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Biomedical subjects

W Sun

Publications and source records attributed to W Sun.

At least 37 records · Page 2Linked to original sources
PubMed1995

[The case-control study of risk factors in primary angle-closure glaucoma].

A case-control study was conducted to determine the risk factors of primary angle-closure glaucoma (PACG). In this study, 103 cases of PACG patients and 95 cases of non-glaucoma controls were investigated. It was found that the risk factors of PACG were family history of glaucoma, irritable temper, short distance between eyes and workplace at work and high blood pressure. The results did not support that cigarette smoking, alcoholic consumption, diabetes mellitus, myopia and blood types were related with PACG.

Adult↗
PubMed1995

Changes in sodium intake and blood pressure in a community-based intervention project in China.

A sodium intervention project was carried out in Tianjin, China, as part of the Tianjin Project, which is a national pilot, community-based intervention programme to reduce non-communicable diseases. The aim of this 3 year sodium project was to evaluate the feasibility and effects of sodium reduction in the population. The evaluation of the programme was based on examinations of independent cross-sectional population samples in 1989 (1719 persons) and in 1992 (2304 persons) in the intervention and matched reference areas. Food weighing and consecutive 3 day food records were used to measure dietary intake. The mean sodium intake fell 22 mmol/day in men and 11 mmol/day in women in the intervention area from 1989 to 1992. The reduction was significant in men (P = 0.001) and near significance in women (P = 0.05). The sodium intake increased significantly in men in the reference area. There was a significant net reduction in sodium intake in men in the intervention area. This reduction was similar in different educational and occupational groups suggesting that the intervention had reached the whole community. The mean systolic blood pressure (SBP) decreased 3 mm Hg for the total population and 2 mm Hg for normotensives in the intervention area. There was a significant net reduction in SBP both in all or in normotensive subjects. These results support the conclusion that community-based sodium intervention is feasible for hypertension prevention. However, the sodium intake is still very high in this population, which warrants further effective intervention.

Adolescent↗
PubMed1994

Domains of Escherichia coli primase: functional activity of a 47-kDa N-terminal proteolytic fragment.

Endoproteinase Asp-N cleaves the 581-amino acid Escherichia coli primase (65,564 Da) into several major fragments. One of these, a 47-kDa fragment containing the complete N terminus and the first 422 amino acids of primase, is capable of primer RNA (pRNA) synthesis in the G4oric/single-stranded DNA binding protein/primase pRNA synthesis system. A cloned 398-amino acid N-terminal fragment of primase can also synthesize pRNA. The sizes of the pRNA synthesized by these N-terminal fragments, however, are smaller than those synthesized by intact primase, suggesting that the C-terminal region of primase plays a role in processivity or regulation of pRNA synthesis. Primase mutants with the last 10 and 40 C-terminal amino acids deleted synthesize pRNA as wild-type primase, indicating that any regulatory sequences must be internal to the C terminus of primase.

Amino Acid Sequence↗
PubMed1994

Biosynthesis of poly-3-hydroxybutyrate in the luminescent bacterium, Vibrio harveyi, and regulation by the lux autoinducer, N-(3-hydroxybutanoyl)homoserine lactone.

Poly-3-hydroxybutyrate (PHB), a biopolymer of important commercial applications, is found in a wide range of Gram-negative and Gram-positive bacteria and cyanobacteria. The present study has resulted in the identification of PHB in the luminescent marine bacteria, Vibrio harveyi, in spite of it being previously classified as PHB-negative. PHB granules with distinct membranes were detected by electron microscopy after fixation and staining of V. harveyi cells with malachite green. Analyses by gas chromatography, nuclear magnetic resonance, infrared, and ultraviolet spectroscopy clearly established the presence of PHB. The synthesis of PHB in V. harveyi was found to be under cell density regulation with the levels increasing from 0 (< 0.2) to 26 mg of PHB/g of dry cell weight during growth in a manner analogous to the induction of luminescence in this bacteria. Moreover, synthesis of PHB in V. harveyi was shown to be controlled by the lux autoinducer, N-(3-hydroxybutanoyl)homoserine lactone, providing not only a potential link between luminescence and PHB production but also showing that the lux autoinducer acts as a general signal transductant. These results have also extended the role of homoserine lactones in metabolic regulation to include the control of synthesis of potential energy reserves.

