This is a report of attempts to compare the growth yields of various species of fastidious mycobacterium inch1ding human pathogens and non-pathogens in the conventional Dubos liquid medium and two simple media formulated recently; one is a medium containing 0.1% hyaluronic acid and 6.0% bovine serum albumin and the other is a semisyntheic medium made of umbilical cord extract supplemented with 10% sheep serum as a final concentration. All mycobacterial strains employed in experiments gave the heaviest growth yields in the hyaluronic acid-bovine serum albumin medium (HAS medium), among the three media. Dubos liquid medium seemed to be inferior to a medium made of umbilical cord extract (UCE medium) in supporting mycobacterial growth. There were three-to seven-fold increases in dry weight of the bacteria grown in the HAS medium as compared with those in the Dubos liquid medium. We also looked for the possible effect of bovine serum albumin (BSA)in the HAS medium on mycobacterial growth. As a result, we found that the amount of BSA in the HAS medium, ranging from zero to 6.0% in the medium, showed no substantial effect on the mycobacteria1 growth.
Comparative Study ; Culture Media/standards* ; Female ; Human ; Hyaluronic Acid/isolation & purification ; Hyaluronic Acid/pharmacology* ; Mycobacterium/growth & development* ; Pregnancy ; Tissue Extracts* ; Umbilical Cord*

No abstract available.
Rickettsia typhi* ; Rickettsia*

No abstract available.
Antibodies* ; Coxiella burnetii* ; Coxiella* ; Korea* ; Prevalence* ; Q Fever*

The 38 kDa protein of Mycobacterium tuberculosis, which was known previously as antigen 5, has been extensively used in the serodiagnosis of tuberculosis. In an attempt to develop and evaluate a serodiagnostic test using the antigen, we expressed the 38 kDa protein in BCG and its seroreactivity was compared to that expressed in Escherichia coli. The coding region of the 38 kDa protein was amplified by PCR, and the gene was cloned into a Mycobacterium-E. coli shuttle expression vector pYMC-his and pQE30 expression vector and expressed in BCG and E. coli, respectively. Both recombinant 38 kDa proteins showed strong seroreactivity against pooled serum from tuberculosis patients. There was no significant difference in seroreactivity between the two recombinant antigens in sera from the far advanced tuberculosis patients. However, of 25 tuberculosis patients graded as ""minimal"" by chest X-ray, 5 (20.0%) were seropositive by r38 kDa expressed in E. coli, while 8 (32.0%) by that expressed in BCG. Likewise, higher seroreactivity by r38 kDa expressed in BCG was found in sera from the moderately advanced tuberculosis. This study thus indicates that the recombinant 38 kDa expressed in BCG is more effective than that expressed in E. coli in detecting antibodies to the native 38 kDa protein of M. tuberculosis in sera from minimally affected tuberculosis patients.
Antibodies ; Clinical Coding ; Clone Cells ; Escherichia coli ; Humans ; Mycobacterium bovis* ; Mycobacterium tuberculosis* ; Mycobacterium* ; Polymerase Chain Reaction ; Serologic Tests* ; Thorax ; Tuberculosis*

The ability to introduce recombinant DNA molecules back into mycobacteria would greatly increase the potential of molecular genetic approaches for the study of mycobacteria as well as for the use in clinical purposes. We have initiated the construction of vectors that facilitates the introduction of recombinant DNA into mycobacteria. The vector was designed to contain replicons for multiplication in mycobacteria and Escherichia coli, a promoter for gene expression, a drug resistant gene for selecting transformants, and a few restriction enzyme sites for convenient cloning. Constructed Mycobacterium-E. coli shuttle vector named p YMC (hsp60) was shown to transform M. smegmatis at high efficiency and maintain plasmid at stable level. The ability of the vector to express cloned foreign gene was also monitored by measuring the expressed level of luciferase gene which was used as a reporter. High level of luciferase activity in M. smegmatis with pYMC (hsp60:luc) was detected confirming successful construction of Mycobacterium-E. coli shuttle vector.
Clone Cells ; Cloning, Organism ; DNA, Recombinant ; Escherichia coli* ; Escherichia* ; Gene Expression ; Genetic Vectors* ; Luciferases ; Molecular Biology ; Mycobacterium* ; Plasmids ; Replicon

No abstract available.
Antibodies* ; Coxiella burnetii* ; Coxiella* ; Korea* ; Prevalence*

