• Title/Summary/Keyword: human-to-human (H2H)

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Characterization of Phosphoinositide-3-kinase, Class 3 (PIK3C3) Gene and Association Tests with Quantitative Traits in Pigs

  • Kim, J.H.;Choi, B.H.;Lim, H.T.;Park, E.W.;Lee, S.H.;Seo, B.Y.;Cho, I.C.;Lee, J.G.;Oh, S.J.;Jeon, J.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.12
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    • pp.1701-1707
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    • 2005
  • This study deals with the characterization of porcine PIK3C3 and association tests with quantitative traits. PIK3C3 belongs to the class 3 PI3Ks that participate in the regulation of hepatic glucose output, glycogen synthase, and antilipolysis in typical insulin target cells such as those in the such as liver, muscle system, and fat. On the analysis of full-length mRNA sequence, the length of the PIK3C3 CDS was recorded as 2,664 bps. As well, nucleotide and amino acid identities between human and pig subjects were 92% and 99%, respectively. Five SNPs were detected over 5 exons. We performed genotyping by using a SNP C2604T on exon24 for 145 F$_2$ animals (from a cross between Korean native boars and Landrace sows) by PCR-RFLP analysis with Hpy8I used to investigate the relationship between growth and fat depot traits. In the total association analysis, which doesn' consider transmission disequilibrium, the SNP showed a significant effect (p<0.05) on body weight and carcass fat at 30 weeks of age as well as a highly significant effect (p<0.01) on back fat. In an additional sib-pair analysis, C allele still showed positive and significant effects (p<0.05) on back fat thickness and carcass fat. Moreover, the effects of C allele on the means of within-family components for carcass fat and back fat were estimated as 2.76 kg and 5.07 mm, respectively. As a result, the SNP of porcine PIK3C3 discovered in this study could be utilized as a possible genetic marker for the selection of pigs that possess low levels of back fat and carcass fat at the slaughter weight.

Development of Purification Process of Recombinant Human Vascular Endotherial Growth Factor (VEGF) using Fusion Protein (융합 단백질을 이용한 재조합 인간 혈관내피세포 성장인자의 정제공정 개발)

  • Sung, Keehyun;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.55 no.3
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    • pp.369-378
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    • 2017
  • Vascular endotherial growth factor (VEGF) is a potent mitogen that stimulates vascular permeability and angiogenesis and has a potential in therapeutic applications. An industrial production method that provides high yield as well as purity is needed. Researches for various factors of mild solubilization with combination of ubiquitin fusion protein to increase solubility were carried out as well as by changing pH and denaturant concentration. Usage of pET28-a bacteral expression vector in BL21 (DE3) host cell was capable of producing approximately 14 g/L VEGF fusion protein in 20L fermentor. A purification process consisting of four chromatography steps including refolding and digestion with UBP1 resulted in mild solublization under the conditions of 2M urea and pH 10.0 due to ubiquitin fusion tag protein that increases in solubility of target protein VEGF. High yield of refolding and dimerization could be obtained between two step Ni-affinity chromatography. Multimeric and misfolded proteins and endotoxin were removed by DEAE anion exchange chromatography. Final monomers were removed from dimers by gel filtration chromatography. Characterization analysis of purified dimeric VEGF was performed using SDS-PAGE and RP-HPLC with a purity of 97%.

Production of ${\beta}-xylosidase$ from Bifidobacterium sp. Int-57 (Bifidobacterium sp. Int-57이 생산하는 ${\beta}-xylosidase$의 생산특성)

  • Kang, Dong-Hyun;Lee, Ke-Ho;Ji, Geun-Eog
    • Korean Journal of Food Science and Technology
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    • v.25 no.2
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    • pp.89-93
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    • 1993
  • In order to study the effect of the intestinal bacteria on the physiology of the human large intestinal tract, we isolated the intestinal bacteria of Koreans and tested the enzymatic patterns. Isolated Bifidobacterium sp. Int-57 showed the higher activity of ${\beta}-xylosidase$ than other intestinal microorganisms. The effect of the carbon sources, nitrogen sources, inorganic salts, initial pH and initial temperature on the production of ${\beta}-xylosidase$ of Bifidobacterium sp. Int-57 was investigated. The most suitable carbon source, nitrogen source and inorganic salt for the production of ${\beta}-xylosidase$ were 1.1% xylose, 0.4% yeast extract and 0.0003% $CoCl_2$ respectively at initial pH 7.0 and temperature $40^{\circ}C$.