4-Butyrolactone↗
PubMed1994

Presence of gonadotropin-releasing hormone mRNA in the rat olfactory piriform cortex.

Gonadotropin releasing hormone (GnRH) neurons are known to be originated from the olfactory placode and to enter the forebrain regions during embryonic development. The present study aims to ascertain whether GnRH is locally expressed in the olfactory cortex. Northern blot hybridization and reverse transcription-polymerase chain reaction revealed that GnRH mRNA was present in the rat olfactory cortex as well as in the olfactory bulb. The predicted size of GnRH mRNA is similar to that detected in the hypothalamus. In situ hybridization histochemistry also showed that GnRH mRNA is highly concentrated in the olfactory piriform cortex. The present data indicate that GnRH is synthesized in the olfactory piriform cortex.

Animals↗
PubMed1994

Production of mouse V/human C chimeric kappa genes by homologous recombination in hybridoma cells. Analysis of vector design and recombinant gene expression.

Homologous recombination between transferred and chromosomal Ig genes in mouse hybridoma cells offers a general method of altering the chromosomal Ig genes in predetermined ways. Recombination is infrequent in hybridoma cells, and we have been interested in improving the methods for identifying and recovering the rare recombinants. We have used vectors that are designed to replace the mouse chromosomal C kappa segment with the human equivalent, so that recombinants produce mouse V/human C chimeric kappa-chains. We describe an enhancerless, replacement type vector that can be used with the herpes thymidine kinase counterselection to provide such enrichment that homologous recombinants constitute 15% of the selected G418-resistant, FIAU-resistant cells. We have also measured the level of chimeric kappa gene expression and found surprisingly that (1) it is very variable among transformants with the same recombinant gene structure, (2) there is no systematic difference in the level of production by recombinants that retain or have lost the J-C kappa intron enhancer, and (3) the amount of chimeric kappa mRNA in even the highest producing transformants is much less than the amount of the corresponding mouse kappa mRNA.

Animals↗
PubMed1994

Primary structure and functional expression of the AMPA/kainate receptor subunit 2 from human brain.

A full-length cDNA clone encoding the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA)/kainate (KA) receptor subunit 2 (HBGR2) was isolated from a human brain cDNA library. The HBGR2 cDNA has an open reading frame of approximately 2.7 kb that codes for an 883-residue protein. At the amino acid level, HBGR2 is 98% identical to its rat counterpart GluR2, and 69% to the AMPA/KA receptor subunit 1 from human brain (HBGR1). Injection of cRNA transcripts from the HBGR2 into oocytes produces barely detectable kainate-activated ionic currents, indicating that the HBGR2 subunit alone weakly expresses homomeric receptor channels. Coexpression of HBGR2 and HBGR1 transcripts, however, evokes kainate-dependent currents which activate at higher agonist concentration than those required by homomeric HBGR1 receptor channels. Coexpressed receptors display a linear current-to-voltage relationship at variance with the inwardly rectifying profile exhibited by HBGR1 homomers. Hence, the HBGR2 subunit coassembles with the HBGR1 subunit to form heteromeric receptor channels akin to the glutamate receptors from rodent brain.

Amino Acid Sequence↗
PubMed1994

Synthesis, surface, and cell-adhesion properties of polyurethanes containing covalently grafted RGD-peptides.

