No Abstract Available.
DNA* ; Interleukin-12* ; Mycobacterium tuberculosis* ; Mycobacterium*

No Abstract Available.
Brucella abortus* ; Brucella* ; Immunity, Humoral* ; Jeju-do* ; Recombinant Proteins*

We purified enolase from Candida albicans KNIH10 strain which was isolated from a clinical specimen in Korea. The purified enolase was used to detect anti-Candida antibodies in sera of patients with invasive candidiasis. For purification of enolase from the crude extract prepared by French pressure at 20,000 PSI, the fast performance liquid chromatography (FPLC) using DEAE-sepharose column was used. The elutes at 0.3-0.4 M NaCl in FPLC was purified with homogenity in SDS-PAGE and its enzymatic activity was confirmed in sera of invasive candidiasis with candidemia patient by immunoblotting. The purified enolase indicated no siggnal (100% specificity) in 40 normal human sera and 75% (6/8) sensitivity in sera of candidemic patients with suspicious invasive candidiasis by immunoblotting.
Antibodies ; Candida albicans* ; Candida* ; Candidemia ; Candidiasis, Invasive ; Chromatography, Liquid ; Electrophoresis, Polyacrylamide Gel ; Humans ; Immunoblotting ; Immunologic Tests* ; Korea* ; Phosphopyruvate Hydratase*

Acinetobacter baumannii strains are emerging pathogens of the nosocomial infection with an increasing frequency in recent years. The therapeutic difficulty due to the wide spread of multiple resistant strains was major problem in A. baumannii infection. It seems likely that high frequency of A. baumannii infection will be increasing epidemiological importance in the future. However, the current limited understanding of the epidemiology of A. baumannii infections is caused by lack of a rapid and practical method for the molecular characterization of A. baumannii strains. This study was undertaken to determine molecular types and genetic similarity among A. baumannii strains isolated from four hospitals by RAPD analysis. Eighty-five strains, including 40 from Chunnam University Hospital, 27 from Dankook University Hospital, 15 from Yonsei University Hospital, and 3 from Seonam University Hospital, were classified into three molecular types. Molecular type II was the most common pattern and included 72 strains. All strains from Dankook University Hospital and 40 strains from Chunnam University Hospital belonged to molecular type I or II. A. baumannii strains form Yonsei University Hospital were very distant similarity values. The range of genetic similarity values among 85 strains of A. baumannii was 0.26 to 1.00. Although phenotypes including biotype and antimicrobial resistance pattern of A. baumannii strains were same or very similar to each other, their RAPD patterns were quite different. Typing with phenotypes was found to be less reliable than molecular typing by RAPD analysis. These results suggest that RAPD analysis provides rapid and simple typing method of A. baumannii strains for epidemiological studies. This work is the first epidemiological report of A. baumannii infections in Korea and it is hoped that results of this work may contribute to a better understanding of the clinical importance and epidemiology of A. baumannii strains.
Acinetobacter baumannii* ; Acinetobacter* ; Cross Infection ; DNA* ; Epidemiology ; Hope ; Korea ; Molecular Typing* ; Phenotype

The non-ELR-containing CXC chemokines Mig and IP-10 have been shown to function as chemotactic cytokines for activated T lymphocytes. In this study, we examined the potential involvement of Mig and IP-10 in antimycobacterial response of mice immunized or infected with M. bovis BCG. The accumulation of Mig and IP-10 mRNA in resident peritoneal monocytes (RPMPHI) was slightly reduced by stimulation with vBCG, and the degree was greater for 24 hr culture even though IFN-gamma was added. Expression of Mig, IP-10, and IFN-gamma in 24 hr delayed-type hypersensitivity (DTH) response was stronger in vBCG-immune mice than in the non-immune. The increase of DTH measured by foot-pad thickness appears to be clearly related to the levels of chemokines Mig and IP10 messages and those of IFN-gamma and IL-12. Stimulation with vBCG for 2 days decreased or completely dropped the levels of Mig message in non-immune or immune splenocytes, respectively, whereas IP-10 message was slightly decreased in 2 days culture. Moreover, messages for IL-12 (p40) showed similar kinetics for Mig. The levels of Mig and IP-10 mRNA during the course of infection with BCG were not readily changed in lungs, livers, and spleens from BCG-infected mice. Although there was no obvious changes of Mig and IP-10 messages in the target organs during infection process, we found that the infection progressed over the first 3 wk before being contained by the emerging immune response suggested from detectable amount of IFN-gamma mRNA around this time. In view of selectivity of chemokines Mig and IP-10 for activated T cells, these data suggest that chemokine Mig and IP-10, especially in collaboration with IL-12 and IFN-gamma, may play a role as T cell recruiters in immune response against mycobacterial infection.
Animals ; Chemokines ; Chemokines, CXC* ; Cooperative Behavior ; Hypersensitivity ; Interleukin-12 ; Kinetics ; Liver ; Lung ; Mice* ; Monocytes ; Mycobacterium bovis* ; RNA, Messenger ; Spleen ; T-Lymphocytes

