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

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Production of hGCSF and GFP Co-Expressed Transgenic Cow Embryo by Somatic Cell Nuclear Transfer Technique (체세포 핵치환 기술을 이용한 hGCSF와 GFP 유전자 동시발현 형질전환 소 배아 생산)

  • Yang, Jung Seok;Joe, So Young;Koo, Bon-Chul;Heo, Young-Tae;Lee, Su Min;Kang, Man-Jong;Song, Hyuk;Ko, Dae Hwan;Uhm, Sang Jun
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.219-224
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    • 2015
  • The purpose of this study is to develop transgenic cell line expressing targeted human granulocyte colony stimulating factor (hGCSF) and green fluorescence protein (GFP) genes as well as production of Somatic Cell Nuclear Transfer (SCNT) embryos derived from co-expressed transgenic donor cells. Constructed pPiggy-mWAP-hGCSF-EF1-GFP vector was chemically transfected into bovine fetus cells and then, only GFP expressed cells were selected as donor cells for SCNT. Cleavage and blastocyst rates of parthenogenetic, SCNT embryos using non-TG cell and hGCSF-GFP dual expressed SCNT embryos were examined (cleavage rate: $78.0{\pm}2.8$ vs. $73.1{\pm}3.2$ vs. $70.4{\pm}4.3%$, developmental rate: $27.2{\pm}3.2$ vs. $21.9{\pm}3.1$ vs. $17.0{\pm}2.9%$). Result indicated that cleavage and blastocyst rates of TG embryos were significantly lower (P<0.05) than those of parthenogenetic and non-TG embryos, respectively. In this study, we successfully produced hGCSF-GFP dual expressed SCNT embryos and cryopreserved to produce transgenic cattle for bioreactor system purpose. Further process of our research will transfer of transgenic embryos to recipients and production of hGCSF secreting cattle.

Liquid Culture of Basidiomycetes on Natural Media (천연물을 이용한 담자균의 균사체 배양)

  • Kim, Seon-Hee;Lee, Jong-Suk;Park, Kyung-Sook;Lee, Jae-Sung;Lee, Hang-Woo;Park, Shin
    • The Korean Journal of Mycology
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    • v.27 no.6 s.93
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    • pp.373-377
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    • 1999
  • This study was designed to develop functional drink from jujube extract through a simple submerged culture of three basidiomycetes species. The optimum Brix and pH of the jujube extract for culturing the Ganoderma lucidum appeared to be 5 Brix and pH 4. Ten days of culture period produced maximum mycelium. The optimum Brix and pH of the jujube extract for culturing the Coriolus versicolor appeared to be 5 Brix and pH 5. Ten days of culture period produced maximum mycelium. The optimum Brix and pH of the jujube extract for culturing the Phellinus igniarius appeared to be 3 Brix and pH 5. For the maximum mycelial production eighteen days of culture period was required for Phellinus igniarius. The antitumor activity of the polysaccharides extracted from the fermented drinks was demonstrated through the tumor cell line experiments. The $IC_{50}$ values of the jujube drinks fermented with Ganoderma lucidum and Phellinus igniarius against stomach cancer cell line appeared to be one fourth that of the jujube drink which was not fermented with basidiomycetes.

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Expression and secretion of CXCL12 are enhanced in autosomal dominant polycystic kidney disease

