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Identification of Mycobacterium avium complex ( MAC ) clinical Strains to a Species Level by Sequencing and PCR - SSCP Analysis of rpoB DNA.

Bum Joon KIM ; Seung Hyun LEE ; Keun Hwa LEE ; Chung Kyu PARK ; Myung Sik CHOI ; Ik Sang KIM ; Sung Bai CHOI ; Eung Su HWANG ; Chang Yung CHA ; Sang Jae KIM ; Gill Han BAI ; Yoon Hoh KOOK

Journal of the Korean Society for Microbiology.1999;34(5):491-500.

A recent study showed that comparative sequence analysis of rpoB DNAs could reveal natural relationships in genus Mycobacterium [J Clin Microbiol. 37 (6). 1999]. rpoB DNAs showed interspecies variation and intraspecies conservation, Based on these data, we developed polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) protocols which enable species differentiation in genus Mycabacterium. When this assay was applied to 24 clinical isolates identified as M. avium complex (MAC) by biochemical test, these were successfully differentiated into M. avium and M. intracellulare. These results were concordant with those obtained by 16s rDNA analysis. It is the first report that PCR-SSCP analysis of rpoB DNA could be used for species differentiation of MAC strains.
DNA* ; DNA, Ribosomal ; Mycobacterium avium Complex* ; Mycobacterium avium* ; Mycobacterium* ; Polymerase Chain Reaction* ; Polymorphism, Single-Stranded Conformational* ; Sequence Analysis

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Expression of Cyclooxygenase - 2 in Intestinal Epithelial Cells in Response to Invasive Bacterial Infection and its Role of Epithelial Cell Apoptosis.

Jung Mogg KIM ; Shin Jae KANG ; Yang Ja CHO

Journal of the Korean Society for Microbiology.1999;34(5):479-489.

Invasion of enteric bacteria, such as Salmonella and invasive E. coli, into intestinal epithelial cells induces proinflammatory gene responses and finally epithelial cell apoptosis. In this study, we asked whether invasive bacterial infection of human intestinal epithelial cells could upregulate cyclooxygenase-2 (COX-2) gene expression and whether increased COX-2 expression could influence intestinal epithelial cell apoptosis. Expression of COX-2 mRNA and prostaglandin (PG) E production were upregulated in HT-29 colon epithelial cells which were infected with S. dublin or invasive E. coli, as examined by quantitative RT-PCR and radioimmunoassay. Inhibition of COX-2 expression and PGE2 production using NS-398, a specific COX-2 inhibitor, showed a significant increase af epithelial cell apoptosis and caspase-3 activation in HT-29 cells infected with invasive bacteria. However, the addition of valerylsalicylate, a specific COX-1 inhibitor, did not change apoptosis in S. dublin-infected HT-29 cells. These results suggest that upregulated COX-2 expression and PGE2 production in response to invasive bacterial infection could contribute to host defense by inhibiting apoptosis of intestinal epithelial cells.
Apoptosis* ; Bacteria ; Bacterial Infections* ; Caspase 3 ; Colon ; Cyclooxygenase 2 ; Dinoprostone ; Enterobacteriaceae ; Epithelial Cells* ; Gene Expression ; HT29 Cells ; Humans ; Prostaglandin-Endoperoxide Synthases* ; Radioimmunoassay ; RNA, Messenger ; Salmonella

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Study on Pathogenicity of Borrelia burgdorferi Sensu Lato Isolated in Korea.

Kyung Hee PARK ; Jung A LIM ; Jong Hyun KIM ; Eui U PARK

Journal of the Korean Society for Microbiology.1999;34(5):471-478.

Spirochetes were isolated from the midgut of Ixodes persulcatus ticks captured at Chungju, Korea and identified as Borrelia afzelii strains by polymerase chain reaction. To determine the pathogenicity of the B. afzelii strains isolated in Korea, the microbiological and pathological features of Lyme disease were observed in C3H/He mice after intraperitoneal inoculation of the fresh isolate of B. afzelii strain. The results are summarized as follows 1) The Borrelia were detected in the tissues of heart, spleen, kidney, urinary bladder and knee joint within 7 days after inoculation of infection by dark field microscopic examination. The isolation rate from heart, urinary bladder and joint was significantly higher than the rate from spleen, kidney, and blood samples. 2) The Borrelia was detected in heart muscle by indirect immunofluorescent antibody test. 3) Antibody to the Borrelia was detected as early as one week after inoculation. 4) The marked tropism of the Borrelia was observed in myocardial, urinary tract and joint tissue. The main pathological features are inflammation in tissues of heart, kidney, joint and urinary bladder. From these results, the Borrelia afzelii strain isolated in Korea were determined as pathogenic strain.
Animals ; Borrelia burgdorferi Group* ; Borrelia burgdorferi* ; Borrelia* ; Chungcheongbuk-do ; Heart ; Inflammation ; Ixodes ; Joints ; Kidney ; Knee Joint ; Korea* ; Lyme Disease ; Mice ; Myocardium ; Pathology ; Polymerase Chain Reaction ; Spirochaetales ; Spleen ; Ticks ; Tropism ; Urinary Bladder ; Urinary Tract ; Virulence*

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Purification Siderophore from Vibrio mimicus ATCC 33653 and its Effect to Bacterial Pathogenecity.

