PubMed Health⌕ Search

Biomedical subjects

S Cheng

Publications and source records attributed to S Cheng.

At least 73 records · Page 4Linked to original sources
PubMed1988

Use of psoralen-modified oligonucleotides to trap three-stranded RecA-DNA complexes and repair of these cross-linked complexes by ABC excinuclease.

A series of site-specifically cross-linked, three-stranded complexes has been prepared using the RecA protein, form I (covalently closed and negatively supercoiled) pUC19 plasmid, and eight psoralen-monoadducted oligonucleotides between 30 and 107 residues in length. Complexes formed much less efficiently if linearized pUC19 was used as the duplex substrate. Quantitative analysis indicates that although RecA is able to utilize a 30-mer in its homologous pairing reaction, incorporation at 50% efficiency requires a single-stranded substrate at least 50 residues long. These three-stranded complexes have been used to study the action mechanism for cross-link repair by ABC excinuclease, and the results are consistent with the recent model of Van Houten, B., Gamper, H., Holbrook, S. R., Hearst, J. E., and Sancar, A. (1986b) Proc. Natl. Acad. Sci. U. S. A. 83, 8077-8081.

Bacterial Proteins↗
PubMed1988

Polyamine metabolism in an obligately alkalophilic Bacillus alcalophilus that grows at pH 11.0.

Bacillus alcalophilus, an obligately alkalophilic bacterium that grows at pH 11.0, has an intracellular pH of 9.5 or less. Unlike all other living organisms, polyamines (putrescine, spermidine and spermine) in B. alcalophilus, if present, will be largely unprotonated. HPLC analysis indicated that spermidine is the major polyamine in B. alcalophilus, accounting for more than 90% of total polyamines, and the level of spermidine varies during growth. Ornithine decarboxylase activity was not detectable in B. alcalophilus under all conditions examined. When [3H]arginine was added to the culture medium, the radioactivity can be recovered from polyamine pool; the distribution is 3% for putrescine, 94% for spermidine, and 3% for spermine, suggesting the presence of arginine pathway for polyamine biosynthesis. The polyamine transport system in B. alcalphilus appears to be Na+-dependent and is highly sensitive to the inhibition of gramicidin S and valinomycin.

Arginine↗
PubMed1988

Study on erythrocyte membrane fluidity by laser Raman spectroscopy.

Differences between patients with cirrhosis of liver, nephrotic syndrome, coronary heart disease and normal subjects in laser Raman spectra of erythrocyte membranes have been found. In regions of 1000-1140 cm-1 and 2840-3000 cm-1, the ratios I1130/I1080 and I2890/I2850 in patient membranes are higher than those in normal ones respectively. These results mean that erythrocyte membrane fluidity of these patients is reduced. This reduction may be attributed to the possibility that erythrocyte membrane of patients get more cholesterol from their plasma and resulted in the modification of the ratio of cholesterol/phospholipids in the membranes.

Coronary Disease↗
PubMed1988

Inbreeding effects on fertility and hatchability associated with the formation of sublines.

The deleterious effect of inbreeding on fertility and hatchability was studied in a population of Leghorns developed primarily for immunogenetics studies. The 1983 population averaged 42% inbreeding and traced back to only two sires and five dams of the 1965 base population. Up to 1975 (Period 1), the population was maintained as a single-unit closed flock, during which time inbreeding increased an average of 1.44% per year. After 1975 (Period 2), the population was split into five sublines. Inbreeding then increased at the average rate of 2.7% per year. The regression of fertility on inbreeding was -.31 +/- .04 in Period 1 and -.17 +/- .06 in Period 2. Corresponding values for hatchability of fertile eggs were +.08 +/- .05 in Period 1 and -.24 +/- .07 in Period 2. The regression of hatchability of total eggs set was -.15 +/- .04 in Period 1 and -.35 +/- .10 in Period 2. The evidence implies that both fertility and hatchability have a heritable basis. As expected, the deleterious effects of inbreeding on reproduction was more severe in Period 2, with sublining.

Animals↗
PubMed1988

Genetic analysis of immunocompetence measures in a White Leghorn chicken line.

