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Biomedical subjects

C Y Yang

Publications and source records attributed to C Y Yang.

At least 289 records · Page 16Linked to original sources
PubMed1984

Primary structure of human class II histocompatibility antigens. Reversed-phase high-performance liquid chromatography for integral membrane proteins.

The amino acid sequence of the human HLA-DR2 class II antigen was determined with the aid of high-performance liquid chromatography. By applying exclusively Hypersil-ODS as the stationary phase in combination with three buffer systems, the amino acid sequences of the extracellular parts of the DR alpha- and DR beta-chains could be established. Moreover, the sensitivity of the procedure allowed the determination of the primary structures of additional class II polypeptides isolated together with the DR2 alpha- and the DR2 beta-chains. Significant insight into the heterogeneity of class II molecules was thereby obtained. In addition, a simple, rapid and selective method of general applicability is described for the isolation of membrane fragments of integral membrane proteins. Using Sep-Pak cartridges with a mobile phase containing 60% formic acid and varying 1-propanol concentrations, the highly hydrophobic peptides can selectively be separated from the hydrophilic fragments.

Amino Acid Sequence↗
PubMed1984

Age-specific prevalence and significance of hepatitis B e antigen and antibody in chronic hepatitis B virus infection in Taiwan: a comparison among asymptomatic carriers, chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma.

The age-specific prevalence of hepatitis B e antigen (HBeAg) and its antibody (anti-HBe) were studied by radioimmunoassay, and compared in a large series of patients with chronic hepatitis B virus (HBV) infection, including 268 asymptomatic carriers, 389 chronic hepatitis, 114 liver cirrhosis, and 278 hepatocellular carcinoma (HCC). The prevalence of HBeAg/anti-HBe in asymptomatic carriers and patients with chronic hepatitis correlated closely with age as HBeAg prevalence decreased and anti-HBe prevalence increased with increasing age (P less than 0.0005), and is probably due to high infection rate at young age in Taiwan. The prevalence of HBeAg in patients with both cirrhosis and HCC are much significantly lower and had no correlation with age. Two peaks of age-specific prevalence of HBeAg and anti-HBe were observed in patients with HCC, implicating two patterns of HBV infection in these patients. The difference in the prevalence of HBeAg and anti-HBe might indicate that asymptomatic carriers, chronic hepatitis, liver cirrhosis, and HCC are sequential sequelae of HBV infection.

Adolescent↗
PubMed1984

Separation of dimethylaminoazobenzenethiohydantoin amino acids by high-performance liquid chromatography at low picomole concentrations.

The separation of all common dimethylaminoazobenzenethiohydantoin (DABTH) amino acids derived from modified Edman sequencing can be achieved by using high-performance liquid chromatography. All derivatives, including DABTH-Ile and DABTH-Leu, can be readily separated in a solvent mixture of sodium acetate buffer and 1% ethylene dichloride in acetonitrile. The high absorbance of the DABTH amino acids at 436 nm makes possible the quantitative determination of these derivatives at picomole concentrations in a relatively short time (30-40 min).

Acetonitriles↗
PubMed1984

Peripheral T-cell subsets in chronic type B hepatitis: correlation with biochemical and histological activities and hepatitis B e antigen/antibody status.

Peripheral T-cell subsets in 77 patients with hepatitis B surface antigen (HBsAg)-positive chronic liver diseases were studied by indirect immunofluorescence using murine monoclonal antibodies against all peripheral T cells (OKT3), T-helper/inducer cells (OKT4), and T-cytoxic/suppressor cells (OKT8). OKT4/OKT8 ratios were significantly reduced in patients with hepatitis B e antigen (HBeAg)-positive chronic liver diseases, including 28 patients with chronic active hepatitis (CAH) (P less than 0.001) and 15 with chronic persistent hepatitis (CPH) (P less than 0.001). OKT4/OKT8 ratios were significantly lower in 21 HBeAg-negative patients with CAH (P less than 0.05), as compared to those of 17 normal controls, while T-cell subsets in 13 patients with HBeAg-negative CPH were essentially normal. Low OKT4/OKT8 ratios significantly correlated with HBeAg positivity (P less than 0.001) and CAH (P less than 0.05), as assessed with multiple regression. There was a significant negative correlation between OKT4/OKT8 ratios and serum glutamic-pyruvic transaminase (SGPT) levels (r = -0.37; P less than 0.01). It was concluded that in chronic hepatitis B virus infection, low OKT4/OKT8 ratios are closely related to active viral replication and more severe histological and biochemical activity.

Adolescent↗
PubMed1984

Determinants for hepatitis B e antigen clearance in chronic type B hepatitis.

