1.Relative risk of virulence factors in Candida-infected mouse.
Donghwa KIM ; Woon Seob SHIN ; Kyoung Ho LEE ; Kyunghoon KIM ; Yoon Sun PARK ; Joo Young PARK ; Choon Myung KOH
Journal of the Korean Society for Microbiology 2000;35(4):317-324
Candida albicans is one of the most frequently isolated fungal pathogens in human. Recently, the prevalence of candida infection has markedly increased, partially due to the increase of immunocompromised hosts. Proposed virulence factors of the pathogenic Candida are the ability to form hyphae to adhere to epithelial cell surfaces, and to secrete acid proteinases and phospholipases. We measured the relative cell surface hydrophobicity (CSH) and the ability of proteinase production (PROT), phospholipase production (PLase), adherence to host epithelium (ADH), and hyphal transition (Germ). The relative risk of virulence factors was analyzed by lethality test in murine model of hematogeneously disseminated candidal infection. According to Cox's proportional hazard analysis, the statistically significant virulence factors were PROT, ADH, and CSH. PROT was the highest risk factor of them. To evaluate the applicability for the diagnosis and treatment of Candidiasis, we examined the protective effect of the active and passive immunizations with the materials purified from virulence factors and antibodies to them in Candia-infected mice model. The mean survival times of active and passive immunized groups were slightly longer than those of non-immunized groups.
Animals
;
Antibodies
;
Candida
;
Candida albicans
;
Candidiasis
;
Diagnosis
;
Epithelial Cells
;
Epithelium
;
Humans
;
Hydrophobic and Hydrophilic Interactions
;
Hyphae
;
Immunization, Passive
;
Immunocompromised Host
;
Mice*
;
Peptide Hydrolases
;
Phospholipases
;
Prevalence
;
Risk Factors
;
Survival Rate
;
Virulence Factors*
;
Virulence*
2.Isolation and identification of Vibrio vulnificus and Vibrio parahaemolyticus from coast of Pusan and Daechon.
Jin Woo JU ; Min Jung PARK ; Moon Soo HEO ; Cho Rok JUNG
Journal of the Korean Society for Microbiology 2000;35(4):309-316
This study was focused on the isolation of pathogenic Vibrio species, V. vulnificus and V. parahaemolyticus from marine environment from May to July of 1999. Isolation sites were coast near by Pusan and Daechon. The results obtained were as follows: 1. Seventy strains of V. parahaemolyticus and 19 strains of V. vulnificus were isolated from a total of 120 specimens. 2. Nineteen strains of V. vulnificus did not fermented arabinose and salicin but fermented lactose and cellobiose. All of V. parahaemolyticus isolates did not fermented lactose and cellobiose. 47 strains of V. parahaemolyticus fermented arabinose but 53 strains did not fermented salicin. 3. V. vulnificus and V. parahaemolyticus isolates showed three different API index numbers with 5046105 and 4346107 dominant. 4. V. vulnificus did not grow on 0% and 8% NaCl containing medium. V. parahaemolyticus grew on 8% NaCl containing medium. 5. V. vulnificus isolates and V. parahaemolyticus revealed different outer membrane protein p rofiles on SDS-PAGE.
Arabinose
;
Busan*
;
Cellobiose
;
Electrophoresis, Polyacrylamide Gel
;
Lactose
;
Membrane Proteins
;
Vibrio parahaemolyticus*
;
Vibrio vulnificus*
;
Vibrio*
3.Analysis of X irradiation related genes in HL60 cells using cDNA microarray.
Keon Uk PARK ; Mi Sun HWANG ; Seong Il SUH ; Min Ho SUH ; Taeg Kyu KWON ; Jong Wook PARK ; Jae We CHO ; Eun Ju CHOI ; Won Ki BAEK
Journal of the Korean Society for Microbiology 2000;35(4):299-308
Recently developed cDNA microarray or DNA chip technology allows expression monitoring of expression of hundreds and thousands of genes simultaneously and provides a format for identifying genes as well as changes in their activity. In order to search for changes in gene expression after X irradiation in HL60 cells, cDNA microarray technique was done. In this study, expression of 588 human genes (including oncogenes, tumor suppressor genes, cell cycle regulator genes, intracellular signal transduction modulator genes, apoptosis related genes, transcription factor genes, growth factors and receptor genes, cytokine genes, etc) were analyzed. For cDNA microarray analysis mRNAs were extracted from control and 8 Gy-irradiated HL60 cells. As a result the changes in expression of several genes were observed. This alteration of gene expression was confirmed by reverse transcription-polymerase chain reaction. The expression of heat shock 60 KD protein, c-jun, erythroid differentiation factor, CPP32, myeloid cell nuclear differentiation antigen, MAP kinase-activated protein kinase, interleukin-8, monocyte chemotactic peptide 1 and RANTES genes was increased, but the expression of p55CDC gene was decreased after X irradiation.
