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

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

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*

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*

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

Several stresses are known to induce synthesis of heat shock protein. The present study was performed to see whether pulmonary ischemia, induced by the bronchial artery occlusion, produced HSP70 in cat lung. To this aim we compared experimental and control groups of cats with respect to the HSP70 production in the lung. Experimental animals were subjected to 10-min bronchial artery occlusion followed by reperfusion. The interval between the end of the occlusion and the end of the reperfusion was 1 hour, 4 hours and 8 hours, whereas control animal was not subjected to any manipulation except anesthesia. According to the interval differences, experimental animals were divided into 1HR, 4HRs and 8HRs groups. To determine the induction of HSP70 in each group, total proteins of lung tissues were extracted and separated by PAGE electrophoresis. Immunoblotting with a mouse monoclonal anti-HSP70 IgG antibody revealed that HSP70 was not detected in the pulmonary tissues resected from control, 1HR or 4HRs groups. In contrast, HSP70 expression in 8HRs group was marked. These results suggest that pulmonary ischemia by the bronchial artery occlusion produces HSP70 in a delayed
Anesthesia ; Animals ; Bronchial Arteries* ; Cats* ; Electrophoresis ; Heat-Shock Proteins ; Immunoblotting ; Immunoglobulin G ; Ischemia ; Lung* ; Mice ; Reperfusion

The aim of this study was to identify antigens for a vaccine or drug target to control rabbit coccidiosis. A combination of 2-dimensional electrophoresis, immunoblotting, and mass spectrometric analysis were used to identify novel antigens from the sporozoites of Eimeria stiedae. Protein spots were recognized by the sera of New Zealand rabbits infected artificially with E. stiedae. The proteins were characterized by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF/TOF-MS) analysis in combination with bioinformatics. Approximately 868 protein spots were detected by silver-staining, and a total of 41 immunoreactive protein spots were recognized by anti-E. stiedae sera. Finally, 23 protein spots were successfully identified. The proteins such as heat shock protein 70 and aspartyl protease may have potential as immunodiagnostic or vaccine antigens. The immunoreactive proteins were found to possess a wide range of biological functions. This study is the first to report the proteins recognized by sera of infected rabbits with E. stiedae, which might be helpful in identifying potential targets for vaccine development to control rabbit coccidiosis.
Coccidiosis* ; Computational Biology ; Eimeria ; Electrophoresis ; HSP70 Heat-Shock Proteins ; Immunoblotting ; Mass Spectrometry ; Rabbits ; Sporozoites