The occurrence of immune complexes in the serum from rats infected with M. leprae-murium and 38 patients with leprosy were studied by the polyethylene glycol precipitation complement consumption (PEG-CC) test and the results were compared in the various forms of the disease. Circulating immune complexes (CIC) were significantly increased in the sera from rats infected with M. lepraemurium compared to normal control rats (P < 0.005). There were no significant differences between the the level of CIC in the sera from lepromatous leprosy patients and that from tuberculoid leprosy patients, but in the sera from patients with erythema nodosum leprosum (ENL) the level of CIC was significantly increased (P < 0.005). And although we couldn't find a corre1ation between the level of CIC and bacterial indices in lepromatous leprosy patients, CIC tends to de-crease after negative conversion of their bacterial indices. These findings suggested that the detection of CIC can be of some practical interest in the early diagnosis of ENL and can be a valuable assessment in following the therapy after negative conversion of their bacterial indices.
Animal ; Antigen-Antibody Complex/analysis* ; Disease Models, Animal ; Human ; Leprosy/immunology* ; Male ; Rats ; Reference Values ; Time Factors

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

PURPOSE: Cyclin D1 plays critical roles in the progression of cells through the G1 phase of the cell cycle, which has been implicated as a putative protooncogene, while p53 protein affects different phase checkpoint pathways in the normal cell cycle, which mutations occur in poor prognosis of cancer. We attempted to determine their significance for tumor behavior and prognosis of transitional cell carcinoma(TCCa) of the bladder in human. MATERIALS AND METHODS: Formalin fixed-paraffin embedded tissue specimens from 102 patients with TCCa of the urinary bladder were examined. Nuclear expression was detected by immunohistochemical analysis with a standard avidin-biotin immunoperoxidase method, using the monoclonal antibody cyclin D1 and p53(DO7). RESULTS: Cyclin D1 and p53 protein immunostaining of the nucleus was observed in 49 tumors(48.0%) and 55 tumors(53.9%) respectively. Overexpression of cyclin D1 showed significant inverse correlation with the histological grade and significant correlation with recurrence. Overexpression of p53 protein showed a significant correlation with the histological grade and stage(p<0.01). 65.3% (32 out of 49 tumors), of the cyclin D1 positive tumors exhibited expression for p53 protein(p<0.05). Patients with TCCa coexpressing cyclin D1 and p53 protein had a significantly poorer prognosis than those expressing neither cyclin D1 nor p53 protein(p<0.001). CONCLUSIONS: Cyclin D1 in bladder TCCa is closely related to early tumor differentiation and associated with recurrence. Simultaneous overexpression of both cyclin D1 and p53 protein is related to more aggressive tumor behavior and poorer prognosis. This indicates that cyclin D1 evaluation may be a further useful element for selecting subgroup of patients who should be treated with more aggressive therapies.
Carcinoma, Transitional Cell* ; Cell Cycle ; Cyclin D1* ; Cyclins* ; Formaldehyde ; G1 Phase ; Humans ; Prognosis ; Recurrence ; Urinary Bladder*

Anti-idiotype antibody (anti-id Ab) which recognizes idiotope in the variable region of immunoglobulin (Ig) can regulate Ab production by B cells in vivo and in vitro. Although it has been reported that anti-id Ab can suppress IgM production by lymphocytes or hybridoma cells without suppression of cell proliferation, the regulatory mechanism of anti-id Ab is not completely understood. We studied the effects of anti-id Ab on the production of IgG class anti-DNA Ab by hybridoma cells, on the proliferation of cells, and on the transcription levels of Ig genes. In contrast to suppressive effect of anti-id Ab on the production of IgM previously reported by others, stimulatory effects of anti-id Ab on the production of IgG by hybridoma cells as well as the proliferation of these .cells were observed. However, little effect of anti-id Ab on the transcription levels of Ig genes was observed. These results suggest that anti-id Ab can increase Ab production by stimulation of cell proliferation. Furthermore, these results suggest that the effect of anti-id Ab on the production of Ab may be determined by the difference in class of Ab produced by hybridoma cells following the treatment with anti-id Ab.
Antibody Formation* ; B-Lymphocytes ; Cell Proliferation ; DNA* ; Genes, Immunoglobulin ; Hybridomas* ; Immunoglobulin G ; Immunoglobulin M ; Immunoglobulins ; Lymphocytes