In an attempt to improve endothelial cell adhesion and growth on a polyurethane copolymer, cell adhesive RGD-containing peptides were grafted to the polymer backbone. Two peptide grafting reaction schemes, including one-step and two-step approaches, were developed. FTIR and amino acid analysis confirmed that coupling of the peptide to the polyurethane backbone was achieved by both the one-step and two-step methods. However, the two-step approach showed a higher peptide coupling efficiency and resulted in better control of the orientation of the grafted peptide. The two-step reaction scheme was used to prepare Gly-Arg-Gly-Asp-Ser-Tyr (GRGDSY), Gly-Arg-Gly-Asp-Val-Tyr (GRGDVY), and Gly-Arg-Gly-Glu-Ser-Tyr (GRGESY) peptide-grafted polyurethanes with two different peptide densities (100 and 250 mumol/g polymer). Dynamic contact angle measurements indicated that the surfaces of the peptide-grafted polyurethanes were more hydrophilic than the starting and carboxylated versions of the precursor polyurethane. In addition, the surface hydrophilicity of the peptide-grafted polymers increased with increasing bulk peptide density. Electron spectroscopy for chemical analysis suggested that the grafted peptide was present at the polymer-air interface, in vacuo, for the peptide-grafted polyurethanes. The surface peptide density appeared to correlate with the incorporated peptide density in the bulk. In vitro endothelial cell adhesion experiments showed that, without the presence of serum in culture medium, the GRGDSY- and GRGDVY-grafted polyurethanes dramatically enhanced cell attachment and spreading compared with the starting, carboxylated, and GRGESY-grafted polymers. Increasing the peptide density from 100 to 250 mumol/g polymer for the GRGDSY- and GRGDVY-grafted polyurethanes resulted in an increase in cell attachment. With approximately the same peptide density (100 or 250 mumol/g polymer), the GRGDVY-grafted polymers supported more adherent cells than did the GRGDSY-grafted polymers. Similar trends were observed in the in vitro endothelial cell growth studies using culture medium containing serum and endothelial cell growth supplement. The GRGDSY- and GRGDVY-grafted polyurethanes promoted more cell growth than did the starting polyurethane. However, the presence of adhesive serum proteins and growth factor diminished the differences between the cell-adhesive peptide grafted polymers and the GRGESY-grafted polymers.

Amino Acid Sequence↗
PubMed1994

Analysis of thymidine kinase gene expression in preimplantation mouse embryos.

Thymidine kinase (TK) activity was examined during the development of preimplantation mouse embryos. TK activity was increased approximately 20-fold from day 2 embryos (2-cell) to day 5 embryos (late blastocyst). TK activity did not change along with the progression into S-phase of the first and the second cell cycles but increased sharply at S-phase of the third cell cycle. Analysis of TK mRNA with a reverse transcription-polymerase chain reaction (RT-PCR) method showed that the level of TK mRNA was low in ovulated eggs and 1-cell embryos and was hardly detectable in day 2 embryos (2-cell), but sharply increased in day 3 embryos (mixture of 5- to 8-cell and morula). The functional role of 5'-flanking sequence of TK gene was also investigated in preimplantation embryos after microinjection with the DNA construct of 5'-flanking sequence of TK (2.4 kb) linked to bacterial lacZ gene (TK2.5lacZ) into the pronucleus of 1-cell and subsequently by histochemical staining with X-gal. beta-Galactosidase activity was first detected in day 3 embryos (8-cell), and 30% of embryos were stained with X-gal in day 4 and day 5 embryos, respectively. These results show that an increase in TK activity occurred after 2-cell stage, and this increase was primarily due to the embryonic activation of TK gene expression. Also, it appears that the 5'-flanking sequence of TK may directly regulate the TK gene expression at the transcriptional level during preimplantation murine development.

Animals↗
PubMed1994

Sodium channel mutations in paramyotonia congenita uncouple inactivation from activation.

Mutations in the adult human skeletal muscle Na+ channel alpha subunit cause the disease paramyotonia congenita. Two paramyotonia congenita mutations, R1448H and R1448C, substitute histidine and cysteine for arginine in the S4 segment of domain 4. These mutations, expressed in a cell line, have only small effects on the activation of Na+ currents, but mutant channels inactivate more slowly with less voltage dependence than wild-type channels and exhibit an enhanced rate of recovery from inactivation. Increase of extracellular pH made the rate of inactivation of R1448H similar to that of R1448C, suggesting that this residue has an extracellular location and that its charge is important for normal inactivation. Analysis of single-channel data reveals that mutant channels inactivate normally from closed states, but poorly from the open state. The data suggest a critical role for the S4 helix of domain 4 in coupling between activation and inactivation.

Base Sequence↗
PubMed1994

Forage and splanchnic tissue mass in growing lambs: effects of dietary forage levels and source on splanchnic tissue mass in growing lambs.

