Eight bacterial strains were examined whether they have phoP/phoQ genes which were known to be involved in the intracellular survival of Salmonella typhimurium. The phoP/phoQ operon were known to sense the stimuli of the genes involved in the adaptation of the environment. Using 514-basepairs EcoRV DNA fragment of phoP region of Salmonella typhimurium as a probe, dot blot hybridization were performed. Chromosomal DNAs of Klebsiella pneumonia, Pseudomonas aeruginosa, Serratia marscescens, Enterobacter cloacae, Salmonella typhimurium, Escherichia coli, Shigella dysenteriae, and Listeria monocytogenes were examined by DNA hybridization assay. Against our expectation, intracellular pathogen, L. monocytogenes, did not have similar DNA sequences to phoP/phoQ of S. typhimurium, while E, coli, S. dysenteriae, and E. cloacae showed the positive signal even though they were not intracellular pathogens. This result suggested that the phoP/phoQ operon was absent in intracellular pathogenic bacterias other than S. typhimurium. Rather it was found in phylogenetically closer bacterias to S. typhimurium, which were not able to survive in intracellular environment. Some different mechanism, which is not dependent on phoP/phoQ operon, could be involved in the intracellular survival of L. monocytogenes.
Bacteria* ; Base Sequence ; Cloaca ; DNA ; Enterobacter cloacae ; Escherichia coli ; Klebsiella ; Listeria monocytogenes ; Operon ; Pneumonia ; Pseudomonas aeruginosa ; Salmonella typhimurium ; Serratia ; Shigella dysenteriae

Epidemiological studies are important in both the prevention and treatment of mycobacterial infections. This study was initiated to establish the pulsed-field gel electrophoresis (PFGE) method, which are not yet extensively studied. The most apprpriate restriction endonucleases included Dral, AsnI, and XbaI. The optimal PFGE condition was different according to the enzymes used. Two stage PFGE was performed, in case of DraI first stage was performed with 10 seconds of initial pulse and 15 seconds of findA pulse, while the second stage was performed with 60 seconds of initial pulse and 70 seconds of final pu',se. The electrophoresis time for DraI-PFGE was 14 hours for each stage. Electrophoresis was performed for 22 hours, in case of XbaI, with 3 seconds of initial pulse and 12 seconds of final pulse. Electrophoresis was performed for 22 hours, in case of AsnI, with 5 seconds of initial pulse and 25 seconds of final pulse. In all cases the voltage of the electrophoresis was maintained constantly at 200 voltage. Standard mycobacterial strains, which included Mycobacterium bovis BCG, M. tuberculosis, and M. fortuitum, could not be differentiated by PFGE analysis. PFGE analysis was performed to differentiate 9 clinically isolated M. fortuitum strains using AsnI. All M. fortuitum strains showed different genotypes except 2 strains. Cluster analysis divided M. fortuitum strains into 2 large groups. PFGE analysis was performed to further differentiate M. fortuitum isolates using XbaI. The undifferentiated 2 M. fortuitum strains showed different PFGE patterns with Xba I. Cluster analysis of the XbaI-PFGE patterns showed more complex grouping than AsnI-PFGE patterns, which showed that XbaI-PFGE analysis was better than AsnI-PFGE in M. fortuitum genotyping. The top dissimilarity values of AsnI-PFGE and XbaI-PFGE were 0.74 and 0.75, respectively. This value was higher than that of arbitrarily primed polymerase chain reaction (AP-PCR) analysis and lower than that of restriction fragment length polymorphism (RFLP) analysis. This suggested that PFGE can be used as a supportive or alternative genotyping method to RFLP analysis.
DNA Restriction Enzymes ; Electrophoresis* ; Electrophoresis, Gel, Pulsed-Field ; Epidemiologic Studies ; Genotype ; Mycobacterium bovis ; Mycobacterium* ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Tuberculosis

