• Korean Journal of Veterinary Service
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• v.17 no.1
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• pp.37-43
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• 1994

The rate of 58 neonatal calves in infection of Theileria sergenti was investigated in random samples on the farms located in Kyunggi, Chonbuk districts of Korea. 1. The criteria used in veryfying infection with T. sergenti included the detection of parasites by giemsa stain and acridine orange stain in the blood smear slides. 2. Further evidence of current or previous exposure to T. sergenti was based on demonstration of T. sergenti specific antibody and antigen by the western immunoblot and the directed immunofluorescent antibody test in the peripherial blood of the calves. 3. The prevalence rates were 35%, 50% in Kyunggi, Chonbuk provinces respectively and the overall prevalence in all the farms was 43.2％ by means of acridine orange stain. 4. The parasites that were observed in the peripherial blood of calves was showen surely by the western immunoblot to the characteristic 34KD antigen among the proteins of T. sergenti (Korean isolate). 5. And the antigen of the neonatal calves reacted at the very highest titer(1 : 2, 560) 6. These data highlight the significances of T. sergenti in the neonatal calf disease in Korea.

• Korean Journal of Veterinary Research
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• v.35 no.1
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• pp.87-94
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• 1995

Western immunoblot analysis of antigen of T sergenti merozoite revealed that the immunodominant proteins of this organism were characterized as the 18KD, 29KD, 34KD, 45KD and 105KD in Korea. The 34KD and 45KD among those immunodominant proteins of the parasite were isolated and their amino acid sequences from the $NH_2$-terminus were determined and synthesized. They respective polypeptides were cationized to enhance their antigenicity, fortified with Freund's adjuvant and tested for immunogenicity in rabbits and cattle. The results obtained were as follows; 1. Theileria sergenti merozoite antigen was shown in 120KD, 100KD, 66KD, 45KD, 34KD and 30KD in western immunoblot using serum of rabbits immunized with 34KD synthetic polypeptide and 70KD, 58KD, 55KD and 45KD using bovine serum. In western immunoblot, 45KD, 34KD and 30KD were recognized by immunized rabbits, and 50KD and 45KD by cattle sera immunized with 45KD synthetic polypeptide, respectively. 2. The ELISA utilizing the synthetic polypeptides demonstrated significant antibody response to the respective peptides. After the 2nd booster injection, an OD of 0.760(preimmunization 0.132) in rabbits and an OD of 0.645(preimmunization 0.488) to 34KD synthetic polypeptide in cattle were observed. In animals immunized with 45KD synthetic polypeptide, after the 2nd booster injection, an OD of 0.640(preimmunization 0.144) in rabbit, and an OD of 0.776 (preimmunization 0.477) in cattle were measured. 3. After the 2nd booster the reciprocal IFA titer was 1:64 in rabbits and 1:512 in cattle immunized with the 34KD synthetic polypeptide. The IFA titre was observed as 1:512 in rabbit and 1:1,024 in cattle in immunized with the 45KD synthetic polypeptide.

• Korean Journal of Veterinary Research
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• v.33 no.4
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• pp.665-671
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• 1993

The rate of 67 neonatal calves's infection of Theileria sergenti was investigated in random samples on the farms located in Kyeongki, Chonbuk and Jeju districts of Korea. The criteria used in verifying infection with T sergenti included the detection of parasites by giemsa's stain and acridine orange stain in the blood smear slides. Further evidence of current or previous exposure to T sergenti was based on the demonstration of T sergenti-specific antibody and antigen by the western immunoblot and the indirect immunofluorescence antibody test in the peripheral blood of the calves. The prevalence rates were 35%, 50% and 100% in Kyeongki, Chonbuk and Jeju provinces respectively and the overall prevalence in all the farms was 43.2% by means of acridine orange stain. The parasites that were observed in the peripheral blood of calves was shown surely by the western immunoblot to the characteristic 34KD antigen among the proteins of T sergenti (Korean Isolate). And the antibody of the neonatal calves reacted at the very highest titer(1 : >2,520). These data highlight the significance of T sergenti in the neonatal calf disease in Korea.