A 6-year (mean 24.5 months) longitudinal study has been undertaken in 237 HBeAg-positive patients with clinicopathologically verified chronic hepatitis. HBeAg clearance occurred in 74 patients at a rate of 16% per year, and a cumulative probability of 67% at the end of a 5-year follow-up. HBeAg clearance was preceded by a temporary "exacerbation" with SGPT greater than 300 IU/L in 62% of the patients. On the other hand, irrespective of the level of SGPT elevation, only 23% of such exacerbations were followed by HBeAg clearance. Further analysis indicates that alphafetoprotein greater than 100 ng/ml, frequently associated with bridging hepatic necrosis, during "exacerbation" predicts HBeAg clearance. In addition, patients with chronic lobular hepatitis and chronic active hepatitis have a higher annual HBeAg clearance rate than patients with chronic persistent hepatitis and nonspecific histologic changes (greater than 15% vs less than 8%, P less than 0.03). Short-term immunosuppressive treatment did not delay HBeAg seroconversion. Male patients had a higher annual HBeAg clearance rate than female patients (18.2% vs 8.7%, P less than 0.05). It was concluded that, in addition to the time factor, the nature of chronic hepatitis, the extent of hepatic damage during "exacerbation", and the sex of the patients are important determinants for HBeAg clearance.

Female↗
PubMed1984

[Primary structure of human class II histocompatibility antigens (HLA-D). II. Separation of alpha from beta-chains of HLA-D from a homozygous lymphoblastoid B cell line, H2LCL (HLA-A3,3;B7,7;Dw2,2;MT1,1;DC1,1;MB1,1), and characterization of the isolated chain].

In a previous communication we described the preparation of the alpha/beta-chain complex of HLA-D membrane antigens from a homozygous lymphoblastoid B cell line, H2LCL (HLA-A3,3; B7,7; Dw2,2; DR2,2; MT1,1; DC1,1; MB1,1). In this paper we present the separation of the alpha- and beta-chains in quantitative scale using hydroxylapatite chromatography with a pH 7.15 phosphate buffer, containing sodium dodecyl sulfate. An alternative isolation procedure is electrophoresis, taking advantage of the different isoelectric points of the alpha- and beta-chains. The relative molecular masses, the isoelectric points and the N-terminal sequences are discussed and compared with the results of other investigators. Remarkable is the polymorphism of both the chains, especially the beta-chains. The importance of the previously described three step purification procedure for the preparation of these antigens for sequence studies has been pointed out.

B-Lymphocytes↗
PubMed1983

The architecture of the animal fatty acid synthetase. II. Separation of the core and thioesterase functions and determination of the N-C orientation of the subunit.

Chicken fatty acid synthetase is cleaved by alpha-chymotrypsin into two fragments of molecular weight 230,000 and 33,000. These fragments may be easily separated by ammonium sulfate fractionation and gel filtration to yield pure preparations. The large 230,000-Da fragment contains all of the core activities of the fatty acid synthetic sequence i.e. acetyl and malonyl transacylases, condensing enzyme, beta-ketoacyl and enoyl reductases, the dehydratase, and the acyl carrier protein. The smaller 33,000-Da fragment retains the thioesterase activity which catalyzes the release of the completed acyl chains from the complex. Antibodies against the purified thioesterase fragment cross-react with analogous (Mr 33,000) peptides released from the complex by other proteases, as well as with all proteolytic intermediates that were predicted by peptide mapping to contain the thioesterase segment (Mattick, J. S., Tsukamoto, Y., Nickless, J., and Wakil, S. J. (1983) J. Biol. Chem. 258, 15291-15299). Amino acid sequence analyses demonstrate that the thioesterase domain is located at the carboxyl terminus of the synthetase monomer, thereby orienting the proteolytic (and functional) sites within the complex with respect to the direction of transcription and translation.

Amino Acid Sequence↗
PubMed1983

The complete primary structure of the allosteric L-lactate dehydrogenase from Lactobacillus casei.

The polypeptide chain of the allosteric L-lactate dehydrogenase (EC 1.1.1.27) of Lactobacillus casei consists of 325 amino acid residues. Despite the strikingly different enzymatic characteristics of the allosteric L-lactate dehydrogenase of L. casei and of the non-allosteric vertebrate enzymes, the sequence of the allosteric enzyme shows a distinct homology with that of the non-allosteric vertebrate enzymes (average identity: 37%). An especially high sequence homology can be identified within the active center (average identity: 70%). A clear deviation of the L. casei enzyme from the vertebrate enzyme is the lack of the first 12 amino acid residues at the N terminus and an additional 7 amino acid residues at the C terminus. The localization of the binding site of the allosteric effector D-fructose 1,6-bisphosphate and pH and effector-induced changes of the spectroscopic properties are discussed on the basis of the primary structure.

Amino Acid Sequence↗
PubMed1983

Blue-sensitive rod input to bipolar and ganglion cells of the Xenopus retina.

