• 제목/요약/키워드: Human Mouse

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인간 Cystathionine ${\gamma}$-Lyase에 의한 Selenomethionine의 Methylselenol로의 직접분해 (Direct Conversion of L-Selenomethionine into Methylselenol by Human Cystathionine ${\gamma}$-Lyase)

  • 조현남;지광환
    • 한국미생물·생명공학회지
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    • 제42권1호
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    • pp.11-17
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    • 2014
  • Selenium(Se)은 사람에게 필수성분이지만 독성이 강한 물질이다. 따라서 생체에서 Se(셀레늄)의 농도조절은 정확하고 효율적으로 이루어져야 한다. 사람은 Se을 음식의 형태로 섭취해야 하고 주로 seleno-L-methionine (L-SeMet)의 형태로 섭취한다. 섭취된 L-SeMet은 methionine 대사와 동일한 효소를 이용하여 Se-adenosyl-L-SeMet으로 대사된다고 알려져 있었다. 그러나 최근 쥐의 간 추출물의 실험에서, L-SeMet이 cystathionine ${\gamma}$-lyase (mouse CGL)의 작용으로 methylselenol ($CH_3SeH$)로 직접적으로 대사된다는 보고가 있었다. CGL은 원래 cystathionine을 L-cysteine과 ${\alpha}$-ketobutyrate, 그리고 $NH_3$로 분해하는 효소로 알려져 있었다. 따라서 본 연구에서는 쥐의 간 추출물 대신에 인간의 CGL을 분리 정제하여 L-SeMet에서 methylselenol의 형성을 확인하고자 하였다. Methylselenol의 표준시료는 dimethyldiselenide를 sodium tetrahydroborate로 환원시켜 준비하였다. 그리고 L-SeMet을 기질로 사용한 효소 반응액 중에서 가스상의 생성물은 GC/MS 스펙트럼으로 분석하였다. 분석 결과 methylselenol의 유도체인 dinitrophenyl selenoether와 일치하였다. 또한 인간 CGL이 L-SeMet에서 methylselenol을 형성하는 반응의 kinetic parameter를 mouse CGL과 비교 분석하였다. 결과적으로 human CGL은 섭취된 L-SeMet의 대사를 책임지고 있는 중요한 효소이다.

Online Digit Recognition using Start and End Point

  • Shim, Jae-chang;Ansari, Md Israfil
    • Journal of Multimedia Information System
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    • 제4권1호
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    • pp.39-42
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    • 2017
  • Communication between human and machine is having been researched from last few decades and still it's a challenging task because human behavior is unpredictable. When it comes on handwritten digits almost each human has their own writing style. Handwritten digit recognition plays an important role, especially in the courtesy amounts on bank checks, postal code on mail address etc. In our study, we proposed an efficient feature extraction system for recognizing single digit number drawn by mouse or by a finger on a screen. Our proposed method combines basic image processing and reading the strokes of a line drawn. It is very simple and easy to implement in various platform as compare to the system which required high system configuration. This system has been designed, implemented, and tested successfully.

Efficient Generation of Human IgG1 Light Kappa Constant Region Knock-in Mouse by CRISPR/Cas9 System

  • Jung, Sundo
    • 대한의생명과학회지
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    • 제25권4호
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    • pp.372-380
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    • 2019
  • Mice with specific modified genes are useful means of studying development and disease. The CRISPR/Cas9 system is a very powerful and effective tool for generating genetically modified mice in a simple and fast manner. To generate human IgG light kappa constant knock-in mice, we tested by microinjection of a mixture of Cas9 protein, single-guide RNA and target homologous recombinant donor DNA into zygotes. We found that the injection of 10 ng/μL of Cas9 protein and crRNA/tracrRNA, rather than single guide RNA, induced the production of knock-in mice more effectively. Thus, our study provides valuable information that will help to improve the production of knock-in mice and contribute the successful generation of humanized Ab-producing mice in Korea.