Inositol 1,4,5-triphosphate(InsP,) is a second messenger for obilizing intracellular Cal'. It can be dephosphorylated by soluble and particulate forms on InsP, 5-phosphatase, or phosphorylated to produce inositol 1,3,4,5-tetrakisphosphate(InsP,) by InsP, 3-kinase. These enzymes represent possible targets for the regulation of the InsP,AnsP. signal. InsP, 3-kinase which catalyses th ATP-dependent phosphorylation of InsP, was purified from bovine brain tissue. All operation were carried out at 41C. Fresh tissure was homogenized and centrifuged. The supernatant was pooled. Proteins were precipitated from 10% polyethylene glycol, and suspended solution was applied to DEAE cellulose column for chromatography. As the result of above procedure, two isozymes of InsP, 3-kinase, I and U were obtained. Each isozyme was applied to Matriz green gel, Calmodulin-Affigel 15 column and subsequent phenyl-TSK HPLC column. Specific activites(SA) and fold of puriety were observed at each purification step of chromatography. At DEAE cellulose chromatography, SA were 1, 0.6 and 11, 4.8 nM/min/mg, and folds
Brain* ; Catalysis ; Chromatography ; Chromatography, DEAE-Cellulose ; Chromatography, High Pressure Liquid ; DEAE-Cellulose ; Inositol* ; Isoenzymes ; Phosphorylation ; Phosphotransferases* ; Polyethylene Glycols ; Second Messenger Systems

No abstract available.
Alzheimer Disease* ; Neurons* ; Neuropeptides*

No abstract available.
Diagnosis* ; DNA* ; Polymerase Chain Reaction*

To evaluate the structural characteristics that might be related to the clinical features noted in preeclamptic patients with focal segmental glomerulosclerosis(FSGS), we analyzed post-partum renal biopsies of 8 preeclamptic patients with FSGS (group 1) by morphometry and glomerular studied the structural-functional relationships. These findings were also compared with those from three postpartum cases with minimal change lesion(group 2) and normal age-matched women(group 3). Mean glomerular volume (MGV) in group 1 and group 2 was (2.64 +/- 0.49) x 10(6) micrometer3 and (2.56+/-0.25)x 10(6) micrometer3, respectively. MGV in both groups was significantly increased compared with that of the control group [(1.11+/-0.22)x10(6) micrometer3](p<0.0005). The volume density of the mesangium/glomerulus [Vv(mes/glom)] in the group 1 patients was significantly increased (p<0.0001) when compared with that of the group 2 and the control group patients. The increment of Vv(mes/glom) was related to both the mesangial cell proliferation and expansion of mesangial matrix. The volume density of the capillary lumen/glomerulus [Vv(cap/glom)] in group I was significantly decreased(p<0.0001) when compared with that of group 2 and the control group. Vv(cap/glom) was directly related to Ccr in group l(r=0.70, p=0.05). These results suggest that reduced capillary luminal area caused by mesangial interposition is related to the decreased glomerular filtration rate in preeclamptic FSGS.
Pregnancy ; Female ; Humans ; Biopsy

No abstract available.
Enterobacteriaceae* ; Starvation*

The DNA fragment containing the phoA of Klebsiella pneumoniae was cloned into pACYC184. The size of the insert. was 4.0 kb and the restriction map showed it contained 3 Pstl sites and 4 PvuLI sites. The nucleotide sequence of the phoA region was determined, which showed strong (80%) sequence similarity with that of Escherichia coli. This suggested that these two species are phylogenetically very close to each other.
Base Sequence ; Clone Cells* ; Cloning, Organism* ; DNA ; Enterobacteriaceae* ; Escherichia coli ; Klebsiella pneumoniae ; Operon*

Epidermoid Cyst is a very rare benign neoplasm of the testis which represents about 1% of all testicular tumors. Ultrasonography findings of epidermoid cyst are so variable in reported cases that it is not eas y to diagnose preoperatively. The echogenecity of mixed echoic. it may show hyperechoic rim, and rarely shows target apperance. Target appearance may be a specific finding of epidermoid cyst, and it may be helpful to diagnose preoperatively. we describe a case of epidermoid cyst of the testis demonstrating double target sign on ultrasonography.
Epidermal Cyst* ; Testicular Neoplasms ; Testis* ; Ultrasonography

No abstract available.
Mandibular Osteotomy*