Apoptosis
;
Cell Cycle
;
Chemokine CCL5
;
DNA, Complementary*
;
Gene Expression
;
Genes, Regulator
;
Genes, Tumor Suppressor
;
HL-60 Cells*
;
Hot Temperature
;
Humans
;
Intercellular Signaling Peptides and Proteins
;
Interleukin-8
;
Monocytes
;
Myeloid Cells
;
Oligonucleotide Array Sequence Analysis*
;
Oncogenes
;
Protein Kinases
;
RNA, Messenger
;
Shock
;
Signal Transduction
;
Transcription Factors
4.Comparison of infrequent restriction site-polymerase chain reaction and pulsed-field gel electrophoresis for molecular typing of Staphylococcus aureus and Escherichia coli.
Wan Shik SHIN ; Tai Gye KIM ; Jung Hyun CHOI ; Dong Gun LEE ; Hee Baeg CHOI ; Jin Hong YOO ; Jong Hyun KIM ; Jin Han KANG ; Woo Sung MIN
Journal of the Korean Society for Microbiology 2000;35(4):289-297
BACKGROUND: Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) are major pathogens in community and hospital. And they sometimes cause the outbreak in hospital in the immunocompromised patients. Pulsed-field gel electrophoresis (PFGE) has been regarded as a standard method for genotyping in epidemiologic studies, but it is laborious and time-consuming. Infrequent restriction site-polymerase chain reaction (IRS-PCR), a new genotyping methods, was performed to compare the applicability with PFGE. METHODS: We performed PFGE and IRS-PCR on S. aurues (n=120) and E. coli (n=117) which were collected clinically in 4 different hospitals. We assessed each method in terms of discriminatory power, quality, and efficiency. RESULTS: In E. coli, the discriminatory power of IRS-PCR was 46.7apprx86.7%, and that of PFGE was 88.9apprx96.7% according to hospital. But in S. aurues, the discriminatory power of IRS-PCR was 20apprx56.7%, and that of PFGE was 40apprx90% according to hospital. The typicality and reproducibility of IRS-PCR were 100% of each. PFGE needed four days to complete the procedure, but IRS-PCR could be performed within one day, IRS-PCR showed better resolution than PFGE. CONCLUSION: In case of gram negative bacteria (like E. coli), IRS-PCR could be a reliable alternative for epidemiologic typing due to better efficiency and comparable discriminatory power. But in the case of gram positive bacteria (like S. aureus), IRS-PCR does not seem to be suitable for the strain-to-strain differentiation. More trials and changes of restriction enzymes or primers could reveal the efficacy of IRS-PCR in the field of molecular typing.
Electrophoresis, Gel, Pulsed-Field*
;
Escherichia coli*
;
Escherichia*
;
Gram-Negative Bacteria
;
Gram-Positive Bacteria
;
Immunocompromised Host
;
Molecular Typing*
;
Staphylococcus aureus*
;
Staphylococcus*
5.A novel PCR primers HPU185 and HPL826 based on 16S rRNA gene for detection of Helicobacter pylori.
Jong Bae KIM ; Geun Hee KIM ; Hong KIM ; Hyun Seok JIN ; Young Sam KIM ; Soo Hyun HA ; Dong Ki LEE
Journal of the Korean Society for Microbiology 2000;35(4):283-288
The PCR primer set JW21-JW22 of Weiss et al. (19), which was reported to amplify a 139-bp fragment of the 16S rRNA gene of Helicobacter pylori, has been recently used for the detection of H. pylori in clinical specimens. However, when we applied JW21-JW22 PCR to other members of the genus Helicobacter and unrelated microorganisms, all of these bacteria produced a 139-bp PCR product. Therefore, we designed a novel primer set, HPU185-HPL826, which produced a 642-bp amplicon of the 16S rRNA gene of H. pylori. Then we further examined the specificity of the novel PCR assay using Southern blot hybridization with an internal probe, HPP225. The PCR assay described in this study was shown to be highly sensitive and specific only to the H. pylori 16S rRNA gene sequences.
