FKBP-12 Exhibits an Inhibitory Activity on Calcium Oxalate Crystal Growth in Vitro

Urolithiasis and calcium oxalate crystal deposition diseases are still significant medical problems. In the course of nephrocalcin cDNA cloning, we have identified FKBP-12 as an inhibitory molecule of calcium oxalate crystal growth. lambdagt 11 cDNA libraries were constructed from renal carcinoma tissues and screened for nephrocalcin cDNA clones using anti-nephrocalcin antibody as a probe. Clones expressing recombinant proteins, which appeared to be antigenically cross-reactive to nephrocalcin, were isolated and their DNA sequences and inhibitory activities on the calcium oxalate crystal growth were determined. One of the clone lambdagt 11 #31-1 had a partial fragment (80 bp) of FKBP-12 cDNA as an insert. Therefore, a full-length FKBP-12 cDNA was PCR-cloned from the lambdagt 11 renal carcinoma cDNA library and was subcloned into an expression vector. The resultant recombinant FKBP-12 exhibited an inhibitory activity on the calcium oxalate crystal growth (Kd=10(-7) M). Physiological effect of the extracellular FKBP-12 was investigated in terms of macrophage activation and proinflammatory cytokine gene induction. Extracellular FKBP-12 failed to activate macrophages even at high concentrations. FKBP-12 seems an anti-stone molecule for the oxalate crystal deposition disease and recurrent stone diseases.

Mutations in the Pre-C and Core Region of Hepatitis B Virus DNA in Chronic HBV Carriers / 대한내과학회지

OBJECTIVES: Causes of diverse clinical sourses of patients with chronic hepatitis B virus(HBV) infection are not fully-known. The host immune response to HBV antigen and the appearance of mutant viruses are believed to be important factors. To determine whether appearance of precore and core mutant viruses are related to the clinical course of the patients, we analysed the entire core region of viral DNA in 7 HBV chronic carriers. METHODS: Serum was obtained from 7 patients(3 chronic active hepatitis, 4 CAH with cirrhosis) and pre-C and core region of HBV were amplified by polymerase chain reaction, then directly sequenced. RESULT: In all 7 HBV DNA there was a point mutation from T to C at nucleotide 2104 of core region, and each DNA also contained 6 to 17 variable point mutations at different nucleotides yielding various amino acid substitution. One of DNA had a point mutation from A to G at nucleotide 1898, converting tryptophan(TGG) to a stop codon(TAG). Two cases of deletion mutations covered C-region segment ranging from nucleotide 2142 to 2306 and one case of deletion covered pre-C region ranging from nucleotide 1815 to 1843. CONCLUSION: Three out of seven DNA contained mutational sites coincided with known immunodominant T cell epitopes and rest of the mutational sites could also affect the antigenecity of the HBV. Therefore, mutant HBV could after the host immune response, and may modulate the clinical course of infection.

DNA-mediated immunization of mice with plasmid encoding HBs antigen

In order to develop an experimental DNA vaccine for the prevention and treatment of hepatitis B virus infection, hepatitis B virus surface antigen (HBsAg) DNA was subcloned into an E. coli-eukaryotic cell shuttle vector and was expressed in the Baculovirus expression system. Intramuscular, intradermal, and intraperitoneal injections of 30 microg of the plasmid DNA expressing HBsAg induced humoral and cellular immune responses in ICR mice. The first IgG antibodies were detected after ten days and specific IgG antibody titers peaked after two months of a single intramuscular DNA injection. Anti-HBs antibody titers gradually increased and peaked at four months following intradermal DNA injection, and in case of intraperitoneal injection they peaked at seven months. Generation of HBs-specific helper T lymphocytes was also investigated through the production of interleukin-2 by T helper cells. Boosting effects of HBs DNA were investigated without much results. In general, DNA-mediated HBs immunization induced humoral and cellular immune responses in mice that appears to simulate immune responses in human during the course of HBV vaccination.