Forty-two crossbred lambs (33.4 kg initial body weight; twenty-four wethers and eighteen ewes) were used in a 42 d experiment with a 2 x 3 factorial treatment arrangement to determine effects of forage level and source on splanchnic tissue mass. Diets were 250 and 750 g/kg of chopped lucerne (Medicago sativa) (A), ryegrass (Lolium multiflorum)-wheat (Triticum aestivum) (RW) or bermudagrass (Cynodon dactylon) (B) hay, with the remainder being maize-based concentrate. Five lambs per treatment were slaughtered at the end of the experiment and measurements made of internal organs and contents of the gastrointestinal tract (GIT). Digestible organic matter intake (DOMI) on the 7 d preceding slaughter was 0.89, 0.83, 0.90, 0.83, 0.77 and 0.61 (SE 0.05) kg/d, and live-weight gain was 0.20, 0.17, 0.18, 0.10, 0.10 and 0.07 (SE 0.02) kg/d for diets A-25, RW-25, B-25, A-75, RW-75 and B-75 respectively. Total GIT mass (fresh) was higher (P < 0.05) for 750 than 250 g forage/kg and for B than RW (4.80, 4.57, 5.55, 5.84, 5.99 and 6.91 kg for diets A-25, RW-25, B-25, A-75, RW-75 and B-75 respectively). Non-fat organic matter was 259, 295, 292, 303, 277 and 264 g for the total GIT; 93, 102, 103, 106, 95 and 97 g for the reticulo-rumen (forage level x type (diet A v. diets RW and B) interaction; P < 0.05); and 204, 196, 202, 177, 156 and 127 g for the liver (SE 10) with diets A-25, RW-25, B-25, A-75, RW-75 and B-75 respectively. In summary, differences in properties of forage A and the grasses at 250 g/kg diet may have influenced GIT mass independent of energy intake and digesta mass. Conversely, with 750 g dietary forage/kg, higher digesta mass for diet B than diet RW appeared responsible for high reticulo-rumen mass relative to DOMI. Greater digesta mass for 750 than 250 g forage/kg may have elevated intestinal tissue mass/DOMI with diets A and B but not with diet RW, for which NDF digestibility was highest.

Animal Feed↗
PubMed1994

Voltage-dependent regulation of modal gating in the rat SkM1 sodium channel expressed in Xenopus oocytes.

The TTX-sensitive rat skeletal muscle sodium channel (rSkM1) exhibits two modes of inactivation (fast vs slow) when the alpha subunit is expressed alone in Xenopus oocytes. In this study, two components are found in the voltage dependence of normalized current inactivation, one having a V1/2 in the expected voltage range (approximately -50 mV, I(N)) and the other with a more hyperpolarized V1/2 (approximately -130 mV, IH) at a holding potential of -90 mV. The I(N) component is associated with the gating mode having rapid inactivation and recovery from inactivation of the macroscopic current (N-mode), while IH corresponds to the slow inactivation and recovery mode (H-mode). These two components are interconvertible and their relative contribution to the total current varies with the holding potential: I(N) is favored by hyperpolarization. The interconversion between the two modes is voltage dependent and is well fit to a first-order two-state model with a voltage dependence of e-fold/8.6 mV and a V1/2 of -62 mV. When the rat sodium channel beta 1-subunit is coinjected with rSkM1, IH is essentially eliminated and the inactivation kinetics of macroscopic current becomes rapid. These two current components and their associated gating modes may represent two conformations of the alpha subunit, one of which can be stabilized either by hyperpolarization or by binding of the beta 1 subunit.

Animals↗
PubMed1994

Common distribution of antigenic determinants and complementation activity on matrix proteins of two vesicular stomatitis virus serotypes.

To compare the antigenic and functional domains of the matrix (M) proteins of vesicular stomatitis virus (VSV) serotypes Indiana (VSV-Ind) and New Jersey (VSV-NJ), deletion mutants and chimeras were cloned in pBSM13 and expressed as in-frame lacZ fusion proteins in Escherichia coli. Non-cross-reactive monoclonal antibodies directed to the two antigenically distinct M proteins were tested by Western blot analysis to map three epitopes of VSV-Ind M protein and four epitopes of VSV-NJ M protein. Epitope 1 of the VSV-Ind M protein and epitope II of the VSV-NJ M protein both mapped to the highly basic N-terminal 34 amino acids of each homotypic M protein. Epitopes 2 and 3 of the VSV-Ind M protein and epitopes III and IV of the VSV-NJ M protein mapped to a region spanning amino acids 35 to 74. Epitope I of the VSV-NJ M protein mapped to a region between amino acid 75 and the C terminus. The similarity in location of the serotypically unique antigenic determinants of the two M proteins suggested that they may have a common functional domain. This hypothesis was substantiated by the finding that the two M proteins and various chimeras expressed in CV-1 cells by a recombinant vaccinia virus system were able to rescue M gene temperature-sensitive mutants of both VSV serotypes.

