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

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Solid-Phase Refolding of Poly-Lysine fusion Protein of hEGF and Angiogenin (Poly-lysine이 연결된 hEGF와 angiogenin의 융합단백질의 고체상 재접힘)

  • Park, Sang-Joong;Ryu, Kang;Suh, Chang-Woo;Chai, Young-Gyu;Kwon, Oh-Byung;Park, Seung-Kook;Lee, Eun-Kyu
    • KSBB Journal
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    • v.17 no.2
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    • pp.153-157
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    • 2002
  • A fusion protein, consisting of a human epidermal growth factor as the recognition domain and human angiogenin as the toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as an inclusion body in recombinant E. coli, yet when the conventional solution-phase refolding process was used the refolding yield was very low due to severe aggregation, probably because of the opposite surface charge resulting from the vastly different pl values of each domain. Accordingly the solid-phase refolding process, which exploits the ionic interactions between a solid matrix and the protein, was tried, however the ionic binding yield was also very low regardless of the resins and pH conditions used. Therefore, to provide a higher affinity toward the solid matrix, six Iysine residues were tagged to the N-terminus of the hEGF domain. When cation exchange resins, such as heparin- or CM-Sepharose, were used as the matrix, the adsorption capacity increased 2.5~3-fold and the subsequent refolding yield increased nearly 15-fold compared to the conventional process. A similat result was also obtained when an Ni-NTA metal affinity resin was used.

Synthesis of Indeno[1.2-c]indenoisoquinoline Derivatives as Potential Topoisomerase I Inhibitors

  • Quynh, Le-Manh;Thanh, Le-Nguyen;Gang, Seong-Gyoung;Chung, Byung-Ho;Cho, Won-Jea
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.340.3-341
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    • 2002
  • During the research for the development of antitumor agents. we found the 3-arylisoquinoline derivatives exhibited potent cytotoxicity against human tumor cell lines. For extending our study on these compounds. indeno[1.2-c]isoquinolines were chosen as the next research target due to previous studied data of the compounds that showed potent topoisomerase I inhibition activity as well as cytotoxicity against many kinds of tumor cell lines. Retrosynthetic consideration of indeno[1.2-d]isoquinolines indicates that the coupling of o-methyltoluamide with o-hydroxymethylbenxonitrile might afford 3-arylisoquinoline which could be translerred to the aldehtde. Indeno [1.2-c]isoquinolines can be formed by and inframolecular ring cyclization method. Various derlvatives of this compound including 11-alkoxy-6-methyl-6H. 11H-indeno[1.2-c]isoquinolin-5-one and biological activity will be presented with the docking model with topisomerase 1 enzyme.

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Apoptosis and Peripheral Benzodiazepin Receptor (PBR) Expression in Human Granulosa-Luteal Cells by GnRH-agonist (GnRH-agonist에 의한 인간 과립-황체화 세포의 세포사멸과 PBR 단백질의 발현)

  • Kim, Sei-Kwang;Youm, Yun-Hee;Yoon, Jeong-Mi;Bai, Sang-Wook;Yang, Hyun-Won;Cho, Dong-Jae;Yoon, Yong-Dal;Song, Chan-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.2
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    • pp.83-94
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    • 2004
  • Objective: To investigate whether GnRH-agonist (GnRH-Ag) using in IVF-ET affects apoptosis of human granulosa-luteal cells and expression of peripheral benzodiazepine receptor (PBR) protein involved in the apoptosis of the cells. Methods: Granulosa-luteal cells obtained during oocyte retrieval were cultured and treated with $10^{-5}M$ GnRH-Ag. Apoptosis of the cells by the treatment was confirmed using DNA fragmentation analysis 24 h after culture. The presence of PBR protein within the cells was examined by immunofluorescence staining and the expression of the protein was analyzed by Western blotting. In addition, it was measured for progesterone and nitric oxide (NO) produced by granulosa-luteal cells after GnRH-Ag treatment. To evaluate the relationship between NO production and PBR expression, sodium nitroprusside (SNP) as a NO donor was added in media and investigated the expression of PBR protein by Western blotting. Results: Apoptosis increased in the granulosa-luteal cells 24 h after GnRH-Ag treatment, whereas the expression of PBR protein significantly decreased. Furthermore, the production of progesterone and nitric oxide (NO) by the cells significantly fell from 12 h after the treatment. In the results of Western blotting after SNP treatment, the expression of PBR protein increased in the treatment with SNP alone to the granulosa-luteal cells, but was suppressed in the treatment with GnRH-Ag and SNP. Additionally, the staining result of PBR protein in the cells showed the even distribution of it through the cell. Conclusion: These results demonstrate that GnRH-Ag treatment induces apoptosis, decreasing expression of PBR protein and NO production in human granulosa-luteal cells. The present study suggests that one of the apoptosis mechanism of human granulosa-luteal cells by GnRH-Ag might be a signal transduction pathway via NO and PBR.

