• 제목/요약/키워드: Single strand

검색결과 286건 처리시간 0.028초

Single Nucleotide Polymorphisms on Peroxisome Proliferator-activated Receptor Genes Associated with Fatness Traits in Chicken

  • Meng, H.;Zhao, J.G.;Li, Z.H.;Li, H.
    • Asian-Australasian Journal of Animal Sciences
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    • 제18권9호
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    • pp.1221-1225
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    • 2005
  • The peroxisome proliferator-activated receptors (PPARs) are members of a superfamily of nuclear hormone receptors. Lots of studies in rodents and humans have shown that PPARs were involved in lipid metabolism and adipocyte differentiation. The main objective of this work was to detect the single nucleotide polymorphisms (SNPs) in whole coding regions of peroxisome proliferator-activated receptor alpha (PPAR-$\alpha$) and gamma (PPAR-$\gamma$) genes with approach of single strand conformation polymorphism (SSCP) in the chicken population of Arber Acres broiler, Hyline layer and three Chinese native breeds (Shiqiza, Beijing You, Bai'r). Two SNPs of C1029T and C297T were found in chicken PPAR-$\alpha$ and PPAR-$\gamma$ genes respectively and each SNP found three genotypes in the experimental populations. The results showed that the distribution frequency of 3 genotypes in Arber Acres broiler, Hyline layer and Chinese native breeds had significant differences on the PPAR-$\alpha$ and PPAR-$\gamma$ gene respectively (p<0.01). Furthermore, in the PPAR-$\alpha$ gene, the results of least square estimation for genotypes and body composition traits showed the BB genotype birds had higher abdominal fat weight (AFW) and percentage of abdominal fat (AFP) than AA genotype birds (p<0.05). From these we conjecture the PPAR-$\alpha$ and PPAR-$\gamma$ genes were suffered intensive selection during the long term commercial breeding and the PPAR-$\alpha$ gene may be a major gene or linked to the major genes that impact chicken fat metabolism and the SNPs could be used in molecular assistant selection (MAS) as a genetic marker for the chicken fatness traits.

Mutation Screening and Association Study of the Folylpolyglutamate Synthetase (FPGS) Gene with Susceptibility to Childhood Acute Lymphoblastic Leukemia

  • Piwkham, Duangjai;Siriboonpiputtana, Teerapong;Beuten, Joke;Pakakasama, Samart;Gelfond, Jonathan AL;Paisooksantivatana, Karan;Tomlinson, Gail E;Rerkamnuaychoke, Budsaba
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권11호
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    • pp.4727-4732
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    • 2015
  • Background: Folylpolyglutamate synthetase (FPGS), an important enzyme in the folate metabolic pathway, plays a central role in intracellular accumulation of folate and antifolate in several mammalian cell types. Loss of FPGS activity results in decreased cellular levels of antifolates and consequently to polyglutamatable antifolates in acute lymphoblastic leukemia (ALL). Materials and Methods: During May 1997 and December 2003, 134 children diagnosed with ALL were recruited from one hospital in Thailand. We performed a mutation analysis in the coding regions of the FPGS gene and the association between single nucleotide polymorphisms (SNPs) within FPGS in a case-control sample of childhood ALL patients. Mutation screening was conducted by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and subsequently with direct sequencing (n=72). Association analysis between common FPGS variants and ALL risk was done in 98 childhood ALL cases and 95 healthy volunteers recruited as controls. Results: Seven SNPs in the FPGS coding region were identified by mutation analysis, 3 of which (IVS13+55C>T, g.1297T>G, and g.1508C>T) were recognized as novel SNPs. Association analysis revealed 3 of 6 SNPs to confer significant increase in ALL risk these being rs7039798 (p=0.014, OR=2.14), rs1544105 (p=0.010, OR= 2.24), and rs10106 (p=0.026, OR=1.99). Conclusions: These findings suggested that common genetic polymorphisms in the FPGS coding region including rs7039789, rs1544105, and rs10106 are significantly associated with increased ALL risk in Thai children.

