No abstract available.
Antibodies* ; Borrelia burgdorferi* ; Borrelia* ; Immunoblotting* ; Lyme Disease*

OBJECTIVES: To investigate the effects of KCl on regulation of circadian gene CLOCK expression, we observed whether induction of CLOCK is influenced by KCl depolarization in B35 rat neuroblastoma cells. METHODS: B35 rat neuroblastoma cells were grown in Dulbecco's modified Eagle's medium supplemented with 10% FBS and 1% penicillin-streptomycin in a 37 degrees C humidified incubator with 5% CO2. Inhibitors including cycloheximide and actinomycin D were pretreated 1 hour before treatment with 50mM KCl. Immunoblotting with anti-CLOCK antibody was done. RESULTS: CLCOK is induced by 50 mM KCl in B35 Rat Neuroblastoma cells, and a maximal induction in CLOCK level reached peak at 8 to 20 hours. The pretreatment of cycloheximide and actinomycin D prevented the induction of CLOCK by 50 mM KCl. CONCLUSION: We suggest that KCl depolarization may play critical roles in several aspects of the circadian gene CLOCK expression.
Animals ; Circadian Clocks ; Cycloheximide ; Dactinomycin ; Immunoblotting ; Incubators ; Neuroblastoma* ; Rats*

Agglutination Tests ; HIV Infections ; diagnosis ; virology ; Immunoblotting ; Immunologic Tests

The aim of this study was to evaluate domestic enzyme immunoassay(EIA) kit ?LG HCD 3.0?(LG) for the detection of antibody to hepatitis C virus(anti-HCV) in comparision with Axsym HCV version 3.0(Axsym), Cobas Core anti-HCV EIA(Cobas). Cobas kit shows better clear distinction between positive and negative by signal/cutoff ratio(S/C), but it also reveal relatively high false positive rate. The concordance rate of test results between LG and Axsym was 96.2%, between LG and Cobas was 95.5%, and total agreement between three EIA kit was 93.9%. LG were relative poor distinction between positive and negative results, but it could be applied clinically as a screening tool for hepatitis C in general population. The S/C of one false negative result by LG was 0.91, and false positive were less than 4.0, therefore we concluded it is necessary to confirm by immunoblotting assay when S/C were between 0.8 and 4.0.
Hepacivirus ; Hepatitis C ; Immunoblotting ; Immunoenzyme Techniques* ; Mass Screening

No abstract available.
Electrophoresis, Polyacrylamide Gel* ; Immunoblotting* ; Korea* ; Rickettsia typhi* ; Rickettsia*

BACKGROUND: Intracellular antibody specific to hepatitis B virus X protein (HBx) might be useful for studying the role of HBx in hepatocellular carcinogenesis and HBV replication. METHODS: With variable region genes for H7 monoclonal anti-HBx Ab, we constructed a vector for bacterial expression of single chain Ab (scFv) and a vector for eukaryotic cell expression of it. The expression of H7 scFv and its binding activity against HBx was examined by immunoblotting and immunofluorescence microscopy. RESULTS: H7 scFv expressed in bacterial cells retained reactivity to HBx. We demonstrated its intracytoplasmic expression in CosM6 eukaryotic cells. CONCLUSION: This is the first study showing the expression of intracellular anti-HBx Ab in eukaryotic cells. H7 scFv may be a good tool to study the function of HBx in HBV infection
Carcinogenesis ; Eukaryotic Cells ; Hepatitis B virus* ; Hepatitis B* ; Hepatitis* ; Immunoblotting ; Microscopy, Fluorescence