Aging and Gasroenterrogi Changes (노화에 따른 위장관 기능의 변화)

  • 조우균
    • The Korean Journal of Food And Nutrition
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    • v.6 no.3
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    • pp.219-230
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    • 1993
  • This research aims to study the changes In gastrointestinal function attributed to aging In human. The thresholds for recognition and detection of flavors became elevated and salivary gland acinar cells decreased in the old age. But most esophageal function remained relatively Intact. Although gastric emptying time has been slowed with aging, the total intestinal transit time did not differ. Atropic gastritis due to H. pylori in old man decreased secretion of acid and Intrinsic factor and absorbability of calcium and iron. Pancreatic secretion is droned in older persons. Prevalence of gallstones rised with age. Liver size and portal blood flow decreased significantly with age. Mucosal surface area has been reported to be slightly diminished in the aging man. Glucose transporters decreased and Insulin tolerance Increased. Absorption of aromatic amino acid is diminished with age. Dietary protein In that aging human increased fecal nitrogen excretion. Vitamin A tolerance increased. Vitamin D receptor concentration decreased and resistance to 1,25-(OH)2D3 action increased. Permeability of aging small Intestine Increased. Zinc balance dirt not differ Copper absorption appeared not to be significantly affected by age. Neurotensin secretion decreased thus slowed colonic peristaltic movements and Intestinal mucosal growth.

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Inhibition of adhesion and biofilm formation in Escherichia coli O157:H7 by diosmin (다이오스민(diosmin)에 의한 병원성 대장균 세포부착 및 생물막 형성 억제)

  • Kim, Hyun Jung;Kim, Seung Min
    • Korean Journal of Food Science and Technology
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    • v.50 no.4
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    • pp.414-419
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    • 2018
  • Escherichia coli O157:H7 is one of the most common foodborne pathogens responsible for outbreaks of hemorrhagic colitis, which can lead to the life-threatening hemolytic-uremic syndrome. In this study, we identified phytochemicals that specifically inhibit the expression of LEE operon in E. coli O157:H7. Among phytochemicals, diosmin decreased the adherence of E. coli O157:H7 towards Caco-2 cells in vitro (p<0.01) and its biofilm formation activity (p<0.05). Quantitative RT-PCR analysis revealed that the transcripts of Ler-regulated genes and genes related to curli production were significantly reduced in the presence of diosmin. However, diosmin does not affect bacterial viability, indicating that the resistance rate to diosmin was remarkably low. Overall, these results provide significant insights into the development of a novel anti-infective agent that is different from conventional antibiotics.

