• Title/Summary/Keyword: mouse assay

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Molecular Cloning and Characterization of Calumenin in Rabbit Skeletal Sarcoplasmic Reticulum

  • Jung, Dai-Hyun;Kim, Do-Han
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.53-53
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    • 2003
  • Calumenin was previously identified as a high affinity Ca$\^$2+/ binding protein in mouse cardiac sarcoplasmic reticulum (SR). For the present study, a 48 kDa skeletal homologue of calumenin was identified by sucrose-density gradient of rabbit skeletal SR membranes, concanavalin A treatment, 2D-gel electrophoresis, $\^$45/Ca$\^$2+/ overlay, Stains-all staining, and MALDI-TOF analysis. We attempted to clone the skeletal calumenin by RT-PCR based on mouse cardiac and human calumenin sequences. The deduced amino acid sequence (315 residues) of the skeletal calumenin showed high identity to mouse cardiac calumenin (90%). As seen in the cardiac calumenin, the deduced sequence contains a 19 amino acid N-terminal signal sequence and a HDEF C-terminal sequence, a putative retrieval signal to ER. Also, the skeletal calumenin contains one N-glycosylation site, three PKC phosphorylation sites, eight casein kinase 2 phosphorylation sites, and 6 EF-hand domains. GST-calumenin showed a conformational change and increased mobility in the presence of Ca$\^$2+/ in SDS-PAGE. Three calumenin interacting proteins (ryanodine receptor 1, glycogen phosphorylase, and phosphofructo kinase) were identified by pull-down assay with GST-calumenin and solubilized SR. All the interactions were Ca$\^$2+/dependent. The present results suggest that calumenin plays an important role in Ca$\^$2+/ homeostasis of muscle cells.

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Antitumor Effect of Natural Products, Purified Tannin from Plants and Screening of BRM function (천연물에서 단리한 식물정제 탄닌의 항암효과 및 생물학적 반응 조절 물질로서의 기능 검색)

  • Lee, Do-Ik;Cho, Jang-Hyun;Lee, Min-Won
    • YAKHAK HOEJI
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    • v.42 no.4
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    • pp.345-352
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    • 1998
  • Praecoxin A, an ellagitannin, purified from Alnus hirsuta var.microphlla was evaluated on the antitumor activity. Praecoxin A had the significant cytotoxicity to s ix tumor cell lines: human chronic myelogenous leukemia K-562, human promyelocytic leukemia HL-60, mouse leukemia P388, mouse lymphocytic leukemia L-1210, sarcoma-l8O, mouse lymphoma L5178Y except L-1210. And the most sensitive cell line was K-562 ($ED_{50}=2.43{\mu}g/ml$). The $ED_{50} of praecoxin A against HL-60, P388, L-1210, sarcoma7l8O and L5178Y were 6.28, 8.66, 10.00, 7.01, $9.32{\mu}g/ml$, respectively. Praecoxin A showed the increasing effect in life span by 36.8% on the 1st day after treatment of 10mg/kg in mice bearing sarcoma-180 tumor cells (ascitic form) via NCI (National Cancer Institute, U.S.A.) protocol in vivo assay. As a result, praecoxin A is considered to show the antitumor activity.

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Differential Expressions of Aquaporin Subtypes in the Adult Mouse Testis

  • Mohamed, Elsayed A.;Im, Ji Woo;Kim, Dong-Hwan;Bae, Hae-Rahn
    • Development and Reproduction
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    • v.26 no.2
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    • pp.59-69
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    • 2022
  • Many efforts have been made to study the expression of aquaporins (AQP) in the mammalian reproductive system, but there are not enough data available regarding their localized expression to fully understand their specific roles in male reproduction. The present study investigated the expression and localization patterns of different AQP subtypes in the adult mouse testes and testicular spermatozoa using an immunofluorescence assay. All the studied AQPs were expressed in the testes and revealed subtype-specific patterns in the intensity and localization depending on the cell types of the testes. AQP7 was the most abundant and intensive AQP subtype in the seminiferous tubules, expressing in Leydig cells and Sertoli cells as well as all stages of germ cells, especially the spermatids and testicular spermatozoa. The expression pattern of AQP3 was similar to that of AQP7, but with higher expression in the basal and lower adluminal compartments rather than the upper adluminalcompartment. AQP8 expression was limited to the spermatogonia and Leydig cells whereas AQP9 expression was exclusive to tails of the testicular spermatozoa and elongated spermatids. Taken together, the abundance and distribution of the AQPs across the different cell types in the testes indicating to their relavance in spermatogenesis, as well as in sperm maturation, transition, and function.

