• Asian-Australasian Journal of Animal Sciences
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• 제16권7호
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• pp.1055-1059
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• 2003

Keratins, the constituents of epithelial intermediate filaments, are precisely regulated in a tissue and development specific manner. There are two types of keratin in bovine. The type I is acidic keratin and the type II is neutral/basic keratin. 1.5 kb of 5' flanking sequence of Korean cattle Keratin IV gene, type II keratin (59 kDa), was cloned and sequenced. A symmetrical motif AApuCCAAA are located in a defined region upstream of the TATA box. Proximal SP1, AP1, E-box and CACC elements as the major determinants of transcription are identified. When it was compared to the bovine sequence from -600 bp to ATG upstream, the homology was 97% in nucleotide sequence. Several A and T sequences, located in the promoter region, are deleted in the Korean cattle. An expression vector consisted of Korean cattle Keratin IV gene promoter/SV40 large T antigen was transfected to HaCaT cell (Epithelial keratinocyte). The transformed HaCaT cells showed active proliferation when treated with PDGF (Platelet-derived growth factor) in 0.3% soft agar compared to control cells. These results indicate that Korean cattle Keratin IVgene promoter can be used as a promoter for transfection into epithelial cell.

• Journal of Microbiology and Biotechnology
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• 제29권6호
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• pp.863-876
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• 2019

Farm animals such as piglets are often affected by environmental stress, which can disturb the gut ecosystem. Antibiotics were commonly used to prevent diarrhea in weaned piglets, but this was banned by the European Union due to the development of antibiotic resistance. However, the use of probiotics instead of antibiotics may reduce the risk posed by pathogenic microorganisms and reduce the incidence of gastrointestinal diseases. Therefore, this study was conducted to investigate the effects of Lactobacillus casei Zhang on the mechanical barrier and immune function of early-weaned piglets infected using Escherichia coli K88 based on histomorphology and immunology. Fourteen-day-old weaned piglets were divided into a control group and experimental groups that were fed L. casei Zhang and infected with E. coli K88 with or without prefeeding and/or postfeeding of L. casei Zhang. The L. casei Zhang dose used was $10^7CFU/g$ diet. Jejunum segments were obtained before histological, immunohistochemical, and western blot analyses were performed. In addition, the relative mRNA expression of toll receptors and cytokines was measured. Piglets fed L. casei Zhang showed significantly increased jejunum villus height, villus height-crypt depth ratio, muscle thickness, and expression of proliferating cell nuclear antigen and tight junction proteins ZO-1 and occludin. The use of L. casei Zhang effectively reduced intestinal inflammation after infection. We found that L. casei Zhang feeding prevented the jejunum damage induced by E. coli K88, suggesting that it may be a potential alternative to antibiotics for preventing diarrhea in early-weaned piglets.

• Ocean and Polar Research
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• 제25권3호
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• pp.249-256
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• 2003

An immunological method was developed in this study to quantify reproductive output of the female butter clam, Saxidomus purpuratus. A clam egg-specific polyclonal antibody was developed using the purified butter clam egg as an antigen. An indirect Enzyme-Linked Immunosorbent Assay (ELISA) was used in quantitative measurement of the eggs. Size of the butter clam eggs ranged from $70.81{\pm}7.52{\mu}m$ in histology or $88.56{\pm}11.31{\mu}m$ in intact eggs. The predominant egg constituent was protein (37.44%), followed by lipids (11.40%) and carbohydrates (9.68%). The SDS-PAGE showed that the egg proteins are composed of several peptides of molecular weights consisting of 247, 200, 99, 91, 54 and 47 kDa. ELISA indicated that the clams collected from Geoje Island in May 2002 produced 8.2 to 26.8% of their body weight as eggs or 9,307,309 to 31,156,333 with a mean of 16,931,893 eggs per individual clam. The results of this study thus suggest that indirect ELISA using rabbit anti-clam egg IgG as a primary antibody is a rapid, affordable and sensitive method to assess reproductive output of 5. purpuratus and possibly other bivalves using a small amount of eggs.