Development of Mono Layer Cell Adsorption Apparatus to Create a Slide for Microscopic Diagnosis (현미경 진단용 슬라이드 제작을 위한 단층세포 흡착장치 개발)

  • Oh, H.Y.;Mun, M.G.;Kim, S.H.;Kim, D.W.;Kang, S.M.;Sung, R.G.;Kim, H.C.
    • Journal of Biomedical Engineering Research
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    • v.36 no.1
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    • pp.1-6
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    • 2015
  • This study aims to design a monolayer cell adsorption apparatus that would help to produce high-quality slides for Liquid-Based Cytology (LBC) of an early cancer diagnosis for human bodies. The LBC collects exfoliated cells of human bodies and spreads the cells on the slides. Through processes of dyeing and cytological examination, the LBC screens for cancers in early stage. In this study, both of a cell suction module and a cell adsorption module, which are the key elements of the monolayer cell adsorption apparatus, were developed, and using those modules, the study set, first, conditions to help both GYN and NON-GYN apply principal cells without de-endothelialization before conducting its own analysis on the utility. As a results, for GYN, apparatus was determined to be able to produce high-quality slides under the condition of 4 and for NON-GYN, the apparatus would come up with other slides of high-quality under the condition of 2. The study carried out a repetitive test on selected conditions and proved 96% of the repetitive success rate. By the results of what has been learned so far, the study presents that the apparatus has a possibility to replace device from South Korea as one of those other currently-applied systems to run the LBC and that the system will also present a new paradigm for cancer diagnosis as it makes a contribution to the improvement in the LBC.

Ex Vivo ${1}^H$ MR Spectroscopy: Normal gastric and cancer tissue (정상 위 조직과 위암 조직의 시험관 내 수소자기공명분광)

  • Cho Ji Youn;Shin Oon Jae;Choi Ki Seung;Kim Su Hyun;Eun Choong Ki;Yang Young Il;Lee Jung Hee;Mun Chi Woong
    • Journal of Gastric Cancer
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    • v.3 no.3
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    • pp.151-157
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    • 2003
  • Purpose: In this study, we attempted to ascertain the proton magnetic resonance spectroscopy (${1}^H$ MRS) peak characteristics of human gastric tissue layers and finally to use the metabolic peaks of MRS to distinguish between normal and abnormal gastric specimens. Materials and Methods: Ex-vivo ${1}^H$ MRS examinations of thirty-five gastric specimens were performed to distinguish abnormal gastric tissues invaded by carcinoma cells from normal stomach-wall tissues. High-resolution 400-MHz (9.4-T) ${1}^H$ nuclear magnetic resonance (NMR) spectra of two gastric layers, a proper muscle layer, and a composite mucosasubmucosa layer were compared with those of clinical 64- MHz (1.5-T) MR spectra. Three-dimensional spoiled gradient recalled (SPGR) images were used to determine the size and the position of a voxel for MRS data collection. Results: For normal gastric tissue layers, the metabolite peaks of 400-MHz ${1}^H$ MRS were primarily found to be as follows: lipids at 0.9 ppm and 1.3 ppm; alanine at 1.58 ppm; N-acetyl neuraminic acid (sialic acid) at 2.03 ppm; and glutathione at 2.25 ppm in common. The broad and featureless featureless spectral peaks of the 64-MHz MRS were bunched near 0.9, 1.3, and 2.0, and 2.2 ppm in human specimens without respect to layers. In a specimen (Borrmmann type III) with a tubular adenocarcinoma, the resonance peaks were measured at 1.26, 1.36 and 3.22 ppm. All the peak intensities of the spectrum of the normal gastric tissue were reduced, but for gastric tumor tissue layers, the lactate peak split into 1.26 and 1.39 ppm, and the peak intensity of choline at 3.21 ppm was increased. Conclusion: We found that decreasing lipids, an increasing lactate peak that split into two peaks, 1.26 ppm and 1.36 ppm, and an increasing choline peak at 3.22 ppm were markers of tumor invasion into the gastric tissue layers. This study implies that MR spectroscopy can be a useful diagnostic tool for gastric cancer.

