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*

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*

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

BACKGROUND: Anti-hepatitis C virus antibody (anti-HCV) assays are recommended for screening HCV-infected persons. The VIDAS Anti-HCV Assay (bioMérieux, France), based on the enzyme-linked fluorescence test principle, was recently introduced in Korea. We evaluated the clinical performance of the VIDAS assay. METHODS: One hundred HCV-positive and 1,002 HCV-negative blood samples confirmed by Architect anti-HCV (Abbott Laboratories, USA) and COBAS TaqMan HCV real-time PCR (Roche Diagnostics, USA) or the Procleix Ultrio Plus Assay (Gen-Probe Incorporated, USA) were obtained from the Human Serum Bank (HSB) and tested by VIDAS. In case of discrepant results, we conducted a recombinant immunoblot assay (RIBA). RESULTS: The agreement rates for known HCV-positive and HCV-negative samples between the VIDAS assay and the HSB testing were 100% (95% confidence interval [CI]: 96.4-100%) and 99.5% (95% CI: 98.8-99.8%), respectively. One of the five discrepant samples was positive for Core 2+ and NS3-2 2+ reactivity, two samples were negative, and the other two were indeterminate regarding NS4 2+ reactivity in RIBA. We observed a significant but weak positive correlation between the titers of VIDAS and Architect assays (r=0.315, P<0.001). CONCLUSIONS: The VIDAS anti-HCV assay, developed on the VIDAS automated immunoassay platform based on the ready-to-use, single-sample test concept may be useful in small-to-medium-sized laboratories. It showed good agreement with Architect anti-HCV and COBAS PCR assays and is therefore useful for detection of HCV infection. Weakly test-positive (ambiguous) samples require additional testing by another anti-HCV, RIBA, or HCV RNA assay.
Automation ; Hepatitis C/*diagnosis ; Hepatitis C Antibodies/*blood ; Humans ; *Immunoassay ; Immunoblotting ; Reagent Kits, Diagnostic ; Sensitivity and Specificity

This study aimed to examine whether the expression of major prostaglandin E2 (PGE2) synthesis enzyme, cyclooxygenase-2 (COX-2), is changed in the kidneys of the rats with lithium-induced nephrogenic diabetes insipidus (Li-NDI). Sprague- Dawley rats treated with lithium for 4 weeks were used as the NDI model and expression of renal COX-2 was determined by immunoblotting and immunohistochemistry. In Li-NDI where urine output was markedly increased and urine osmolality was significantly decreased, COX-2 expression in the inner medulla was decreased (28% of control), while it increased 18-fold in the cortex and outer medulla. Consistent with this, labeling intensity of COX-2 in macula densa region was increased, whereas it was decreased in the interstitial cells in the inner medulla, indicating a differential regulation of COX-2 between the cortex and inner medulla in Li-NDI. Accordingly, urinary PGE2 excretion was significantly increased in Li-NDI. In conclusion, there is a differential regulation of COX-2 between cortex and inner medulla in Li- NDI and urinary PGE2 excretion is increased in Li-NDI, possibly due to an increased renal production. This may suggest that increased renal production of PGE2 could play a role in modulating water reabsorption in the renal collecting duct in Li-NDI.
Animals ; Aquaporins ; Cyclooxygenase 2* ; Diabetes Insipidus, Nephrogenic* ; Dinoprostone ; Immunoblotting ; Immunohistochemistry ; Kidney ; Lithium ; Osmolar Concentration ; Prostaglandins ; Rats*

BACKGROUND: The human mic2 gene is a pseudoautosomal gene that encodes a cell surface antigen, CD99. High levels of CD99 constitute a tumor marker in Ewing s sarcoma (ES). We have recently demonstrated that CD99-induced apoptosis occurs only in undifferentiated ES cells, not in differentiated ES cells, raising the possibility of the involvement of CD99 in neural ontogeny. METHODS: To elucidate the relations between the expression of CD99 and the differentiation of neural cells and the mechanism by which the expression of CD99 is regulated, we analyzed the differential patterns of CD99 expression in SH-SY5Y by treatment of 12-O-tetradecanoyl- 13-phorbol acetate (TPA) and retinoic acid. In addition, to explore the transcriptional activity of CD 99 during neural cell differentiation, SH-SY5Y cells were transiently transfected with a CD99 promoter-driven luciferase construct, and treated with the inducers. RESULTS: In immunoblotting and flow cytometry, the expression level of CD99 was increased on differentiated SH-SY5Y cells induced by TPA and retinoic acid. The luciferase activity was elevated by the treatment with TPA, known to mature SH-SY5Y cells toward a sympathetic neuronal lineage, whereas retinoic acid inducing a sympathetic chromaffin lineage displayed little effect. CONCLUSION: The result indicates that CD99 might be expressed only on cells maturing toward a neuronal lineage among differentiating primitive neuronal cells. In addition, the expression of CD99 seems to be regulated at the transcriptional level during the differentiation.
Antigens, Surface ; Apoptosis ; Cell Differentiation ; Cell Line* ; Flow Cytometry ; Humans ; Immunoblotting ; Luciferases ; Neuroblastoma* ; Neurons ; Sarcoma ; Tretinoin

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

We investigated the expression of aquaporin 1 (AQP1) in tissues from canines with an inherited anomaly that causes their erythrocytes to have high K+. Northern blot analysis revealed abundant AQP1 expression in lung and kidney, though little expression was found in spleen. Using anti-C-terminus for dog AQP1, abundant expression was shown in kidney, trachea, and eye, but little expression was shown in pancreas and cerebrum, indicating that AQP1 expression in canine tissues is similar to that noted in other mammals.
Animals ; Aquaporin 1/*metabolism ; Blotting, Northern ; Dogs ; Erythrocytes/*chemistry ; Immunoblotting ; Potassium/*analysis ; Viscera/metabolism