not affected by this drug. The concentrations for nystatin and its structural analog, amphotericin B, required to produce half maximal inhibition (IC50) of the current were estimated to be about 3 and 60 microM, respectively. The effects of nystatin on the amplitude and inactivation of Kv1.3 currents were not voltage-dependent. In inside-out patches, tetraethylammonium (TEA) produced a rapid block of Kv1.3 currents upon the onset of a voltage pulse, while the inhibition by nystatin developed slowly. When co-applied with TEA, nystatin potentiated the extent of the TEA-dependent block, and the kinetic effect of nystatin was slowed by TEA. In summary, nystatin, a compound frequently used in perforated patch recordings to preserve intracellular dialyzable components, specifically inhibited the potassium channel Kv1.3 at concentrations well below those required for perforation. The site of this inhibition may be different from that for TEA and is readily accessible from the cytoplasmic side of the membrane.

Amphotericin B↗

PubMed1996

Virus encoding an encephalitogenic peptide protects mice from experimental allergic encephalomyelitis.

The association of viral infections with autoimmune central nervous system (CNS) diseases such as post-infectious encephalomyelitis and possibly multiple sclerosis (MS) prompted the investigation to understand how virus infection could modulate autoimmune responses. Recombinant vaccinia viruses encoding an encephalitogenic portion of myelin basic protein (MBP) were evaluated in an animal model for human demyelinating disease, experimental allergic encephalomyelitis (EAE). We have determined that mice vaccinated with recombinant viruses encoding an encephalitogenic region of MBP were protected from EAE. In vivo depletion of CD8+ T cells did not abrogate this protection, suggesting lack of regulation by this cell type. These studies demonstrate that virus infection may be a means to modulated immune responsiveness to CNS disease.

Amino Acid Sequence↗

PubMed1996

Immunohistochemical quantitation of 4-aminobiphenyl-DNA adducts and p53 nuclear overexpression in T1 bladder cancer of smokers and nonsmokers.

An immunoperoxidase method, using a monoclonal antibody which recognizes 4-aminobiphenyl (4-ABP)-DNA adducts, was developed for the detection and quantitation of DNA damage in bladder tissue and applied to stored paraffin blocks of transurethral resection specimens of 46 patients with T1 bladder cancer. Mean relative staining intensity for 4-ABP-DNA adducts was significantly higher in current smokers (275 +/- 81, n = 24) compared to nonsmokers (113 +/- 71, n = 22) (P < 0.0001). There was a linear relationship between mean levels of relative staining and number of cigarettes smoked with lower levels in the 1-19 cig/day group (205 +/- 30, n = 5), compared to the 20-40 (289 +/- 40, n = 7) and the >40 cig/day group (351 +/- 57, n = 3)(P < 0.001). Nuclear overexpression of p53, analyzed by immunoperoxidase staining, was observed in 27 (59%) of the 45 stage T1 tumors analyzed. There was a significant correlation between p53 overexpression and recurrence of disease (odds ratio = 12.3, P < 0.01). Nuclear staining of p53 was also correlated with smoking status, cig/day and 4-ABP-DNA adducts. This work demonstrates that the immunohistochemical method has sufficient sensitivity for detection of 4-ABP-DNA adducts in human bladder samples. The method has several advantages including small sample size, the possibility of retrospective analysis of stored paraffin blocks, the ability to analyze binding in specific cell types, and a relatively low cost.

Aged↗

PubMed1996

Longitudinal study of intrahepatic portal flow: a novel approach using Doppler technique.

The present study investigates whether the application of the summation of portal blood flow (SPBF), measured from the left umbilical portion and the right anterior branch by Doppler, could be of use in the longitudinal study of intrahepatic portal flow or not. Forty-five subjects were divided into two groups. Group I subjects included males and post-menopausal females. Thirty-seven subjects (19 normal volunteers, 12 subjects with chronic hepatitis, six cirrhotics) were investigated by the more experienced doctor A at 9 day intervals for an intra-observer study. Among them, seven subjects (four normal volunteers, three with chronic hepatitis) were also investigated consecutively at 10 min apart by doctor B on days 1 and 10 for an interobserver study. Group II subjects included eight normal subjects with a regular menstrual cycle. They were investigated by doctor A during menstruation, mid-follicular, ovulatory and mid-luteal phases of the menstrual cycle to evaluate the possible impact of the menstrual cycle on SPBF measurement. The reproducibility of measurements was assessed by the coefficient of variation (CV) and the reproducibility coefficient. Data are expressed as mean +/- s.d. The CV of the SPBF for intra-observer and interobserver study were 4.88 +/- 3.36 and 4.19 +/- 2.53, respectively. All differences of SPBF were within their corresponding reproducibility limits. The results of SPBF measurements obtained from different phases of the menstrual cycle were variable. Only the comparison between the data obtained from mid-follicular and mid-luteal phases could demonstrate acceptable CV values (3.98 +/- 3.12%) and all the differences in SPBF measurements were within the repeatability limit. In conclusion, Doppler SPBF measurement may be useful in the longitudinal evaluation of intrahepatic portal flow changes.

Adult↗

PubMed1996

Percutaneous intravascular ethanol injection of the supplying tumor vessel in the treatment of hepatocellular carcinoma larger than 3 cm.

Percutaneous intravascular ethanol injection of the supplying vessel of the tumor using color Doppler imaging was performed in three patients (three lesions) with hepatocellular carcinoma measuring more than 3 cm. Intravascular injection was achieved in six out ot seven attempts. The total amount of ethanol injected intravascularly was small (10, 15,0 and 26 ml for each patient). One attempt failed and the ethanol was injected perivascularly. No serious complication was noted. After treatment, two patients showed remarkable decrease in tumor size and one showed massive tumor necrosis. In conclusion, ethanol injection using the intravascular approach may be of value in the palliative management of large hepatocellular carcinomas.

Aged↗

PubMed1996

Changes in intrahepatic portal hemodynamic in early stage hepatic abscesses.

Color Doppler sonography was used to investigate the changes in intrahepatic portal hemodynamics in early stage hepatic abscesses (21 lesions). At time of patients' admission, 10 lesions measuring over 6 cm showed reverse flow from at least one of their corresponding segmental portal branches, but 11 lesions measuring 6 cm or less did not show this phenomenon. All portal branches with reverse flow eventually returned to a normal direction of flow after successful medication. In conclusion, a large early stage hepatic abscess may show temporary reverse portal flow on color imaging. Thus, a differential diagnosis of hepatic tumors with reverse portal flow should include early stage hepatic abscesses.

Adult↗

PubMed1996

Treatment of hepatocellular carcinoma smaller than 5 cm by transcatheter arterial chemoembolization.

To assess the effectiveness of transcatheter arterial chemoembolization (TACE) in the treatment of small hepatocellular carcinoma (HCC) and to analyze the prognostic factors, a total of 77 patients with histologically proven HCC, < or = 5 cm in diameter, were enrolled for the study and followed for more than 2 years, The overall cumulative 1-, 2-, and 3-year survival rates were 79.2%, 50.6% and 36.7%, respectively. The median survival time of Child-Pugh's A patients was 990 +/- 146 days, which was significantly better than the median survival time of Child-Pugh's B patients (450 +/- 82 days) Furthermore, positive HBeAg and alpha-Fetoprotein (AFP) were factors close to statistically significant. In contrast, sex, age, HBsAg, Anti-HCV, tumor type, tumor size and tumor number were not related to the prognosis in small HCC who received TACE.

Adult↗