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Molecular Characterization of an H5N3 Influenza Virus Isolated from Spot-Billed Duck

  • Lee, Jin Hwa;Kwon, Hyuk Moo;Sung, Haan Woo
    • Korean Journal of Poultry Science
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    • v.40 no.3
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    • pp.243-252
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    • 2013
  • Among the 16 hemagglutinin (HA) subtypes of avian influenza virus (AIV), only the H5 and H7 subtypes have caused highly pathogenic avian influenza (HPAI) in poultry. However, most H5 or H7 subtype viruses are categorized as low pathogenic avian influenza (LPAI). Some AIVs, including the H5 and H7 HPAI viruses, have shown the ability to infect humans directly. In this study, we describe the biological and molecular characterization of an H5N3 AIV (SBD/KR/KNU SYG06/06) isolated from spot-billed duck (Anas poecilorhyncha) in Korea. A phylogenetic analysis of the eight viral genes showed that the SBD/KR/KNU SYG06/06 isolate belongs to the Eurasian lineage and that the SBD/KR/KNU SYG06/06 isolate was clearly different from HPAI H5N1 strains, including human isolates and the Italian HPAI H5N2 strains. Additionally, no relationship was found between SBD/KR/KNU SYG06/06 and the Korean HPAI H5N1 isolates. The SBD/KR/ KNU SYG06/06 isolate had avian specific receptor binding site residues in the HA protein and the four C-terminal amino acids in the NS1 protein. The HA protein of the SBD/KR/KNU SYG06/06 isolate exhibited the typical LPAI motif at the cleavage site and this virus produced no cytopathic effects in MDCK cells without trypsin. Given these results, we suggest that the H5N3 AIV isolated from the spot-billed duck should be considered an LPAI virus and should have no pathogenic effect in humans.

Mouse In Vitro Fertilization (마우스 난자(卵子)의 체외수정(體外受精)에 관(關)한 연구(硏究))

  • Lim, Y.T.;Choi, S.H.;Kim, J.G.;Moon, S.Y.;Lee, J.Y.;Chang, Y.S.
    • Clinical and Experimental Reproductive Medicine
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    • v.11 no.2
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    • pp.51-57
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    • 1984
  • The success of human in vitro fertilization (IVF) & embryo transfer (ET) has focused attention on the culture conditions that can provide optimal development of the preimplantation embryo. Studies of in vitro fertilization using mouse have direct implications to human IVF, since similar conditions are used for both species. Mouse IVF as a quality control system for human IVF & ET was studied since Feb., 1984. The results were as follows: 1. Egg retrieval following superovulation in IeR mice was l5.1${\pm}$5.3 eggs ovulated/mouse (Mean${\pm}$ S.D.) 2. In vitro cleavage rate was 61.7% (1146 eggs cleaved/l858 eggs inseminated) and % blastocyst was 42.6%. 3. In comparison with two media of Ham's F-10 and m-KRB, in vitro cleavage rate were 40.9%/63.l% and %blastocyst were 44.3%/61.2% (P<0.05). 4. It was concluded that mouse IVF system has a valuable place in human IVF & ET as a quality-control system and in human reproductive physiology as a research model.

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Study on the Antitumor Activity of Tripterygium Regelii Sprague (미역줄나무의 항암활성에 관한 연구)

  • Park, Wan-Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.2
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    • pp.441-445
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    • 2005
  • Tripterygium regelii has been used as an oriental medicine, especially antiparasitic, anti-inflammatory and detoxifying agents in East asia. During our research to develop new antitumor agents from natural products, MeOH ext. and CH2Cl2 ext. of Tripterygium regelii showed the potent antitumor activity. In order to purify active compounds from Tripterygium regelii, activity-guided fractionation was carried out. Silica gel and RP-18 column chromatography for the active fraction led to the isolation of two compounds and their antitumor activities were studied. Those two compounds didn't show potent antitumor activity against human tumor cell lines. The structure of two compounds were determined by $^1H-NMR$, $^{13}C-NMR$, DEPT, $^1H-^{13}C$ COSY and IR spectrum. Compound I and Compound II were turned out to be Celastrol, and ${\beta}-sitosteryl-3-o-{\beta}-D-glucopyranoside$ respectively.