• Title/Summary/Keyword: Expression Concentration

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Effects of Cordyceps militaris(CM) on Osteoclastogenesis and Gene Expression (동충하초가 파골세포의 분화와 유전자 발현에 미치는 영향)

  • Choi, Kyung-Hee;Yoo, Jeong-Eun;Hwang, Gwi-Seo;Yoo, Dong-Youl
    • The Journal of Korean Obstetrics and Gynecology
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    • v.25 no.3
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    • pp.16-26
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    • 2012
  • Objectives: This study was performed to evaluate effects of Cordyceps militaris(CM) on osteoclast differentiation and its related gene expression. Methods: We used mouse myeloid cells RAW 264.7 stimulated by receptor activator of nuclear factor kappa-B ligand(RANKL) to induce osteoclast differentiation. There are four groups of which RAW 264.7 cells are not stimulated by RANKL (Normal), stimulated by RANKL without CM(Control), stimulated by RANKL with 0.1 ${\mu}g/ml$ of CM(CM 0.1), stimulated by RANKL with 1 ${\mu}g/ml$ of CM(CM 1). Osteoclastogenesis was measured by counting Tartrate-resistant acid phosphatase-positive multinucleated cells [TRAP(+) MNC]. RT-PCR was performed to evaluate the inhibitory effect of CM on gene expression(TRAP, AKT1, JNK1, NFATc1, c-Fos, MITF). Results: 1. CM decreased the number of TRAP(+) osteoclast in RANKL-stimulated RAW 264.7 cell at the concentration of 0.1 ${\mu}g/ml$ and 1 ${\mu}g/ml$. 2. CM decreased the expression of TRAP in osteoclast at the concentration of 1 ${\mu}g/ml$. 3. CM decreased the expression of AKT1, JNK1 in osteoclast at the concentration of 1 ${\mu}g/ml$. 4. CM didn't affect the expression of NFATc1, c-Fos, MITF in osteoclast. Conclusions: Cordyceps militaris has inhibitory effects on osteoclast differentiation and its related gene expression. These results suggest that Cordyceps militaris has a potential as a treatment of osteoporosis.

Gene Gun-Mediated Human Erythropoietin Gene Expression in Primary Cultured Oviduct Cells from Laying Hens

  • Ochiai, H.;Park, H.M.;Sasaki, R.;Okumura, J.;Muramatsu, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.1
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    • pp.9-14
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    • 1999
  • Factors affecting gene gun-mediated expression of the human erythropoietin gene were investigated in primary cultured oviduct cells from laying hens. The human erythropoietin gene was transfected by a gene gun method at $1.25{\mu}g$ per dish, and cultured in a synthetic serum-free medium for 72 hrs. The concentration of human erythropoietin mRNA was determined by RNA : RNA solution hybridization. In experiment 1, the effect of changing the shooting pressure of DNA-coated microparticles with nitrogen gas was tested at 20 and $60kgf/cm^2$. The results showed that the erythropoietin mRNA concentration was significantly higher at 60 than $20kgf/cm^2$. In experiment 2, the effects of supplementing the medium with fetal calf serum at 10%, and raising the shooting pressure from 60 to $80kgf/cm^2$ on the cell number and erythropoietin gene expression were examined. Although supplementation with fetal calf serum significantly increased the cell numbes compared with no supplemented controls (p < 0.05), erythropoietin mRNA concentration per $10^3$ cells was not affected. Raising the shooting pressure from 60 to $80kgf/cm^2$ did not affect either of the parameters, In experiment 3, the effect of supplementing ascorbate 2-phosphate at 0.5 mM was tested. The results indicated that the ascorbate supplementation significantly increased the cell number (p < 0.05), and tended to increase erythropoietin mRNA concentration (p < 0.1). Thus, for human erythropoietin gene expression by using the gene gun method, shooting pressure with nitrogen gas should be sufficient at $60kgf/cm^2$ and supplementation with ascorbate phosphate would be useful to enhance not only the cell proliferation but also erythropoietin gene expression.

