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Biomedical subjects

S Jin

Publications and source records attributed to S Jin.

At least 199 records · Page 11Linked to original sources

Phenotypic expression of Pseudomonas syringae avr genes in E. coli is linked to the activities of the hrp-encoded secretion system.

The specific recognition of elicitors produced by plant pathogenic bacteria carrying avirulence (avr) genes is postulated to initiate cellular defense responses in plants expressing corresponding resistance genes. The biochemical functions of most avr genes, however, are not known. A heterologous system was developed to phenotypically express Pseudomonas syringae avr genes in Escherichia coli cells that required the P. syringae hrp cluster. E. coli MC4100 transformants carrying the plasmic-borne P. syringae pv. syringae Pss61 hrp cluster and p. syringae pv. glycinea avrB expressed from a triple lacUV5 promoter gained the ability to elicit the hypersensitive response in soybean cultivars expressing Rpg1 and in an Arabidopsis thaliana accession expressing RPM1. Inactivation of energy transducing or outer membrane components of the hrp-encoded secretion system blocked phenotypic expression expression of avrB in E. coli, but deletions abolishing harpinPSS production had little effect on the production of the AvrB phenotype by the E. coli transformants. Phenotypic expression of avrA, AvrPto, avrRpm1, avrRpt2, and avrPph3 in E. coli was also shown to require the hrp cluster. The results indicate that generation of the Avr phenotype in P. syringae strains is specifically dependent on the secretion activities of the hrp cluster.

Base Sequence↗

Trends in hospitalizations for diarrhea in United States children from 1979 through 1992: estimates of the morbidity associated with rotavirus.

OBJECTIVES: To examine trends in the hospitalizations of children for diarrheal disease in the U.S. and to provide estimates for the burden of disease associated with rotavirus diarrhea. METHODS: Data for diarrheal hospitalizations among U.S. children ages 1 month through 4 years were compiled from the National Hospital Discharge Survey for the years 1979 through 1992. Between 1979 and 1992, 12% of all hospitalizations of U.S. children 1 month through 4 years of age had an International Classification of Diseases code for diarrhea listed in one of the top three positions on the discharge diagnosis. RESULTS: The annual rate of diarrheal hospitalizations, 97 per 10 000 persons (average, 185 742 per year), did not change substantially during the 14-year study period and accounted annually for 724 394 inpatient days (3.9 days per hospitalization). For most diarrheal hospitalizations (75.9%) no causative agent was specified in the National Hospital Discharge Survey records; of the remaining 24.8%, viruses were most commonly reported (19.3%), followed by bacteria (5.1%) and parasites (0.7%). The proportion of hospitalizations associated with viral diarrheas rose from 13% to 27% during the 14-year study period, whereas the proportion of hospitalizations for noninfectious diarrhea declined from 79% to 60%. Every year the number of hospitalizations peaked from November through April, the "winter" months, among children ages 4 through 35 months; this peak began in the West during November and December and reached the Northeast by March. CONCLUSIONS: Diarrhea continues to be a common cause of hospitalization among children in the United States and the winter seasonality estimated to be caused in large part by rotavirus would be expected to decrease if rotavirus vaccines currently being developed were introduced. Our analysis of temporal trends in diarrheal hospitalizations provides a unique surrogate with which to estimate the disease burden associated with rotavirus diarrhea.

Bacterial Infections↗

Purification, characterization, and primary structure of Clostridium perfringens lambda-toxin, a thermolysin-like metalloprotease.