FtsH is a membrane-bound, ATP-dependent metalloprotease that is involved in a variety of cellular functions including the regulation of responses to heat and stress shock. Previously, we had cloned and sequenced pneumococcal ftsH gene whose deduced amino acid sequence was very similar to those of several gram-positive bacteria and Escherichia coli, except for the N-terminal domain that was responsible for membrane anchoring. In order to better understand the role of Streptococcus pneumoniae FtsH, we expressed pneumococcal ftsH gene in Escherichia coli. When it was expressed from a strong promoter, Ptac, a considerable amount of the recombinant FtsH was produced, although the prolonged induction resulted in not only accumulation of breakdown products but also ceasing of the further growth of E. coli host. This indicated that the expression of the exogenous ftsH gene was tightly regulated since the excessive FtsH appeared detrimental to bacterial cells. In Western blotting, the pneumococcal FtsH protein, whether native or recombinant, was reactive to anti-E. coli FtsH serum. The observation that FtsH proteins were well conserved throughout the bacterial kingdom and its expression level was fine-tuned suggests an important role for this protein in the stress adaptation which may be related to infecting process by pneumococci.
Amino Acid Sequence ; Blotting, Western ; Clone Cells ; Escherichia coli ; Gram-Positive Bacteria ; Hot Temperature ; Membranes ; Shock ; Streptococcus pneumoniae* ; Streptococcus*

Two-dimensional gel electrophoresis (2-DE) is an essential tool of proteomics to analyse the entire set of proteins of an organism and its variation between organisms. Helicobacter pylori was tried to identify differences between strains. As the first step, whole H. pylori was lysed using high concentration urea contained lysis buffer (9.5 M Urea, 4% CHAPS, 35 mM Tris, 65 mM DTT, 0.01% SDS and 0.5% Ampholite (Bio-Rad, pH 3-10)). The extract (10 mug) was rehydrated to commercially available immobilised pH gradient (IPG) strips, then the proteins were separated according to their charges as the first dimensional separation. The IPG strips were placed on Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) to separate according to molecular mass of the proteins as the second dimension. The separated protein spots were visualised by silver staining in order to compare different expression of proteins between strains. Approximately 120 spots were identified in each mini-protein electrophoresised gel, furthermore about 65 to 75 spots were regarded as identical proteins in terms of pI value and molecular weight between strains used. In addition, distinct differences were found between strains, such as 219-1, Y7 and Y14, CH150. Two representative strains were examined using strips which had pH range from 4 to 7. This strips showed a number of isoforms which were considered large spots on pH range 3-10. Furthermore, the rest of spots on pH 4-7 IPG strips appeared very distinctive compared to broad range IPG strips. 2-DE seems to be an excellent tool for analysing and identifying variations between H. pylori strains.
Electrophoresis ; Electrophoresis, Gel, Two-Dimensional* ; Helicobacter pylori* ; Helicobacter* ; Hydrogen-Ion Concentration ; Molecular Weight ; Protein Isoforms ; Proteomics ; Proton-Motive Force ; Silver Staining ; Sodium ; Urea

The viruses of Herpes Simplex Virus 1 (HSV-1), Herpes Simplex Virus 2 (HSV-2) and Varicella-Zoster virus (VZV) which belong to the alpha herpes subfamily are important human pathogens. When eruptions were not fully developed from these viral infections, clinical diagnosis was not always easy and required virological confirmation test. The above viruses were reactivated in individuals who were compromised in immune competence for one reason or another. Polymerase chain reaction (PCR) enables rapid and sensitive detection of HSV and VZV DNAs. Its sensitivity was largely influenced by choice of primers. Authors conducted a study to detect of those three viruses in human specimens including vesicle fluid and joint fluid and serum using PCR methods. Primers used for this study were the general primer pair GPHV-RU which was known to amplify within the genes enjoying the highest degree of homology between UL15 of HSV and UL42 of VZV. PCR with primers hybridized pair GPHV-RU amplifies a 396 bp with HSV-1 and HSV-2 standard stain DNA and 405 bp with VZV standard strain DNA. Restriction enzyme cleavage with HpaII and DdeI were used to detect and distinguish DNAs of HSV-1 and HSV-2 and VZV. The purpose of this study was a rapid and easy detection of VZV and HSV-1 or HSV-2 from various clinical specimens (vesicle fluid, serum and joint fluid) by PCR method. Used methods were: HSV PCR with primer 1, 2 and HpaII RE digestion; VZV nested PCR; HSV PCR with primer A, B and BssHII RE digestion. 1) In 33 cases (33/42, 78.6%) VZV was detected single or mixed infection from 42 clinical specimens which included vesicle fluid (5), serum from respiratory infected children (10), serum from immune suppressed adult cancer patients (7) and joint fluid from arthritis patients (20). 2) In 20 cases (20/42, 42.6%) HSV was detected singly or mixed infection and 19 of the cases were HSV-2 and 1 case was HSV-1. 3) In 19 cases (19/42, 45.2%) VZV was singly detected which included serum from respiratory infected children (6 cases), joint fluid from arthritis patients (9 cases), vesicle fluid (2 cases) and serum form immunosuppressed cancer patients (2 cases). 4) HSV was singly detected in 6 cases (6/42, 14.3%) which included joint fluid from arthritis patients (5 cases) and serum form respiratory infected children (1 cases). 5) 14 cases of VZV and HSV mixed infection (14/42, 33.3%) were detected. They included vesicle fluid (3 cases), serum form immunosuppressed cancer patients (4 cases), serum from respiratory infected children (2 cases) and joint fluid from arthritis patients (5 cases). 6) HSV-1 and HSV-2 detection and typing by HSV PCR with primer A, B and BssHII RE digestion method was more sensitive and the results were easier to detect than on other method.
Adult ; Arthritis ; Child ; Coinfection ; Diagnosis ; Digestion ; DNA ; Herpes Simplex* ; Herpesvirus 1, Human* ; Herpesvirus 2, Human ; Herpesvirus 3, Human* ; Humans ; Joints* ; Mental Competency ; Polymerase Chain Reaction* ; Simplexvirus*