Human placental extract suppresses lipopolysaccharide-induced expressions of cyclooxygenase-2 and inducible nitric oxide synthase in mouse BV2 microglial cells

  • Yang, Sang-Eun;Kim, Yong-Suk;Park, Dong-Suk
    • Journal of Acupuncture Research
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    • 제22권2호
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    • pp.111-121
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    • 2005
  • Human placental extract (HPE), which is prepared from the placenta of healthy pregnant females, has been widely used in clinical field. HPE is known to possess anti-inflammatory, anti-viral, anti-oxidative, anti-mutagenic, and analgesic properties. In this study, the effect of HPE against lipopolysaccharide (LPS)-induced inflammation was investigated. From the present results, HPE was shown to suppress prostaglandin E2 synthesis (PGE2) and nitric oxide (NO) production by inhibition on the LPS-stimulated enhancement of the cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) expressions in mouse BV2 microglial cells. These results suggest that HPE may offer a valuable mean of therapy for the treatment of brain inflammatory diseases by attenuating LPS-induced PGE2 and NO production.

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The use of animal models in rheumatoid arthritis research

  • Jin-Sun Kong;Gi Heon Jeong;Seung-Ah Yoo
    • Journal of Yeungnam Medical Science
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    • 제40권1호
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    • pp.23-29
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    • 2023
  • The pathological hallmark of rheumatoid arthritis (RA) is a synovial pannus that comprises proliferating and invasive fibroblast-like synoviocytes, infiltrating inflammatory cells, and an associated neoangiogenic response. Animal models have been established to study these pathological features of human RA. Spontaneous and induced animal models of RA primarily reflect inflammatory aspects of the disease. Among various induced animal models, collagen-induced arthritis (CIA) and collagen antibody-induced arthritis (CAIA) models are widely used to study the pathogenesis of RA. Improved transplantation techniques for severe combined immunodeficiency (SCID) mouse models of RA can be used to evaluate the effectiveness of potential therapeutics in human tissues and cells. This review provides basic information on various animal models of RA, including CIA and CAIA. In addition, we describe a SCID mouse coimplantation model that can measure the long-distance migration of human RA synoviocytes and cartilage destruction induced by these cells.

Development and Characterization of a Specific Anti-Caveolin-1 Antibody for Caveolin-1 Functional Study in Human, Goat and Mouse

  • Ke, Meng-Wei;Jiang, Yan-Nian;Li, Yi-Hung;Tseng, Ting-Yu;Kung, Ming-Shung;Huang, Chiun-Sheng;Cheng, Winston Teng-Kuei;Hsu, Jih-Tay;Ju, Yu-Ten
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권6호
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    • pp.856-865
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    • 2007
  • Caveolin-1 of the caveolin family of proteins regulates mammary gland development and has been shown to play a contradictory role in breast tumor progression. A specific anti-Caveolin-1 antibody will be useful for functional study of Caveolin-1 in different tissues. In this study, we generated a rabbit polyclonal antibody that specifically recognizes the N-terminal amino acids 50-65 of Caveolin-1. This polyclonal antibody specifically reacted with Caveolin-1 extracted from cells of different species, including human epithelial A431 cells, goat primary mammary epithelial cells and mice fibroblast NIH 3T3 cells, by Western blotting. Endogenous Caveolin-1 protein expressing in cells and normal human tissues was detected by this polyclonal antibody using immunocytofluorescent and immunohistochemical staining, respectively. Furthermore, an apparent decrease in Caveolin-1 expression in tumorous breast and colon tissues was detected by this polyclonal antibody. In conclusion, we have identified amino acids 50-65 of Caveolin-1, which contains an epitope that is specific to Caveolin-1 and is conserved in the human, goat and mouse. In future, this anti-Caveolin-1 antibody can be used to examine the progression of breast and colon cancers and to study functions of Caveolin-1 in human, goat and mouse cells.

인간양수에 의한 생쥐 난자 투명대의 정자수용능력 억제의 관찰 (Human Amniotic Fluid Induces Spontaneous Hardening of the Zona Pellucida of Mouse Immature Oocytes During Maturation In Vitro)