  • Kim, Hyunho;Sung, Jinmo;Kim, Hyunsuk;Ryu, Hyunjin;Park, Hayne Cho;Oh, Yun Kyu;Lee, Hyun-Seob;Oh, Kook-Hwan;Ahn, Curie
    • BMB Reports
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    • v.52 no.7
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    • pp.463-468
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    • 2019
  • Autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic diseases (frequency of 1/1000-1/400), is characterized by numerous fluid-filled renal cysts (RCs). Inactivation of the PKD1 or PKD2 gene by germline and somatic mutations is necessary for cyst formation in ADPKD. To mechanistically understand cyst formation and growth, we isolated RCs from Korean patients with ADPKD and immortalized them with human telomerase reverse transcriptase (hTERT). Three hTERT-immortalized RC cell lines were characterized as proximal epithelial cells with germline and somatic PKD1 mutations. Thus, we first established hTERT-immortalized proximal cyst cells with somatic PKD1 mutations. Through transcriptome sequencing and Gene Ontology (GO) analysis, we found that upregulated genes were related to cell division and that downregulated genes were related to cell differentiation. We wondered whether the upregulated gene for the chemokine CXCL12 is related to the mTOR signaling pathway in cyst growth in ADPKD. CXCL12 mRNA expression and secretion were increased in RC cell lines. We then examined CXCL12 levels in RC fluids from patients with ADPKD and found increased CXCL12 levels. The CXCL12 receptor CXC chemokine receptor 4 (CXCR4) was upregulated, and the mTOR signaling pathway, which is downstream of the CXCL12/CXCR4 axis, was activated in ADPKD kidney tissue. To confirm activation of the mTOR signaling pathway by CXCL12 via CXCR4, we treated the RC cell lines with recombinant CXCL12 and the CXCR4 antagonist AMD3100; CXCL12 induced the mTOR signaling pathway, but the CXCR4 antagonist AMD3100 blocked the mTOR signaling pathway. Taken together, these results suggest that enhanced CXCL12 in RC fluids activates the mTOR signaling pathway via CXCR4 in ADPKD cyst growth.

White mineral trioxide aggregate mixed with calcium chloride dihydrate: chemical analysis and biological properties

  • Ahmed, Hany Mohamed Aly;Luddin, Norhayati;Kannan, Thirumulu Ponnuraj;Mokhtar, Khairani Idah;Ahmad, Azlina
    • Restorative Dentistry and Endodontics
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    • v.42 no.3
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    • pp.176-187
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    • 2017
  • Objectives: This study aimed to evaluate the chemical and biological properties of fast-set white mineral trioxide aggregate (FS WMTA), which was WMTA combined with calcium chloride dihydrate ($CaCl_2{\cdot}2H_2O$), compared to that of WMTA. Materials and Methods: Surface morphology, elemental, and phase analysis were examined using scanning electron microscope (SEM), energy dispersive X-ray microanalysis (EDX), and X-ray diffraction (XRD), respectively. The cytotoxicity and cell attachment properties were evaluated on human periodontal ligament fibroblasts (HPLFs) using methyl-thiazoldiphenyltetrazolium (MTT) assay and under SEM after 24 and 72 hours, respectively. Results: Results showed that the addition of $CaCl_2{\cdot}2H_2O$ to WMTA affected the surface morphology and chemical composition. Although FS WMTA exhibited a non-cytotoxic profile, the cell viability values of this combination were lesser than WMTA, and the difference was significant in 7 out of 10 concentrations at the 2 time intervals (p < 0.05). HPLFs adhered over the surface of WMTA and at the interface, after 24 hours of incubation. After 72 hours, there were increased numbers of HPLFs with prominent cytoplasmic processes. Similar findings were observed with FS WMTA, but the cells were not as confluent as with WMTA. Conclusions: The addition of $CaCl_2{\cdot}2H_2O$ to WMTA affected its chemical properties. The favorable biological profile of FS WMTA towards HPLFs may have a potential impact on its clinical application for repair of perforation defects.

Removal and inactivation of bovine herpes virus and murine encephalomycarditis virus by a chromatography, pasteurization, and lyophilization during the manufacture of urokinase from human urine

  • Choe, Yong-Un;Lee, Seong-Rae;Park, Dae-Han;Lee, Gyeong-Myeong;Gu, Bon-Mok;Kim, In-Seop;U, Han-Sang;Lee, Seong-Min
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.615-618
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    • 2000
  • The purpose of present study was to examine the efficacy of PAB (para-amino benzamidine) affinity column chromatography, pasteurization ($60^{\circ}C$ heat treatment for 10 h), and lyophilization steps, employed in the manufacture of urokinase from human urine, in the removal and/or inactivation of urine-born viruses. Bovine herpes virus (BHV) and Murine encephalomyocarditis virus (EMCV) were selected for this study. Samples from the relevant stages of the production process were spiked with the viruses and the amount of virus in each fraction was quantified by 50% tissue culture infectious dose ($TCID_{50}$). BHV and EMCV were effectively partitioned from urokinase during PAB chromatography with the log reduction factors of 6.71 and 5.27, respectively. Pasteurization was a robust and effective step in inactivating BHV and EMCV, of which titers were reduced from initial titers of $8.65\;log_{10}\;TCID_{50}$ and $7.81\;log_{10}\;TCID_{50}$, respectively, to undetectable levels within 1 hour of treatment. The log reduction factors achieved during lyophilization were 2.06 for BHV and 4.54 for EMCV. These results indicate that the production process for urokinase has sufficient virus reducing capacity to achieve a high margin of virus safety.

