1.Immunoblotting analysis of antibodies against borrelia burgdorferi, the lyme disease agent, in sera from the Korean residents.
Sang Nae CHO ; Tae Yoon LEE ; Mi Kyeong LEE ; Dook Soon KIM ; Joo Deuk KIM
Journal of the Korean Society for Microbiology 1991;26(3):263-272
No abstract available.
Antibodies*
;
Borrelia burgdorferi*
;
Borrelia*
;
Immunoblotting*
;
Lyme Disease*
2.Regulation of the Circadian Gene CLOCK Expression by KCl Depolarization in B35 Rat Neuroblastoma Cells.
Won Je JEON ; Se Hyun KIM ; Myoung Suk SEO ; Ung Gu KANG ; Yong Sik KIM ; Yong Min AHN
Journal of Korean Neuropsychiatric Association 2006;45(1):21-27
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
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Circadian Clocks
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Cycloheximide
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Dactinomycin
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Immunoblotting
;
Incubators
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Neuroblastoma*
;
Rats*
3.Comparison of Three Third-generation Anti-HCV Enzyme Immunoassay Tests.
Hee Soon CHO ; Jin Young MOON ; Chae Hoon LEE ; Kyung Dong KIM
Yeungnam University Journal of Medicine 1998;15(1):143-150
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
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Hepatitis C
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Immunoblotting
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Immunoenzyme Techniques*
;
Mass Screening
4.Antigen analysis of rickettsia typhi isolated in Korea: SDS-PAGE and immunoblotting characters.
Min Kee CHO ; Chang Soon YOON ; Yoon Won KIM ; Hee Sook KIM ; Chang Hong MIN
Journal of the Korean Society for Microbiology 1992;27(5):427-434
No abstract available.
Electrophoresis, Polyacrylamide Gel*
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Immunoblotting*
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Korea*
;
Rickettsia typhi*
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Rickettsia*
6.The increased expression of CD99 in a differentiated neuroblastoma cell line.
Immune Network 2001;1(1):53-60
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
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Apoptosis
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Cell Differentiation
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Cell Line*
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Flow Cytometry
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Humans
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Immunoblotting
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Luciferases
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Neuroblastoma*
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Neurons
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Sarcoma
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Tretinoin
7.Changes of allergenecity of salted and fermented shrimp.
Sang Woon BAE ; Soo Young CHOI ; Hyun Sun JIN ; Cheol Woo KIM ; Kwang Eun LEE ; Eun Seok KANG ; Jun Yong PARK ; Sung Pil HONG ; Hoon Young CHOI ; Jae Hun JUNG ; Yong Soo KIM ; Chein Soo HONG
Journal of Asthma, Allergy and Clinical Immunology 2003;23(1):44-52
BACKGROUND AND OBJECTIVE: Shrimp is one of the most common food allergens. The salted and fermented shrimp (S/F shrimp) is a traditional food and ingested frequently in the daily life of many Korean people. But few studies have been investigated on the allergenicity of S/F shrimp. The aim of our study is to observe the allergenicity of S/F shrimp and to compare it with the allergenicity of shrimp. MATERIAL AND METHODS: Crude extracts were made from three kinds of S/F shrimps and four kinds of shrimps after boiling. The extracts were used for SDS-PAGE, ELISA, ELISA inhibition test, immunoblotting, and immunoblotting inhibition test with sera from 10 subjects who have shrimp specific IgE by radioallergosorbent (RAST) (3+ or 4+). RESULTS: The protein concentrations of S/F shrimp extracts were less than half of the extracts of shrimps (1.9 mg/ml, 1.2 mg/ml, 1.1 mg/ml vs 4mg/ml, 3.83 mg/ml, 6.8 mg/ml, 6.6 mg/ml). The extracts of S/F shrimp showed weaker and more diffusely distributed protein bands in SDS-PAGE and showed less than half of the O.D. value in IgE-ELISA compared to those of shrimp extracts. The specific IgE immunoblotting of S/F shrimp extracts showed 9 bands (36, 33, 30, 29, 28, 26, 23, 20, 19 kDa). Dose dependent inhibition was observed between shrimps and S/F shrimp in ELISA-inhibition and immnunoblotting inhibition test. The allergen concentration of S/F shrimp needed to inhibit 50% of shrimp-lgE ELISA was five times higher than that of shrimp. CONCLUSION: The crude extracts of S/F shrimps showed less than half the amount of protein compared to crude extracts of shrimps have and also showed reduced allergenicity by salting and fermentation.
Allergens
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Complex Mixtures
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Electrophoresis, Polyacrylamide Gel
;
Enzyme-Linked Immunosorbent Assay
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Fermentation
;
Immunoblotting
;
Immunoglobulin E
8.A Case of Pemphigus Foliaceus Developed During Pregnancy.
Jin Hyouk CHOI ; Sung Wook LIM ; Yeon Jin KIM ; Jang Seok BANG ; Jeong Woo LEE ; Moo Kyu SUH ; Tae Hoon KIM ; Jung Ran KIM ; Soo Chan KIM ; Bong Koo LEE ; Jin Chun SUH
Korean Journal of Dermatology 2001;39(12):1449-1451
Pemphigus foliaceus is characterized by superficial blisters with acantholysis in the upper portion of the epidermis. Pemphigus foliaceus in pregnancy is rare. We report a case of pemphigus foliaceus in a 26-year-old pregnant woman who showed erythematous bullae, erosions and crusts on the whole body for 2 months. She was diagnosed by clinical and histopathological findings, direct immunofluorescent test and immunoblotting analysis. She was treated with prednisolone for 28 days and skin lesions were markedly improved.
Acantholysis
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Adult
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Blister
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Epidermis
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Female
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Humans
;
Immunoblotting
;
Pemphigus*
;
Prednisolone
;
Pregnancy*
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Pregnant Women
;
Skin
9.Role of Integrin, FAK (Focal Adhesion Kinase) and ERK (Extracellular Signal Regulated Kinase) on the Suppressed Cell Proliferation of Endometrial Cancer Cells by GnRH (Gonadotropin-Releasing Hormone).
Jong Rak CHOI ; Dong Wook PARK ; Dong Soon CHOI ; Churl K MIN
Korean Journal of Fertility and Sterility 2006;33(2):115-123
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*
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Endometrial Neoplasms*
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Female
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Gonadotropin-Releasing Hormone*
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Humans
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Immunoblotting
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Integrin beta3
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Phosphorylation
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Signal Transduction
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Thymidine
10.Expression of Intracellular Single Chain Antibody Specific to Hepatitis B Virus X Protein.
Young Hee JIN ; Hyung Il KIM ; Sun PARK
Immune Network 2003;3(1):23-28
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
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Eukaryotic Cells
;
Hepatitis B virus*
;
Hepatitis B*
;
Hepatitis*
;
Immunoblotting
;
Microscopy, Fluorescence