단세포 전기영동법을 이용한 인체, 마우스 및 랫드 림프구의 방사선에 의해 유발된 DNA 손상 측정 (Analysis of gamma-ray-induced DNA damage in human, mouse and rat peripheral blood lymphocytes using single-cell gel electrophoresis)

  • 오헌;정우희;박혜란;김성호;조성기
    • 대한수의학회지
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    • 제44권1호
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    • pp.41-47
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    • 2004
  • The alkaline single-cell gel electrophoresis (SCGE) assay, called the comet assay, has been applied to detect DNA damage induced by a number of chemicals and biological factors in vivo and in vitro. The DNA damage was analysed by tail moment (TM) and tail length (TL), which were markers of DNA strand breaks in SCGE. Human, mouse and rat peripheral blood lymphocytes (PBLs) were irradiated with different doses of $^{60}Co$ ${\gamma}$-rays, e.g. 1, 2, 4, and 8 Gy at a dose rate of 1 Gy/min. A dose-dependent increase in TM (p<0.01) and TL (p<0.01) was obtained at all the radiation doses (1-8 Gy) in human, mouse and rat PBLs. Mouse PBLs were more sensitive than human PBLs which were in turn more sensitive than rat PBLs when the treated dosages were 1 and 2 Gy. However, human PBLs were more sensitive than mouse PBLs which were in turn more sensitive than rat PBLs when the irradiation dosages were 4 and 8 Gy. Data from all three species could be fitted to a linear-quadratic model. These results indicated that there may be inherent differences in the radio-sensitivity among PBLs of mammalian species.

Effects of Reactive Oxygen Species on DNA Stability in Humnn Spermatozoa

  • Kang, Hee-Gyoo;Kim, Tai-Jeon;Bae, Hyung-Joon;Moon, Hi-Joo;Kim, Myo-Kyung;Kim, Dong-Hoon;Sungwon-Han;Lee, Ho-Joon;Yang, Hye-Young
    • 대한의생명과학회지
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    • 제7권4호
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    • pp.181-190
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    • 2001
  • This study was designed to investigate the effects of reactive oxygen species (ROS) on DNA stability in human spermatozoa. To verify human spermatozoa were incubated with xanthine-xanthine oxidase (X 100$\mu$M-XO 50 mlU ~ 400 mIU), $H_2O_2$ (125 $\mu$M ~ 1 mM), sodium nitroprusside (SNP 0.1 $\mu$M ~ 100 $\mu$M) or lymphocyte. Otherwise, spermatozoa were incubated under low $O_2$ (5%) condition. Damage of sperm DNA was analyzed by single cell electrophoresis (Comet assay) and flow cytometry after acridine orange staining. In the presence of ROS, there was increase in DNA damage. The rate of DNA single strand breakage (9.0$\pm$1.0% ~ 46.0$\pm$4.6%) and DNA fragmentation (7.51$\pm$1.0% ~ 29.5$\pm$4.6%) were similar regardless of the kinds of ROS and exposure time. DNA damage in the lower $O_2$ condition (5%) was lower than ambient $O_2$ condition (20%). Taken together, it suggested that sperm DNA might be damaged by ROS. In the presence of ROS, increase in DNA damage and chromatin instability was obvious in spite of short exposure. Although present study reconfirmed that sperm incubation in the low concentration of ROS have the benefit m the induction of capacitation and Ah, the increase in DNA damage by ROS and possible genetic problem should be considered before the human trials.