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Induction of Apoptosis Signaling by a Glycoprotein of Capsosiphon fulvescens in AGS Cell (매생이 (Capsosiphon fulvescens) 당단백질에 의한 인간 위암세포 사멸기전)

  • Kim, Young-Min;Kim, In-Hye;Nam, Taek-Jeong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.3
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    • pp.216-224
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    • 2011
  • Capsosiphon fulvescens is well-known green sea algae that, in recent years, has been proposed as a potential anticancer drug. In this study, we found that C. fulvescens glycoprotein (Cf-GP) had pro-apoptotic effects on human gastric carcinoma cells. By SDS-PAGE, we confirmed that C. fulvescens extract contained a glycoprotein. Using H33342 staining, we found that the Cf-GP caused cell death in a does-dependent manner, while an MTS assay showed decreased cellular viability due to induction of apoptosis. To determine the effect of Cf-GP on apoptosis-related cellular events, cells were treated with Cf-GP and the expression of several apoptosis-related protein was determined by Western blotting. Our results indicate that Cf-GP activated both a caspase cascade and PARP, which is a substrate of caspase-3, caspase-8 and the Bcl-2 family proteins. In addition, we assessed caspase-3, and -8 activation and annexin V staining. Our results revealed a cell cycle arrest, itself leading to an increased percentage of sub-G1 cells. Our findings indicate that Cf-GP may be a source of bio-functional material with therapeutic effects on human gastrointestinal cancer.

Factors Affecting in Vitro Activity of LB20304, a New Flu-oroquinolone

  • Paek, Kyoung-Sook;Ahn, Mi-Jeong;Kim, Mu-Yong;Kim, In-Chull;Kwak, Jin-Hwan
    • Archives of Pharmacal Research
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    • v.19 no.2
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    • pp.143-147
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    • 1996
  • LB20304 is a novel fluoroquinolone that exhibits a potent broad spectrum antibacterial activity against both gram-positve and gram-negative bacteria. The MICs (Minimal Inhibitory Concentration) of LB20304 were determined against both gram-positve and gram-negative bacteria under various conditions including several media, pHs, and inoculum concentrations. The in vitro activity of LB20304 was not significantly affected by the changes in testing conditions such as components of media and inoculum concentrations, but it was slightly reduced by acid condition. The MICs and MBCs (Minimal Bactericidal Concentration) of LB20304 against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa were hardly affected by the presence of 50 % human serum, mouse serum, guinea pig serum or horse serum, and the MBCs were equal to or at most four-times higher than the MiCs. The activities of LB20304 were decreased by the presence of high concentraion of $Mg^{++}$ or human urine (pH, 5.5) in the test media. The frequencies of mutants resistant to LB20304 were similar to or lower than those found in ciprofloxacin and sparfloxacin.

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Molecular Size and Distribution of Zinc-binding Ligands in Rat Pancreatic Tissue

  • Kwun, In-Sook;Donald Oberleas
    • Preventive Nutrition and Food Science
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    • v.2 no.3
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    • pp.219-224
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    • 1997
  • The pancreas is an important organ in the maintenance of zine homeostasis. The pancreatic tissue used in this study was obtained from rats fed varying levels of dietary Ca nd phytate followed by intraperitoneal {TEX}${65}^Zn${/TEX} injection. THe objective of this study was to determine the molecular size and distribution of compounds that may represent zinc-binding complexes in pancreatic tissue homogenates. The supernatant of the homogenized pancreatic tissue was separated using a Sephadex G-75 column with Tris buffer at pH 8.1. All subfractions were assayed for zinc, protein and {TEX}${65}^Zn${/TEX} activity. The elution of subfractions from pancreatic tissue homogenates showed a prominent peak corresponding to the high molecular weight protein standard (>66kd). A sall molecular weigth protein (<6.5kd), that was absorbed at 280nm, was also present: prominently in low Ca group, however not much as in high Ca group. These small compounds may combine weakly with zinc in pancreatic tissue an serve as zinc-binding ligands in pancreatic/biliary fluid. In the duodenum, these ligands dissociate zinc into an ionic form which becomes vulnerable to phytate complexation.