Soo Jung PARK ; Seong A JU ; Moon Soo HEO ; Cho Rock JUNG ; Jin Woo JU

Journal of the Korean Society for Microbiology.1999;34(5):461-470.

Growth under conditions of iron-restriction and the production of siderophore was examined in Vibrio mimicus ATCC 33653. This strain grew and multiplied in the presence of the high-affinity iron chelators ethylenediamine-di (o-hydroxyphenylacetic acid). Chrorne azurol S (CAS) agar and solution were used to detect the production of siderophore under these condition. Siderophore could be detected in the iron-rcstricted culture supernatants. The siderophore was extracted from iron-restricted culture supernatants by phenol-chloroform-ether method and purified by Dowex ion-exchange and Sephadex G-25 gel filtracton chromatography. The purified siderophore was confirmed by paper chromatography and HPLC. The Purified siderophore enhanced the growth of V. mimicus when the bacterium was grown in iron limited medium. Injection of both the siderohore and the bacteria to mice resulted in more rapid death than that of the only bacteria. However, the siderophore did not show lethality to mice and any toxicity to cell line like HeLa and U937.
Agar ; Animals ; Bacteria ; Cell Line ; Chelating Agents ; Chromatography ; Chromatography, High Pressure Liquid ; Chromatography, Paper ; Iron ; Mice ; Vibrio mimicus* ; Vibrio*

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Detection of Astrovirus Infection from Hospitalized Young Children Feces by Reverse Transcription - Polymerase Chain Reaction.

Hae Kyung PARK ; So Youn WOO ; Ju Young SEOH ; Young Hae CHONG ; Jeong Wan SEO

Journal of the Korean Society for Microbiology.1999;34(5):453-459.

Astrovirus is frequently associated with diarrhea in children. It can not be readily isolated by cell culture, and an electronmicroscope is usually used for detection of this agent. Recently in 1995 a combined method of reverse transcription-polymerase chain reaction (RT-PCR) was designed for easier detection of astrovirus, which is based on the conserved sequence in 3'-end of genomes of the 7 known serotypes of human astrovirus. As of yet there has not been any report of astrovirus data in Korea using the RT-PCR methods. The purpose of this study was to detect astrovirus incidence, severity of symptoms, seasonal variation and coinfection rate with rotavirus in Korean children inpatients with diarrhea. Fecal specimens from 61 young children hospitalized with gasteroenteritis Korea from Jan. 1996 through Mar. 1997. They were examined for astroviurs infection by RT-PCR method. Results are as follows: 1. Astrovirus was detected at 9.8% (6/61) from fecal specimens of children with severe diarrhea by EIA using monoclonal antibody coated plates. 2. Astorvirus was detected at 29.5% (18/61) from fecal specimens of children with severe diarrhea by RT-PCR. 3. The age of the 18 children affected by astrovirus ranged from 2 monthes to 7 years with mean of 3.0 years. 4. Mean hospital stay of the 1S children was 6.1 days. 5. Five (27.8%) astrovirus RT-PCR positive strains were confirmed in November and in December, respectively out of 18 specimens in total. 6. Astrovirus coinfection with rotavirus type G1 was confirmed in 15/16 specimens (93.8%), and with type G2 was in 1/16 specimens (6.3%).
Cell Culture Techniques ; Child* ; Coinfection ; Conserved Sequence ; Diarrhea ; Feces* ; Genome ; Humans ; Incidence ; Inpatients ; Korea ; Length of Stay ; Mamastrovirus ; Polymerase Chain Reaction* ; Reverse Transcription* ; Rotavirus ; Seasons

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Plasmid Profile and B - Lactamase Type of Multidrug - Resistant Salmonella typhi Isolated from Korea, 1997.

Jung Sik YOO ; Young Hack SHIN ; Kyung Soo OH ; Jeom Kyu LEE ; Ki Sang KIM

Journal of the Korean Society for Microbiology.1999;34(5):445-452.

Eight strains of multidrug-resistant (MDR) Salmonella typhi were isolated from Kyonggi area during January-February,1997. They were resistant to ampiciUin, amoxicillin, carbeniciillin, tetracycline, chloramphenicol, trimethoprim/sulfamethoxazole, trimethoprim. Eight strains had one plasmid respectively which size was approximately M.W 220 kb and showed same restriction pattern by endonuclease HindIII. The plasmid was similar to the plasmid in size that was related to multidrug resistant S. typhi isolated from southeast Asia. It were transferred by conjugation to recipient E, coli K-12 in frequency of 2.43 x10-4 - 1.73 x 10-2 and transconjugant showed same drug-resistant pattem with donor cells. All of 8 strains produced B-lactamase that was assummed to TEM-1 type by isoelectric focusing and PCR.
Amoxicillin ; Asia, Southeastern ; Chloramphenicol ; Deoxyribonuclease HindIII ; Gyeonggi-do ; Humans ; Isoelectric Focusing ; Korea* ; Plasmids* ; Polymerase Chain Reaction ; Salmonella typhi* ; Salmonella* ; Tetracycline ; Tissue Donors ; Trimethoprim

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Infectivity of Orientia tsutsugamushi to Various Eukaryotic Cells and Their Cellular Invasion Mechanism.