• 대한수의학회지
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• 제43권1호
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• pp.129-137
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• 2003

Porcine reproductive and respiratory syndrome (PRRS) is characterized by reproductive failures in sows and respiratory problems in piglets. The nucleocapsid(N) protein, encoded by the open reading frame 7 (ORF7) gene, is known to be the most abundant and antigenic protein in PRRS virus. Therefore, it was suggested that the N protein could be a suitable candidate for the detection of PRRS virus-specific antibodies and diagnosis of PRRS. In the present study, the ORF7 gene encoding the N protein was cloned and expressed as a fusion protein with the glutathione S-transferase (GST) in Escherichia coli. The resulting GST-N recombinant protein was used as an antigen for an indirect sandwich enzyme-linked immunosorbent assay (i-ELISA). Expressed GST-N recombinant protein was migrated at 41 kDa and reacted with ORF7-specific monoclonal antibody by Western blotting. In order to increase the specificity of the ELISA for the detection of PRRS virus-specific antibodes, an i-ELISA was developed using an anti-GST antibody as a capture antibody. The sensitivity and specificity of developed i-ELISA were 92% and 96%, respectively. Based on these results, it was suggested that the i-ELISA is a simple and rapid test for screening a large number of swine sera for the anti-PRRS virus antibodies.

• Parasites, Hosts and Diseases
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• 제31권1호
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• pp.43-48
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• 1993

요꼬가와흡충증은 주로 은어회를 먹어 감염되는 흡충증으로서 소장염(소장염)을 일으켜 피로 감, 복통, 설사 등 중상이 나타나며 말초혈액 호산구증가를 동반한다. 이 흡충중은 대변에서 충란을 검출하여 진단하나 급성 장염으로 발현하는 환자의 일부에서는 대변검사로 충란을 발견 할 수 없어 보조적인 진단으로 특이항체검사가 필요한 경우가 있다. 이 실험은 요꼬가와흡충증 의 특이항체검사의 가치를 검토하고 검사에서 항원으로 사용하는 피낭유충 생리식염수 추출액 내 특이항원 단백질을 찾기 위하여 실시하였다. 항원으로는 요꼬가와흡충 피낭유충의 추출액을 사용하였고 감염 및 대조혈청은 인상적으로 급성 요로가와흡충증으로 진단한 환자 11명, 간질 (간질) 환자 5명, 간흡충 환자 5명, 폐흡충 환자 5명과 대조군 11명의 것을 사용하였다. 항체 검사로 효소면역측정 법으로 특이 IgG항체를 검사한 결과 임상적으로 진단한 요꼬가와흡충증 환자 11명중 10명이 양성반응을 나타내었다. 요꼬가와흡충 항원에 대하여 다른 흡충 감염자 혈청도 교차반응을 보였으나 동종항원에 대하여 가장 높은 흡광도를 나타내었다. 따라서 특이항체검사에서는 여러가지 흡충항원에 대한 항체검사를 동시에 검사할 필요가 있다고 판단하였다. 항원 구성 단백질을 관찰하기 위하여 7.5-15% gel에서 SDS-PAGE한 결과 항원에서 주(주)단백질대 14개 이상을 구별할 수 있었다. SDS-PAGE/immunoblot을 실시하여 요꼬가와흡충증에서만 반응하는 특이항원대를 검색한 바, 항원 단백질대 대부분이 다른 흡충증 환자 혈청과 교차반응을 나타내었으나 66 및 22 kDa 단백질대는 교차반응이 제일 적어 특이항원 단백질이라고 판단하였다.

