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Biomedical subjects

S Jin

Publications and source records attributed to S Jin.

At least 217 records · Page 12Linked to original sources
PubMed1995

Studies on crystal structures, active-centre geometry and depurinating mechanism of two ribosome-inactivating proteins.

Two ribosome-inactivating proteins, trichosanthin and alpha-momorcharin, have been studied in the forms of complexes with ATP or formycin, by an X-ray-crystallographic method at 1.6-2.0 A (0.16-0.20 nm) resolution. The native alpha-momorcharin had been studied at 2.2 A resolution. Structures of trichosanthin were determined by a multiple isomorphous replacement method. Structures of alpha-momorcharin were determined by a molecular replacement method using refined trichosanthin as the searching model. Small ligands in all these complexes have been recognized and built on the difference in electron density. All these structures have been refined to achieve good results, both in terms of crystallography and of ideal geometry. These two proteins show considerable similarity in their three-dimensional folding and to that of related proteins. On the basis of these structures, detailed geometries of the active centres of these two proteins are described and are compared with those of related proteins. In all complexes the interactions between ligand atoms and protein atoms, including hydrophobic forces, aromatic stacking interactions and hydrogen bonds, are found to be specific towards the adenine base. The relationship between the sequence conservation of ribosome-inactivating proteins and their active-centre geometry was analysed. A depurinating mechanism of ribosome-inactivating proteins is proposed on the basis of these results. The N-7 atom of the substrate base group is proposed to be protonated by an acidic residue in the active centre.

Amino Acid Sequence↗
PubMed1995

Krebs cycle function is required for activation of the Spo0A transcription factor in Bacillus subtilis.

Expression of genes early during sporulation in Bacillus subtilis requires the activity of the transcription factor encoded by spo0A. The active, phosphorylated form of Spo0A is produced through the action of a multicomponent pathway, the phosphorelay. A mutant defective in the first three enzymes of the Krebs citric acid cycle was unable to express early sporulation genes, apparently because of a failure to activate the phosphorelay. Cells that produce an altered Spo0A protein that can be phosphorylated by an alternative pathway were not dependent on Krebs cycle function for early sporulation gene expression. These findings suggest that Krebs cycle enzymes transmit a signal to activate the phosphorelay and that B. subtilis monitors its metabolic potential before committing itself to spore formation.

Bacillus subtilis↗
PubMed1995

Distribution of medical insurance in China.

This paper investigates factors related to the distribution of medical insurance coverage in China, using information from an eight-province household survey of almost 16,000 individuals, conducted in 1989. Results of bivariate analyses show that medical insurance coverage, defined very broadly, varies considerably by individual and regional characteristics. Age, gender, education, occupation, employment sector, urbanization, level of industrial and commercial development, and province are all related to being insured or not. In addition, we find that the type of insurance program available to people varies by these same factors, and that the benefits provided by the seemingly uniform public and worker programs also vary, especially by province and degree of urban development. When the individual and regional variables are considered together in logistic regression analyses, the factors most strongly statistically related to the likelihood of being insured are where one works and where one lives. The distribution of insurance benefits in China appears to result in a pattern in which the rural and the poor, who are often at great risk of illness, are less likely to have medical insurance.

Adolescent↗
PubMed1995

Identification of a plant-encoded analog of PKR, the mammalian double-stranded RNA-dependent protein kinase.

Plant virus or viroid infection stimulates the phosphorylation of a plant-encoded protein of M(r) 68,000 to 70,000 (now termed pPKR) that is associated with double-stranded RNA-stimulated protein kinase activity. Using various biochemical and immunological comparisons, we have demonstrated that this plant protein is an analog of the mammalian PKR enzymes. pPKR is both cytosolic and ribosome associated, similar to mammalian PKR, and appears to be capable of phosphorylating exogenous histones. Monoclonal anti-serum to the human PKR as well as antiserum to a conserved double-stranded RNA-binding domain present on mammalian PKR demonstrated cross-reactivity with pPKR. Likewise, polyclonal antiserum to the pPKR detected the mouse and human PKR in western blot analysis. Northern blot analysis of a mammalian PKR cDNA detected a specific 2.5-kb transcript present in plant poly (A)+ RNA.

Animals↗
PubMed1995

An Agrobacterium virulence factor encoded by a Ti plasmid gene or a chromosomal gene is required for T-DNA transfer into plants.

