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PCR-RFLP for the Identification of Mammalian Livestock Animal Species
Han, Sang-Hyun,Park, Seon-Mi,Oh, Hong-Shik,Kang, Geunho,Park, Beom-Young,Ko, Moon-Suck,Cho, Sang-Rae,Kang, Yong-Jun,Kim, Sang-Geum,Cho, In-Cheol The Korean Society of Embryo Transfer 2013 한국동물생명공학회지 Vol.28 No.4
Precise, rapid and simple methods for species identification in animals are among the most important techniques in the livestock industry and research fields including meat classification. In this study, polymerase chain reaction (PCR) based molecular identification using inter species polymorphisms were examined by PCR-restriction fragment length polymorphism (RFLP) analysis for mitochondrial DNA (mtDNA) cytochrome b (CYTB) gene sequences among four mammalian livestock animals (cattle, horse, goat and pig). The results from PCR-RFLP analysis using the AluI restriction enzyme were also provided for the species-specific band patterns among CYTB gene sequences in these four species. The AluI-digestion for CYTB genes provided interesting migration patterns differentially displayed according to each species. Cattle and horse had one AluI-recognition site at different nucleotide positions and their AluI-digested fragments showed different band patterns on the gels. Pig had two AluI-recognition sites within the amplified CYTB sequences and produced three bands on the gels. Goat had no AluI-recognition site and was located at the same position as the uncut PCR product. The results showed the species-specific band patterns on a single gel among the four livestock animal species by AluI-RFLP. In addition, the results from blind tests for the meat samples collected from providers without any records showed the identical information on the species recorded by observing their phenotypes before slaughter. The application of this PCR-RFLP method can be useful and provide rapid, simple, and clear information regarding species identification for various tissue samples originating from tested livestock species.
아토피피부염 환자에서 국소 피메크로리무스의 효과와 순응도에 관한 다기관 설문조사
김범준 ( Beom Joon Kim ),김명남 ( Myeung Nam Kim ),김규한 ( Kyu Han Kim ),김도원 ( Do Won Kim ),노영석 ( Young Suck Ro ),박천욱 ( Chun Wook Park ),이승철 ( Seung Chul Lee ),이애영 ( Ai Young Lee ),최지호 ( Jee Ho Choi ),이광훈 ( K 대한피부과학회 2008 대한피부과학회지 Vol.46 No.10
Background: Atopic dermatitis (AD) is a chronic, relapsing inflammatory skin disease that`s influenced by many genetic and environmental factors. Various treatment modalities are being applied for AD, including phototherapy, topical applicants and systemic agents. However, there has been no mass survey in Korea concerning the compliance of patients` to use their prescribed medication, which might influence the clinical efficacy of the physicians` treatment. Objective: The purpose of the present study is to evaluate the efficacy, safety and patient compliance with using topical pimecrolimus to treat AD. Methods: We reviewed the medical recordings, laboratory profiles, clinical severity scoring and photographs of AD patients at 9 general hospitals in Seoul and the local area of Korea. Interviews with the patients and a telephone survey were also done. Those patients who weren`t perscribed topical applicants or those who did not use topical pimecrolimus or steroid within 6 months were excluded from this study. Results: Topical pimecrolimus cream effectively controlled AD with a reduction of the EASI score from baseline 13.75 to 11.39 at 2 weeks and 4.46 at 10 weeks of topical pimecrolimus application. When topical pimecrolimus cream was applied for more than 12 months it significantly suppressed the recurrence and reactivation of AD (p<0.05). Although 22.6% of the patients complained of adverse effects, these were all transient and they did not evoke significant medical problems. Using topical pimecrolimus cream did not show significant adverse effects or complications. Conclusion: Topical pimecrolimus might well be an effective treatment modality for treating AD when patients show good compliance for applying the cream. (Korean J Dermatol 2008;46(10):1357∼1361)
Choi, Suck-Chei,Kim, Beom-Su,Yoon, Kwon-Ha,Song, Moon-Young,Oh, Hyun-Mee,Han, Weon-Cheol,Kim, Tae-Hyeon,Kim, Eun-Cheol,Jun, Chang Duk The Korean Society for Integrative Biology 2002 Korean journal of biological sciences Vol.6 No.2
Nitric oxide has high affinity for iron, and thus it can cause intracellular iron loss. We tested the idea that intracellular iron can be the primary target of NO toxicity by comparing the signaling mechanisms involved in cell death caused by iron depletion and that caused by NO. Treatment of HL-60 cells with a NO donor, S-nitroso-N-acetyl-DL-penicillamine (SNAP), decreased the intracellular iron level rapidly as that observed with the iron chelator deferoxamine (DFO). Iron chelators such as DFO and mimosine could induce death of human leukemic HL-60 cells by a mechanism requiring activation of p38 kinase, c-Jun N-terminal kinase, caspase-3 and caspase-8. DFO and SNAP also caused release of cytochrome c from mitochondria. Inhibition of p38 kinase by a selective inhibitor, SB203580, abolished the NO and DFO-induced cell death, release of cytochrome c, and activation of caspase-3 and caspase-8, thus indicating that p38 kinase lies upstream in the cell death processes. In a parallel situation, the cells that are sensitive to NO showed similar sensitivity to DFO. Moreover, simultaneous addition of ferric citrate, an iron-containing compound, inhibited the SNAP and DFO-induced activation of caspases and also blocked the NO-mediated cell cycle arrest at $G_1$ phase. Collectively, our data implicate that the NO-induced cell death of tumor cells including HL-60 cells is mediated by depletion of iron and further suggest that activation of p38 kinase lies upstream of cytochrome c release and caspase activation involved in this apoptotic process.