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Functional Brain Mapping Using $H_2^{15}O$ Positron Emission Tomography ( II ): Mapping of Human Working Memory ($H_2^{15}O$ 양전자단층촬영술을 이용한 뇌기능 지도 작성(II): 작업 기억의 지도 작성)

  • Lee, Jae-Sung;Lee, Dong-Soo;Lee, Sang-Kun;Nam, Hyun-Woo;Kim, Seok-Ki;Park, Kwang-Suk;Jeong, Jae-Min;Chung, June-Key;Lee, Myung-Chul
    • The Korean Journal of Nuclear Medicine
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    • v.32 no.3
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    • pp.238-249
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    • 1998
  • Purpose: To localize and compare the neural basis of verbal and visual human working memory, we performed functional activation study using $H_2^{15}O$ PET. Materials and Methods: Repeated $H_2^{15}O$ PET scans with one control and three different activation tasks were performed on six right-handed normal volunteers. Each activation task was composed of 13 match-ing trials. On each trial, four targets, a fixation dot and a probe were presented sequentially and subject's task was to press a response button to indicate whether or not the probe was one of the previous targets. Short meaningful Korean words, simple drawings and monochromic pictures of human faces were used as matching objects for verbal or visual memory. All the images were spatially normalized and the differences between control and activation states were statistically analyzed using SPM96. Results: Statistical analysis of verbal memory activation with short words showed activation in the left Broca's area, promoter cortex, cerebellum and right cingulate gyrus. In verbal memory with simple drawings, activation was shown in the larger regions including where activated with short words and left superior temporal cortex, basal ganglia, thalamus, prefrontal cortex, anterior portion of right superior temporal gyrus and right infero-lateral frontal cortex. On the other hand, the visual memory task activated predominantly right-sided structures, especially inferior frontal cortex, supplementary motor cortex and superior parietal cortex. Conclusion: The results are consistent with the hypothesis of the laterality and dissociation of the verbal and visual working memory from the invasive electrophysiological studies and emphasize the pivotal role of frontal cortex and cingulate gyrus in working memory system.

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Larval Gnathostomes and Spargana in Chinese Edible Frogs, Hoplobatrachus rugulosus, from Myanmar: Potential Risk of Human Infection

  • Chai, Jong-Yil;Jung, Bong-Kwang;Ryu, in-Youp;Kim, Hyun-Seung;Hong, Sung-Jong;Htoon, Thi Thi;Tin, Htay Htay;Na, Byoung-Kuk;Sohn, Woon-Mok
    • Parasites, Hosts and Diseases
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    • v.58 no.4
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    • pp.467-473
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    • 2020
  • Chinese edible frogs, Hoplobatrachus rugulosus, were examined to estimate the potential risks of human gnathostomiasis and sparganosis in Myanmar. A total of 20 frogs were purchased in a local market of Yangon and examined with naked eyes and the artificial digestion method after skin peeling in June 2018 and June 2019. Larvae of gnathostomes and Spirometra (=spargana) were detected in 15 (75.0%) and 15 (75.0%) frogs with average intensities of 10.5 and 6.3 larvae per infected frog, respectively. Gnathostome larvae were 2.75-3.80 (av. 3.30) mm long and 0.29-0.36 (0.33) mm wide. They had a characteristic head bulb with 4 rows of hooklets, a muscular long esophagus, and 2 pairs of cervical sac. The mean number of hooklets were 41, 44, 47, and 50 on the 1st, 2nd, 3rd, and 4th row, respectively. Collected spargana were actively moving, particularly with the scolex part, and have ivory-white color and variable in size. Conclusively, it has been first confirmed that Chinese edible frogs, H. rugulosus, are highly infected with larval gnathostomes and spargana in this study. Consuming these frogs is considered a potential risk of human gnathostomiasis and sparganosis in Myanmar.