Mutagenesis of the vir region on the Ti plasmid of Agrobacterium tumefaciens revealed a new locus, virJ, that is induced by the plant-wound signal molecule, acetosyringone (AS). virJ lies between virA and virB, and is transcribed in the same direction. The amino acid sequence of virJ is similar to a region of a previously characterized chromosomal gene, acvB, required for virulence. virJ can complement the avirulent phenotype of an acvB mutant, indicating that virJ and acvB encode the same factor required for tumorigenesis. Southern analysis revealed that virJ is present on the Ti plasmid of an octopine but not a nopaline strain whereas acvB is present on the chromosomes of both octopine and nopaline strains. While virJ is regulated by AS under the control of the virA/virG two-component regulatory system, acvB is not induced by AS. VirJ possesses a putative signal peptide and was found predominantly in the periplasmic fraction. The strain lacking both acvB and virJ had an impaired ability to transfer T-DNA into plant cells, suggesting that the factor encoded by virJ or acvB is required for T-DNA transfer from A. tumefaciens to plant cells. acvB is the first chromosomal gene implicated in T-DNA transfer, but whether it functions specifically for this process is not clear. We hypothesize that virJ evolved from acvB, presumably for a more specialized role in tumorigenesis.

Acetophenones↗
PubMed1995

Use of Electroporation To Generate a Thiobacillus neapolitanus Carboxysome Mutant.

Two cloning vectors designed for use in Escherichia coli and the thiobacilli were constructed by combining a Thiobacillus intermedius plasmid replicon with a multicloning site, lacZ(prm1), and either a kanamycin or a streptomycin resistance gene. Conditions necessary for the introduction of DNA into T. intermedius and T. neapolitanus via electroporation were examined and optimized. By using optimal electroporation conditions, the gene encoding a carboxysome shell protein, csoS1A, was insertionally inactivated in T. neapolitanus. The mutant showed a reduced number of carboxysomes and an increased level of CO(inf2) necessary for growth.

Journal Article↗
PubMed1995

[Textual research on diagnosis by lingual vessels].

Being a unique Chinese diagnostic measure, this method focussing its observation on the ventral vessels of the tongue, is first recorded in Neijing, saying that it is indicated when the vessels are varicose by puncturing for bloodletting in malaria, mad, and sadness. Ge Hong and Cao Yuanfan applied it for juandice, also for prognosis. Yimenfang predicts by this method the diagnosis for prognosis of the mother and foetus in difficult delivery, while Chen Zhiming popularized it in his Chan Nan sheng Si Jue. Inherited from the theory of Collateral Vessel Diagnosis, this approach pays attention to the observation of the forms and colors of collateral vessel, claiming that dark green color and congestion have bad diagnosis. Though not well developed, it was applied as extra-acupoints for swelling tongue, sore throat and jaundice. While Zhou Xuehai elaborated Liu Shouzhen's idea and put forward a theory of minute-vessel-stasis which is wonderfully conincided with modern microciroulation theory. In 1964, Zhang Zanchen again advocated its diagnostic value in tumor, hepatopathy and cor pulmonale.

China↗
PubMed1995

[The regulation of calmodulin in the cell cycle].

In order to study the role of calmodulin (CaM) in the cell cycle, RC 3 cells carring the CaM expression vectors which was constructed by joining the CaM cDNA with a plasmid of mouse mammary tumor virus (MMTV), were used in this experiment. The CaM expression vectors transcription is regulated by a dexamethasone (DXM) inducible MMTV LTR promoter. Upon addition of DXM, cells have transiently increased CaM mRNA and protein levels. Increased CaM caused a acceleration of proliferation. Flow cytometric analysis showed that progression though G1, G2 and metaphase was accelerated by increase in CaM levels, while treatment with the CaM antagonist trifluoperazine (TFP) blocked cell cycle progression at G1/S boundary and during G2/M and metaphase. The studies have shown that CaM is important in controlling progression at three points in the cell cycle: (1) The G1/S boundary to permit the initiation of DNA synthesis; (2) The G2/M boundary to permit the initiation of mitosis; (3) At the metaphase/anaphase transition of mitosis to permit chromosome segration and the completion of mitosis. This study indicates that the RC 3 cell is a useful experimental cell model for studing the effect of a transient increase of intracellular CaM levels on control of cell cycle.