OBJECTIVE: To investigate new signal transduction cascade through integrin, FAK and ERK in the suppressed cell proliferation by GnRH-I and -II. METHOD: Human endometrial cancer cells (HEC1A) were cultured under the following condition: DMEM/F12 (10% FBS). GnRH-I and -II were treated time (0, 5, 10, 15, 20, 30 min; 100 nM) and dose (10 nM or 100 nM; 20 min) dependent manner according to experimental purposes. Cell proliferation was measured using [3H] thymidine incorporation assay. Immunoblotting was utilized to detect proteins. RESULTS: GnRH-I and -II inhibited proliferation of HEC1A cells and induced expression of integrin beta3. Phosphorylation of FAK and ERK were induced by GnRH-I and -II. CONCLUSION: GnRH inhibited cell proliferation via the expression of integrin and FAK, ERK phosphorylation.
Cell Proliferation* ; Endometrial Neoplasms* ; Female ; Gonadotropin-Releasing Hormone* ; Humans ; Immunoblotting ; Integrin beta3 ; Phosphorylation ; Signal Transduction ; Thymidine

No abstract available.
Antibody Formation* ; Chickenpox* ; Child* ; Female ; Herpesvirus 3, Human* ; Humans ; Immunoblotting* ; Immunoglobulin G* ; Pregnant Women*

PURPOSE: We tried to evaluate whether the detection rate of Helicobacter pylori in gastric biopsy specimens could be improved by using pre-embedding immunoelectron microscopy. METHODS: A total of 119 children who complained of upper gastrointestinal symptoms were endoscoped at the Gyeongsang National University Hospital from July, 1996 to July, 1999. Five biopsy specimens(three for urease test, one for hematoxylin-eosin(H and E) staining, and one for pre- embedding immunoelectron microscopy) were obtained from each antrum and body. Immunoblotting analysis were also performed. RESULTS: Among the 119 patients, H. pylori were found in 116 patients(97.5%) by the immunoelectron microscopy. Among three patients who were found H. pylori negative in immunoelectron microscopy, two patients showed H. pylori in H and E stained slides and one patient was urease test positive(color change within six hours). Urease tests were positive in 107 patients(89.9 %). The positive rate of immunoblotting tests was 81.5%. However, only 13 patients(10.9%) showed H. pylori on the H and E stained antrum or body tissue. CONCLUSION: In this study, we found H. pylori histopathologically in most of the pediatric patients who complained of upper gastrointestinal symptoms. This study showed that pre-embedding immunoelectron microscopic examinations can be used as a gold standard in the diagnosis of childhood H. pylori infection. However, this method also has limited capacity to detect widely scattered H. pylori compared to the other histopathologic diagnostic methods.
Biopsy ; Child ; Diagnosis ; Helicobacter pylori* ; Helicobacter* ; Humans ; Immunoblotting* ; Microscopy, Immunoelectron* ; Urease

BACKGROUND/AIMS: Astaxanthin is a carotenoid pigment that has antioxidant, antitumoral, and anti-inflammatory properties. In this in vitro study, we investigated the mechanism of anticancer effects of astaxanthin in gastric carcinoma cell lines. METHODS: The human gastric adenocarcinoma cell lines AGS, KATO-III, MKN-45, and SNU-1 were treated with various concentrations of astaxanthin. A cell viability test, cell cycle analysis, and immunoblotting were performed. RESULTS: The viability of each cancer cell line was suppressed by astaxanthin in a dose-dependent manner with significantly decreased proliferation in KATO-III and SNU-1 cells. Astaxanthin increased the number of cells in the G0/G1 phase but reduced the proportion of S phase KATO-III and SNU-1 cells. Phosphorylated extracellular signal-regulated kinase (ERK) was decreased in an inverse dose-dependent correlation with astaxanthin concentration, and the expression of p27(kip-1) increased the KATO-III and SNU-1 cell lines in an astaxanthin dose-dependent manner. CONCLUSIONS: Astaxanthin inhibits proliferation by interrupting cell cycle progression in KATO-III and SNU-1 gastric cancer cells. This may be caused by the inhibition of the phosphorylation of ERK and the enhanced expression of p27(kip-1).
Adenocarcinoma ; Cell Cycle* ; Cell Line* ; Cell Survival ; Humans* ; Immunoblotting ; Phosphorylation ; Phosphotransferases ; S Phase ; Stomach Neoplasms*