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Detection of DNA Damage in Carp Using Single-Cell Gel Electrophoresis Assay for Genotoxicity Monitoring

  • Jin, Hai-Hong;Lee, Jae-Hyung;Hyun, Chang-Kee
    • Journal of Microbiology and Biotechnology
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    • 제14권2호
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    • pp.268-275
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    • 2004
  • To investigate the potential application of the single-cell gel electrophoresis (SCGE) assay to carp as an aquatic pollution monitoring technique, gill, liver, and blood cells were isolated from carp exposed to a direct-acting mutagen, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), or indirect mutagen, $benzo[\alpha]pyrene$ $(B[\alpha]P)$, then the DNA strand breakage was analyzed using the assay. Based on testing 5 different cell isolation methods and 6 electrophoretic conditions, the optimized assay conditions were found to be cell isolation by filter pressing and electrophoresis at a lower voltage and longer running time (at 0.4 V/cm for 40 min). In preliminary experiments, gill and liver cells isolated from carp exposed to MNNG in vitro exhibited DNA damage signals even with 0.5 ppb exposure, which is a much higher dose than previously reported. In the gill cells isolated from carp exposed to 0.01-0.5 ppm MNNG in vivo, significant dose-and time-dependent increases were observed in the tail for 4 days. As such, the linear correlation between the relative damage index (RDI) values and time for each dose based on the initial 48-h exposure appeared to provide effective criteria for the genotoxicity monitoring of direct-acting mutagenic pollution. In contrast, the in vivo exposure of carp to 0.25-1.0 ppm of $B[\alpha]P$ for 7 days resulted in dose-and time-dependent responses in the liver cells, in which 24-h delayed responses for metabolizing activation and gradual repair after 48 h were also observed. Thus, the negative-sloped linear correlation between the RDI and time at each dose based on the initial 48 h appeared to provide more effective criteria for the genotoxicity monitoring of indirect mutagenic pollution.

조선시대 도검 패용 광다회의 제작기법 분석 (Analysis of the Manufacturing Techniques for the KwangDahoe Tying on the Sword in Joseon Dynasty)

  • 백지선;정광용
    • 헤리티지:역사와 과학
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    • 제50권3호
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    • pp.64-87
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    • 2017
  • 조선시대 끈을 일컫는 다회는 우리의 복식사, 문화사, 생활사가 녹아있는 산물로서 지역과 계층을 불문하고 다양한 분야에서 전반적으로 사용되었다. 국내외 다수의 유물이 현존하고 있으나 연구의 주요대상으로 선정되지 못하여 연구가 미흡하고 기법 전승 등에서도 접근성이 낮아 연구에 어려움이 많다. 본 연구에서는 도검의 패용(佩用)을 위해 사용된 광다회의 제작기법을 분석하여 다회의 비파괴적인 제작기법 분석 방안을 마련하고, 다회의 올바른 보존 및 계승에 기여하고자 하였다. 패용 광다회는 물건의 패용을 위한 용도로 사용되는 끈이며, 매듭을 맺기 위해 사용되기도 하여 현재는 매듭장에 의해 전승되고 있다. 제작원리는 원다회와 동일하고 다회틀 상판의 심지가 지지대 역할을 하기 때문에 제작된 다회의 중앙에 빈 공간이 존재하는 것이 특징이다. 이와 같은 다회는 3차원적으로 제작되어 제작기법이 복잡하고 현재까지 올의 개수와 배열에 따라 제작되는 다회에 관한 기초자료가 마련되어있지 않아 제작기법의 규명에 어려움이 많았다. 특히 제작기법의 추정을 위해 올을 해체하는 경우가 존재하지만 제작기법의 규명에는 어려움이 있고, 본래의 상태로 되돌릴 수 없다는 문제점도 있다. 따라서 다회의 비파괴적인 제작기법 분석방안 마련과 실제 유물에 대한 적용 가능성 평가의 필요성이 크다. 유물의 제작기법 분석은 다회 제작기법의 DB구축 결과를 바탕으로 하였고, DB구축을 위하여 다회의 도식화 및 복원샘플 제작, 형태적 특징 분석을 진행하였다. 또한 DB의 신뢰도를 높이기 위하여 유물의 X-ray CT 촬영으로 분석결과를 뒷받침할 근거를 마련하였으며, 촬영결과는 다회치기기법에 근거하여 제작기법을 해석하였다. 제작기법 분석의 대상으로 선정한 패용 광다회는 경인미술관 소장 도검(녹칠어피갑금동장별운도, 어피갑금동장곡병환도)의 패용을 위한 것으로 다회의 예술성, 기능성과 상징성을 겸비한 유산이며 광다회가 온전히 남아있어 띠돈을 비롯한 패용장식이 모두 온전한 유물이다. 유물의 제작기법 분석결과를 토대로 짜임형태를 관찰하였고 2점 모두 좌우측면에서 1회 교차된 것임을 알 수 있었다. 짜임형태 길이비 측정을 통해 20개의 올을 사용하여 제작된 것으로 확인되었다. X-ray CT 촬영결과 DB의 분석결과와 동일한 결과가 도출되어 차후 국내외에 산재되어있는 다회의 분석 및 연구를 위한 기초자료로서 활용이 가능할 것으로 사료된다. 이번 연구를 통해 미력하게나마 소중한 전통문화의 보존 및 계승, 발전에 도움이 되기를 바란다.