The lambda-toxin of Clostridium perfringens type B NCIB10691 was purified by ammonium sulfate precipitation, followed by size exclusion, anion-exchange, and hydrophobic interaction chromatography. The purified toxin had an apparent molecular mass of 36 kDa, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The toxin possessed casein-hydrolyzing activity, which was inhibited specifically by metal chelators, indicating that the toxin is a metalloprotease. The gene encoding the lambda-toxin (lam), which was shown by Southern analysis to be located on a 70-kb plasmid, was cloned into Escherichia coli cells. Nucleotide and N-terminal amino acid sequencing revealed that the lam gene encodes a 553-amino-acid protein, which is processed into a mature form, the molecular mass of which was calculated to be 35,722 Da. The deduced amino acid sequence of the mature enzyme contains an HEXXH motif characteristic of zinc metalloproteases and is homologous to other known enzymes belonging to the thermolysin family. The purified toxin degraded various biologically important substances, such as collagen, fibronectin, fibrinogen, immunoglobulin A, and the complement C3 component. It caused an increase in vascular permeability and hemorrhagic edema on injection into the dorsal skin of mice. These results suggest that the toxin contributes to the pathogenesis of histolytic infection by lambda-toxin-producing C. perfringens.

Amino Acid Sequence↗

Characterization of the major citrate synthase of Bacillus subtilis.

The major citrate synthase of Bacillus subtilis (CS-II) was purified to near homogeneity and shown to correspond to the product of the citZ gene. Accumulation of CS-II during exponential growth and stationary phases paralleled expression of the citZ gene. The physical and kinetic properties of CS-II were similar to those of citrate synthase enzymes from Bacillus megaterium and from eukaryotic cells but differed from those of citrate synthases from many gram-negative bacteria.

Amino Acid Sequence↗

Two-step secretion of the Serratia marcescens extracellular nuclease.

The extracellular nuclease of Serratia marcescens is one of a wide variety of enzymes secreted into the growth medium. Its appearance occurs late in the growth of a culture, and its gene, nucA, is transcriptionally regulated in a complex fashion by growth phase and other factors. Pulse-labeling studies reveal that extracellular secretion of nuclease occurs as a two-step process. In the first step, nuclease is rapidly translocated across the cytoplasmic membrane into the periplasm, where it accumulates as a mature active nuclease. A precursor protein, nuclease still carrying its signal sequence, was detected in the presence of carbonyl cyanide m-chlorophenylhydrazone or sodium azide, suggesting that this initial translocation and signal processing step involves an energy-dependent and Sec-dependent pathway in S. marcescens. The second step of secretion across the outer membrane is a slow process requiring between 30 to 120 min, depending on growth conditions.

Biological Transport↗

A Bacillus subtilis malate dehydrogenase gene.

A Bacillus subtilis gene for malate dehydrogenase (citH) was found downstream of genes for citrate synthase and isocitrate dehydrogenase. Disruption of citH caused partial auxotrophy for aspartate and a requirement for aspartate during sporulation. In the absence of aspartate, citH mutant cells were blocked at a late stage of spore formation.

Amino Acid Sequence↗

Mutations in two Ku homologs define a DNA end-joining repair pathway in Saccharomyces cerevisiae.

DNA double-strand break (DSB) repair in mammalian cells is dependent on the Ku DNA binding protein complex. However, the mechanism of Ku-mediated repair is not understood. We discovered a Saccharomyces cerevisiae gene (KU80) that is structurally similar to the 80-kDa mammalian Ku subunit. Ku8O associates with the product of the HDF1 gene, forming the major DNA end-binding complex of yeast cells. DNA end binding was absent in ku80delta, hdf1delta, or ku80delta hdf1delta strains. Antisera specific for epitope tags on Ku80 and Hdf1 were used in supershift and immunodepletion experiments to show that both proteins are directly involved in DNA end binding. In vivo, the efficiency of two DNA end-joining processes were reduced >10-fold in ku8Odelta, hdfldelta, or ku80delta hdf1delta strains: repair of linear plasmid DNA and repair of an HO endonuclease-induced chromosomal DSB. These DNA-joining defects correlated with DNA damage sensitivity, because ku80delta and hdf1delta strains were also sensitive to methylmethane sulfonate (MMS). Ku-dependent repair is distinct from homologous recombination, because deletion of KU80 and HDF1 increased the MMS sensitivity of rad52delta. Interestingly, rad5Odelta, also shown here to be defective in end joining, was epistatic with Ku mutations for MMS repair and end joining. Therefore, Ku and Rad50 participate in an end-joining pathway that is distinct from homologous recombinational repair. Yeast DNA end joining is functionally analogous to DSB repair and V(D)J recombination in mammalian cells.