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Antioxidant and Antiallergic Activity of Compounds from the Aerial Parts of Scutellaria baicalensis Georgi (황금 지상부의 항산화 및 항 알러지 활성 성분)

  • Cha Ja-Hyun;Kim Hyun-Wok;Kim Sun-Gun;Jung Sung-Hee;Whang Wan-Kyunn
    • YAKHAK HOEJI
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    • v.50 no.2
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    • pp.136-143
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    • 2006
  • Roots of Scutellaria baicalensis have been used for fever remedy; diuresis, antiphlogistic. For the investigation of the active component from aerial parts of Scutellaria baicalensis, MeOH extracts from aerial parts of Scutellaria baicalensis were suspended with $H_2O$, and partitioned by $CHCl_3$. In order to investigate the efficacy of antioxidative activity the activity guided fraction and isolation of physiologically active substance were peformed. Its $H_2O,\;30\%,\;60\%$ MeOH and MeOH fractions were examined on antioxidative activity using DPPH method and TBARS assay; It was revealed that $30\%\;and\;60\%$ MeOH fractions have significant anti-oxidative activity. its fractions testing type I allergy, compound 48/80 induced systemic anaphylaxis was applied. As a result, compared with reference (cromolygate), these fraction significantly inhibited systemic anaphylaxis by $71\%\;and\;57\%$, respectively. From $30\%,\;60\%$ MeOH fraction, five compounds were isolated and elucidated apigenin 6-C-${\alpha}$-L-arabinopyranosyl-8-C-${\beta}$-D-glucopyranoside (isoschaftside, I), scutellarein 7-O-${\beta}$-D-glucuronopyranoside (scutellarin, II), apigenin 7-O- ${\beta}$-D-glucuronopyranoside (III), isoscutellarein 8-O-${\beta}$-D-glucuronopyranoside (IV), kaempferol 3-O-${\beta}$-D-glucopyranoside (V) through their physicochemical data and spectroscopic methods. We measured radical scavenging activity with DPPH method and anti-lipid peroxidative efficacy on human LDL with TBARS assay. [$I] showed antioxidant activities in order. Type I allergy compound 48/80 induced systemic anaphylaxis was applied. $[V inhibited systemic anaphylaxis in order.

Analysis of Quantitative Trait Loci (QTLs) for Unsaturated Fatty Acid Contents in Soybean Seed Using Recombinant Inbred Lines (콩에서 microsatellite marker를 이용한 불포화지방산 함량의 양적형질 유전자좌의 분석)

  • Kim, Hyeun-Kyeung;Im, Moo-Hyeog;Choung, Myoung-Gun
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1665-1670
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    • 2008
  • Soybean oil is an important source of vegetable oil for human food and nonfood applications and accounts for approximately 22% of the world's total edible oil production. Improvement of the quality and quantity of soybean seed oil constituents is one of the most important objectives in soybean breeding. The objective of this study was to identify quantitative trait loci (QTLs) that control oleic, linoleic, and linolenic acid contents in soybean. The 117 $F_{2:10}$ recombinant inbred lines (RIL) developed from a cross of 'Keunolkong' and 'Shinpaldalkong' were used. Narrow-sense heritability estimates based on a plot mean on seed weight, protein and oil content were 0.85, 0.82 and 0.81, respectively. Eight independent QTLs for oleic acid content were identified from linkage group (LG) A2, C1, D2, F, G, L, and O. Seven QTLs for linoleic acid content were located on LG D1b, E, H, I and L. Oil content was related with five QTLs located on LG C1, H, J, K, and L. Oleic, linoleic, and linolenic acid have two common QTLs on LG C1 and L. Thus, we identified major loci improving soybean oil quality.