The Effects of Dietary Zinc Deficiency on the Expression of Metallothionein, Absorption and Distribution of Cadmium in Rats (아연결핍이 흰쥐에서 metallothionein의 발현과 카드뮴의 흡수 및 분포에 미치는 영향)

  • Jeon Yong-Wook;Choi Byung-Sun;Park Jung-Duck
    • Environmental Analysis Health and Toxicology
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    • v.19 no.2
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    • pp.191-200
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    • 2004
  • Zinc (Zn) is an essential element in biological process, however inadequate Zn status in general population have been recognized. To update the knowledge for Zn-cadmium (Cd) interaction, we studied the intestinal uptake and transport, and the expression of metal transporter proteins (divalent metal transporter 1, DMT1 ; metal transporter protein 1, MTP1 ; zinc transporter 1, ZnTl ; metallothionein 1 , MT1) in duodenum after Cd exposure using Zn deficient animal model. Rats were led Zn deficient (ZnD, 0.5-1.0 mgZn/kg) or Zn supplemented (ZnS, 50mg Zn/kg) diet for 4 weeks, and followed single administration of $^{109}$ CdCl$_2$orally. The body Zn flatus and tissue Cd concentration were determined at 24 hrs after Cd administration. Total body burden of Cd and Cd absorption index (AI, %) were estimated based on the tissue Cd analyzed. DMT1, MTP1, ZnTl and MT1 mRNA were analyzed by using RT-PCR method. Feeding of Zn deficient diet for 4 weeks produced a reduced body weight gain and a depletion of body Zn. Tissue Cd concentration, body burden of Cd and Cd absorption index were higher in the ZnD diet fed rats than the ZnS diet red rats. Especially, Cd concentration in the small intestine (duodenum, jejunum and ileum) and the colon of FeD diet fed rats were higher markedly than in the FeS diet group. The expression levels of DMT1, MTP1 and ZnT1 mRNA in FeD diet fed rats were similar to the FeS diet. The level of MT1 mRNA expression was significantly lower in the FeD than the FeS diet fed rats. Taken together, theses results indicate that Zn deficiency in diet induce an increased intestinal absorption and tissue retention of Cd, and down -regulate the MT1 expression in the intestine which might be play a part of role in Cd absorption and transport in mammalian. These findings suggest that deficiency of essential metal could be enhanced the toxicity of toxic, non-esstial metals through the metal-metal interaction.

Induction of the T7 Promoter Using Lactose for Production of Recombinant Plasminogen Kringle 1-3 in Escherichia coli

  • Lim, Hyung-Kwon;Lee, Shi-Uk;Chung, Soo-Il;Jung, Kyung-Hwan;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.225-230
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    • 2004
  • A plasminogen kringle domain 1 to 3, rKl-3, was expressed in Escherichia coli under the control of T7 promoter. For the cost-effective production of rKl-3, the induction process was analyzed and optimized. Induction characteristics with lactose were analyzed in terms of induction time and inducer concentration in various culture conditions including batch and high-cell-density fed-batch cultures. In the fed-batch culture, the induction around 6 h after initiation of the DO-stat fed-batch culture resulted in the highest expression level of rKI-3 among the induction points examined. The highest demand of oxygen at this point was crucial for the maximum expression level of rKI-3. As the lactose concentration increased, the expression level also increased, though the expression level showed a plateau above a concentration of 14 mM of lactose. Lactose acted less specifically than IPTG since most of it was hydrolyzed to glucose and galactose. However, using lactose, the cell growth and the maximum expression level of rKl-3 increased by 20% and 24%, respectively, compared with those using IPTG in the fed-batch culture. The lactose seemed to be hydrolyzed by intracellular and extracellular $\beta$-galactosidase liberated by cell lysis at the same time. Residual concentration of glucose was maintained to a a limit of detection by high performance liquid chromatography, and galactose was not consumed by the host strain Escherichia coli BL2l(DE3).

Overexpression and Secretory Production of Endoxylanase from Recombinant Bacillus subtilis (재조합 Bacillus subtilis로부터 endoxylanase의 과발현 및 분비생산)