To investigate resistance to lamivudine (3TC), we examined the incidence of M184V in 20 HIV-1 patients treated with 3TC for 13.1 +/- 9 months. Fourteen of 20 patients had been exposed to zidovudine (ZDV) or didanosine (ddl) prior to 3TC therapy. Nested PCR targeting to reverse transcriptase (RT) and direct sequencing were performed for peripheral blood mononuclear cells sampled serially. There were resistance mutations to ZDV in at least 9 patients at baseline, although there was no resistance mutation to 3TC. We could detect M184V in 6 (30%) out of 20 patients. The incidence of M184V increased as the duration of therapy prolongs (13% in samples<12 months; 47% in samples gtoreq 12 months). The frequency of mutation M184V was higher in patients with previous mutation to ZDV than in patients with wild type. Resistance mutation was not detected in 7 patients. This study shows that resistance to 3TC tends to develop rapidly in patients with baseline mutations or two drugs combination therapy than in those treated simultaneously with triple drugs. This report is the first on resistance to 3TC in Korean AIDS patients.
Didanosine ; HIV-1* ; Humans ; Incidence ; Lamivudine* ; Polymerase Chain Reaction ; RNA-Directed DNA Polymerase ; Zidovudine

Ninety two strains of Streptococcus pyogenes were isolated from patients with pharyngitis, scarlet fever, skin infection, and invasive streptococcal infections in Seoul, Korea from January to December, 1998. All isolates were epidemiologically characterized by T protein serotype, and serum opacity factor (OF) detection to phenotypes. To analyze the genetic relationship, fifty two isolates including 32 erythromycin-clindamycin (Em-Cm) resistant strains, 20 antimicrobial susceptible strains were attempted to the pulsed-field gel electrophoresis (PFGE). T protein serotype showed 16 kinds in distribution including T12 and T4. Among the total isolates, 40 strains (43.5%) belonged to the T12 serotype and twenty strains (21.7%) to T4 serotype. On the other hand, when infection aspect of S. pyogenes isolates were analysed by T serotype distribution, T12 type was predominant for pharyngitidis which contributed to 21 strains (53%) and for skin infection isolates which contributed to 11 strains (28%), respectively. In case of T4 type, it was the most predominant pharyngitidis isolates which contributed to 8 strains (40%). In T serotype distribution of Em-Cm resistant strains, 27 strains (84%) of the thirty two showed T12 serotype. In minimum inhibitory concentration (MIC) values of Em-Cm resistance isolates, thirty two isolates showed resistant to erythromycin 27 strains (84%), had high MIC of >128 mug/ml. And also to clindamycin, twenty two strains (69%) had high MIC of >128 mug/ml. When OF detection of Em-Cm resistance of S. pyogenes isolates were analyzed by T serotype distribution, T12 serotype isolates revealed that all of the isolates except one strain were OF negative. In PFGE profile analysis to Em-Cm resistance isolates, of the twenty seven, Em-Cm resistance of T12 serotype isolates, 26 strains showed identical PFGE profile and all of these isolates revealed that OF negative. Eighty four percent of Em-Cm resistance S. pyogenes isolates had identical phenotype and PFGE profile. These results strongly suggested that the Em-Cm resistant S. pyogenes isolates from Seoul area showed close genetic correlation and PFGE could be available tool for molecular epidemiology.
Clindamycin ; Electrophoresis, Gel, Pulsed-Field* ; Erythromycin ; Hand ; Humans ; Korea* ; Microbial Sensitivity Tests ; Molecular Epidemiology ; Pharyngitis ; Phenotype ; Scarlet Fever ; Seoul ; Skin ; Streptococcal Infections ; Streptococcus pyogenes* ; Streptococcus*