양파(Allium cepa) 추출물의 간보호 및 항산화 효과 (The Hepatotprotective and Antioxidative Effects of Onion (Allium cepa) Extracts in Rat Hepatocyte Primary Culture)

  • 임상철;임태진
    • 한국자원식물학회지
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    • 제18권3호
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    • pp.470-478
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    • 2005
  • 본 연구의 목적은 양파추출물의 간보호 및 항산화 효과를 조사하기 위함이다. 간 손상을 유발시키는 t-BHP (1.5 mM) 존재하에 간세포를 0, 0.01, 0.05, 0.1 및 0.3 mg/ml의 다양한 농도의 양파추출물로 1시간 동안 일차배양하였다. t-BHP는 GOT와 LDH 활성 및 TBARS 농도를 증가시켰으며 MTT값은 감소시켰다. 0.05 mg/ml 농도의 양파추출물 첨가는 t-BHP에 의해 증가된 GOT 및 LDH 활성을 감소시켰으며 0.1 mg/ml 농도의 양파추출물은 t-BHP에 의해 감소된 MTT 값을 증가시켰다. 또한 0.01 mg/ml 농도의 양파추출물 첨가는 t-BHP에 의해 증가된 TBARS 농도를 감소시켜 양파추출물이 t-BHP에 의해 유발된 간손상과 지질과산화를 억제시켰다. t-BHP 처리는 간세포의 catalase, GSH-Px 및 GSH-Rd 활성을 현저히 감소시켰다. 그러나 0.1 mg/ml 농도의 양파추출물 첨가는 t-BHP에 의해 감소된 catalase GSH-Px 및 GSH-Rd 활성을 증가시켰으며 특히 catalase 활성은 t-BHP 무첨가군 수준까지 증가시켰다. 또한 hydroxyl radical을 생성하는 Fenton 시약의 존재하에 plasmid DNA를 양파추출물과 함께 배양한 결과 양파추출물은 농도 의존적으로 hydroxyl radical에 의해 유도된 single-strand 절단을 억제하였다. 이상과 같이 간세포 일차배양에서 양파추출물은 t-BHP에 의해 유발된 간독성, 간세포 생존율 감소, 지질과산화를 농도 의존적으로 억제시켰고 또한 t-BHP에 의해 억제된 GSH-Px, GSH-Rd 및 catalase의 활성을 증가시켰다. 이와 같이 양파추출물의 간보호 및 항산화 효과는 항산화 효소 특히 catalase의 활성 증가와 hydroxyl radical에 의해 유도된 산화억제 및 이에 따른 지질과산화 억제에 기인하는 것으로 사료된다.