Immunocompetence of the Iowa State University S1 White Leghorn chicken line was studied. This line was divided into eight sublines based upon erythrocyte antigen B (Ea-B) allele (B1B1 or B19B19), antibody response to glutamic acid60-alanine30-tyrosine10 (GAT) (high or low), and response to Rous sarcoma virus-induced tumors (progression or regression). Antibody responses to Pasteurella multocida (PM), Mycoplasma gallisepticum (MG), and infectious bursal disease virus vaccines were evaluated by enzyme-linked immunosorbent assay. Phagocytic activity and T cell-mediated response were measured by carbon clearance and phytohemagglutinin (PHA) injection assays, respectively. Significant haplotype (subline) differences and sire family differences were observed in all three measurements. Significant sex differences were observed in phagocytic activity and T cell-mediated responses. Haplotypes with high antibody responses to GAT had significantly higher antibody titers to PM and MG vaccines than haplotypes with low antibody responses. Significant positive correlations were observed between antibody levels to the two vaccines. A significant negative correlation was seen between phagocytic activity and T cell-mediated response of females. The data suggest that the total immunocompetence profile of an individual must be considered to select for optimum immune responsiveness.

Animals↗
PubMed1987

Isolation of the structural genes for the Rieske Fe-S protein, cytochrome b and cytochrome c1 all components of the ubiquinol: cytochrome c2 oxidoreductase complex of Rhodopseudomonas capsulata.

The structural genes for the Rieske Fe-S protein (petA), cytochrome b (petB) and cytochrome c1 (petC) subunits of the ubiquinol:cytochrome c2 oxidoreductase (bc1 complex) of Rhodopseudomonas capsulata have been cloned by complementation, using a mutant defective in this complex. The location of these genes on the obtained plasmid, pR14A, was determined using synthetic mixed oligonucleotide probes corresponding to highly conserved amino acid sequences of these proteins from various organisms. Their correct identity was established by partial sequencing. The petA, petB and petC genes were found to lie close to each other in this order, spanning two adjacent EcoRI fragments of 2.7 X 10(3) and 1.3 X 10(3) base-pairs, respectively. An insertion-deletion mutation, covering most of petB and all of petC and an insertion mutation, located in petB were constructed in vitro and were introduced into the chromosome of an otherwise wild-type strain by gene transfer agent-mediated genetic crosses. The bc-1 mutants obtained were defective in photosynthesis but, as expected, they could grow by respiration because of a branched respiratory pathway. Therefore, in R. capsulata a functional bc1 complex is essential in vivo for photosynthesis but not for respiration. Further, in the respiratory pathway the branch point must be before the bc1 complex, most likely at the quinone pool. These mutants were also proficient in anaerobic growth in the presence of dimethylsulfoxide, indicating that a functional bc1 complex is not required for this pathway. Several other insertions and deletions, located outside of the pet gene cluster, were also constructed. The ability of these latter mutants to grow photosynthetically suggested that no other gene essential for photosynthesis is located in the proximity of the pet cluster. The plasmid pR14A was shown to complement in trans the bc-1 insertion or insertion-deletion mutants, indicating that the pet genes were expressed in R. capsulata. Cross-hybridization experiments showed that the pet cluster was quite distinct from other known genes involved in photosynthesis.

Amino Acid Sequence↗
PubMed1987

Effects of washing and gel filtration on the ultrastructure of human platelets.

The ultrastructure of washed and gel-filtered platelets has been examined by scanning- and transmission-electron microscopy. Three different washing procedures previously established for functional studies have been employed. Each washing method resulted in typical morphologic alterations of the platelets. Gel filtration of platelet-rich plasma appeared less harmful than washing. Specimens fixed immediately after elution contained numerous platelets with typical shape changes which were restored almost completely after incubation at 37 degrees C. Our study emphasizes the need for cautious interpretation of results obtained with separated platelets and mandates morphologic controls for such experiments.

Blood Platelets↗
PubMed1986

Characterization of a membrane-associated 3,3',5-triiodo-L-thyronine binding protein by use of monoclonal antibodies.