• Tuberculosis and Respiratory Diseases
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• v.50 no.5
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• pp.549-556
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• 2001

Background : IGF-I is an important mitogen in many types of malignancies. Tumors also express many IGF binding proteins, which modulate IGF action. The propose of this study was to evaluate the effect of IGF-I and IGFBP on cell proliferation in mouse lung cancer cells (3LL). Methods : The cellular proliferation of 3LL with the treatment of growth factors was evaluated using MTT assay. Western ligand blot was performed in order to determine whether 3LL cells secrete IGFBPs and we evaluated the effect of IGFBP on cellular proliferation. Results : The treatment of 3LL cells with IGF-I increased cellular proliferation in a serum free media. Western ligand blot of conditioned medium of 3LL with $^{125}I$-IGF-I demonstrated one single major band with an estimated molecular mass of 24 kDa. This band was identified as IGFBP-4 with immunoblot analysis using antisera. The addition of anti-IGFBP-4 antibody to abrogate the effect of IGFBP-4 resulted in increased cellular proliferation suggesting that IGFBP-4 inhibits cell growth. Conclusion : IGF-I increases cellular proliferation, however the secreted IGFBP-4 has an inhibitory function on cell growth in 3LL. These findings suggest that IGF-I and IGFBP are involved in the cell proliferation.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.05a
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• pp.145-146
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• 2000

The presence of cross-reacting muramidase in Lactobacillus delbrueckii ssp. bulgaricus ULl2 was shown by using monoclonal antibodies raised against an muramidase-2 of Enterococcus hirae ATCC 9790. The separation of protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by Western immunoblot confirmed the presence of one cross-reacting band in Enterococcus hirae with an estimated molecular mass of 80 kDa, L. bulgaricus cultured cells harvested after 4 and 12 h were submitted to different autolysin releasing procedures and the liberated products were allowed to cross-react with muramidase-2 antibodies in order to estimate the efficiency of each treatment. Although the cultured cells harvested after 4 h yielded only a slight immune-reaction in Western immunoblots against these enzyme monoclonal antibodies, a strong signal was observed for the cell walls obtained from the same experimental conditions and treated with Triton X-100 surfactant. The same phenomenon was also observed by light fluorescence microscopy. Immune-labelling followed by optical and electron microscopy have shown that the muramidase-2 of L. bulgaricus ULl2 was essentially localized in the innermost part of the cell wall.(omitted)

• Journal of Microbiology and Biotechnology
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• v.4 no.3
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• pp.176-182
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• 1994

The light-organ symbiont of pine cone fish, Vibrio fischeri, senses its presence in the host and responds to environmental changes by differentially expressing its symbiosis-related luminescence genes. The V. fischeri luminescence genes are activated by LuxR protein in the presence of an autoinducer. In an effort to elucidate the mechanism of regulation of luxR, a plasmid containing luxR was mutagenized in vitro with hydroxylamine and a luxR mutant plasmid was isolated by its ability to activate luminescence genes cloned in E. coli in the absence of the autoinducer. The specific base change identified by DNA sequencing was only single base transition at ＋78 from the transcriptional start of luxR. Based on a Western immunoblot analysis, the nucleotide change directed the synthesis of much higher level of LuxR protein without any amino acid substitutions. The results suggest that the region including the ＋78th base is presumably internal operator required for autorepression of luxR, and the increased cellular level of LuxR results in activation of luminescence genes by autoinducer independent fashion.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1992.05a
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• pp.54-54
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• 1992

