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

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Rapid Detection of Growth factors of intestinal Lactic Acid Bacteria (장내유산균 증식인자의 신속한 검색)

  • 한명주;임혜영;김동현
    • Journal of Food Hygiene and Safety
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    • v.8 no.2
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    • pp.91-95
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    • 1993
  • The growth of Bifidobacterium and Lactobacillus isolated from human interstinal bacteria were induced by water extract and U-step extract of soybean and carrot and the pH of these bacteria-cultured media were decreased. The increasing growth rates of these bacteria are related to the decrease of the pH of these bacteria-cultured media. When human intestinal flora as starter were inoculated into the medium containing water extract of soybean and carrot. the growth of lactic bacteria were also induced and the pH of the media were decreased. By measuring the pH of the media which were inoculated and cultured intestinal bacteria as a starter, it is possible to determine whether the food are the growth factors of intestinal lactic acid bacteria or not. By this method, the food which decreased pH of the medium were soybean, turnip, carrot. leek, garic, dropwork, wonnwood and onion. 'These foods may induce lactic acid bacteria in human in1estlne.

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Study on Apoptosis Effect and Mechanism by Bojungikki-tang on Human Cancer Cell Line H460 (폐암세포주(肺癌細胞株) H460에 대(對)한 보중익기탕(補中益氣湯)의 세포고사효과(細胞枯死效果) 및 기전연구(機轉硏究))

  • Lee, Seung-Eon;Hong, Jae-Eui;Lee, Si-Hyeong;Shin, Jo-Young;Ro, Seung-Seok
    • The Journal of Internal Korean Medicine
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    • v.25 no.4
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    • pp.274-288
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    • 2004
  • Objectives : This study was designed to evaluate the effect on cytotoxicity of Bojungikki-tang(BIT) in human lung cancer H460 cells. Methods : BIT-induced cell death was confirmed as apoptosis characterized by chromatin condensation and increase of the $sub-G_1$, DNA content. It was tested whether the water extract of BIT affects the cell cycle regulators such as, p2l/Cipl, p27/Kipl, cyclin $B_1$. Results : The data showed that treatment of BIT decreased the viability of H460 cells in a dose-dependent manner. p2l/Cip1 is gradually decreased by the addition of the cells with BIT extract. Interestingly, p27/Kip1 is not detected for 24 hr after the addition of BIT extract, however, after 24 hr, p27/Kipl markedly increased. In addition, cyclin $B_1$, decreased in a time dependent manner after the addition of the water extract. The activation of caspase -3 protease was further confirmed by degradation of procaspase-8 protease andpoly(ADP-ribose) polymerase(P ARP) by BIT in H460 cells. Moreover, BIT induced the increase of Bak expression. Conclusion : These results suggest that the extract of BIT exerts anticancer effects to induce the death of human lung cancer H460 cells via down regulation of cell cycle regulators such as p2l/Cip1, and cyclin B1 or up regulation of cell cycle regulators such as p27/Kip1. Moerover results suggest that BIT induces an apoptosis in H460 cells via activation of intrinsic caspase cascades.

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Expression of the EPO-like Domains of Human Thrombopoietin in Escherichia coli

  • Koh, Yeo-Wook;Koo, Tai-Young;Ju, Sang-Myoung;Kwon, Chang-Hyuk;Chung, Joo-Young;Park, Myung-Hwan;Yang, Jai-Myung;Park, Seung-Kook
    • Journal of Microbiology and Biotechnology
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    • v.8 no.6
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    • pp.553-559
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    • 1998
  • cDNA of human thrombopoietin (hTPO) amplified by polymerase chain reaction from a cDNA library of human fetal liver was cloned. EPO-like domains ($hTPO_{153} \;or\; hTPO_{l63})\; of\; hTPO(hTPO_{332}$) were expressed in Escherichin coli using several kinds of expression systems, such as ompA secretion, thioredoxin fusion, and the $P_L$ and T7 expression systems. To obtain $hTPO_{153}$ in soluble form, $hTPO_{153}$ cDNA was fused in-frame behind the gene encoding ompA signal sequence and thioredoxin protein. When fused with either of the genes, $hTPO_{153}$ was not expressed to the detectable level. However, a high level expression of the EPO-like domain of hTPO was obtained using the PL and T7 expression system. $hTPO_{153} \;or\; hTPO_{l63} cDNA were subcloned into the pLex and pET-28a(+) vectors under the control of the inducible$ P_L\;T_7$ promoter, respectively. Proteins expressed using pl.ex vector and pET-28a(+) detected in insoluble forms with an expression level of about 14% and 9% of total cellular proteins, respectively, and the level of expression was rapidly diminished in 2 h after the maximum level of expression was reached.