  • 김종현;남수완
    • Journal of Life Science
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    • v.10 no.2
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    • pp.125-130
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    • 2000
  • To overproduce endoxylanase from a recombinant Bacillus subtilis harboring the pJHKJ4 plasmid, the effects of carbon and nitrogen sources on the cell growth and expression level of endoxylanase were investigated in the flask cultures. Among the various carbon and nitrogen sources tested, glucose and maltose as carbon source and yeast extract as nitrogen source were found to be the most effective for the cell growth and the endoxylanase expression. When the concentration of glucose was increased from 0.5% to 5%, the highest activity of extracellular endoxylanse, 166 unit/$m\ell$, was observed at 2% glucose. In case of maltose, the endoxylanase was stably produced at the level of 180 unit/$m\ell$, regardless of the concentration of maltose. The higher the concentration of yeast extract, the greater cell growth and endoxylanase expression were obtained. However, the highest endoxylanase activity per unit cell mass was observed with 1% yeast extract. With the optimized medium (2% glucose, 1% yeast extract, etc), about 630 unit/$m\ell$ of endoxylanse was expressed through the batch fermentation in a fermentor, which expression level corresponded to about 0.7 g-endoxylanase protein /$\ell$. It was also found that the plasmid was stably maintained above 70% level, and more than 90% of endoxylanase activity was detected in the extracellular medium.

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Effect of White Ginseng on the Function of Mouse Peritoneal Macrophages and their Gene Expression (백삼성분이 마우스 복강 탐식세포의 기능 및 유전자 발현에 미치는 영향)

  • 배지현
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.6
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    • pp.1252-1257
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    • 1997
  • In order to investigate the immunomodulatory mechanism of white ginseng, the effects of total saponin of Ginsenoside Rb$_2$component on the phagocytosis and reactive oxygen intermediate(ROI) production of mouse peritoneal macrophages were studied. Both phagocytosis assay nitrobluetetrazolium reduction test showed 20$\mu\textrm{g}$/ml concentration of total saponin significantly increased the activity of phagocytosis and production of ROI. Also cytokine gene expression of the macrophages was analyzed using reverse transcription polymerase chain reaction. In the RT-PCR assay, 20$\mu\textrm{g}$/ml concentration of either total saponin or Ginsenoside Rb$_2$increased IL-1 and TNF expression of the macrophages.

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Effect of Cordyceps militaris mycelia containing improved cordycepin on expression gene in the melanin biosynthesis pathway

  • Si Young Ha;Ji Young Jung;Jae-Kyung Yang
    • Journal of Mushroom
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    • v.21 no.1
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    • pp.8-14
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    • 2023
  • This study aimed to verify the whitening effect of Cordyceps militaris, which is distributed in several countries worldwide, including Korea, Japan, and China, and has various medical effects. To screen the efficacy of C. militaris, the inhibitory activity of tyrosinase, which was 66% at a concentration of 1 mg/mL, was measured. Thereafter, the survival rate of melanoma cells was measured, and cell experiments were conducted at a concentration of 90% or more in which C. militaris was not toxic to cells. After measuring the inhibitory effect of TRP-1, TRP-2, tyrosinase protein, and mRNA expression, which are factors influencing melanin synthesis, C. militaris was found to decrease in all factors, with an expression level that was significantly lower compared to quercetin. This confirmed that C. militaris stimulated with LED has excellent whitening activity and can be used as a functional whitening cosmetics material.

Association of Endocrine Factors (Insulin-Like Growth Factor-II and Binding Protein-3) with Litter Size in Pigs

  • Yun, J.S.;Kang, W.J.;Seo, D.S.;Park, S.S.;Hong, K.C.;Lee, C.Y.;Ko, Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.3
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    • pp.307-315
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    • 2001
  • Litter size has been one of the important economic traits in porcine reproduction. The insulin-like growth factor (IGF) system has been shown to mediate actions of the steroid hormone or to synergize with other endocrine factors so that it consequently plays roles in reproductive processes, including ovulation, implantation, maintenance of pregnancy, and fetal development. However, the effect of the serum IGF system on porcine litter size has not been deeply studied. Therefore, this study was conducted to relate serum IFG-II concentration and IGF binding protein-3 (IGFBP-3) expression with porcine litter size. Moreover, the possible association of those with estrogen receptor (ER) as a candidate gene for litter size was investigated. Swine were separated into two groups showing high and low litter sizes, and sera were collected from sows in the estrous cycle to postnatal growth of their female progeny. Serum IFG-II concentration was measured by radioimmunoassay and IGFBP-3 expression was detected by Western ligand blotting. During the estrous cycle, IGFBP-3 expression in both groups decreased moderately from metestrus to estrus, but IFG-II concentration showed a reverse pattern. Also, IFG-II concentration and IGFBP-3 expression decreased gradually as pregnancy proceeded. Unlike IGFBP-3, IFG-II decreased moderately as newborn pigs grew. Significant differences in serum IFG-II amount between the two groups were detected at 60 (p<0.01), 75, 90, and 105 d (p<0.05) of pregnancy and at 60 (p<0.01), 45, and 105 d (p<0.05) of postnatal growth. Furthermore, based on ER genotypes, a high litter size group with genotypes AB and BB showed lower IFG-II concentration than a low litter size group with a genotype AA during pregnancy. Taken together, the results indicate that the serum IFG-II and IGFBP-3 are correlated with the litter size in pigs.