3T3 Cells↗
PubMed1994

Sequences of the Serratia marcescens rplS and trmD genes.

The trmD and rplS genes from the Serratia marcescens trmD operon are presented. These genes are about 85% similar to those of Escherichia coli and encode very nearly identical proteins. However, there are an additional 22 nt around the beginning of trmD in S. marcescens which in addition uses a GUG start codon.

Amino Acid Sequence↗
PubMed1994

The binding site of the transcriptional activator VirG from Agrobacterium comprises both conserved and specific nonconserved sequences.

Virulence genes of Agrobacterium tumefaciens are transcriptionally activated in response to phenolic compounds and certain sugars. The transcription activator VirG specifically binds to fragments containing the conserved vir box sequence present in the promoter region of all vir genes. This study shows that both the vir box as well as specific nonconserved sequences downstream of the vir box are required for VirG binding and transcriptional activation. Insertion of the identified VirG binding site into the lac promoter resulted in transcriptional activation of this heterologous promoter in response to the plant phenolic signal molecule acetosyringone.

Agrobacterium tumefaciens↗
PubMed1994

Efficient fuzzy control strategies for the application of pH-stat to fed-batch cultivation of genetically engineered Escherichia coli.

In the cultivation of genetically engineered Escherichia coli it is very important to control the substrate concentration at an appropriate level in order to avoid the accumulation of acetate, thereby elevating the expression level of plasmid-encoded protein. In this paper, a pH-stat mode of fuzzy control was considered for the overexpression of beta-galactosidase in the fed-batch cultivation of recombinant E. coli. In the simple pH-stat fuzzy control, the response of pH change in the culture broth to the feeding rate of glucose was used to estimate the glucose consumption rate. In the modified pH-stat fuzzy control, the glucose consumption rate was accurately estimated by using pH change and the change in the carbon dioxide content of the exhaust gas. With this control strategy, the cell density could be increased to 72 g DCW dm-3, which was twofold higher than that attained in the cultivation with the simple pH-stat fuzzy control. The bulk beta-galactosidase concentration was increased to 4150 U cm-3, which was threefold higher than when the simple pH-stat control was used.

Carbon Dioxide↗
PubMed1994

Role of NMDA, AMPA and kainate receptors in mediating glutamate- and 4-AP-induced dopamine and acetylcholine release from rat striatal slices.

Striatal slices, preincubated with [3H]dopamine and [14C]choline, were superfused continuously and subjected to electrical field stimulation (3 Hz) and perfused with amino acid analogues or 4-amino pyridine (4-AP). The released radioactivity was used to monitor release of the neurotransmitters dopamine (DA) and acetylcholine (ACh). Glutamate, NMDA (in the absence of Mg2+), AMPA, kainic acid, domoate and 4-AP all induced DA and ACh release in a concentration-dependent manner. The DA and ACh release induced by NMDA (15 microM) and glutamate (1 mM) was essentially abolished by Mg2+ (1.15 mM), whereas release induced by AMPA (100 microM), kainic acid (100 microM) or 4-AP (30 microM) was not reduced. Tetrodotoxin (1 microM) essentially abolished the effects of NMDA, markedly reduced the effects of glutamate, AMPA and 4-AP, whereas the effect of kainic acid was only modestly affected. MK-801 (30 nM) reduced NMDA-induced DA release by some 70% and ACh release by 30%. MK-801 reduced 4-AP-induced DA release by 40% but not ACh release. CNQX in a concentration (10 microM) that scarcely affected NMDA-induced ACh release, but blocked that induced by AMPA, kainic acid or domoate, reduced the ACh release induced by 4-AP. In summary, DA and ACh release from rat striatum can be stimulated by activation of NMDA and non-NMDA glutamate receptors, and this mechanism is activated by the potassium channel blocker 4-AP.

4-Aminopyridine↗
PubMed1994

Equity and the utilization of health services: report of an eight-province survey in China.

