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R Yao

Publications and source records attributed to R Yao.

At least 55 records · Page 3Linked to original sources
PubMed1997

[The method for studying kinetics of formation reaction of complex in dimeric system].

The method for studying kinetics of formation reaction of complex in aggregation system was reported in this paper. The concentration of more than one species was calculated by improved method for calculating dimeration constants. Kinetics of reaction between Cu (II) and tri-sulfophthalocyanine (S3Pc) was studied in this way. It shows that the rate of reaction between Cu(II) and S3Pc can be expressed as follows: v = K[Cu][S3Pc]

Copper↗
PubMed1996

Human gamma-glutamyl hydrolase: cloning and characterization of the enzyme expressed in vitro.

A cDNA encoding human gamma-glutamyl hydrolase has been identified by searching an expressed sequence tag data base and using rat gamma-glutamyl hydrolase cDNA as the query sequence. The cDNA encodes a 318-amino acid protein of Mr 35,960. The deduced amino acid sequence of human gamma-glutamyl hydrolase shows 67% identity to that of rat gamma-glutamyl hydrolase. In both rat and human the 24 amino acids preceding the N terminus constitute a structural motif that is analogous to a leader or signal sequence. There are four consensus asparagine glycosylation sites in the human sequence, with three of them conserved in the rat enzyme. Expression of both the human and rat cDNA in Escherichia coli produced antigenically related proteins with enzyme activities characteristic of the native human and rat enzymes, respectively, when methotrexate di- or pentaglutamate were used as substrates. With the latter substrate the rat enzyme cleaved the innermost gamma-glutamyl linkage resulting in the sole production of methotrexate as the pteroyl containing product. The human enzyme differed in that it produced methotrexate tetraglutamate initially, followed by the triglutamate, and then the diglutamate and methotrexate. Hence the rat enzyme is an endopeptidase with methotrexate pentaglutamate as substrate, whereas the human enzyme exhibits exopeptidase activity. Another difference is that the expressed rat enzyme is equally active on methotrexate di- and pentaglutamate whereas the human enzyme has severalfold greater activity on methotrexate pentaglutamate compared with the diglutamate. These properties are consistent with the enzymes derived from human and rat sources.

Amino Acid Sequence↗
PubMed1996

Growth factor-dependent survival of rodent fibroblasts requires phosphatidylinositol 3-kinase but is independent of pp70S6K activity.

A variety of mammalian cells undergo apoptosis when deprived of growth factors, but the intracellular signaling pathways responsible for cell survival remain to be characterized. In the present study, we have investigated the role of PI 3-kinase and pp70S6K in growth factor-dependent survival of rodent fibroblasts. As previously reported for PC12 pheochromocytoma cells, Rat-1 and REF52 cells underwent apoptosis following either serum-deprivation or treatment with the PI 3-kinase inhibitors wortmannin and LY294002. In contrast, NIH3T3 and BALB 3T3 cells were resistant to apoptosis induced by either serum-deprivation or PI 3-kinase inhibition. It thus appears that PI 3-kinase is specifically required to prevent apoptosis of fibroblast cell lines that are dependent upon growth factors for their survival. Consistent with this role of PI 3-kinase, serum and growth factors maintained steady state levels of PI 3-kinase activity that rapidly decreased following serum-deprivation. Serum and growth factors similarly maintained a steady state level of pp70S6K, which is thought to be activated downstream of PI 3-kinase. However, inhibition of pp70S6K activation by rapamycin failed to induce apoptosis in either Rat-1 or PC12 cells. Cell survival thus appears to require a PI 3-kinase signaling pathway that is independent of pp70S6K activation.

3T3 Cells↗
PubMed1996

Identification, cloning, and sequencing of a cDNA coding for rat gamma-glutamyl hydrolase.

Purified gamma-glutamyl hydrolase secreted from rat H35 hepatoma cells has been characterized as a diffuse band of 55 kDa on SDS-polyacrylamide gel electrophoresis that is converted to bands of 35 and 33 kDa after enzymatic removal of N-linked carbohydrate. Polyclonal antibodies against 55-kDa gamma-glutamyl hydrolase captured the enzyme activity and recognized the glycosylated and both deglycosylated forms of gamma-glutamyl hydrolase. A complete cDNA sequence of gamma-glutamyl hydrolase was obtained using degenerate oligonucleotides derived from peptide sequences, screening of a rat hepatoma cDNA library, and reverse transcription polymerase chain reaction. Based upon the deduced amino acid sequence the peptide component of gamma-glutamyl hydrolase had a molecular weight of 33,400. The results of amino acid analysis of the purified protein agreed with the deduced amino acid sequence in which there are seven potential asparagine-containing glycosylation sites.

Amino Acid Sequence↗
PubMed1996

Developmental changes in glycoconjugate composition during chick lens morphogenesis.