Kyung Soo IHN ; Seung Hoon HAN ; Hang Rae KIM ; Seung Yong SEONG ; Ik Sang KIM ; Myung Sik CHOI

Journal of the Korean Society for Microbiology.1999;34(5):435-443.

Orientia tsutsugamushi is obligate intracellular bacterium that grows within the cytoplasm of the eukaryotic host cells. Therefore capability of the attachment, entry into the host cell and intracellular survival should be critical process for oriential infection. In this study we investigated the cellular invasion mechanism of Orientia tsutsugamushi and the role of transmembrane heparan sulfate proteoglycan, which binds diverse components at the cellular microenvironment and is implicated as host cell receptors for a variety of microbial pathogens. First of all Orientia tsutsugamushi can invade a wide range of nonprofessional phagocytic cells including fibroblast, epithelial cells a#nd endothelial cells of various host species, including B and T lymphocytes. Thus, it was postulated that the attachment of O. tsutsugamushi requires the recognition of ubiquitous surface structures of many kinds of host cells. Treatments with heparan sulfate and heparin inhibited the infection of Orientia tsutsugamushi in dose-dependent manner for L cell, mouse fibroblast, whereas other glycosaminoglycans such as chondroitin sulfate had no effect. Collectively, these findings provide strong evidence that initial interaction with heparan sulfate proteoglycan is required for the oriential invasion into host cells.
Animals ; Cellular Microenvironment ; Chondroitin Sulfates ; Cytoplasm ; Endothelial Cells ; Epithelial Cells ; Eukaryotic Cells* ; Fibroblasts ; Glycosaminoglycans ; Heparan Sulfate Proteoglycans ; Heparin ; Heparitin Sulfate ; Mice ; Orientia tsutsugamushi* ; Phagocytes ; T-Lymphocytes

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Characterization of a Vibrio parahaemolyticus Phage Isolated from Marine.

Sun Ok YOON ; Seong A JU ; Moon Soo HEO ; Cho Rok JUNG ; Jin Woo JU

Journal of the Korean Society for Microbiology.1999;34(5):423-433.

A novel bacteriophage, designated as VPP97, that infects the strains of Vibiro parahaemolyticus (hallophilic, Gram-negative bacterium) isolated most commonly from marine environments, has been discovered, and several of its properties have been determined. The plaques were clear and sized 0.6-1.0 mm in diameter. The virion forms a single band on 70% sucrose gradient and p1.50 CsC1 gradient by sucrose gradient centrifugation and CsCI gradient centrifugation respectively. It has a hexagonal head and a relatively long tail, as shown by electron microscopy. Vibrio alginolyticus, Vibrio fluvialis and Vibrio furnissii were also sensitive to this phage It was almost totally inactivated at 70 degree C and at pH below 5 or over 10. The nucleic acid of VPP97 is composed of DNA. The VPP97 had 9 specific structural proteins sized between 21.5 kDa and 97.4 kDa on SDS-PAGE. When V. parahaemolyticus cultures were treated with either phage VPP97 or one of the several antibiotics for 2 hours, the viable number of V. parahaemolyticus treated with the phage VPP97 is lower than that treated with chloramphenicol, erythromycin or penicillin, but not lower than that treated with tetracycline. Mice that have responded to the phage treatment revealed the lower numbers of V. parahaemolyticus in small intestine and less damage on small intestine compared to the untreated mice. Therefore, we suggest that the phage treatment appears effective to the infection by V. parahaemolyticus.
Animals ; Anti-Bacterial Agents ; Bacteriophages* ; Centrifugation ; Chloramphenicol ; DNA ; Electrophoresis, Polyacrylamide Gel ; Erythromycin ; Head ; Hydrogen-Ion Concentration ; Intestine, Small ; Mice ; Microscopy, Electron ; Penicillins ; Sucrose ; Tail ; Tetracycline ; Vibrio alginolyticus ; Vibrio parahaemolyticus* ; Vibrio* ; Virion

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Inhibition of IL-2 dependent DTLL-2 proliferation by immune complex from patient with ovarian cancer.

Sang Deuk CHUNG ; Chang Hwan PARK ; Yong Hoon CHUNG ; Kyung Hee KIM ; Yang Ja CHO

Journal of the Korean Society for Microbiology.1993;28(4):331-327.

No abstract available.
Antigen-Antibody Complex* ; Humans ; Interleukin-2* ; Ovarian Neoplasms*

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Generation of monoclonal antibodies reactive to human interleukin 2(IL 2).

Youn Mun HA ; Jun Hyung LEE ; Seung Min YOO ; Jeong Je CHO ; Soon Tae HO ; Jae Kyung PARK ; Sang Yun NAM

Journal of the Korean Society for Microbiology.1993;28(4):313-319.

No abstract available.
Antibodies, Monoclonal* ; Humans* ; Interleukins*

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