Effect of Water Extract from Artemisiae Argi Folium on Hydrogen Peroxide Production within Mouse Macrophage Raw 264.7 Cells Treated with Gallic acid, EtOH, LPS, and Acetaminophen (Gallic acid, EtOH, LPS, Acetaminophen으로 유발된 마우스 대식세포 내 hydrogen peroxide 생성억제에 대한 애엽 물추출물의 영향 연구)

  • Park, Wan-Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.6
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    • pp.1495-1499
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    • 2008
  • The purpose of this study is to investigate the effect of water extract from Artemisiae Argi Folium (WAAF) on hydrogen peroxide production within mouse macrophage Raw 264.7 Cells treated with gallic acid, EtOH, LPS, and acetaminophen. WAAF (0${\sim}$400 ug/mL) was treated with gallic acid, EtOH, LPS, acetaminophen. And the intracellular productions of hydrogen peroxide were measured by dihydrorhodamine 123 (DHR) assay. WAAF showed the restoration of the intracellular productions of hydrogen peroxide which were reduced by gallic acid, EtOH, LPS, and acetaminophen in Raw 264.7 Cells. WAAF could be supposed to have the immunological activity related with macrophage's oxidative burst.

Effect of Oroxylin A on Hydrogen Peroxide Production in Polyinosinic-Polycytidylic acid-induced TM4 Mouse Testis Sertoli Cells (Oroxylin A가 polyinosinic-polycytidylic acid로 유발된 생쥐 서톨리세포 TM4의 hydrogen peroxide 생성증가에 미치는 영향)

  • Park, Wan Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.4
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    • pp.384-389
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    • 2014
  • The purpose of this study is to investigate the modulatory effect of oroxylin A on hydrogen peroxide production in TM4 mouse testis sertoli cells induced by the synthetic analog of double-stranded RNA [polyinosinic-polycytidylic acid]. Hydrogen peroxide production was measured by dihydrorhodamine 123 (DHR) assay. Oroxylin A significantly inhibited the polyinosinic-polycytidylic acid (PIC)-induced production of hydrogen peroxide for 0.5, 2, 12, 18, and 24 hr incubation at the concentrations of 5, 10, 25, and $50{\mu}M$ in TM4 (P < 0.05) in dose dependent manner. These results suggest that oroxylin A has a protective effect against PIC-induced cellular toxicity with its inhibition of hydrogen peroxide production in PIC-induced sertoli cells.

Isolation and Evaluation of an Antitumor Constituent from Pyrolae Herba (녹수초의 항암활성물질의 분리 및 항암력 평가)

  • Bae, Ki-Hwan;Kim, Hwan-Mook;Lee, Sang-Myung
    • YAKHAK HOEJI
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    • v.40 no.2
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    • pp.225-229
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    • 1996
  • The cytotoixic effect of Pylorae Herba (Pyrola japonica Klenze) against L1210 and K-562 cells was studied in vitro. The methanolic extract of Pylorae Radix was add ed to the culture of L1210 cells and K-562 cells for the cytotoxic activity and the ED50 values of hexane, ethylacetate, buthanol and water fractions from methanolic extract were determined using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazoliumbromide) assay. The active constituent isolated by bioassay guided fractionation followed by purification gave rise to a yellow needle crystal and was clarified to be chimaphilline by the comparison with the published data. The average life spans with it were not prolonged significantly on tumor growth in hybrid female mouse (BDF1-KIST) inoculated subcutaneously with P388 cells.