The following lectins: Con A, WGA, sWGA, PNA, RCA and UEA were used to study developmental changes in the expression of glycoconjugates during chicken lens morphogenesis. Con A, WGA, sWGA binding epitopes were observed in the lens placode and vesicle. Once the fiber mass was formed, the glycoconjugates were mainly found at the epithelial-fiber-cell junction, on epithelial cell membranes but only weakly on fiber-cell membranes. The PNA reaction was restricted to the apical surface of cells of the lens placode and vesicle and to the epithelial-fiber junction throughout the rest of lens development. The RCA reaction was mainly localized to the apical plasma membrane and moderately at the lateral plasma membrane of cells of the lens placode and vesicle and maintained this staining pattern in the lens epithelial cell during the progressive development of lens. UEA binding was initially localized along the posterior elongating cells of the lens vesicle, and then expressed in both epithelial and fiber cells. Subsequently UEA binding was restricted to the epithelium and central fiber mass. The extensive distribution of glycoconjugates on the surface of the invaginating placode cells suggests a role during invagination and subsequent detachment of the placode from the surface ectoderm. The capsule was labelled by Con A, WGA, sWGA, RCA and PNA but not by UEA. The posterior capsule was more intensely reactive with Con A, RCA and PNA than the anterior capsule.

Animals↗
PubMed1996

Electron microscopic detection of glycoconjugates in the chicken lens.

The cytochemical localization of glycoconjugates in the 14-day old embryonic chick lens was analysed by lectin-gold labelling. Con A/HRP gold particles, specific for D-mannose labelled the interior of the rough endoplasmic reticulum, membranes of the Golgi complex, secretory vesicles and the plasma membranes of the lens epithelial cell. The lens capsule was heavily labelled. Lens fiber cell membranes were also labelled. In contrast LFA, specific for neuraminic acid, did not bind to the endoplasmic reticulum or nuclear membrane. Labelling of the Golgi complex, secretory vesicles and capsule was observed. The plasma membranes of epithelial and fiber cells were extensively labelled, and probably reflects the presence of glycolipids such as gangliosides.

Animals↗
PubMed1996

An environmentally regulated pilus-like appendage involved in Campylobacter pathogenesis.

Examination of strains of Campylobacter jejuni, Campylobacter coli, and Campylobacter fetus by electron microscopy revealed that they produced peritrichous pilus-like appendages when the bacteria were grown in the presence of bile salts. Various bile-salt supplements were used and it was found that deoxycholate and chenodeoxycholic acid caused a significant enhancement of pilus production and resulted in a highly aggregative phenotype. Morphologically, the pili were between 4 and 7 nm in width and were greater than 1 micron in length. A gene, termed pspA, which encodes a predicted protein resembling protease IV of Escherichia coli, was identified in C. jejuni strain 81-176. A site-specific insertional mutation within this gene resulted in the loss of pilus synthesis as determined by electron microscopy. Insertions upstream and downstream of the gene had no effect on pilus production. The non-piliated mutant of strain 81-176 showed no reduction in adherence to or invasion of INT 407 cells in vitro. However, this mutant, while still possessing the ability to colonize ferrets, caused significantly reduced disease symptoms in this animal model.

Amino Acid Sequence↗
PubMed1996

Perinatal expression of adenylyl cyclase subtypes in rat brown adipose tissue.

The ability of norepinephrine to stimulate adenylyl cyclase (AC) activity increases during the perinatal period in rat brown adipose tissue (BAT), and this increase is associated with changes in the activities of both GS alpha and AC. The purpose of this study was to determine which AC subtypes are present in neonatal BAT and to examine whether the perinatal increase in AC activity corresponds to an increase in the expression of a particular AC subtype. Analysis of AC mRNAs by nuclease protection assay demonstrated the presence of mRNAs encoding AC-III, AC-IV, AC-VI, and AC-IX in embryonic and postnatal BAT. Of the subtypes detected, only AC-III mRNA levels increased substantially during the perinatal period. The increase in AC-III expression was paralleled by an increase in isoproterenol-stimulated AC activity. Treatment of neonates was the sympathetic neurotoxin 6-hydroxydopamine abolished the perinatal increase in both AC activity and AC-III mRNA levels but had no effect on the expression of other AC subtypes. These results strongly indicate that the increase in AC activity during the perinatal period is due to an increase in the expression of AC-III.

Adenylyl Cyclases↗
PubMed1995

Regulation of the Ras signaling pathway by GTPase-activating protein in PC12 cells.