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Simple Purification of Escherichia coli-Derived Recombinant Human Interleukin-2 Expressed with N-terminus Fusion of Glucagon

  • Won Hye-Soon;Lee Jeewon;Kim In-Ho;Park Young-Hoon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.1
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    • pp.13-16
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    • 2000
  • Simple procedures have been devised for purifying recombinant human interleukin-2 (hIL-2), which was expressed in Escberichia coli using sequences of glucagon molecules and enterokinase cleavage site as an N-terminus fusion partner. The insoluble aggregates of recombinant fusion protein produced in E. coli cytoplasm were easily dissolved by simple alkaline pH shift $(8\rightarrow12\rightarrow8)$. Following enterokinase cleavage, the recombinant hIL-2 was finally purified by one-step reversed-phase HPLC with high purity. The ease and high efficiency of this simple purification process seem to mainly result from the role of used glucagon fusion partner, which could be applied to the production of other therapeutically important proteins.

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Anti-Helicobacter pylori, Cytotoxic, and Anti-inflammatory Activities of White Ginseng Extract

  • Jee, Hee-Sook;Chang, Kyung-Hoon;Moon, Sun-Hee;Park, Se-Ho;Paik, Hyun-Dong
    • Food Science and Biotechnology
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    • v.17 no.5
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    • pp.1106-1109
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    • 2008
  • The anti-Helicobacter pylori activity, cytotoxicity, and anti-inflammatory activity of white ginseng extract (WGE) were investigated in vitro in this study. The antimicrobial effects of WGE toward H. pylori strains 52 J99, SSI, and 51 were tested using the disk diffusion method. Among these H. pylori strains, H. pylori 52 was the most sensitive, having the largest inhibition zone (19 mm), followed by J99, SSI, and 51. The zone of inhibition due to WGE increased significantly with increasing dosage. The cytotoxicity of WGE toward the human cancer cell lines A-549 (human lung carcinoma), HEC-1-B (human endometrial adenocarcinoma), HeLa (human uterin adenocarcinoma), and SW-156 (human kidney carcinoma) was measured using the 3-(4,5-dimethylthizol-2-yl)-2,5-diphenylate-tetrazolium bromide (MTT) assay. WGE exhibited an inhibitory effect on cell growth at 2.0 mg/mL for all tumor cell lines. An analysis of anti-inflammatory activity using the RAW 264.7 cell line showed that the inhibition of nitric oxide (NO) production increased as the WGE content increased. These results demonstrate the potential of WGE to be used as a health-promoting substance.

Development of Refolding Process to Obtain Active Recombinant Human Bone Morphogenetic Protein-2 and its Osteogenic Efficacy on Oral Stem Cells

  • Lee, Ji-Hye;Jang, Young-Joo
    • International Journal of Oral Biology
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    • v.42 no.2
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    • pp.71-78
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    • 2017
  • BMP-2 is a well-known TGF-beta related growth factor, having a significant role in bone and cartilage formation. It has been employed to promote bone formation in some clinical trials, and to differentiate mesenchymal stem cells into osteoblasts. However, it is difficult to obtain this protein in its soluble and active form. hBMP-2 is expressed as an inclusion body in the bacterial system. To continuously supply hBMP-2 for research, we optimized the refolding of recombinant hBMP-2 expressed in E. coli, and established an efficient method by using detergent and alkali. Using a heparin column, the recombinant hBMP-2 was purified with the correct refolding. Although combinatorial refolding remarkably enhanced the solubility of the inclusion body, a higher yield of active dimer form of hBMP-2 was obtained from one-step refolding with detergent. The refolded recombinant hBMP-2 induced alkaline phosphatase activity in mouse myoblasts, at $ED_{50}$ of 300-480ng/ml. Furthermore, the expressions of osteogenic markers were upregulated in hPDLSCs and hDPSCs. Therefore, using the process described in this study, the refolded hBMP-2 might be cost-effectively useful for various differentiation experiments in a laboratory.

Effect of Inhibitor of Glycogen Synthase Kinase 3 on Self-Renewal of Human Embryonic Stem Cells

  • Lee Eunyoung;Rho Jeung-yon;Yu Kwon;Paik Sang-Gi;Lee Kyung-Kwang;Han Yong-Mahn
    • Reproductive and Developmental Biology
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    • v.29 no.2
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    • pp.93-99
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    • 2005
  • Human embryonic stem cells (hESCs) derived from the inner cell mass of blastocysts have the ability to renew themselves and to differentiate into cell types of all lineage. The present study was carried out to investigate whether the Wnt signaling pathway is related to maintaining self-renewal of hESCs. Glycogen Synthase Kinase 3 (GSK-3) inhibitor, BIO ((2'Z,3'E)-6-Bromoindirubin-3'-oxime) was treated to Miz-hES1 line for activation of Wnt signaling pathway. BIO-nontreated hESCs (control) and BID-treated hESCs were cultured for 5 days in the modified feeder-free system. During the culture of hESCs, differences were observed in the colony morphology between 2 groups. Controls were spread outwards whereas BIO-nontreated hESCs were clumped in the center and the differentiated cells were spreading outwards in the edges. The results of stem cell specific marker staining indicated that control were differentiated in large part whereas BIO-treated hESCs maintain self-renewal in the center of the colony. The results of lineage marker staining suggested that outer cells of the hESC colony were differentiated to the neuronal progenitor cells in both control and BIO-treated hESC. These results indicate that Wnt signaling is related to self-renewal in hESCs. In addition, control group showed higher composition of apoptotic cells $(23.76\%)$ than the BID-treated group $(5.59\%)$. These results indicate that BIO is effective on antapoptosis of hESCs.