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Effects of Fructus Schisandrae Water Extract on Cultured Mouse Myocardial Cells Induced by Xanthine Oxidase/Hypoxanthine (오미자 추출물이 산소지유기에 의하여 손상된 생쥐의 배양 심근세포에 미치는 영향)

  • 주은정
    • Journal of Nutrition and Health
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    • v.33 no.7
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    • pp.739-744
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    • 2000
  • The purpose of this study was to elucidate protective effect of Fructus Schsandrae(FS) water extract against xanthine oxidase/hypoxanthine(XO/HX)-induced cardiotoxicity in myocardial cells this experiment was performed. Cardiotoxicity of XO/HX was examined by MTT(MTT [3-(4,5-dimethylthiazol-2-yl)-2.5,-diphenyl tetrazolium bromide) assay. XO/HX induced the decrease of cell viability. Also XO/HX induced the increase of LDH activity and the decrease of beating rate on cultured myocardial cells in a dose-dependent manner. To investigate cardioprotective effect of FS water extract cultures were preincubated with FS water extract for 3 hours. Cultures were then exposed to XO/HX for 72 hours. FS water extract have an efficacy in decreaasing LDH activity and increasing heart beating rate on cultured myocardial cells damaged by XO/HX. From the results it is suggested that XO/HX may show toxic effect in cultured myocardial cells derived from neonatal mouse and FS water extract is effective in the prevention of XO/HX-induced cardiotoxicity.

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Anti-idiotypic Antibodies against Bovine Growth Hormone

  • Verma, N.K.;Sodhi, R.;Rajput, Y.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.5
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    • pp.732-737
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    • 2003
  • Anti-antibodies against three mouse monoclonal antibodies viz. IIB5D6, VIA6E8 and VIC1F9 (specific to bovine growth hormone) in rabbits have been generated and characterized. Ammonium sulfate fractionated and affinity-purified monoclonal antibodies were used for producing anti-antibodies. The generated anti-antibodies were against common as well as uncommon antigenic determinants present in mouse monoclonal antibodies. The raised anti-antibodies replaced [$I^125$ ]bGH bound to goat liver microsomes indicating production of anti-idiotypic antibodies against bovine growth hormone. These antibodies can have profound implications in vivo in lactating bovines for enhancing milk yield.

Effect of Chestnut Bark Extracts on Tyrosinase Gene Expression (율피 추출물이 티로시나아제 유전자 발현에 미치는 효과)

  • Chin Jong-Eon;Kim Kwan-Chun
    • Journal of environmental and Sanitary engineering
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    • v.20 no.3 s.57
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    • pp.10-16
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    • 2005
  • Chestnut bark extract by methanol repressed the expression of tyrosinase gene of B16 mouse melanoma cell containing tyrosinase promoter. $10{\mu}g/ml{\ell}\;100{\mu}g/m{\ell}$, $1mg/m{\ell}$ of the extract repressed expression of tyrosinase gene about $38\%,\;47\%,\;and\;78\%$, respectively, compared with control. In the MTT assay, the same extract exhibited very low cytotoxicity at $1{\mu}g/m{\ell},\;10{\mu}g/m{\ell},\;100{\mu}g/m{\ell},\;and\;1mg/m{\ell}$, respectively. The fractions of Methylene chloride and ethyl acetate did not showed the repressive effect on the expression of tyrosinase gene, but the fraction of butyl alcohol repressed highly at $10{\mu}g/m{\ell}\;and\;100{\mu}g/m{\ell}$.

Effects of Glycine maxim Extract on the Activity of Tyrosinase Promoter (대두(Glycine maxim) 추출물이 티로시나아제 프로모터 활성에 미치는 효과)

  • Chin, Jong-Eon;Kim, Kwan-Chun
    • The Korean Journal of Food And Nutrition
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    • v.21 no.3
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    • pp.386-390
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    • 2008
  • The methanolic extract of Glycine maxim increased the expression of the promoter in B16 mouse melanoma cells harboring a tyrosinase promoter. Extract concentrations of 10 ${\mu}g/m{\ell}$ and 100 ${\mu}g/m{\ell}$ resulted in tyrosinase promoter expression rates of approximately 113% and 184%, respectively, as compared to the control. The fraction layers consisting of butyl alcohol and methylene chloride improved expression effects on the tyrosinase promoter. In particular, the butyl alcohol fraction evidenced a high expression rate at 100 ${\mu}g/m{\ell}$. In the MTT assay, the methanolic extract did not evidence cytotoxicity at concentrations under 500 ${\mu}g/m{\ell}$. Therefore, the results observed with the extract of Glycine maxim showed that the substance exerted a positive effect on the tyrosinase promoter.