Vibrio mimicus, marine bacteria pathogenic for fish, can causes acute gastroenteritis in human. Iron limmited condition like in human body, may change the surface structure of V. mimicus. In this study we obse'rved the effect of iron limmited condition on outer membrane protein of V. mimicus. Ethylenediamine-di (O-hydroxy-phenylacetic) acid (EDDA), an iron chelator, delayed the time to reach expotential growth of V. mimicus in brain heart infusion medium from 3 hours to 20 hours. Outer membrane protein of V. mimicus-CON (cultured in BHI) and V. mimicus-EDDA (cultured in BHI contain EDDA) were seperated by 1% sarcosine from total cell envelop. SDS-PAGE of V. mimicus-EDDA and V. mimicus-CON showed similar protein profiles contain 37 kDa major protein but 86 and 90 kDa protein were induced differently. Immunological properties of above protein were determined by ELISA and western blotting. 86 kDa EDDA- specific OMP was induced in V. mimicus (isolate 96-1), V. parahaemolyticus (serotype 09), V. alginolyticus (isolate 95-1), E. coli (human isolate) and V. vulnificus ATCC 27562 in iron limmited condition.
Bacteria ; Blotting, Western ; Brain ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Gastroenteritis ; Heart ; Human Body ; Humans ; Iron* ; Membrane Proteins ; Membranes* ; Sarcosine ; Vibrio mimicus* ; Vibrio*

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*

No abstract available.

The objective of the present study was to identify the characteristics of phospholipase C (PLC) isozymes purified from bovine brain and to investigate their interrelationship with G protein. The purified PLC isozymes β, γ and δ were obtained and the characteristics of PLC activity on various concentrations of free Ca²⁺ were observed. The activity of PLC was increased with increasing Ca²⁺ concentration and the activity PLC δ was increased higher in the presence of phosphatidyl choline (PC) than in the absence of PC. For vesicle formation as the structure of cell membrane, cholic acid and deoxycholic acid as detergent on phosphatidylinositol bisphosphate (PIP₂) substrate containing PC were used, and then the activity of PLC isozymes were increased with increasing concentration of cholate, from 0.2% to 1% and were increased slightly in deoxycholate. In the PIP₂ containing phospholipid and glycolipid as brain extract, the activity of PLC isozymes were checked in 0.2-1% cholic acid. The activities of PLC isozymes were continuously increased up to 1% cholic acid. The quantitation of PLC isozymes from several bovine organs by radioimmunoassay was made. Brain was the most sufficient organ in terms of amount of PLC β and δ. A large amount of PLC δ was existed in adrenal gland. The binding capacity of GTPrS and G protein was observed and other observations of the binding effect of GTPrS-G protein and PLC monoclonal Ab-Protein A from tissue homogenate with PLC were made. From the observation the binding capacity was revealed the range of 0.11-1.49%. The effects of each type of G protein on the percent activity of purified PLC isozymes were observed. From the observation, activities of isozymes were increased in Goa & Gmix, and the activities of PLC β and δ were increased in Gβγ and Gia. Activities of PLC β and γ were decreased in Gta but PLC δ increased.
Adrenal Glands ; Brain* ; Cell Membrane ; Cholates ; Cholic Acid ; Choline ; Deoxycholic Acid ; Detergents ; GTP-Binding Proteins* ; Isoenzymes ; Phosphatidylinositols ; Phospholipases* ; Radioimmunoassay ; Type C Phospholipases*

Control of tuberculosis is threatened by widesread emergence of drug resistant Mycobacterium tuberculosis. Rifampin is a key component among therapeutic regimens for the tuberculosis; therefore patients in whom resistance to this drug develop have a poor outlook, particularly if rifampin resistance is associated with resistance to other tuberculosis drugs. The purpose of this study was to detect the mutation in rpoB gene of rifampin resistant M. tuberculosis in Korea and to evaluate the usefulness of the method in clinical aspects. A sample of 80 M. tuberculosis was studied, and it included 40 rifampin resistance isolates and 40 rifampin sensitive isolates by conventional methods. The detection method involved the amplification by polymerase chain reaction (PCR) of the Rif' region and the identification of mutations by single-strand DNA conformation polymorphism analysis (SSCP) of the amplification products (157 bp). Mutation were identified in 39 of 40 rifampin resistant isolates, and in 1 of 40 rifampin sensitive isolates.
Humans ; Korea ; Mycobacterium tuberculosis* ; Mycobacterium* ; Nucleic Acid Conformation ; Polymerase Chain Reaction* ; Rifampin ; Tuberculosis

