• Title/Summary/Keyword: Human Mouse

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

  • Cho, Hyun-Nam;Jhee, Kwang-Hwan
    • Microbiology and Biotechnology Letters
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    • v.42 no.1
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    • pp.11-17
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    • 2014
  • Selenium is an essential trace element for mammals, but it is very toxic. Therefore, the control of selenium concentrations should be precisely and effectively monitored. Selenium is naturally obtained through foods and seleno-L-methionine (LSeMet) is a major form of selenium. It has been reported that L-SeMet is only converted into Se-adenosyl-L-SeMet. However, a recent study suggested that L-SeMet was directly metabolized into methylselenol ($CH_3SeH$) in mouse liver extract by the reaction of cystathionine ${\gamma}$-lyase (CGL). The canonical reaction of CGL was known to catalyze the cleavage of L-cystathionine to L-cysteine, ${\alpha}$-ketobutyrate and $NH_3$. In the present study, we found that L-SeMet could be directly converted to $CH_3SeH$ using purified homogenous human CGL instead of mouse liver cytosol. Authentic $CH_3SeH$ was prepared by reduction of dimethyldiselenide with sodium tetrahydroborate. The gaseous product of the enzymatic reaction with L-SeMet was analyzed by GC/MS spectrometry. The GC/MS data was identical to that of authentic dinitrophenyl selenoether. We also analyzed the kinetic parameters for the formation of $CH_3SeH$ from L-SeMet by human and mouse CGL. These results suggest that human CGL is a critical enzyme which is responsible for L-SeMet metabolism.

Online Digit Recognition using Start and End Point

  • Shim, Jae-chang;Ansari, Md Israfil
    • Journal of Multimedia Information System
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    • v.4 no.1
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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
    • Biomedical Science Letters
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    • v.25 no.4
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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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    • v.22 no.2
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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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    • v.40 no.1
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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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    • v.20 no.6
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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.

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

  • Park, Kee-Sang;Lee, Taek-Hoo;Song, Hai-Bum;Chun, Sang-Sik
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.1
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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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Effects of Water Extract from fermented Chaga Mushroom(Inonotus obliquus) on the Proliferation of Human Cancer Cell Lines. (발효 차가버섯 추출물이 인체 종양세포주 증식에 미치는 영향)

  • Cha, Jae-Young;Park, Sang-Hyun;Heo, Jin-Sun;Cho, Young-Su
    • Journal of Life Science
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    • v.17 no.5 s.85
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    • pp.671-677
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    • 2007
  • This study was performed to investigate the effect of the water-extract from non-fermented or fermented Chaga mushrooms (Inonotus obliquus) on the proliferation and apoptosis of the NIH3T3 mouse normal fibroblast cells and various human cancer cell lines including HCT-15 human colon carcinoma, AGS human gastric carcinoma, MCF-7 human breast adenocarcinoma, Hep3B human hepatocellular carcinoma and HeLa human cervical carcinoma using MTT(3-[4,5-dimethylthiazol-2-yl] -2,5-diphenyl tetrazolium bromide) assay and DNA fragmentation. In an anti-cancer test using various human cancer cells, fermented Chaga mushroom extract showed higher antiproliferating effect than that of non-fermented Chaga mushroom extract. Mouse normal NIH3T3 cells were exhibited 80% above survival under fermented or non-fermented Chngn mushroom extract of various concentrations(0, 0.5 and 1 mg/ml). Fermented Chaga mushroom extract significantly inhibited cell growth on HCT-15 cells in a dose-dependent manner. HCT-15 cells treated with non-fermented or fermented Chaga mushrooms extract produced a distinct oligonucleosomal ladder with different sizes of DNA fragments, a typical characteristic of cells undergoing apoptosis. These results suggest that fermented Chaga mushroom extract suppresses growth of HCT-15 human colon carcinoma cells through apoptosis.

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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    • v.4 no.3
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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.

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

  • 박지영;이준호
    • Journal of KIISE:Software and Applications
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    • v.29 no.11
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    • pp.851-859
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    • 2002
  • This paper discusses the design of a working system that visually recognizes hand gestures for the control of a window based user interface. We present a method for tracking the fingertip of the index finger using a single camera. Our method is based on CAMSHIFT algorithm and performs better than the CAMSHIFT algorithm in that it tracks well particular hand poses used in the system in complex backgrounds. We describe how the location of the fingertip is mapped to a location on the monitor, and how it Is both necessary and possible to smooth the path of the fingertip location using a physical model of a mouse pointer. Our method is able to track in real time, yet not absorb a major share of computational resources. The performance of our system shows a great promise that we will be able to use this methodology to control computers in near future.