  • 박기상;이택후;송해범;전상식
    • Clinical and Experimental Reproductive Medicine
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    • 제27권1호
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    • pp.23-29
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    • 2000
  • Objective: Zona pellucida (ZP) has been thought to be the barrier of egg to sperm penetration before and after fertilization. The phenomenon of ZP hardening has been considered as a post-fertilization event until now, and it is generally accepted that it is caused by the secretory products of cortical granules released during the cortical reaction. Hardening of ZP could occur "spontaneously" in mammalian oocytes in standard culture conditions, and that it is probably not a consequence of cortical reaction. The purpose of our study was to investigate the effect of human amniotic fluid (HAF) on nuclear maturation (NM) and fertilization ability of mouse immature oocytes. Methods: HAF was obtained from patients undergoing amniocentesis at $16{\sim}20$ weeks of gestation. HAF from five to ten patients was centrifuged and the supernatants was pooled. Cumulusenclosed mouse immature oocytes were incubated in the medium containing HAF, and examined to confirm NM and fertilization. Female ICR mice (about 3 weeks old) were stimulated with 7.5 IU PMSG. Immature oocytes were isolated at $48{\sim}52$ hrs post PMSG injection and cultured in TCM-199 supplemented with 20% HAF for 18 hrs. FBS was used as a control for the examination. Matured oocytes (MII) were fertilized with sperms collected from the epididymis of male mice (over 10 weeks old). Fertilization was in conducted T6 medium containing 15 mg/ml BSA, and confirmed at 6 hrs post-insemination. Fertilization rate was assessed in zona-intact or zona-free oocytes (denuded by trypsin). Evaluation of NM and fertilization was carried out by rapid staining method. ZP hardening was evaluated by incubating cumulus cell-free mature oocytes in 0.001% chymotrypsin at $37^{\circ}C$ for 10 min. Results: There was no significant difference between the effects of HAF (86.6%) and FBS (87.7%) supplements on NM of immature oocytes. When maturation medium was supplemented with HAF, total fertilization rates (7%) were significantly lower (p<0.01) than that of FBS (85.1%). In HAF group, fertilization rate was increased (p<0.01) in zona-free oocytes (7% versus 100%). The resistance of mouse oocyte ZP to digestion by chymotrypsin after maturation in vitro was significantly higher (p<0.01) in HAF group (86.7%) than in FBS (6.7%). To culture oocytes in FBS were very effective in preventing ZP hardening. However cultured oocytes in HAF showed high rate of ZP hardening (p<0.01). Conclusions: These results suggest that HAF can be used as a supplement for the NM of mouse immature oocytes in vitro. However, HAF induces spontaneous hardening of ZP of mouse immaure oocytes during maturation in vitro.

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발효 차가버섯 추출물이 인체 종양세포주 증식에 미치는 영향 (Effects of Water Extract from fermented Chaga Mushroom(Inonotus obliquus) on the Proliferation of Human Cancer Cell Lines.)

  • 차재영;박상현;허진선;조영수
    • 생명과학회지
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    • 제17권5호
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    • pp.671-677
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    • 2007
  • 발효 및 비발효 차가버섯 수용성 추출물이 정상 세포주 NIH3T3 mouse normal fibroblast cell 및 인체 종양 세포주 AGS human gastric cancer cell(위암), HCT-15 human colon cancer cell(대장암), Hep3B human hepatoma cancer cell(간암), MCF-7 human breast cancer cell(유방암), HeLa human cervical cancer cell(자궁경부암)에서 MTT(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay 방법에 의한 세포 증식 억제와 암세포 증식억제의 기전 연구의 일환으로 apoptosis가 일어날 때 나타나는 DNA fragmentation을 agarose gel electrophoresis 방법으로 검토하였다. 인체 종양 세포주의 생육저해 효과가 발효 차가버섯 추출물이 비발효 차가버섯 추출물보다 강한 것으로 나타났다. 그러나 동일한 실험조건하에서 마우스 정상 세포주 NIH3T3은 80% 이상의 생존율을 나타내어 정상 세포주에는 큰 영향을 미치지 않는 것으로 나타났다. 특히, 발효 및 비발효 차가버섯 추출물에서 본 실험에 사용한 세포주 중에서 대장암 세포주 HCT-15에 대해 가장 세포 증식 억제효과가 뛰어났으며, 이러한 효과는 첨가 농도 의존적 이였다. 발효 및 비발효 차가버섯 추출물에 의한 암세포 증식억제가 기전 연구로 apoptosis가 일어날 때 나타나는 DNA fragmentation을 세포로부터 genomic DNA를 분리하여 agarose gel electrophoresis 방법으로 조사한 결과, 정상세포인 NIH3T3 세포는 DNA fragmentation이 거의 일어나지 않아 세포 생존율 결과와 유사한 경향을 보였으나, 특히 대장암 세포주인 HCT-15에서는 발효 차가버섯뿐만 아니라 비발효 차가버섯 추출물에서도 DNA fragmentation이 많이 일어나는 것이 관찰되어 암세포 증식억제 효과가 높다는 결과를 뒷받침 해주고 있다.

