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

OBJECTIVE: To determine the effect of magnesium sulfate (MgSO4) on the apoptosis and invasion of cytotrophoblasts in vitro under normal and hypoxic condition as assessed immunoblot analyses of Bcl-2/Bax, invasion assay and immunohistochemical staining of integrin alpha1. METHODS: Normal cytotrophoblasts were isolated from second trimester placentas and cultured in several physiologically relevant concentrations of MgSO4 under control tissue culture condition (20% O2) or hypoxic condition (1-2% O2). Apoptosis of cytotrophoblasts was estimated by immunoblotting for Bcl-2/Bax, invasiveness was estimated by invasion assay and immunohistochemical staining of Integrin alpha1. RESULTS: The expression of Bcl-2 did not change under standard condition, but it decreased under hypoxic condition with increasing of MgSO4 concentrations. The expression of Bax did not change under both standard condition and hypoxic condition with increasing MgSO4 concentrations. The invasiveness of cytotrophoblasts significantly decreased under both control and hypoxic conditions with increasing of MgSO4 concentrations. The expression of Integrin alpha1 immumohistochemical staining significantly decreased under control condition and showed decreasing tendency under hypoxic condition with increasing of MgSO4 concentrations. CONCLUSION: MgSO4 might induce cytotrophoblasts to the apoptosis and inhibit invasion of cytotrophoblasts under hypoxic condition.
Apoptosis* ; Female ; Humans ; Immunoblotting ; Integrin alpha1 ; Magnesium Sulfate* ; Magnesium* ; Placenta ; Pregnancy ; Pregnancy Trimester, Second ; Trophoblasts*

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

OBJECTIVES: The aim of the present study is to explore the effect of fluoxetine on transcription, translation and activity of tryptophan hydroxylase (TPH), and intracellular level of serotonin. METHODS: The expression level of the TPH mRNA and the protein, the TPH enzyme activity, and the intracellular level of serotonin were explored at the fluoxetine-treated RBL-2H3 cells. Real-time RT-PCR and immunoblotting analysis confirmed changes in the expression of TPH mRNA and protein. The activity of TPH was measured using [3H]tryptophan. The intracellular level of serotonin was measured by HPLC. RESULTS: The TPH activity was gradually increased on time from 24hr to 72hr. The real-time RT-PCR also revealed that the TPH mRNA was increased at 12, 24 and 72hr in the fluoxetine-treated RBL-2H3 cells. The immunoblotting analysis also revealed that the TPH protein was decreased at 72hr in the fluoxetine-treated RBL-2H3 cells. The intracellular level of serotonin was increased at 48hr after treatment of fluoxetine. CONCLUSION: Fluoxetine induced the increases of the TPH mRNA, the TPH enzyme activity and intracellular level of serotonin, and the decrease of the TPH protein expression at the RBL- 2H3 cells.
Chromatography, High Pressure Liquid ; Fluoxetine ; Immunoblotting ; RNA, Messenger ; Serotonin ; Tryptophan Hydroxylase* ; Tryptophan*

BACKGROUND: Currently, serological assay, immunoblotting, and nucleic acid amplification test (NAT) are required as reentry tests for deferred donors with hepatitis C virus (HCV) or human immunodeficiency virus (HIV) screening reactive result. However, immunoblotting must be performed even for serological nonreactive donors. In this study, the efficacy of immunoblot applications for serological nonreactive donors in donor reentry procedures was examined. METHODS: We analyzed non-qualified donors with immunoblot results from 2011 to 2015 in Korea and investigated reentry procedures related with HCV or HIV in other countries. RESULTS: Percentages of donors who could not be released due to immunoblot results even with serological nonreactive results were 54.2% (1,824/3,367) for HCV and 35.9% (4,300/11,964). In the case of 662 donors, their results were considered to be different using other assay kits or based on other criteria. In other countries, immunoblotting is not required as a donor reentry test. CONCLUSION: Indeterminate or reactive immunoblotting results in serological nonreactive donors were due to nonspecific reactions. It is not reasonable to apply immunoblotting to serological nonreactive donors. Therefore, we suggest that immunoblot assays be excluded from the reentry test.
Hepacivirus ; HIV* ; Humans ; Immunoblotting ; Korea ; Mass Screening* ; Nucleic Acid Amplification Techniques ; 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