PCR-RFLP for the Identification of Mammalian Livestock Animal Species
Sang-Hyun Han,Seon-mi Park,Hong-Shik Oh,Geunho Kang,Beom-Young Park,Moon-Suck Ko,Sang-Rae Cho,Yong-Jun Kang,Sang-Geum Kim,In-Cheol Cho 韓國受精卵移植學會 2013 한국동물생명공학회지 Vol.28 No.4
Precise, rapid and simple methods for species identification in animals are among the most important techniques in the livestock industry and research fields including meat classification. In this study, polymerase chain reaction (PCR) based molecular identification using inter species polymorphisms were examined by PCR-restriction fragment length polymorphism (RFLP) analysis for mitochondrial DNA (mtDNA) cytochrome b (CYTB) gene sequences among four mammalian livestock animals (cattle, horse, goat and pig). The results from PCR-RFLP analysis using the AluI restriction enzyme were also provided for the species-specific band patterns among CYTB gene sequences in these four species. The AluI-digestion for CYTB genes provided interesting migration patterns differentially displayed according to each species. Cattle and horse had one AluI-recognition site at different nucleotide positions and their AluI-digested fragments showed different band patterns on the gels. Pig had two AluI-recognition sites within the amplified CYTB sequences and produced three bands on the gels. Goat had no AluI-recognition site and was located at the same position as the uncut PCR product. The results showed the species-specific band patterns on a single gel among the four livestock animal species by AluI-RFLP. In addition, the results from blind tests for the meat samples collected from providers without any records showed the identical information on the species recorded by observing their phenotypes before slaughter. The application of this PCR-RFLP method can be useful and provide rapid, simple, and clear information regarding species identification for various tissue samples originating from tested livestock species.
비육돈의 도체형질과 A-FABP Promoter 유전자형의 연관 분석
한상현,조인철,이종언,고문석,성필남,박범영,Han, Sang-Hyun,Cho, In-Cheol,Lee, Chong-Eon,Ko, Moon-Suck,Seong, Pil-Nam,Park, Beom-Young 한국축산식품학회 2007 한국축산식품학회지 Vol.27 No.3
본 연구는 비육돈에서 지방세포지방산결합단백질(A-FABP) 유전자의 유전자형과 도체형질의 상관관계를 조사하였다. 유전자 A-FABP의 프로모터 영역에 대해 HinfI-RFLP를 이용하여 유전자형을 조사한 결과 두 가지 유전자형(-406T/T T/C)들이 발견되었고 C/C는 발견되지 않았다. 유전자형 중 -406C/C 동형접합자가 발견되지 않는 이유는 비육돈 생산에 관여하는 돼지 품종들 중에서 Duroc 품종만이 이 유전자에 대한 고도의 다형성을 나타내기 때문으로 추정되었다. 유전자형 -406C/-를 나타내는 돼지들은 근내지방과 도체중이 통계적으로 유의적인 증가함을 나타내었으나 (p<0.05), 그 외의 도체형질들(조지방, 육색, 조직감, 수분 삼출도, 근육분리도)은 유의한 차이를 나타내지 않았다 (p>0.05). 본 연구를 통해 돼지 A-FABP 유전자의 -406C/- 유전자형이 근내지방에 대한 유용한 분자 마커가 될 수 있을 뿐만 아니라 다형성이 비육돈의 근내지방에 대한 유용한 분자 마커가 될 수 있으며, 특히 Duroc 품종을 최종 종모돈으로 이용할 경우 이 유전자형을 보유한 돼지를 이용하는 분자육종체계에서 marbling이 잘 된 돼지의 생산을 통한 육질 개선에 기여할 것으로 사료된다. The association between the promoter region genotypes of the porcine adipocyte fatty acid binding protein (A-FABP) gene and carcass traits in commercial pigs was examined. Interestingly, only two different genotypes (-406T/T and T/C) for the A-FABP gene were detected using Hinf-RFLP for the substitution mutation T-406C in the A-FABP promoter in commercial pigs, and no -406C/C homozygotes were detected. The reason for the lack of the -406 C/C genotype is due to the breeding system in which only Duroc, which has a high frequency of -406C/- in this locus among the three breeds involved in commercial pig production, is typically used as a terminal sire. The pigs containing the genotype -406C/- were significantly associated with an increase in intramuscula. fat content and carcass weight (p<0.05), but there was no association with the other carcass traits tested (fat composition, color score, texture score, moisture, and separation score between muscles). This study suggests that the -406C/- genotype of the porcine A-FABP gene may not only be a useful molecular marker for intramuscular fat, but may also contribute to the improvement of meat quality by the production of well-marbled pigs by breeding animals containing this genotype, especially Duroc, as a terminal sire for commercial pigs.