We have investigated the role of Ras GTPase-activating protein (GAP) in NGF-induced neuronal differentiation by overexpressing both wild-type and membrane-targeted GAP in PC12 cells. Extension of neurites in response to NGF was completely blocked in cells expressing the highest level of membrane-targeted GAP and significantly inhibited in cells expressing either wild-type GAP or lower levels of membrane-targeted GAP. Overexpression of membrane-targeted GAP similarly inhibited induction of differentiation by src, but not by ras or raf oncogenes, indicating that GAP inhibits differentiation of PC12 cells by downregulating Ras function. GAP overexpression also inhibited stimulation of mitogen-activated protein (MAP) kinase and induction of immediate-early genes in response to NGF. In cells expressing wild-type GAP or lower levels of membrane-targeted GAP, the initial activation of MAP kinase and immediate-early gene expression were only partially inhibited. However, GAP expression in these cells resulted in substantial inhibition of sustained MAP kinase activity following NGF treatment, consistent with the inhibition of neurite extension in these cell lines. These results indicate that GAP acts as a negative regulation, rather than an effector, of Ras signaling in PC12 cells.

Animals↗
PubMed1995

Requirement for phosphatidylinositol-3 kinase in the prevention of apoptosis by nerve growth factor.

Nerve growth factor (NGF) induces both differentiation and survival of neurons by binding to the Trk receptor protein tyrosine kinase. Although Ras is required for differentiation, it was not required for NGF-mediated survival of rat pheochromocytoma PC-12 cells in serum-free medium. However, the ability of NGF to prevent apoptosis (programmed cell death) was inhibited by wortmannin or LY294002, two specific inhibitors of phosphatidylinositol (Pl)-3 kinase. Moreover, platelet-derived growth factor (PDGF) prevented apoptosis of PC-12 cells expressing the wild-type PDGF receptor, but not of cells expressing a mutant receptor that failed to activate Pl-3 kinase. Cell survival thus appears to be mediated by a Pl-3 kinase signaling pathway distinct from the pathway that mediates differentiation.

Androstadienes↗
PubMed1995

Effects of gamma-glutamyl hydrolase on folyl and antifolylpolyglutamates in cultured H35 hepatoma cells.

A subline of H35 hepatoma cells (H35D cells) that have been made resistant to 5,10-dideazatetrahydrofolate exhibits an increase in gamma-glutamyl hydrolase (GH) activity. GH is a lysosomal enzyme in H35 and H35D cells on the basis of comparison of the distribution of enzyme activity with other known lysosomal enzymes. The hydrolysis rate of methotrexate polyglutamate with isolated, intact lysosomes is 4-5-fold greater in H35D cells than in H35 cells. GH activity in isolated lysosomes is in part dependent on the presence of a reducing agent such as mercaptoethanol. Permeabilization of lysosomal preparations from both cell types by Triton X-100 causes a 10-fold enhancement in GH activity. The result of the enhanced activity of GH in H35D cells is a marked reduction in antifolylpolyglutamate concentration, with the parent antifolate being the predominant intracellular species found under all conditions tested. Unlike antifolates, the total intracellular folate concentration is nearly identical in both cells under standard culture conditions up to 10 microns folic acid. However, the chain length of folylpolyglutamates consists of predominantly triglutamates and tetraglutamates in H35D cells with increased GH, whereas it consists of pentaglutamates and hexaglutamates in the parental cells. At 50 and 100 microns folic acid, the folate accumulation in H35D cells is less than half that of H35 cells, and the predominant polyglutamate species in the H35D cells are the diglutamates through the tetraglutamates. The results demonstrate that the two H35 cell lines having equal folylpolyglutamate synthetase but that one with enhanced lysosomal GH activity exhibits a marked reduction in the amount and gamma-glutamyl chain length of folylpolyglutamates and antifolylpolyglutamates.

Animals↗
PubMed1995

Socio-economic status and micro-environmental factors in relation to the risk of Japanese encephalitis: a case-control study.

In a population-based case-control study in Southern Henan Province, central China, children suffering from Japanese encephalitis (JE) were compared with neighborhood controls matched by age and sex in terms of several social and environmental variables. Factors found by crude analysis to be associated with an increased risk of JE included lower family income, lower parental education, living in houses near the periphery of villages and poor quality of houses. When adjustment was made for confounding variables, only the association of house location within village remained of borderline significance (OR = 0.51, 95% CI = 0.15 approximately 1.03). It is suggested that the beneficial effect of higher family income and parental education was partly due to the fact that those parents might be more conscious about having their children vaccinated in the situation where there was a shortage of JE vaccine in the study area.

Animals↗
PubMed1995

Prognostic factors of early sequelae and fatal outcome of Japanese encephalitis.