This paper investigates equity with respect to one component of welfare in China--the provision and use of health services. Based upon a large-scale survey of almost 16,000 individuals in eight provinces in China, we examine a sub-sample of working-age adults who have identified themselves as injured or ill during the four weeks prior to being interviewed. We found that, beyond the level of severity of the reported condition, very few individual-level factors are related to the use of services when ill or injured. Only gender (female) and employment in state-run enterprises are associated with higher patterns of use. These results suggest that China has achieved a very wide distribution of clinics and other services at the local level, and that they are widely used by those who identify need for them. It is rare to be over half an hour away by bike from some form of care and the majority of care appears to be reasonably inexpensive. This broad availability of services contrasts with recent reports from China stressing declining accessibility, and paints a picture of relatively equal access to health care.

Adult↗
PubMed1994

The chromatographic behaviour of cephalosporins in gel filtration chromatography, a novel method to separate high molecular weight impurities.

The interaction between cephalosporins and the dextran matrix of Sephadex gel in gel filtration chromatography has been thoroughly studied. Twelve cephalosporins with specific structures were examined under different chromatographic conditions, including 12 different mobile phases comprising inorganic or organic compounds of different charge or/and density of electrons on their negative ions, different types of Sephadex gel (Sephadex G-10 and Sephadex G-50) and different flow rates. It was found that the more negative the charge or/and density of electrons on the negative ions of buffer components, the more was the adsorption of cephalosporins on the solid phase; this indicated tht the mobile phase played an important rôle in gel filtration chromatography for cephalosporins. By choice of suitable chromatographic conditions, optimum separation of high molecular weight impurities from cephalosporins could be achieved. The novel method could be used as a routine method for the quality control of cephalosporin preparations.

Cephalosporins↗
PubMed1994

PilR, a transcriptional regulator of piliation in Pseudomonas aeruginosa, binds to a cis-acting sequence upstream of the pilin gene promoter.

The PilR protein of Pseudomonas aeruginosa is a transcriptional activator of the pilin gene and belongs to a two-component sensor-regulator family. PilR was overproduced by fusing pilR to the gene for the maltose-binding protein (malE), yielding a MalE-PilR hybrid protein. The plasmid with the malE-pilR fusion, when introduced into a non-piliated pilR mutant strain of P. aeruginosa, restored piliation, indicating that the hybrid protein retains PilR function in vivo. The MalE-PilR protein was purified from Escherichia coli and used in a series of DNA-binding studies. A specific pilin promoter-binding activity of MalE-PilR was observed in a gel retardation assay. Subsequent DNase I footprinting analysis revealed a 40 bp PilR-binding site located at the -120 to -80 region, relative to the transcriptional start site of the pilin gene. This PilR-binding region consists of a nine-base sequence and three consensus sequences of 5'-(N)4-6C/GTGTC-3', in a tandem array in which the first 7-9 bp are bound by the PilR on the non-coding strand, leaving the last two nucleotides (TC) unbound. On the coding strand, PilR binds to sequences complementary to the two middle consensus sequences of the non-coding strand. A sequence similar to the NifA recognition site (5'-TGT-(N)11-ACA-3') is also found within the PilR-binding region. Deletion analysis and disruption of the individual consensus PilR-binding sequences by site-directed mutagenesis revealed that all four PilR-binding sites are absolutely required for the PilS/PilR-mediated pilin gene expression.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Outer Membrane Proteins↗
PubMed1994

Identification of two distinct Bacillus subtilis citrate synthase genes.

Two distinct Bacillus subtilis genes (citA and citZ) were found to encode citrate synthase isozymes that catalyze the first step of the Krebs cycle. The citA gene was cloned by genetic complementation of an Escherichia coli citrate synthase mutant strain (W620) and was in a monocistronic transcriptional unit. A divergently transcribed gene, citR, could encode a protein with strong similarity to the bacterial LysR family of regulatory proteins. A null mutation in citA had little effect on citrate synthase enzyme activity or sporulation. The residual citrate synthase activity was purified from a citA null mutant strain, and the partial amino acid sequence for the purified protein (CitZ) was determined. The citZ gene was cloned from B. subtilis chromosomal DNA by using a PCR-generated probe synthesized with oligonucleotide primers derived from the partial amino acid sequence of purified CitZ. The citZ gene proved to be the first gene in a tricistronic cluster that also included citC (coding for isocitrate dehydrogenase) and citH (coding for malate dehydrogenase). A mutation in citZ caused a substantial loss of citrate synthase enzyme activity, glutamate auxotrophy, and a defect in sporulation.

Aconitate Hydratase↗