• Tuberculosis and Respiratory Diseases
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• 제47권3호
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• pp.311-320
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• 1999

연구 배경 : 최근 전세계적으로 결핵이 증가하며 새로운 문제로 대두되고 있으며, 특히 결핵의 유병률이 높은 우리나라의 경우에는 민감도가 높으며 간편한 검사방법 이 필요하다. M. tuberculosis 항원을 사용한 rapid membrane based assay 검사 방법으로 최근에 폐결핵 환자의 결핵항원 특이 항체를 검출한다고 알려진 상용화된 2 가지 kit를 이용하여 폐결핵의 혈청학적 진단 가능성을 알아보고자 하였다. 방법 : 1998년 3월부터 9월까지 충남대학교 병원에서 입원 치료를 받은 호흡기 환자 총 45예를 대상으로 하였다. 객담 도말 양성의 활동성 폐결핵 22예와 비활동성 폐결핵 14예 그리고 대조군으로 비결핵성 호흡기 질환 9예를 대상으로 하여 STAT-PAK ULTRA FAST$^{(R)}$ 및 ICT tuberculosis$^{(R)}$ 검사를 실시하여 결핵 혈청 검사의 유용성을 알아보고 자하였다. 결과: STAT-PAK ULTRA FAST$^{(R)}$의 경우에는 폐결핵과 비활동성 폐결핵의 비교에서 민감도는 77.3%, 28.6%, 양성 63.0%, 음성 예측도는 44.4%, 위양성률은 71.4%, 위음성률은 22.7% 이었고, 활동성 폐결핵과 비결핵성 호흡기 질환의 비교에서는 민감도는 77.3%, 특이도는 33.3%, 양성 예측도는 73.9%, 음성 에측도는 37.5%, 위양성률은 66.7%, 위음성률은 22.7% 이었다. ICT tuberculosis$^{(R)}$의 경우에는 활동성 폐결핵과 비활동성 폐결핵의 비교에서는 민감도는 54.5%. 특이도는 57%, 양성 예측도는 66.7%, 음성 예측도는 44.4%, 위양성률은 42.9%, 위음성률은 45.4 %이었고, 활동성 폐결핵과 비결핵성 호흡기 질환에서는 민감도는 54.5%, 특이도는 100%, 양성 예측도는 100%, 음성 예측도는 47.4%. 위양성률은 0%, 위음성률은 45.4%이었다. 폐결핵의 중증정도(extent) 와 양성율은 상기 2가지 kit에서 큰 차이가 없었다. 결론: 따라서 상기 2가지의 kit는 항체 반응이 다양하여 임상적으로 폐결핵의 진단에 이용하는 것에는 문제가 있다고 생각되며, 일단 양성반응을 보인 환자의 경우, 항결핵제 투여에 따른 추적 검사에서 항체가의 변화가 있는지에 대한 연구가 필요하리라 생각된다.

• 사상체질의학회지
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• 제22권3호
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• pp.141-151
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• 2010

1. Objectives: Three herbal formulas (Yuldahanso-tang, Chungsimyonja-tang, and Taeeumjowi-tang) for Taeeumin were applied to investigate the immunological activities on antigen (Ag)-specific or Ag-non-specific immune responses in murine macrophage cell line (RAW 264.7) and ovalbumin (OVA)-immunized mice. 2. Methods: This study was carried out in nitric oxide (NO) synthesis in RAW 264.7 cells and cellular proliferation in mouse splenocytes according to three herbal formulas. C57BL/6 mice were immunized intraperitonially with OVA/aluminium ($100\;{\mu}g/200\;{\mu}g$/mouse) on day 1, 8, and 15. Three herbal formulas were administrated to mice orally for 3 weeks from day 1. On day 22, OVA-, lipopolysaccharide (LPS)-, and concanavalin A (Con A)-stimulated splenocyte proliferation and antibodies (OVA-specific antibodies of the IgG, IgG1, and total IgM classes) in plasma were measured. 3. Results: Yuldahanso-tang and Chungsimyonja-tang increased NO synthesis in RAW 264.7 cells. Three herbal formulas significantly enhanced cellular proliferation by LPS and Con A in splenocytes from OVA-immunized mice (p<.001). Three herbal formulas for Taeeumin also significantly enhanced plasma OVA-specific IgG, IgG1, and total IgM levels compared with the OVA/Alum group. 4. Conclusion: These results suggested that three herbal formulas for Taeeumin could be used as stimulator of immune response.