Bacteria
;
Blotting, Southern
;
Genes, rRNA*
;
Helicobacter pylori*
;
Helicobacter*
;
Polymerase Chain Reaction*
;
Sensitivity and Specificity
6.Overexpression, purification and truncation analysis of RmlC protein of Mycobacterium tuberculosis.
Jong Seok LEE ; Tae Yoon LEE ; Jae Ho PARK ; Jong Sun KIM ; Tae Jin LEE ; Jai Youl LEE ; Sung Kwang KIM
Journal of the Korean Society for Microbiology 2000;35(4):273-282
dTDP-rhamnose provides L-rhamnose to the bridge-like structure between mycolyl arabinogalactan and peptidoglycan of the mycobacterial cell wall. dTDP-rhamnose is composed of glucose-1-phosphate and dTTP by four enzymes encoded by rmlA-D. To determine the region(s) of RmlC protein essential for its dTDP-4-keto-6-deoxyglucose epimerase activity, we overexpressed both whole (202 amino acids) and three different truncated (N-terminal 106 or 150 or C-terminal 97 amino acids) RmlC proteins of Mycobacterium tuberculosis. The RmlC enzyme activity in the soluble lysates of DELTArmlC E. coli strain SPHI874 (DE3 PlysS) expressing the wild type or truncated rmlC genes was initially analyzed by three sequential reactions from dTDP-glucose to dTDP-rhamnose in the presence of purified RmlB and RmlD. All three soluble lysates containing the truncated RmlC proteins showed no enzyme activity, while that containing the wild type RmlC was active. This wild type RmlC was then overexpressed and purified. The incubation of the purified RmlC enzyme so obtained with dTDP-4-keto-6-deoxyglucose resulted in the conversion of dTDP-4-keto-rhamnose. The results show that the truncated regions of the RmlC protein are important for the RmlC enzyme activity in M. tuberculosis.
Cell Wall
;
Mycobacterium tuberculosis*
;
Mycobacterium*
;
Peptidoglycan
;
Tuberculosis
7.Detection of Multidrug Resistant Patterns and Associated - genes of Methicillin Resistant Staphylococcus aureus ( MRSA ) Isolated from Clinical Specimens.
Eun Gyoung LIM ; Young Hee KIM ; Ji Yung MUN ; Yung Bu KIM ; Yang Hyo OH
Journal of the Korean Society for Microbiology 2000;35(5):356-356
No Abstract Available.
Methicillin Resistance*
;
Methicillin*
;
Methicillin-Resistant Staphylococcus aureus*
;
Staphylococcus aureus*
;
Staphylococcus*
8.Virulence Factors and Genotyping of Vibrio parahaemolyticus.
Eun Gyoung LIM ; Young Hee KIM ; Ji Yung MUN ; Yang Hyo OH ; Yung Bu KIM
Journal of the Korean Society for Microbiology 2000;35(5):355-355
No Abstract Available.
Vibrio parahaemolyticus*
;
Vibrio*
;
Virulence Factors*
;
Virulence*
9.Detection of Multidrug Resistant Patterns and Associated - genes of Methicillin Resistant Staphylococcus aureus ( MRSA ) Isolated from Clinical Specimens.
Eun Gyoung LIM ; Young Hee KIM ; Ji Yung MUN ; Yung Bu KIM ; Yang Hyo OH
Journal of the Korean Society for Microbiology 2000;35(5):356-356
No Abstract Available.
Methicillin Resistance*
;
Methicillin*
;
Methicillin-Resistant Staphylococcus aureus*
;
Staphylococcus aureus*
;
Staphylococcus*
10.Virulence Factors and Genotyping of Vibrio parahaemolyticus.
Eun Gyoung LIM ; Young Hee KIM ; Ji Yung MUN ; Yang Hyo OH ; Yung Bu KIM
Journal of the Korean Society for Microbiology 2000;35(5):355-355
No Abstract Available.
Vibrio parahaemolyticus*
;
Vibrio*
;
Virulence Factors*
;
Virulence*
Result Analysis
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