Ginseng Gintonin Activates the Human Cardiac Delayed Rectifier K+ Channel: Involvement of Ca2+/Calmodulin Binding Sites

  • Choi, Sun-Hye;Lee, Byung-Hwan;Kim, Hyeon-Joong;Jung, Seok-Won;Kim, Hyun-Sook;Shin, Ho-Chul;Lee, Jun-Hee;Kim, Hyoung-Chun;Rhim, Hyewhon;Hwang, Sung-Hee;Ha, Tal Soo;Kim, Hyun-Ji;Cho, Hana;Nah, Seung-Yeol
    • Molecules and Cells
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    • v.37 no.9
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    • pp.656-663
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    • 2014
  • Gintonin, a novel, ginseng-derived G protein-coupled lysophosphatidic acid (LPA) receptor ligand, elicits $[Ca^{2+}]_i$ transients in neuronal and non-neuronal cells via pertussis toxin-sensitive and pertussis toxin-insensitive G proteins. The slowly activating delayed rectifier $K^+$ ($I_{Ks}$) channel is a cardiac $K^+$ channel composed of KCNQ1 and KCNE1 subunits. The C terminus of the KCNQ1 channel protein has two calmodulin-binding sites that are involved in regulating $I_{Ks}$ channels. In this study, we investigated the molecular mechanisms of gintonin-mediated activation of human $I_{Ks}$ channel activity by expressing human $I_{Ks}$ channels in Xenopus oocytes. We found that gintonin enhances $I_{Ks}$ channel currents in concentration- and voltage-dependent manners. The $EC_{50}$ for the $I_{Ks}$ channel was $0.05{\pm}0.01{\mu}g/ml$. Gintonin-mediated activation 1 of the $I_{Ks}$ channels was blocked by an LPA1/3 receptor antagonist, an active phospholipase C inhibitor, an $IP_3$ receptor antagonist, and the calcium chelator BAPTA. Gintonin-mediated activation of both the $I_{Ks}$ channel was also blocked by the calmodulin (CaM) blocker calmidazolium. Mutations in the KCNQ1 $[Ca^{2+}]_i$/CaM-binding IQ motif sites (S373P, W392R, or R539W)blocked the action of gintonin on $I_{Ks}$ channel. However, gintonin had no effect on hERG $K^+$ channel activity. These results show that gintonin-mediated enhancement of $I_{Ks}$ channel currents is achieved through binding of the $[Ca^{2+}]_i$/CaM complex to the C terminus of KCNQ1 subunit.

Antioxidant Activity and Inhibitive Effects on Human Leukemia Cells of Edible Mushrooms Extracts (식용버섯 추출물의 항산화 활성 및 혈액암세포에 대한 저해효과)

  • Kim Hyun Jeong;Bae Joon-Tae;Lee Ji-Won;Hwang Bo Mi-Hyang;Im Hyo Gwon;Lee In-Seon
    • Food Science and Preservation
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    • v.12 no.1
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    • pp.80-85
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    • 2005
  • The effect of 12 edible mushroom species on the antioxidant and cytotoxicity on cancer cells were studied Methanol extract of Lyophyllum ulmarium, Cordyceps militaris and Sarcodon aspratus showed $30\~60\%$ DPPH radical scavenging activity and $39\~53\%$ protective effects against the cytotoxicity of $B_2O_2$. Methanol extracts of Sarcodon aspratus, Lyophyllum ulmarium, Cordyceps militaris, Agaricus blazei and Ganoderma lucidum revealed high inhibitive activities in cytotoxicity on human leukemia tells such as promyelocytic leukemia cell (HL60) and histiocytic lymphoma cell (U937). Highest toxicity was observed against HL60 cells in Sarcodon aspratus methanol extract showing $70.5\%$ inhibition at 1mg/mL whereas Cordyceps militaris methanol extract showed $81.5\%$ inhibition against U937 cells. Most water extracts of edible mushrooms exhibited the lowest effect against HL60 and U937 cells compared to methanol extract. These extract did not show cytotoxic effects against human lymphocyte. Results revealed 5 kinds of edible murshroom (Cordyceps militaris, Agaricus blaxei, Lyophyllum ulmarium, Ganoderma lucidum and Sarcodon aspratus) have strong antioxidative and in vitro anticancer efforts.

In vitro Anti-oxidant and Anti-inflammatory Effects of Moringa Folium and Eucommiae Cortex 2:1 (g/g) Mixed Formula (모링가:두충 2:1 (g:g) 복합 조성물의 in vitro 항산화 및 항염 효능 연구)