Four mouse hybridoma cell lines have been isolated which secrete antibodies to the membrane-associated thyroid hormone binding protein (Mr 55,000) from human epidermoid carcinoma A431 cells. J6 is rat specific; J2 is human and monkey specific; J8 and J9 have a wider specificity and react with similar thyroid hormone binding proteins (p55) from human, monkey, rat, and hamster. None of these antibodies reacts with mouse cells. J2, J6, and J9 are of the IgG1k class, and J8 is an IgAk antibody. p55 was characterized by using these monoclonal antibodies. It is not posttranslationally processed by glycosylation, phosphorylation, or sulfation. It has a cellular degradation rate t1/2 approximately equal to 3.2 h. Using immunofluorescence and electron microscopic immunocytochemistry, p55 was found to be associated with the lumenal face of the endoplasmic reticulum and nuclear envelope. When cell homogenates were prepared, significant amounts of p55 were released into the 110000g supernatant, indicating that p55 is loosely associated with the endoplasmic reticulum and nuclear envelope.

Animals↗
PubMed1985

High failure rates in outpatient treatment of salpingitis with either tetracycline alone or penicillin/ampicillin combination.

Eight hundred twenty-five ambulatory women with a clinical diagnosis of salpingitis were randomized to a 10-day course of either penicillin/ampicillin or tetracycline. Forty-four percent of women had gonococcal salpingitis and 56% nongonococcal salpingitis. Overall, both regimens cured equal proportions of women: At 30 days, 81% were cured by penicillin/ampicillin and 82% by tetracycline. However, the proportion of women with gonococcal salpingitis cured by 30 days was significantly greater than that of women with nongonococcal salpingitis. By 30 days, 14% of women with gonococcal salpingitis and 21% of women with nongonococcal salpingitis were not cured by either regimen. These data suggest that both regimens were only marginally acceptable for women with gonococcal salpingitis and that neither regimen was acceptable for nongonococcal salpingitis.

Adolescent↗
PubMed1985

Mechanisms of cholesterol synthesis inhibition by D-glucosamine.

The amino sugar D-glucosamine possesses antitumor activity which is thought to depend in part upon its ability to impair cholesterol biosynthesis and damage cellular membranes. The present study examined the effect of glucosamine on acetate utilization for lipid and sterol synthesis in rat C6 glial tumor cells. At cytotoxic concentrations, the amino sugar inhibited [14C]acetate incorporation into nonesterified sterols and lipids but increased the flow of label into cholesteryl esters. A comparison of the rates of acetate utilization for glucosamine metabolism (N-acetylation) and sterol and lipid synthesis suggested that glucosamine might act by competing for a common cytosolic pool of acetyl CoA. The inhibition of lipid and sterol synthesis, however, remained constant over a wide range of extracellular acetate concentrations. These results suggest that, if glucosamine acts by restricting the supply of acetate for these biosynthetic processes, it probably inhibits a step prior to the formation of acetyl CoA. Alternative mechanisms are discussed.

Acetates↗
PubMed1985

Characterization of the 3,3',5-triiodo-L-thyronine-binding site on plasma membranes from human placenta.

The binding of [125I]T3 to sites on human placenta plasma membranes was characterized, and the binding site was solubilized after affinity labeling with N-bromoacetyl-[125I]T3 (BrAc[125I]T3). Two classes of T3-binding sites were detected. One class has a high affinity (Kd = 2.0nM) and a low capacity (approximately 320 fmol/mg protein); the other has a low affinity (Kd = 18.5 microM) and a high capacity (approximately 2.2 pmol/mg protein). The binding sites were found to be specific for T3 in that other thyroid hormone analogs (D-T3, rT3, D-T4, and L-T4) were less effective or ineffective in displacing the bound [125I]T3. The affinity labeling ligand BrAc[125I]T3 was found to specifically label a protein with an apparent mol wt of 65,000, as determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The BrAc[125I]T3-labeled protein was solubilized with 2 mM 3-[( 3-cholamidopropyl)dimethylammonio]1-propane sulfonate. The apparent mol wt of the labeled protein was between 140,000 and 150,000 by Sephadex-G-200 gel filtration. These data demonstrate that a high affinity binding site specific for T3 is present on plasma membranes from human placenta and that the binding site is a protein, most likely a dimer, with a native mol wt between 140,000 and 150,000.