• Animal cells and systems
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• 제12권1호
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• pp.15-21
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• 2008

Nitric oxide(NO) has been known to play important roles in numerous physiologic processes including neurotransmission, vasorelaxation, and cellular apoptosis. Using a mouse cDNA gene chip, we examined expression patterns and time course of NO-dependent genes in mouse macrophage RAW264.7 cells. Genes shown to be upregulated more than two fold or at least at two serial time points were further selected and validated by RT-PCR. Finally, 81 selected genes were classified by function as signaling, apoptosis, inflammation, transcription, translation, ionic homeostasis and metabolism. Among those, genes related with signaling, apoptosis and inflammation, such as guanylate cyclase 1, soluble, alpha3(Gucy1a3); protein kinase C, alpha($Pkc{\alpha}$); lymphocyte protein tyrosine kinase(Lck); BCL2/adenovirus E1B 19 kDa-interacting protein(Bnip3); apoptotic protease activating factor 1(Apaf1); X-linked inhibitor of apoptosis(Xiap); cyclin G1(Ccng1); chemokine(C-C motif) ligand 4(Ccl4); B cell translocation gene 2, anti-proliferative(Btg2); lysozyme 2(Lyz2); secreted phosphoprotein 1(Spp1); heme oxygenase(decycling) 1(Hmox1); CD14 antigen(Cd14); and granulin(Grn) may play important roles in NO-dependent responses in murine macrophages.

• 사상체질의학회지
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• 제22권4호
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• pp.77-84
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• 2010

1. Objectives Yanggyuksanhwa-tang for Soyangin was applied to investigate the immunological activities on antigen (Ag)-specific or Ag-non-specific immune responses on murine macrophage cell line (RAW 264.7) and ovalbumin/aluminium (OVA/Alum)-immunized mice. 2. Methods This study were carried out in nitric oxide (NO) synthesis on RAW 264.7 cells and cellular proliferation on mouse splenocytes. C57BL/6 mice were immunized intraperitonially with OVA/Alum (100 ${\mu}g$/200 ${\mu}g$) on day 1, 8, and 15. Yanggyuksanhwa-tang was administrated to mice orally for 3 weeks from day 1. On day 22, OVA-, lipopolysaccharide (LPS)-, and concanavalin A (Con A)-stimulated splenocyte proliferation and antibodies (OVA-specific antibodies of the IgG, IgG1, and total IgM classes) in plasma were measured. 3. Results Yanggyuksanhwa-tang significantly enhanced cellular proliferation by LPS and Con A on splenocytes from OVA/Alum-immunized mice (p<.001). Yanggyuksanhwa-tang also significantly enhanced plasma OVA-specific IgG (p<.001), IgG1 (p<.001), and total IgM (p<.01) levels compared with the OVA/Alum group. 4. Conclusions These results suggested that Yanggyuksanhwa-tang for Soyangin could be used as immunopotent.

• 대한한의학회지
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• 제31권5호
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• pp.112-123
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• 2010

Objectives: Three yin-tonifying formulae (Ssanghwa-tang, Yukmijihwang-tang and Jaeumganghwa-tang) were applied to investigate their immunological activities on antigen (Ag)-specific or Ag-non-specific immune responses in the murine macrophage cell line (RAW 264.7) and in ovalbumin (OVA)-immunized mice. Methods: This study was carried out in nitricoxide (NO) synthesis in RAW 264.7 cells and cellular proliferation in mouse splenocytes in association with three herbal formulas. C57BL/6 mice were immunized intraperitoneally with OVA/aluminum ($100\;{\mu}g/200\;{\mu}g$/mouse) on days 1, 8, and 15. Three herbal formulas were administrated to mice orally for 3 weeks from day 1. On day 22, OVA-, lipopolysaccharide (LPS)-, and concanavalin A (Con A)-stimulated splenocyte proliferation and antibodies (OVA-specific antibodies of the IgG, IgG1, and total IgM classes) in plasma were measured. Results: All three yin-tonifying formulas significantly enhanced cellular proliferation by LPS and Con A in splenocytes from OVA-immunized mice (p<0.001). Also, these herbal formulas all significantly enhanced plasma OVA-specific IgG, IgG1, and total IgM levels compared with the OVA/Alum group. Conclusion: These results suggested that the three yin-tonifying formulae could be used as stimulators of immune response.