약물, hormone, 독성물질등의 대사능과 발암 가능성등이 간장 장해시 및 ketosis시에 달라지는 원인과 기전, 독성물질 대사효소의 변동과 그 작용기전을 규명하고자, 대표적인 간장장해 물질인 carbon tetrachloride를 rat에 투여하여 간장 장해를 일으키고, 당뇨병, starvation, high-fat diet처리하여 ketosls상태를 만든 후에, specific cytochrome P45O polyclonal antibodies와 cDNA probes를 사용하여, enzyme activitieg, Western immunoblot analysis와 mRNA Northern blot analysis 등을 실험하여, 간장 장해와 ketosis시 cytochrome P45O의 변동과 그 작용기전, regulation을 규명하고자 하였다. 실험 결과, $CCl_4$투여후 P450IIE enzyme (aniline hydroxylase) 활성이 시간 의존적으로 급격히 떨어졌고, P450IIE protein양이 똑같은 방식으로 감소되었으나 mRNA level은 변화가 없었다. $CCl_4$에 의해서 P450IIE는 protein의 특이적인 파괴에 의한 post-translational reduction됨을 알 수 있었다. 반면에 당뇨병, starvation, high-fat diet등 ketosis시에는 P450IIE 효소활성이 2-3배 증가되었고, P450IIE protein양도 같은 수준으로 증가되었으며, mRNA도 증가 되었다. Ketosis시에는 P450IIE가 pretranslational activation됨을 알 수 있었다.

• Toxicological Research
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• v.9 no.2
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• pp.153-158
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• 1993

Pyridine and acetone are efficacious inducers of CYP2E1 in both rats and rabbits. The response in the elevation of CYP2E1 levels, changes in CYP2E1 mRNA levels, and enhanced translational processing of hepatic CYP2E1 mRNA during the early phase of CYP2E1 induction by the solvents pyridine and acetone were examined. Time-depen-dent incease in CYP2E1 levels occurred at early times (6-24h) following a single dose of pyridine treatmene, as assessed by Western immunoblot analysis, whereas the levels in CYP2E1 mRNA transiently decreased at 12h post-treatment, returning to the level present in untreated animals.

• Parasites, Hosts and Diseases
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• v.33 no.2
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• pp.101-106
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• 1995

Soluble protein of purified Pneumocvstis cnrinii was prepared from experimentally infected rats. SDS-PAGE of the crude antigen resolved about 20 protein bands from 20 to 200 kDa. Out of thenl, 116 kDa band strongly reacted and 45-55 and 100 kDa bands reacted weakly to the positive reference human serum from U.S.A. Western blot analysis with sera of 130 normal children and 15 newborns in Korea revealed specific IgG antibody reaction to 40-55 and 116 kDa protein bands. Forty percent (40.0%) of the 145 sera were positive with any of the antigenic protein bands of R corinii. The positive rate was 56% in 50 males and 33.3% in 48 females. The protein bands 40-55 and 116 kDa from rat P. carinii were confirmed to cross-react with human sera in Korea.

• Journal of fish pathology
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• v.12 no.2
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• pp.89-99
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• 1999

New sixteen heat shock proteins (Hsps) and ten ethanol shock proteins were appeared on the analysis with SDS-PAGE when cultivation temperature for the Vibrio vulnifrcus ATCC 27562 strain was shifted-up to $42^{\circ}C$ from $30^{\circ}C$ for 20 mins and treated with of 6% ethanol for 10 mins, respectively. Even the induction of thermotolerance in V. vulnificus was coincided with the induction of Hsps if the pre-shock was adjusted to thermal temperature. Outer membrane proteins (OMPs) that were purified from the membrane of cells after heat shock showed more immunodominant pattern to the immunized rabbit anti-V. vulnificus O serum in enzyme-linked immunosorbent assay (ELISA). On the western immunoblot analysis it was confirmed that both 62 kDa IMP and 69 kDa OMP in the Hsps and 48 kDa IMP a major OMP in the ethanol shock proteins were reacted with rabbit anti-V. vulnificus O sera. Agglutination titer of the heat shocked V. vulnificus with rabbit anti-V. vulnificus O serum was higher than that of the untreated bacteria.