Antibodies, Monoclonal↗
PubMed1994

Membrane-binding domains and cytopathogenesis of the matrix protein of vesicular stomatitis virus.

The membrane-binding affinity of the matrix (M) protein of vesicular stomatitis virus (VSV) was examined by comparing the cellular distribution of wild-type (wt) virus M protein with that of temperature-sensitive (ts) and deletion mutants probed by indirect fluorescent-antibody staining and fractionation of infected or plasmid-transfected CV1 cells. The M-gene mutant tsO23 caused cytopathic rounding of cells infected at permissive temperature but not of cells at the nonpermissive temperature; wt VSV also causes rounding, which prohibits study of M protein distribution by fluorescent-antibody staining. Little or no M protein can be detected in the plasma membrane of cells infected with tsO23 at the nonpermissive temperature, whereas approximately 20% of the M protein colocalized with the membrane fraction of cells infected with tsO23 at the permissive temperature. Cells transfected with a plasmid expressing intact 229-amino-acid wt M protein (M1-229) exhibited cytopathic cell rounding and actin filament dissolution, whereas cells retained normal polygonal morphology and actin filaments when transfected with plasmids expressing M proteins truncated to the first 74 N-terminal amino acids (M1-74) or deleted of the first 50 amino acids (M51-229) or amino acids 1 to 50 and 75 to 106 (M51-74/107-229). Truncated proteins M1-74 and M51-229 were readily detectable in the plasma membrane and cytosol of transfected cells as determined by both fluorescent-antibody staining and cell fractionation, as was the plasmid-expressed intact wt M protein. However, the expressed doubly deleted protein M51-74/107-229 could not be detected in plasma membrane by fluorescent-antibody staining or by cell fractionation, suggesting the presence of two membrane-binding sites spanning the region of amino acids 1 to 50 and amino acids 75 to 106 of the VSV M protein. These in vivo data were confirmed by an in vitro binding assay in which intact M protein and its deletion mutants were reconstituted in high- or low-ionic-strength buffers with synthetic membranes in the form of sonicated unilammelar vesicles. The results of these experiments appear to confirm the presence of two membrane-binding sites on the VSV M protein, one binding peripherally by electrostatic forces at the highly charged NH2 terminus and the other stably binding membrane integration of hydrophobic amino acids and located by a hydropathy plot between amino acids 88 and 119.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1994

Coordinated expression of a 45 kD protein and ozone toxicity in a human bronchial epithelial cell line.

The human bronchial epithelial cell line, BEAS-2B, which was immortalized by transformation with SV40 virus, when grown biphasically between 0.1 and 1.0 ppm of ozone and liquid medium showed increased release of Cr, decreased synthesis of various macromolecules, and decreased cell viability. Cell injury was a function of the concentration of ozone to which the cells were exposed. Furthermore, in proportion to the extent of cell injury, ozone exposure also induced and/or enhanced synthesis of a 45 kD protein but not any of the well-characterized heat shock proteins, e.g., HSP 70. Actinomycin D prevented enhanced synthesis of the 45 kD protein in cells exposed to ozone, suggesting transcriptional regulation of expression of the 45 kD protein. Enhanced synthesis of the 45 kD protein was not observed in cells treated with heat, cigarette smoke condensate, hydrogen peroxide, or bleomycin. High concentrations of glutathione added to the culture medium reduced ozone toxicity and ozone-enhanced synthesis of the 45 kD protein. These results suggest that ozone injury and enhanced expression of a gene encoding a 45 kD protein of as yet unknown function are coordinated in the SV40-immortalized bronchial epithelial cells.

Bleomycin↗