Control of tuberculosis is threatened by widespread emergence of drug resistance in Mycobacterium tuberculosis. Understanding the molecular basis of resistance might lead to development of novel rapid methods for diagnosing drug resistance. Rifampin is a key component among therapeutic regimens for the tuberculosis; therefore, patients who have drug resistance do not convalesce satisfactorily. The molecular mechanism of resistance to rifampin in M. tuberulosis has been elucidated. Substitutions of a limited number of highly conserved amino acids encoded by the rpoB gene are responsible for the ""single-step"" high-level resistance of M. tuberculosis to rifampin. Currently, two genotype-based protocols allow drug test from minimally grown cultured materials: (i)mutation identification by direct sequencing of PCR-amplified material. and (ii)mutation screening by PCR-SSCP. The purpose of this study is to evaluate the usefulness of the both methods. A sample of 75 isolates of M. tuberculosis was studied, and it inculded 36 rifampin-resistant strains and 39 rifampin-sensitive strains by conventional methods. Mutaions were identified in 36 rifampin-resistant isolates but in none of 39 sensitive isolates. All mutations were clustered within a region of 23 amino acids. Both methods allow detection of rifampin resistance in 2 to 3 days and will thus help in the early management of infection by M. tuberculosis.
Amino Acids ; Drug Resistance ; Humans ; Mass Screening ; Mycobacterium tuberculosis* ; Mycobacterium* ; Rifampin ; Tuberculosis

Pulmonary endometriosis is a rare disease which is characterized by hemoptysis during menstruation (catamenial hemoptysis). We report a case of pulmonary endometriosis in a 33-year-old housewife. She has had regular menses with moderate flow and minimal dysmenorrhea. She had undergone curettage in May 1995 for artificial abortion. In July 1995, she experienced the first episode of hemoptysis. A chest CT scan revealed a 2.0 1.0 cm sized ill-defined soft tissue density in the periphery of anterior segment of the left upper lobe with a surrounding irregular ground-glass opacity. A left upper lobectomy was done under the diagnosis of pulmonary endometriosis. Cut section of the resected lung showed a round red-brownish solid lesion, measuring 2.0x1.0cm in cross. Microscopically a focus of the endometrial tissue, which was composed of endometrial glands and stroma, was found in the lung parenchyme and many hemosiderin laden macrophages were seen in the surrounding alveoli. The postoperative course was favorable with no further episodes of hemoptysis.
Adult ; Curettage ; Diagnosis ; Dysmenorrhea ; Endometriosis* ; Female ; Hemoptysis ; Hemosiderin ; Humans ; Lung ; Macrophages ; Menstruation ; Rare Diseases ; Tomography, X-Ray Computed

Porokeratosis are characterized by distinct clinical findings of a keratotic ridge that corresponds to the cornoid lamella on histology and has well defined potential for malignancy. We report a case of basal cell carcinoma(BCC) arising in linear porokeratosis in a 77-yearold man.
Carcinoma, Basal Cell* ; Porokeratosis*

Despite the continually improving results of total knee arthroplasty, infection remains the most debilitating complication. The treatments of infected total knee arthroplasty were variable, but initially we removed infected implants and inserted antibiotic containing cemented spacer. Postoperatively, patients were mobilized in a 30 knee flexion state and treated with parenteral antibiotics. After control of infection was clinically and radiologically determined, we inserted PCL substitute total knee prosthesis. Five days postoperatively, patient began touch down standing exercise. We observed two cases in whom infected total knee arthroplasty had been salvaged successfully with two-stage implantation using cement spacers containing antibiotics.
Anti-Bacterial Agents* ; Arthroplasty* ; Humans ; Knee Prosthesis ; Knee* ; Replantation*

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