  • Heo, Seok-Mo;Yang, Jin-Won;Kim, Jong-Lae;Park, Mi-Ryeong;Kim, Tae Gi;Ku, Sae-Kwang;Park, Soo-Jin
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.36 no.4
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    • pp.125-129
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    • 2022
  • The purpose of study is to evaluate in vitro anti-oxidant and anti-inflammatory effects of Moringa Folium and Eucommiae Cortex 2:1 (g/g) mixtures (MEMix). HaCaT and human normal dermal fibroblast were treated with 0.01-1 mg/mL of MEMix to monitor cytotoxicity. Radical scavenging activities of MEMix were examined by DPPH assay. To explore anti-inflammatory effect, Raw 264.7 cells were pretreated with MEMix for 1h and subsequently exposed to LPS for 18h. NO release and cytotoxicity of Raw 264.7 cells were measured by adding Griess and MTT reagents, respectively. TNF-α, IL-1β, IL-6, and PGE2 productions were examined by ELISA. Immunoblot analysis was conducted to examine COX-2 expression in MEMix pretreated Raw 264.7 cells. Up to 1 mg/mL concentration, treatment of MEMix for 24 h did not affect normal dermal fibroblast viability and significantly reduced cell viability of HaCaT cells with no concentration dependency. MEMix increased DPPH radical scavenging activity with concentration dependency. Radical scavenging activities by 1 mg/mL of MEMix was comparable with 30 µM of trolox. Pretreatment of MEMix did not change the reduction of Raw 264.7 cell viability. Exposure of LPS in Raw 264.7 cells significantly increased NO, TNF-α, IL-1β, IL-6, and PGE2 productions, and MEMix pretreatment attenuated these productions by LPS concentration dependently. However, pretreatment with MEMix did not change COX-2 expression by LPS in Raw 264.7 cells. MEMix showed in vitro anti-oxidant and anti-inflammatory activities. MEMix would be useful candidate agent against inflammation.

Effects of the Fraction of Sambucus Williamsii, NNMBS 246, on Osteoblastic Differentiation

  • Kang, Soon-Il;Park, Jaesuh;Kwon, Il-Keun;Kim, Eun-Cheol
    • CELLMED
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    • v.8 no.3
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    • pp.13.1-13.8
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    • 2018
  • In the field of osteoporosis, there has been growing interest in anabolic agents that enhance bone formation. The purpose of this study was to examine the effects of NNMBS 246 osteoblastic differentiation with associated signaling pathways. NNMBS 246 markedly increased alkaline phosphatase (ALP) activity and calcium nodule formation. Stimulation with NNMBS 246 not only increased the differentiation markers (ALP, OPN, OCN) level and transcription markers (RUNX2, Osterix) mRNA expression but also upregulated the ECM molecules and OPG mRNA expression. Treatments of NNMBS 246 downregulated MMPs (MMP-1, MMP-2, MMP-9), but RANKL mRNA expression. Furthermore, NNMBS 246 activated osteoblastic differentiation markers and formed calcium nodules in human periodontal ligament cells (hPDLCs) and cementoblast cells. NNMBS 246 induced phosphorylation of MAPKs, Akt, nuclear p65 and IkB-${\alpha}$. BMP-2/Smad and ${\beta}$-catenin signaling pathways were activated by NNMBS 246. Sirtinol (SIRT1 inhibitor) inhibited NNMBS 246-induced osteoblastic differentiation markers mRNA expression. These results suggested that NNMBS 246 has the potential to enhance osteoblastogenesis probably through the activation of BMP/Smad and ${\beta}$-catenin signal pathways, and SIRT1 plays as critical mediator in bone anabolic effect of NNMBS 246.

Heterogeneity of Solitary Behaviors in 4 year old Children Related to Child Inhibition and Parenting Behaviors (4세 아동의 단독행동 하위유형들과 행동억제 및 양육행동의 관련성)

  • Park, Seong-Yeon;Song, Ju-Hyun;Rubin, Kenneth H.
    • Korean Journal of Child Studies
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    • v.29 no.4
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    • pp.97-113
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    • 2008
  • In order to examine the antecedents of socially reticent, solitary passive and solitary active behaviors in preschoolers, 38 2 year olds were observed in Kagan's(1989) traditional behavioral inhibition paradigm and thereafter, at age four, in a same age, same sex quartet of peers. Correlation analysis revealed the independent and unique nature of solitary passive, solitary active and reticence behaviors at age 4 suggesting the heterogeneity of the three forms of solitude during the preschool period. Regression analysis indicated that associations between 2 year old inhibition and shyness and 4 year old reticence were positive when mothers demonstrated a high frequency of over protectiveness; similarly, associations between 2 year old inhibition and shyness and 4 year old solitary active behavior were significant and positive when mothers demonstrated high frequencies of authoritarian behavior.

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