A Case of Isolated-organ Tuberculosis Causing Common Bile Duct Obstruction: Tuberculous periductal lymphadenitis / 대한소화기내시경학회지

The explanation of isolated-organ tuberculosis rests on the assumption that in the course of the lymphatic or hematogenous dissemination of bacilli, organisms may be rapidly destroyed in all other sites save for the particular tissue involved in the isolated tuberculous process. Tuberculosis can arise in all tissues having lymphatics or blood supply, but the disease causing biliary tract obstruction has been known to be rare. Recently, we experi-enced a case of isolated-organ tuberculosis causing common bile duct obstruction and periductal lymph node enlargement in a 46-year-old Korean male. An ultrasonography-guided percutaneous needle biopsy revealed a granulomatous inflammation of the lymph node. After 7 months of anti-tuberculous medication, the common bile duct obstruction and periductal lymph node enlargement disappeared completely in a follow up abdominal CT and ERCP.

Respiratory Variations of Doppler Echocardiographic Parameters in Cardiac Tamponade

BACKGROUND: Cardiac tamponade is associated with the expiratory increase and the expira-tory decrease in left ventricular filling flow. With Doppler echocardiography, we analyzed the respiratory variations of mitral and tricuspid inflows, and pulmonary and hepatic venous flows in patients with cardiac tamponade. METHODS: Respiratory hemodynamic changes in mitral and tricuspid inflows and pulmonary and hepatic venous flows were evaluated using Doppler echocardiography in 13 patients (6 men and 7 women; mean age 51+/-13 years) with large pericardial effusion and clinical cardiac tamponade, and compared the results with those of 11 control subjects (3 men and 8 women, mean age 53+/-13 years). Doppler examination was repeated after pericardiocentesis in 6 patients. RESULTS: Peak velocity of early diastolic mitral inflow (E velocity) decreased during inspiration compared with expiratory increase; the mean percentage change was 40%. Peak velocity of late diastolic mitral inflow (A velocity) decreased 13% during inspiration. E/A ratio decreased 31% during inspiration. Deceleration time (DT) and isovolumic relaxation time (IVRT) increased by 26% and 44%, respectively, during inspiration. But respiratory variations of tricuspid inflow were opposite to those of mitral inflow. Tricuspid E velocity increased during inspiration and decre-ased during expiration. The mean percentage change was 123%, which was larger than thte 40% of mitral inflow. The most characteristic findings of pulmonary venous flow during respiration were the expiratory increases of peak diastolic velocity (DV) and diastolic time-velocity integral (D-TVI). The mean percentage changes of peak systolic velocity (SV), DV and D-TVI during respiration were 27%, 45% and 53% respectively. In contrast, the SV and DV of hepatic venous flow increased during inspiration and decreased during expiration. The respiratory variations of peak systolic reverse flow velocity (SR) and peak diastolic reverse flow velocity (DR) were opposite to those of SV and DV. DR notably increased during expiration, and the mean percentage change was 61%. The ratio of RFI (Inspiratory reverse flow integral) to FFI (forward flow integral) of the tamponade group was 270%. The mean percentage changes of each parameters decreased after pericardiocentesis. CONCLUSION: Patients with cardiac tamponade showed inspiratory increases of diastolic tri-cuspid filling flow and hepatic forward flow. Expiratory increases included diastolic mitral filling flow, pulmonary venous systolic and diastolic flow, and hepatic venous reverse flow. Such res-piratory variations decreased after pericardiocentesis.

Identification of Acinetobacter calcoaceticus - Acinetobacter baumannii Complex by Ribotyping

Acinetobacter species encounters frequently with clinical specimens and now accounts for a substantial proportion of endemic nosocomial infections in Korea. Recent trends indicate that the antimicrobial resistant strains of Acinetobacter species are increasing. Sixty-one strains were isolated from specimens of patients suspected of nosocomial infections during 1991 to 1996. At present, phenotypic identification of Acinetobacter using biochemical test may not be reliable and resulted in the difficulty to clarify the source of infections and epidemiological study of hospital-acquired infections. Aware of the importance of rational taxonomic proposal for these isolates, correct species identification of these organisms by molecular typing method was carried out. A total of fifty-four strains of A. calcoaceticus-A. baumannii complex species which were identified to genospecies 2 and 13 by biochemical characteristics was subjected to identify by ribotyping using restriction endonuclease EcoRI, ClaI, and SalI. Of fifty-four strains, twenty-five strains were identified as A. baumannii (genospecies 2) and twenty-one strains as genospecies 13, and six strains changed to genospecies 3, and the rest two strains were confirmed as A. haemolyticus (genospecies 4). This result suggests that the ribotyping may be of value for identification of genospecies and epidemiological information of Acinetobacter strains.