Study of The Apoptotic Mechanisms of Gunbibosinhangam-tang on Human Neuroblastoma Cell Line BE2 (Human Neuroblastoma Cell Line BE2에 대한 건비보신항암탕(健脾補腎抗癌湯)의 세포고사 기전 연구)

  • Cho, Young-Kee;Moon, Mi-Hyun;Lee, Seong-Kyun;Jeong, Hyun-Ae;Lee, Jung-Sub;Nam, Sang-Kyu;Moon, Goo;Shin, Sun-Ho;Kim, Dong-Woung
    • The Journal of Internal Korean Medicine
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    • v.27 no.3
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    • pp.725-736
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    • 2006
  • Objective: In order to investigate cell death mechanisms by Gunbibosinhangam-Tang(G.B.H) in cancer cells, the activities of apoptosis signaling pathway were tested in human neuroblastoma cell line BE2. Methods: Viability of BE2 cells was markedly decreased by treatment of the water extract of G.B.H in a dose-dependent manner. G.B.H-induced cell death was confirmed as apoptosis characterized by chromatin condensation, We tested whether the water extract of G.B.H affects the anti-apoptotic proteins such as Bcl-$X_L$ Results: Bcl-$X_L$ was uneffected by the addition of the water extract of G.B.H in a time-dependent manner. Cleavage of PARP(poly-ADP-ribose polymerase) by activation of caspase-8 protease was also observed in BE2 cells by the treatment of the water extract of G.B.H. Conclusion: These results suggest that the water extract of G.B.H exerts anti-cancer effects on human neuroblastoma BE2 cells by inducing the apoptotic death via activation of intrinsic caspase cascades.

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A Study of Sex Difference by Chemical Determination of Dentin in Human Teeth (치아경조직의 화학적 판정에 의한 성차 연구)

  • 장세활;김종열
    • Journal of Oral Medicine and Pain
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    • v.8 no.1
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    • pp.127-142
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    • 1983
  • This thesis was to investigate sex defferencd by employing the methodes of Bernadskij's and it's modification by means of the evaluation of pH of dentin in 162 extracted teeth. The obtained were as follows: 1. In 72% of all experinments, pH of human dentin substance was higher in female than in male. Mean pH was 13.175±0.031 in male and 13.245±0.030 in female. 2. After comparative studies on each pH in anatomical region of tooth arrangement, the sex difference of pH was most obviously showed in molars. 3. In pH change according to ages, the 30's was the highest in male and the 20's in female, also pH showed decresing tendency by the increasing of age in both male and female. 4. There was no pH difference in anatomical regions of tooth arrangment. 5. Mean pH was 13.282±0.013 in under 50 years of age and 13.282±0.022 over 50. It proved that pH was higher in under 50 years of age than in over 50.

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Cytotoxicity Evaluation of Elastomeric Impression Materials Using Different Fibroblasts Cell Lines

  • Kwon, Jae-Sung;Kim, Kyoung-Nam
    • Journal of Korean Dental Science
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    • v.7 no.2
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    • pp.80-86
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    • 2014
  • Purpose: Current common method of cytotoxicity evaluation for elastomeric impression materials use animal based cell lines, which the clinical relevance has been often questioned. Hence, the purpose of this study was to examine the difference in results with both human based and animal based fibroblast cell line. Materials and Methods: Three types of fibroblast cells were used in this study; conventional mouse fibroblasts of L929, human gingival fibroblasts (HGF-1), and immortalized human oral fibrobalsts (hTERT-hNOF). Test on extract and test by direct contact using different commercially available elastomeric impression materials were carried out according to the international standards. Result: There was significant difference in cell viability between types of fibroblasts cell used, where HGF-1 showed highest cell viability and L929 the lowest. Conclusion: Within the limitation of this study, careful consideration must be given when selecting the cells and interpreting the results for cytotoxicity evaluation of elastomeric impression materials, where use of human based cell lines such as hTERT-hNOF would be appropriate for both ease of cytotoxicity test and clinical relevance.