Intracellular recordings were obtained from chromatic and non-chromatic bipolar cells, identified by Lucifer yellow injection in the Xenopus retina. The chromatic cells, which lacked center-surround organization, were short wavelength hyperpolarizing (lambda max 445 nm) and long wavelength depolarizing. Under photopic conditions the depolarizing component was driven by 612 nm cones, but under mesopic conditions it appeared that 524 nm rods also constituted an input to the response. The non-chromatic bipolars encountered were of the off-center (hyperpolarizing) variety, with an active antagonistic surround, and peak spectral sensitivity in the red portion of the spectrum. Extracellular recordings were obtained from color-coded ganglion cells classified as type 1 or 2 in frog retina by Maturana et al. (1966) [J. gen. Physiol. 43, 129-175] and Bäckström and Reuter (1975) [J. Physiol. 246, 79-107]. The spectral sensitivity of the long latency "on" component was matched by the density spectrum of the 445 nm rod. This response component lacked center-surround organization and showed a relatively broad area of spatial integration. In contrast, a short latency component had a spectral sensitivity matched by the 612 nm cone pigment under photopic conditions, was either "on" or "off" center, showed center-surround organization and had a relatively small area of spatial integration. We speculate that in Xenopus retina, both chromatic and non-chromatic bipolar cells provide synaptic input to the class 1,2 ganglion cell.

Action Potentials↗
PubMed1983

Retinal hexokinase: kinetic properties and the effect of cyclic 3',5'-adenosine monophosphate.

We measured hexokinase (EC 2.7.1.1) activity in particulate and soluble fractions isolated from bullfrog (Rana catesbeiana) retinas. Seventy-three percent of the hexokinase (HK) activity was associated with the particulate fraction, 27% with the soluble fraction. Both HK fractions could phosphorylate fructose, glucose, 2-deoxy-D-glucose, and mannose, but not galactose. The Km for glucose was 0.14 mM, for 2-deoxy-D-glucose, 3.6 mM. With glucose as substrate, the Vmax for particulate HK was 125-148 microM retina-1 min-1, for soluble HK, 37 microM retina-1 min-1. Product inhibition of particulate HK activity by glucose 6-phosphate was marked, whereas 2-deoxy-D-glucose 6-phosphate did not inhibit the activity. Cyclic AMP stimulated the HK activity of both retinal fractions nearly twofold at concentrations of 0.2-0.8 mM; AMP was much less effective in this regard.

Animals↗
PubMed1982

Uptake and localization of 3H-2 deoxy-D-glucose by retinal photoreceptors.

Following dark incubation of isolated retinas of Xenopus laevis in Ringer solution supplemented with 3H-2-Deoxy-D-glucose (2DG), virtually all of the uptake of the label was by the inner segments and synaptic bases of the photoreceptor cells. Autoradiographs prepared from conventionally fixed tissue showed the same cellular distribution of label as those prepared from identically incubated, unfixed, freeze-dried retinas. However, fixation removed about 77% of the total counts. This fixation-labile, soluble fraction was identified as being primarily 2DG-6 phosphate by thin-layer chromatography. The remaining insoluble fraction corresponded in distribution to glycogen grains. In cones, glycogen is stored primarily in the paraboloid, whereas in rods it is distributed throughout the inner segment and synaptic base. EM autoradiographs illustrated that these were the sites over which fixation-resistant 2DG label was localized. Measurements of radioactivity associated with extracts of retinal glycogen following 2DG incubation demonstrated that a disproportionately high fraction of total counts were associated with the glycogen fraction. We conclude that in the amphibian retina 2DG may be incorporated into glycogen.

Animals↗
PubMed1982

Transport and phosphorylation of 2-deoxy-D-glucose by amphibian retina. Effects of light and darkness.

We studied the uptake of 2-deoxy-D-glucose (2DG) and the synthesis of its phosphorylated product 2DG-6-phosphate (2DG-6P) by the retinas of the clawed frog (Xenopus laevis) and the bullfrog (Rana catesbeiana). Autoradiographs showed that most of the retinal 2DG uptake is by the photoreceptor layer. The 2DG accumulation by isolated Xenopus retinas was time and concentration dependent. The Kt for transport was 5.05 mM; Vmax was 6.99 X 10(-10) mol . mg-1 tissue wet weight min-1. The Km for 2DG-6P formation was estimated to be 2-3 mM and Vmax to be approximately 4 x 10(-9) mol . mg-1 min-1. 2DG uptake was inhibited competitively by glucose with a Ki of 2.29 mM. Exposure to light reduced 2DG uptake by no more than 10% as compared with dark uptake. Low sodium or ouabain (10(-4)-10(-7) M) treatment did not significantly alter 2DG uptake as compared with control retinas. In experiments upon intact, anesthetized bullfrogs, light reduced both the total amount of radioactivity acquired by the retina and the fraction of 2DG-6P present. The results are discussed in terms of the fraction of energy consumed by the retina required to maintain the photoreceptor dark current.

Animals↗