Study on the Production and Function of Macrophage Migration Inhibitory Factor ( MIF ) : Effect of Recombinant MIF and MIF cDNA on the Induction of Cytokine mRNA / 대한면역학회지

In order to study the functions of migration inhibitory factor (MIF) as macrophage activating cytokine and to investigate the possibility of MIF cDNA as gene therapeutic agent or adjuvant, we produced recombinant MIF (rMIF), anti-MIF antibody and pcDNA I plasmid containing mMIF cDNA (mMIF plasmid). We have investigated the effects of recombinant mMIF or mMIF plasmid on the expression of immune response-related gene in the mouse peritoneal macrophage or splenocyte. Recombinant mMIF produced by Baculovirus expression system was biologically active; it increased mRNA expression of tumor necrosis factor (TNF)-a, Interleukin (IL)-1, IL-6, granulocyte monocyte-colony stimulating factor (GM-CSF), nitric oxide synthase (NOS), Fas and Bcl-x when applied to the cultures of mouse peritoneal macrophage. Anti-mMIF antibody blocked these effects of mMIF on macrophage. Plasmid DNA carrying MIF cDNA inoculated into mouse peritoneal cavity also increased mRNA transcriptions from TNF, IL-1, IL-6, IL-12, GM-CSF, NOS genes of peritoneal macrophage. It enhanced proliferation of splenocyte stimulated with phorbol myristate acetate and IL-2 mRNA expression of splenocytes. Frorn these results, we conclude that rMIF is a strong macrophage activating factor and especially MIF plasmid can be used as an immune potentiating DNA drug in gene therapy for cancer or DNA adjuvant in vaccination in future.

Peripheral Hemodynamic Responses Induced during Dipyridamole Infusion and the Relationships to the Coronary Artery Disease

BACKGROUND: Perfusion scintigraphy with dipyridamole have been reported to be useful for diagnosis of coronary artery disease and the assessment of the presence and extent of myocardium at ischemic risk, especially in patients who can not undergo dynamic exercise testing. Dipyridamole, pharmacologic coronary vasodilator, also induces fall in blood pressure and rise in heart rate. The purpose of this study was to answer the question if dipyridamole induced peripheral hemodynamic responses were related to chest pain, ST changes on EKG, scintigraphic defect or extent of coronary stenosis. METHODS: Dipyridamole 99mTc-MIBI myocardial scintigraphy and coronary angiography on 43 subjects who were suspected to have coronary artery disease. The peripheral hemodynamic response was graded as absent(group 0) if there was a 10 but 10 but 20mm fall in SBP and/or >20 beats/min rise in HR. RESULTS: The overall diagnostic sensitivity and specificity for coronary artery disease of dipyridamole perfusion scintigraphy were 68%, 83% while per vessel sensitivity and specificity for coronary artery disease were 66%, 97%. The numbers of induced chest pain and ischemic ST changes among hemodynamic subgroups, were 40%, 40% in group 0, 33%, 27% in group 1 and 50%, 40% in group 2 without significant difference in each hemodynamic subgroups. Either the numbers of diseased coronary arteries or the numbers of patients demonstrationg reversible scintigraphic defects were not statically different among each subgroups. CONCLUSION: Although the peripheral hemodynamic response dose not always correlate with its central coronary effect but dipyridamlole 99mTc-MIBI myocardial perfusion scintigraphy is an useful test for diagnosis of coronary artery disease.

A comparative study of anemia in hemodialysis patients and CAPD patients / 대한신장학회잡지

No abstract available.