Amino Acid Sequence↗

Differential gene expression by Pseudomonas aeruginosa during interaction with respiratory mucus.

Pseudomonas aeruginosa is a common respiratory tract pathogen that causes serious infections in patients with cystic fibrosis. A number of putative virulence factors have been characterized in several laboratories, and some have been implicated in human infections, based on criteria such as the phenotype of isolates from infected patients, an immune response to a particular antigenic factor, and the effect of a virulence factor on infectivity in an animal model. We have developed a series of genetic tools to study the selective regulation of expression of P. aeruginosa genes during interactions of the pathogen with host tissues. These tools are based on direct enrichment of bacteria, when a particular promoter is induced or repressed. We have found that interaction of bacteria with mucus from patients with cystic fibrosis results in marked induction of expression of several genes, including one that encodes a lipopolysaccharide biosynthetic enzyme, a gene for a protein responsible for uptake of the ferric pyochelin siderophore, and a new gene homologous with a class of iron-responsive repressors. The tools described here are useful for identification of induced or repressed genes in various animal models of infection or in controlled laboratory conditions that mimic natural infections of humans. Such genes might not be detectable when bacteria are cultured in laboratory conditions, and these tools are therefore useful for general probing of a bacterial genome for genes regulated during different stages of infection.

Animals↗

[Sixty years of study on history of diseases].

The study of history of diseases includes research on archeology, ancient medical works and systematic study on the history of diseases. By reviewing the process of 60 years, the paper points out that the process includes two approaches. On the one hand, it is defined by modern disease titles, offering historical background to cosmopolitan medicine and integrated Chinese and western medicine, and retifying the positive role of Chinese ancient medical workers' contribution to mankind. On the other hand, it also defined by traditional Chinese disease-syndrome titles, offering experiences for the development of Chinese medical academy and realization of modernization of Chinese medicine. Both approaches are mutually promoted, infiltrated, thus stimulated the deeper development for the study of history of diseases.

China↗

Determinants of suicide ideation: a comparison of Chinese and American college students.

A LISREL model that incorporates both social and psychological factors was used to explain Chinese and American college students' suicide ideation. Questionnaire data were obtained from one Chinese sample (N = 320) from four universities in Beijing and one American sample (N = 452) from one university in the Rocky Mountain area. As in the American sample, Chinese females score higher on the ideation scale than Chinese males, but the overall rate is lower for the Chinese than for the American college students. The findings in the American data support previous literature that family cohesion and religiosity are inversely related to suicide ideation, while the Chinese data suggest a positive correlation between religiosity and suicide ideation. This article offers a comparison of different cultural environments for Chinese and American adolescent development.

Adolescent↗

Gallium-67 scanning for detection of alveolitis in idiopathic pulmonary fibrosis and sarcoidosis.

OBJECTIVE: To investigate 67Gallium scanning for the detection of alveolitis in idiopathic pulmonary fibrosis and sarcoidosis. PATIENTS AND METHODS: 67Ga scintigraphy was performed in 14 patients with IPF (age: mean +/- s = 58.3 +/- 10.3 years), 13 patients with sarcoidosis (age: mean +/- s = 46.9 +/- 9.0 years) and 11 controls (age: mean +/- s = 44.0 +/- 10.5 years). RESULTS: Lung/thigh ratio was respectively 3.18 +/- 0.07 in IPF, 3.12 +/- 0.94 in sarcoidosis and 2.11 +/- 0.26 in controls. IPF and sarcoidosis groups had a significantly higher lung/thigh ratio comparing with the control group (P < 0.05). Three patients received 67Ga scanning examination after corticosteroid therapy and the 67Ga uptake decreased. CONCLUSIONS: The 67Ga scintigraphy is useful in monitoring the activity and extent of alveolitis.