Affinity Labels↗
PubMed1985

Structural similarities between the plasma membrane binding sites for L-thyroxine and 3,3',5-triiodo-L-thyronine in cultured cells.

Using 125I-labeled L-thyroxine (T4), the binding of [125I]T4 to GH3 rat pituitary tumor cells was studied. At 15 degrees C, the binding of [125I]T4 to cells is saturable and specific. Least squares analysis of binding data showed two classes of binding sites with apparent dissociation constants of 4.3 +/- 0.3 nM and 350 +/- 30nM and binding capacities of (3.8 +/- 0.5) X 10(4) and (9.1 +/- 0.35) X 10(6) sites/cell, respectively. Affinity labeling of cells or purified plasma membranes with N-bromoacetyl-[125I]T4 (BrAc[125I]T4) showed a major specifically labeled protein band with an apparent molecular mass of 55 kilodaltons (kDal). Digestion of the 55-kDal protein from cells and plasma membrane by Staphylococcus aureus V8 protease or elastase gave similar peptide fragments. Thus, the 55-kDal protein labeled from intact cells is the same protein as that from purified plasma membranes. Peptide mapping was further used to compare the 55-kDal protein specifically labeled by either N-bromoacetyl-3,[125I]3',5-triiodo-L-thyronine (BrAc[125I]T3) or BrAc[125I]T4 in intact cells and highly purified plasma membranes. Very similar patterns were obtained. These results indicate that plasma membrane T3 and T4 binding sites have similar hormone binding domains. In addition the plasma membrane T3 and T4 binding sites of Swiss 3T3-4 mouse fibroblasts and A431 human epithelioid carcinoma cells are structurally similar to the T3 and T4 binding sites of GH3 cells.

Affinity Labels↗
PubMed1984

Antibodies against the plasma membrane 3,3',5-triiodo-L-thyronine binding protein of rat pituitary GH3 cells: partial characterization and cross-species immunoreactivity.

To develop antibodies against the plasma membrane 3,3',5-triiodo-L-thyronine (T3) binding protein (M.W. 55,000), rabbits were immunized with formalin-fixed GH3 cells or highly purified plasma membranes from these cells. Antibodies were screened by immunoprecipitation using detergent solubilized N-bromoacetyl-[125I]T3-labeled 55K protein. Among the nine detergents tested, 0.18% CHAPS was found to be the best in its solubilization efficiency and its ability to maintain the integrity of the antigenicity of the 55K protein. The N-bromoacetyl-[125I]T3-labeled 55K protein was also immunoprecipitated by anti-T3 antibodies. The anti-55K protein antibodies cross-reacted with plasma membrane T3 binding proteins from cultured cells and tissues of human and rodent origin. These results indicate that structural similarities exist in human and rodent plasma membrane T3 binding proteins. These antibodies should provide a powerful tool in the characterization and in probing the function(s) of the plasma membrane T3 binding protein in cells.

Animals↗
PubMed1983

Synthesis of lysosomal alpha-mannosidase in normal and mannosidosis fibroblasts.

The biosynthesis and secretion of lysosomal alpha-mannosidase was studied in metabolically labelled fibroblasts from controls and two patients with mannosidosis. Normal fibroblasts secrete alpha-mannosidase as a 110kDa polypeptide. Intracellularly alpha-mannosidase is represented by several polypeptides with apparent Mrs ranging from 40 to 67kDa. In two mannosidosis cell lines none of intra- and extracellular polypeptides of alpha-mannosidase were detectable. The mannosidosis fibroblasts secreted acid alpha-mannosidase activity at one third of the normal rate. In contrast to normal cells the secretion was not enhanced by NH4C1 and the secreted activity was not immunoprecipitable, indicating that the acid alpha-mannosidase activity secreted by mannosidosis fibroblasts is not related to the lysosomal alpha-mannosidase.

Cells, Cultured↗