열목어 생식선자극호르몬의 cDNA cloning 및 CHO 세포를 이용한 발현검토

  • 최은주;손영창
    • 한국양식학회:학술대회논문집
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    • 한국양식학회 2003년도 추계학술발표대회 논문요약집
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    • pp.21-21
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    • 2003
  • 경골어류의 뇌하수체에서는 두 종류의 생식선자극호르몬 (FSH, LH)이 생산되며, 이 호르몬들은 공통적인 α 쇄와 특이적인 β 쇄를 가진다. 연어과 어종들에서, FSH는 난황형성과 정자형성의 역할을 하며, LH는 배우자의 최종성숙을 조절한다. 냉수성 고유어종인 열목어 (Brachymystax lenok)의 멸종을 방지하고 종묘생산을 원활히 하기 위하여 먼저 열목어의 GTHα, FSHβ 및 LHβ 쇄를 cloning하여 염기서열을 결정하였다. 열목어 GTHα, FSHβ 및 LHβ의 cDNA는 산천어의 해당 cDNA와 높은 상동성 (각각 84, 95, 98%)을 보였다. 다음으로 기능적인 생식선자극호르몬을 제작하기 위해서 2개의 쇄를 single-strand로 연결하여 진핵세포를 숙주로하는 시스템에서 생식선자극호르몬을 생산할 수 있는 구조체인 FSHβ-GTHα (235 amino acids) 와 LHβ-GTHα (240 amino acids)를 각각 재조합하였다. 또한 각각의 융합단백질 생산용 구조체의 3'-말단에는 단백질추출이 용이하도록 histidine×6 구조를 첨가하였다. 이상의 단일쇄 FSH와 LH 유전자재조합 산물을 포유동물 유래의 세포 (CHO-K1)에 liposome chemical을 사용하여 유전자도입 후 세포에서 분비되는 단백질을 모니터링하였다. 배지를 부분정제한 후 SDS-PACE로 조사한 결과, LH 재조합 유전자를 도입한 후 48-60 시간째에 약 25 kDa의 단백질로서 관찰되었다. 현재 FSH 재조합 유전자에 대해서도 조사중이며, 향후 이를 재료로 하여 기능형 생식선자극호르몬을 생산하고 추출하기 위한 연구가 계속적으로 수행 될 것이다.

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Vitamin B6 Deficiency, Genome Instability and Cancer

  • Wu, Xia-Yu;Lu, Lin
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권11호
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    • pp.5333-5338
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    • 2012
  • Vitamin B6 functions as a coenzyme in >140 enzymatic reactions involved in the metabolism of amino acids, carbohydrates, neurotransmitters, and lipids. It comprises a group of three related 3-hydroxy-2-methyl-pyrimidine derivatives: pyridoxine (PN), pyridoxal (PL), pyridoxamine (PM) and their phosphorylated derivatives [pyridoxal 5'-phosphate (PLP) and pyridoxamine 5'-phosphate (PMP)], In the folate metabolism pathway, PLP is a cofactor for the mitochondrial and cytoplasmic isozymes of serine hydroxymethyltransferase (SHMT2 and SHMT1), the P-protein of the glycine cleavage system, cystathionine ${\beta}$-synthase (CBS) and ${\gamma}$-cystathionase, and betaine hydroxymethyltransferase (BHMT), all of which contribute to homocysteine metabolism either through folate-mediated one-carbon metabolism or the transsulfuration pathway. Folate cofactors carry and chemically activate single carbons for the synthesis of purines, thymidylate and methionine. So the evidence indicates that vitamin B6 plays an important role in maintenance of the genome, epigenetic stability and homocysteine metabolism. This article focuses on studies of strand breaks, micronuclei, or chromosomal aberrations regarding protective effects of vitamin B6, and probes whether it is folate-mediated one-carbon metabolism or the transsulfuration pathway for vitamin B6 which plays critical roles in prevention of cancer and cardiovascular disease.