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*

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

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

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

OBJECTIVE: Systemic Lupus Erythematosus(SLE) is characterized by various autoantibodies to a variety of nuclear antigens. Certain extractable nuclear antigens(ENA) have been served as extremly useful aids in differentiation of clinical subset, diagnostic marker and in the detection of early forms of systemic rheumatic diseases. This study was to employ immunoblot to determine the prevalences of antibodies to ENA in SLE compared with immunodiffusion. METHODS: Sera were obtained from 127 SLE patients. Antibodies to ENA were assessed by double immunodiffusion (DID) and immunoblot method. RESULTS: Using the immunblot method, the prevalences of antibodies to ENA were as follows: The antibody to Sm was 27%, UIRNP 33.6%, Ro 41.8%, La 14%, ribosomal P 14% and Ku 4%. The prevalences of antibodies to ENA by DID were as follows: The antibody to Sm was 15%, RNP 24.5%, Ro 54.3% and La 9.4%. CONCLUSIONS: Compared with immunodiffusion, results using immunoblot showed greater sensitivity in the detection of autoantibodies to ENA in SLE.
Antibodies* ; Antigens, Nuclear ; Autoantibodies ; Humans ; Immunoblotting* ; Immunodiffusion* ; Lupus Erythematosus, Systemic* ; Prevalence ; Rheumatic Diseases

OBJECTIVE: We observed the developmental pattern of activation of MAPK signal transduction pathways known to be activated by electroconvulsive shock(ECS) in young rat hippocampus after kainic acid(KA)-induced seizure. METHODS: We used the method of immunoblotting for examining the basal protein amount and basal level of phosphorylation of MAPK kinase(SAPK/ERK kinase -1, SEK-1), MAPK(c-Jun N terminal protein kinase, JNK), transcription factor(c-Jun) and immediate early gene proteins(Fos) in rat hippocampus at postnatal day 7, 14, and 21, respectively. We also examined the changes of phosphorylation of those proteins after kainic acid-induced seizure in the same way. RESULTS: The basal protein amounts of SEK-1, JNK, and c-Jun did not show age-dependent changes and basal level of phosphorylation of JNK and c-Jun remains unchanged throughout the early developmental period. The basal level of phosphorylation of SEK-1 was peaked at postnatal 7 days and then decreased with aging. After kainic acid-induced seizure, the change of phosphorylation of JNK was not observed but those of SEK-1 and c-Jun increased after postnatal day 14. The expression of Fos was observed at postnatal day 7 and also increased with aging. CONCLUSION: These results show that the MAPK signal transduction system in rat hippocampus matures in accordance with aging, but the process of maturation differs depending specific proteins. This study suggests the signal transduction cascade(SEK-1 - JNK - c-Jun - Fos) which is well established in cell line studies may not be applied to rat hipposcampus because we could not observe the activation of JNK after KA-induced seizure in young rat hippocampus.
Aging ; Animals ; Cell Line ; Hippocampus* ; Immunoblotting ; Kainic Acid ; Phosphorylation ; Phosphotransferases ; Protein Kinases ; Rats* ; Seizures* ; Signal Transduction*

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

This study was conducted to identify the protein antigens reacting with IgG antibodies in the sera of variaus stages of syphilis before treatment, the antigens common to both Treponema pallidum and Treponema phagedenis, and the antigens specific only to T. pzllidum, employing sodium dodecylsulfate polyacrylamide gel electrophoresis(SDS PAGE) and immunoblotting. Before treatment, the most strongly reacting antigens of T. pallidum precipitated by IgG antibodies in the sera of patients were polypeptides of molecular weights 47, 000, 36,500, 15,500 and 14,000. Eleven antigens common to both T. palbdum and T. phagedenis were observed, and antigens of molecular weights 86,500, 68,500,15,500 and 14,000 showed to be specific only to T. pallidum. From the results obtained, it could be concluded that of the major antigens of T. pallidum, the antigens of molecular weights 15,500 and 14,000 could serve to develop newer serologic tests for syphilis.
Antibodies* ; Humans ; Immunoblotting ; Immunoglobulin G* ; Molecular Weight ; Peptides ; Serologic Tests ; Sodium ; Syphilis* ; Treponema ; Treponema pallidum

A newly developed third generation enzyme immunoassay(Lucky HCD 3.0 EIA) for hepatitis C virus(HCV) antibodies was added with the envelope(E1E2)/NS4 fusion proteins and expanded NS5 proteins as well as the core/NS3 fusion proteins. Authors evaluated the HCD 3.0 EIA with the previously available second generation EIA(HCD 2.0) in 10,435 Red Cross blood donors. Among 10,435 donors who were screened for the presence of HCV antibodies by HCD 2.0 assay, 22(0.21%) sera were repeatedly reactive. All of these sera were tested for further testing. Only 13 of all tested sera were reactive by HCD 3.0 EIA, and nine sera were not reactive. Nine of 13 HCD 3.0 positive sera were reactive by recombinant immunoblot assay(Lucky-Confirm). Also seven of these 13 sera had detectable HCV genomic RNA by reverse transcriptase-polymerase chain reaction(RT-PCR). None of nine HCD 3.0 negative samples had detectable immunoblot assay and HCV genomic RNA. It is concluded that the new HCV EIA can decrease a significant false positivity of second generation EIA in a blood donor population. This new assay correlates well with detection of HCV-RNA by RT-PCR and identifies donors who are truly infected.
Antibodies ; Blood Donors* ; Hepatitis C ; Hepatitis C Antibodies ; Humans ; Immunoblotting* ; Red Cross ; RNA ; Tissue Donors