A clinical case control study to identify prognostic factors present at hospital admission associated with early sequelae and fatal outcome of acute Japanese encephalitis (JE) was carried out in Gusi county, Henan Province, central China from June to September 1991. A total of 70 patients with laboratory-confirmed acute JE were studied, of whom 3 cases died and 33 cases had neurological or psychiatric sequelae at the end of three months follow-up. The results showed that acute JE at younger age, with higher body temperature, high white cell count in CSF, and deep coma present at hospital admission were markers for unfavorable outcomes (sequelae or fatal). A history of the vaccination was not correlated with the early sequelae and fatal outcome of the disease. The paper suggests that early diagnosis and treatment and universal JE vaccination for all susceptible populations are keys for decreasing incidence of sequelae and fatal outcome of acute JE.

Acute Disease↗
PubMed1994

An archaeal S-layer gene homolog with repetitive subunits.

An S-layer protein gene homolog of the slgA gene of two Methanothermus species was found in the genome of another methanogenic archaeon of a different family, Methanosarcina mazei S-6. The new gene (slgB) encodes a molecule (SlpB) with the characteristics of S-layer proteins. The N-terminal half of SlpB is 44% identical to that encoded by SlgA, but the other half shows distinctive features: four 56 amino acid long tandem repeats, and Trp-Xaa-Trp clusters located six amino acids from the N-terminus of each repeat.

Amino Acid Sequence↗
PubMed1994

Isolation of motile and non-motile insertional mutants of Campylobacter jejuni: the role of motility in adherence and invasion of eukaryotic cells.

A method of insertional mutagenesis for naturally transformable organisms has been adapted from Haemophilus influenzae and applied to the study of the pathogenesis of Campylobacter jejuni. A series of kanamycin-resistant insertional mutants of C. jejuni 81-176 has been generated and screened for loss of ability to invade INT407 cells. Eight noninvasive mutants were identified which showed 18-200-fold reductions in the level of invasion compared with the parent. Three of these eight show defects in motility, and five are fully motile. The three mutants with motility defects were further characterized to evaluate the method. One mutant, K2-32, which is non-adherent and non-invasive, has an insertion of the kanamycin-resistance cassette into the flaA flagellin gene and has greatly reduced motility and a truncated flagellar filament typical of flaA mutants. The adherent non-invasive mutants K2-37 and K2-55 are phenotypically paralysed, i.e. they have a full-length flagellar filament but are non-motile. All three mutants show an aberration in flagellar structure at the point at which the filament attaches to the cell. Mutants K2-37 and K2-55 represent overlapping deletions affecting the same gene, termed pflA (paralysed flagella). This gene encodes a predicted protein of 788 amino acid residues and a molecular weight of 90,977 with no significant homology to known proteins. Site-specific insertional mutants into this open reading frame result in the same paralysed flagellar phenotype and the same invasion defects as the original mutants. The differences in adherence between the two classes of flagellar mutant suggest that flagellin can serve as a secondary adhesion, although other adhesins mediate a motility-dependent internalization process. Characterization of the mutants at the molecular level and in animal models should further contribute to our understanding of the pathogenicity of these organisms.

Amino Acid Sequence↗
PubMed1994

Studies on the extracellular polysaccharide from Agrobacterium radiobacter biovar I S-1231.

A strain S-1231 isolated from specimen of soil around Beijing area is gram-negative, non-sporing, motile by peritrichous flagella. It produces exopolysaccharide succinoglycan from carbohydrates as its carbon source but not starch and cellulose. Acid is produced during fermentation of glucose. Growing for 12-24 hr, the cells are rods 0.7-0.8 x 1.3-1.5 microns, round ended, single or in pairs. Colonies on nutrient agar plate are unpigmented, circular, raised, smooth and moist-glistening, edge entire. The organism produces 3-ketolactose and is unable to invade sunflower tissue. The G+C content of DNA is 62.8-63.4 mol%. The organism is referred to as Agrobacterium radiobacter. Moreover, the strain is oxidase-positive, catalase-positive, H2S-produce and can grow at 35 degrees C and 2% NaCl also. Litmus milk is alkalified. Thus, the organism was renamed Agrobacterium radiobacter biovar I. Component analyses showed that the exopolysaccharide (Agran-S) from A. radiobacter biovar I S-1231 consisted of D-glucose (69.1%), D-galactose (8.6%), pyruvic acid (9.5%) and succinic acid (10.5%). Methylation analyses revealed that the polysaccharide Agran-S contained following main structural units: (1-->3)-linked D-glucose (21.2%), (1-->3)-linked D-galactose (11.4%), (1-->6)-linked D-glucose (10.5%), (1-->4)-linked D-glucose (30.4%), (1-->4, 1-->6)-linked D-glucose (22.2%) and terminal D-glucose (4.3%). The -1H-NMR spectrum of the polysaccharide indicated that the linkages in the polymer are all beta-glycosidic. The IR spectra of the polysaccharide revealed the presence of ester linkage in polysaccharide Agran-S.

Bacterial Typing Techniques↗