Aged↗

[APC mutation analysis in sporadic colorectal cancer].

APC gene, responsible for familial adenomatous polyposiscoli, is one of the tumor suppressor genes involved in tumorigenesis of sporadic colorectal cancer. Because the majority of APC mutations are located in the 5' end of exon 15 and 95% of them generate stop codon, a coupled in vitro transcription and translation system (IVSP) was used to detect somatic APC mutation of exon 15 in sporadic colorectal cancer. Forty-nine mutated sporadic cases containing 59 truncated bands (9 cases with 2 bands) were identified. The results of the study showed: 1. APC frameshift mutations were more frequent than point mutations (60% vs 40%). Most of the frameshift mutations (32/35) were deletion or insertion of 1-2 bp, whereas 16 of all 23 point mutations presented C-to-T transitions. Mutations (57/59) were mostly but unevenly distributed in segment 2 and 3 of exon 15, while in segment 3, 69% (45/59) of all the mutations were predominantly located in mutation cluster region (codon 1286-1531). In seven of the 9 cases with 2 mutations their close proximity with each other is suggestive of the possibility of mutations at 2 independent alleles. The similarities between APC somatic and germline mutations suggest that APC gene might well have some common functions during the tumorigenesis of sporadic and hereditary coloretal cancer.

Colonic Neoplasms↗

[Testicular torsion: report of 12 patients].

12 cases of testicular torsion were stadieded. It is helpful to get diagnosis for early testicular torsion using radionuclide testicular imaging, B-ultrasound and color Doppler flow imaging. A peculiar type of testicular torsion was unilateral defect of radionuclide distribution. The causes, diagnoses and treatments of testicular torsion were discussed in detail.

Adolescent↗

Genetic evidence for direct sensing of phenolic compounds by the VirA protein of Agrobacterium tumefaciens.

The virulence (vir) genes of Agrobacterium tumefaciens are induced by low-molecular-weight phenolic compounds and monosaccharides through a two-component regulatory system consisting of the VirA and VirG proteins. However, it is not clear how the phenolic compounds are sensed by the VirA/VirG system. We tested the vir-inducing abilities of 15 different phenolic compounds using four wild-type strains of A. tumefaciens--KU12, C58, A6, and Bo542. We analyzed the relationship between structures of the phenolic compounds and levels of vir gene expression in these strains. In strain KU12, vir genes were not induced by phenolic compounds containing 4'-hydroxy, 3'-methoxy, and 5'-methoxy groups, such as acetosyringone, which strongly induced vir genes of the other three strains. On the other hand, vir genes of strain KU12 were induced by phenolic compounds containing only a 4'-hydroxy group, such as 4-hydroxyacetophenone, which did not induce vir genes of the other three strains. The vir genes of strains KU12, A6, and Bo542 were all induced by phenolic compounds containing 4'-hydroxy and 3'-methoxy groups, such as acetovanillone. By transferring different Ti plasmids into isogenic chromosomal backgrounds, we showed that the phenolic-sensing determinant is associated with Ti plasmid. Subcloning of Ti plasmid indicates that the virA locus determines which phenolic compounds can function as vir gene inducers. These results suggest that the VirA protein directly senses the phenolic compounds for vir gene activation.

Agrobacterium tumefaciens↗

Opposing effects of TGF-beta 2 on the Th1 cell development of naive CD4+ T cells isolated from different mouse strains.