Effects of Korean Medicine Music Therapy on Anxiety, Anger Expression, and Concentration of Smart phone Addicted Adolescents (한방음악치료가 스마트폰 중독 청소년의 불안과 분노표현 및 주의집중력 향상에 미치는 영향)

  • Lee, Mi-hee;Lee, Seung-hyun
    • Journal of Korean Medical classics
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    • v.30 no.3
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    • pp.133-141
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    • 2017
  • Objectives : As of 2013, South Korea ranked number one in the world in smartphone penetration rate with 67.6%. Unfortunately, with the rise of smart phone use, side effects such as fraud, loss of personal information, identity theft, formation and widening of a mobile information gap, and last, but not least, smart phone addiction, also increased. Methods : Seventh (7th)graders from a girls' junior high school in Seoul, the capital of South Korea, we reevaluated the Smart phone Addiction Proneness Scale (S-scale). Among the 9 students classified as high risk users, 5 were randomly selected for a study group, and the remaining 4 students, meanwhile, were designated as a control group. The study group received Korean medicine music therapy (KMMT) in a total of 8 sessions, each session lasting 50 minutes. Both pre- and post-interventions, the two groups completed the S-scale, Harris and Harris' concentration grid exercise, STAI, and STAXI-K. We analyzed the data by t-test and paired t-test using IBM SPSS Statistics. Results : KMMT had a significant influence on proneness to smart phone addiction (t=3.130, p=.035). KMMT had positive effects on concentration improvement (t=-2.935, p=.043), trait-anxiety symptoms improvement (t=2,992, p=.040), and anger-expression control (t=4.416, p=.012). Conclusions : KMMT had positive, improving effects on trait anxiety and anger expression, which in turn enhanced the improvement of concentration as well as the decrease of smart phone-addiction proneness.

Vaccinium angustifolium Root Extract Suppresses FcεRI Expression in Human Basophilic KU812F Cells.

  • Shim, Sun Yup;Lee, Kyung Dong;Lee, Mina
    • Preventive Nutrition and Food Science
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    • v.22 no.1
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    • pp.9-15
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    • 2017
  • Vaccinium angustifolium, commonly known as the lowbush blueberry, is a rich source of flavonoids, with which various human physiological activities have been associated. The present study focuses on the investigation of the effect of the methanolic extract of V. angustifolium root extract (VAE) on high affinity immunoglobulin E receptor ($Fc{\varepsilon}RI$) ${\alpha}$ chain antibody (CRA-1)-induced allergic reaction in human basophilic KU812F cells. The total phenolic content of VAE was found to be $170{\pm}1.9mg$ gallic acid equivalents/g. Flow cytometry analysis revealed that the cell surface expression of $Fc{\varepsilon}RI$ was suppressed in a concentration-dependent manner upon culture with VAE. Reverse-transcriptase polymerase chain reaction analysis showed that the mRNA level of the $Fc{\varepsilon}RI$ ${\alpha}$ chain was reduced in a concentration-dependent manner as a result of VAE treatment. Western blot analysis revealed that the protein expression of $Fc{\varepsilon}RI$ and the phosphorylation of extracellular signal-regulated kinases (ERK) 1/2 were concentration-dependently inhibited by VAE. We determined that VAE inhibited anti-CRA-1-induced histamine release, in addition to the elevation of intracellular calcium concentration ([$Ca^{2+}$]i), in a concentration-dependent manner. These results indicate that VAE may exert an anti-allergic effect via the inhibition of calcium influx and histamine release, which occurs as a result of the down-regulation of $Fc{\varepsilon}RI$ expression through inhibition of ERK 1/2 activation.