Anticancer activity of subfractions containing pure compounds of Chaga mushroom (Inonotus obliquus) extract in human cancer cells and in Balbc/c mice bearing Sarcoma-180 cells

  • Chung, Mi-Ja;Chung, Cha-Kwon;Jeong, Yoon-Hwa;Ham, Seung-Shi
    • Nutrition Research and Practice
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    • 제4권3호
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    • pp.177-182
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    • 2010
  • The Chaga mushroom (Inonotus obliquus) has been used in folk medicine to treat cancers. However, limited information exists on the underlying anticancer effects of the major component of I. obliquus in vivo. We hypothesize that the pure compounds ($3{\beta}$-hydroxy-lanosta-8,24-dien-21-al, inotodiol and lanosterol, respectively) separated from I. obliquus would inhibit tumor growth in Balbc/c mice bearing Sarcoma-180 cells (S-180) in vivo and growth of human carcinoma cells in vitro. To test this hypothesis, the growth inhibition of each subfraction isolated from I. obliquus on human carcinoma cell lines (lung carcinoma A-549 cells, stomach adenocarcinoma AGS cells, breast adenocarcinoma MCF-7 cells, and cervical adenocarcinoma HeLa cells) was tested in vitro. Then, after S-180 implantation, the mice were fed a normal chow supplemented with 0, 0.1 or 0.2 mg of subfraction 1, 2 or 3 per mouse per day. All of the subfractions isolated from I. obliquus showed significant cytotoxic activity against the selected cancer cell lines in vitro. Subfraction 1 was more active than subfraction 2 and subfraction 3 against the A549, AGS and MCF-7 cancer cell lines in vitro. In in vivo results, subfraction 1 isolated from I. obliquus at concentrations of 0.1 and 0.2 mg/mouse per day significantly decreased tumor volume by 23.96% and 33.71%, respectively, as compared with the control. Subfractions 2 and 3 also significantly inhibited tumor growth in mice bearing S-180 as compared with the control mouse tumor. Subfraction 1 isolated from I. obliquus showed greater inhibition of tumor growth than subfractions 2 and 3, which agrees well with the in vitro results. The results suggest that I. obliquus and its compounds in these subfractions isolated from I. obliquus could be used as natural anticancer ingredients in the food and/or pharmaceutical industry.

휴먼마우스 구현을 위한 효율적인 손끝좌표 추적 및 마우스 포인트 제어기법 (Efficient Fingertip Tracking and Mouse Pointer Control for Implementation of a Human Mouse)

  • 박지영;이준호
    • 한국정보과학회논문지:소프트웨어및응용
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    • 제29권11호
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    • pp.851-859
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    • 2002
  • 본 연구는 마우스의 입력을 사람의 손동작으로 대체하는 휴먼마우스 시스템을 위한 새로운 손끝 좌표 추적기법과 마우스 포인터 움직임 결정법을 제안한다. 손끝좌표 추적에는 얼굴영역 추적을 위해 제안되었던 CAMSHIFT 알고리즘을 개선하여 적용하였다. 정확한 손 영역 검출을 위하여 각 사용자의 환경에 최적화된 피부영역 컬러 정보를 얻을 수 있는 실시간 피부영역 학습과 손의 자유로운 움직임으로 인하여 발생하는 손 영역의 크기 및 방향변화를 고려한 영역제한 기법을 적용하였다. 또한 손의 주축을 이용한 손끝좌표 계산법을 통해 빠르고 정확하게 손끝의 위치를 찾을 수 있다. 실시간 손끝좌표 검출에는 처리속도의 한계가 있어 얻어지는 좌표의 연속성이 결여되어 마우스 포인터의 움직임이 연결되지 않는 문제점이 있다. 연속적인 마우스 포인터의 움직임 표현을 위해 손끝좌표의 이동거리를 통해 마우스 포인터의 속도와 가속도를 계산하여 마우스 포인터의 운동방정식을 정의하고 이를 이용하여 마우스 포인터의 위치를 결정한다. 제안한 알고리즘을 적용하여 실험한 결과 빠르고 정확한 손끝좌표 추적과 마우스 포인터의 자연스러운 움직임이 가능함을 보였다.