The development of naive dense CD4+ T cells from different mouse strains toward Th1 cells, as monitored by measuring secondary IFN-gamma production, was affected by TGF-beta 2 in a differential way. Th1 cell development of naive CD4+ T cells from strains C57Bl/6, BALB/c, and NMRI primed by immobilized anti-CD3 mAb was strongly inhibited in the presence of TGF-beta 2. Even when the Th1 cell-inducer IL-12 was added, the same effect of TGF-beta 2 was observed. In contrast, Th1 development was substantially promoted by TGF-beta 2 with T cells from C3H/He and CBA/J mice. Further analyses using CD4+ T cells from (C57Bl/6xCBA/J)F1 hybrids or DBA/1 mice showed that Th1 development was inhibited by TGF-beta 2 if the T cells were activated by anti-CD3 mAb, but it was enhanced upon costimulation with anti-CD28 mAb. Determination of primary IL-2 production revealed that T cells from (C57Bl/6xCBA/J)F1 and DBA/1 mice produced low amounts of IL-2 following stimulation by anti-CD3 mAb alone and comparatively high amounts after coactivation by anti-CD28 mAb. In the presence of TGF-beta 2, the production of IL-2 was completely suppressed if such T cells were activated solely by anti-CD3 mAb, but it was only partially inhibited after costimulation by anti-CD28 mAb. Furthermore, TGF-beta 2-promoted Th1 development of such T cells was strongly inhibited after neutralization of endogenously produced IL-2 and completely restored by the addition of human IL-2. Thus, our results indicate that the TGF-beta 2-mediated stimulation of Th1 cell development requires the presence of relatively high concentrations of IL-2. Therefore, the opposing effect of TGF-beta 2 on the Th1 cell development of naive CD4+ T cells from different mouse strains appears to be the result of the variable potency of the respective CD4+ T cells to produce IL-2 in the presence of TGF-beta 2.

Animals↗

Polyomavirus persists in CD4/8 double-knockout, but not in CD4 or CD8 single-knockout mice.

The effects of incomplete immunocompetence on possible persistence and reactivation of polyomavirus in adult mice were investigated by a polyomavirus-specific polymerase chain reaction (PCR). The presence of virus DNA was followed between 4 days and 2 months postinfection (p.i.) in polyomavirus-infected normal adult A/Sn mice and CD4-/- and CD8-/- single-knockout, as well as CD4-/-8-/- double-knockout BALB/c or C57BL/6 mice. The same study was performed in A/Sn mice immunosuppressed by thymectomy (THX), cytosine-beta-D-arabinofuranoside (Ara-C) treatment, and total body irradiation (TBI). Primary polyomavirus infection of CD4-/- or CD8-/- single-knockout mice was similar to that obtained in normal adult mice when followed by PCR. Viral DNA was detected in a limited number of organs during 4 weeks p.i., but was no longer observed after 1-2 months. In contrast, the virus could be detected in most organs of CD4-/-8-/- double-negative mice and in THX-, Ara-C-, and TBI-treated adult mice and was still present 1-2 months p.i. In polyomavirus-infected normal adult mice a later immunosuppression did not lead to reactivation of the virus. Furthermore, if a second challenge of polyomavirus was administered 4 weeks after primary infection in both normal or recently immunosuppressed mice no viral DNA could be detected by PCR.

Animals↗

The Multicenter AIDS Cohort Study: retention after 9 1/2 years.

In a longitudinal, multicenter study of 4,954 men at risk for human immunodeficiency virus infection and acquired immunodeficiency syndrome, data from the first 9.5 years of follow-up (April 1984 through September 1993) were used to determine differences between those who remained in the study and those who dropped out. Demographic variables (age, race, education, employment, and study center), health status (human immunodeficiency virus type 1 serostatus and depression), and behavioral characteristics (alcohol drinking, drug use, and anal-receptive intercourse) were analyzed. Strategies for promoting retention included having frequent contact with participants, generating trust, keeping participants well-informed, utilizing multiple resources for follow-up, and providing flexible methods of participation. After 9.5 years of follow-up, vital status was known for 4,385 (88.5%) of the participants. Results from multiple logistic regression showed that race, age, education, and smoking were each significantly associated with nonparticipation (p < 0.001). A high level of retention was maintained in this well-educated and highly motivated cohort of homosexual/bisexual men. Extensive follow-up methods may improve case-finding. Nonwhite race, younger age, less education, and smoking were important predictors of dropping out. These findings identify specific groups for targeting follow-up efforts to reduce potential bias due to dropout.

Acquired Immunodeficiency Syndrome↗