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Kö,nig, Alexander,Yang, Jaewon,Jo, Eunji,Park, Kyu Ho Paul,Kim, Hyun,Than, Thoa Thi,Song, Xiyong,Qi, Xiaoxuan,Dai, Xinghong,Park, Soonju,Shum, David,Ryu, Wang-Shick,Kim, Jung-Hee,Yoon, Seung Kew,P Elsevier 2019 Journal of hepatology Vol.71 No.2
<P><B>Background & Aims</B></P> <P>As hepatitis B virus (HBV) spreads through the infected liver it is simultaneously secreted into the blood. HBV-susceptible <I>in vitro</I> infection models do not efficiently amplify viral progeny or support cell-to-cell spread. We sought to establish a cell culture system for the amplification of infectious HBV from clinical specimens.</P> <P><B>Methods</B></P> <P>An HBV-susceptible sodium-taurocholate cotransporting polypeptide-overexpressing HepG2 cell clone (HepG2-NTCPsec+) producing high titers of infectious progeny was selected. Secreted HBV progeny were characterized by native gel electrophoresis and electron microscopy. Comparative RNA-seq transcriptomics was performed to quantify the expression of host proviral and restriction factors. Viral spread routes were evaluated using HBV entry- or replication inhibitors, visualization of viral cell-to-cell spread in reporter cells, and nearest neighbor infection determination. Amplification kinetics of HBV genotypes B-D were analyzed.</P> <P><B>Results</B></P> <P>Infected HepG2-NTCPsec+ secreted high levels of large HBV surface protein-enveloped infectious HBV progeny with typical appearance under electron microscopy. RNA-seq transcriptomics revealed that HBV does not induce significant gene expression changes in HepG2-NTCPsec+, however, transcription factors favoring HBV amplification were more strongly expressed than in less permissive HepG2-NTCPsec−. Upon inoculation with HBV-containing patient sera, rates of infected cells increased from 10% initially to 70% by viral spread to adjacent cells, and viral progeny and antigens were efficiently secreted. HepG2-NTCPsec+ supported up to 1,300-fold net amplification of HBV genomes depending on the source of virus. Viral spread and amplification were abolished by entry and replication inhibitors; viral rebound was observed after inhibitor discontinuation.</P> <P><B>Conclusions</B></P> <P>The novel HepG2-NTCPsec+ cells efficiently support the complete HBV life cycle, long-term viral spread and amplification of HBV derived from patients or cell culture, resembling relevant features of HBV-infected patients.</P> <P><B>Lay summary</B></P> <P>Currently available laboratory systems are unable to reproduce the dynamics of hepatitis B virus (HBV) spread through the infected liver and release into the blood. We developed a slowly dividing liver-derived cell line which multiplies infectious viral particles upon inoculation with patient- or cell culture-derived HBV. This new infection model can improve therapy by measuring, in advance, the sensitivity of a patient’s HBV strain to specific antiviral drugs.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Cell culture system that mimicks complete HBV life cycle from entry to egress. </LI> <LI> Efficient <I>in vitro</I> infection with crude HBV patient sera. </LI> <LI> Up to 50- and 1,300-fold net amplification of patient- and cell culture-derived input HBV in the supernatant. </LI> <LI> Polyethylene glycol-independent HBV spread to adjacent cells, forming infected cell clusters. </LI> <LI> Evaluation of patient- and cell culture-derived HBV amplification w/wo antivirals over 8 weeks. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>
유방암 환자의 말초혈액에서 역전사효소연쇄중합반응을 이용한 Human Mammaglobin 측정의 임상적 유용성
김재홍,강석윤,송정엽,최태영,임홍석,김선경,김영진,박준성,김현수,최진혁,임호영,김효철 대한조혈모세포이식학회 2001 대한조혈모세포이식학회지 Vol.6 No.2
Background: The mammaglobin gene encodes a novel protein that is secreted from the mammalian epithelium of normal breast tissue as well as malignant breast cancer tissues. In order to ascertain the prognostic value of mammaglobin gene in breast cancer patients, we measured the expression of human mammaglobin (hMAM) by RT-PCR method in various stages of breast cancer patients. Methods: Peripheral blood samples from forty healthy volunteers and 114 breast cancer patients were obtained. Peripheral blood stem cells (PBSC) collected for the purpose of autologous stem cell transplantation in five patients with metastatic breast cancer and ten patient with high risk for relapse and no evidence of disease were used for hMAM assay. Results: All samples from peripheral blood of forty healthy individuals (twenty males and twenty females) were negative for hMAM, whereas 43 of 114 samples (38%) from breast cancer patients were positive for hMAM mRNA. All the normal breast tissues were positive for hMAM mRNA. hMAM mRNA expression was detected in 11 of 42 (26%) in breast cancer patients who underwent for curative resection and had no evidence of disease, in 8 of 25 (34%) with chemo-sensitive relapsed disease, and in 16 of 32 (53%) with chemo-refractory progressive disease. Eight (53%) samples from peripheral blood of 15 breast cancer patients with metastatic disease at diagnosis were positive for hMAM. Three (20%) samples from peripheral blood stem cells of 15 breast cancer patients for high dose chemotherapy were positive for hMAM. Conclusion : In contrast to healthy volunteers, hMAM transcripts were detected in the peripheral blood of breast cancer patients. The frequency of hMAM expression in peripheral blood was correlated with the clinical stages of disease, but, was not significant. The contamination of hMAM expressing cells in the stem cell pool warrants additional effective purging method before the transplantation. The clinical relevance of hMAM RT-PCR-based tumor cell detection in the peripheral blood of breast cancer patients should be further evaluated in prospective studies.
Song, Man-K.,Li, Xiang-Z.,Oh, Young-K.,Lee, Chang-Kyu,Hyun, Y. Asian Australasian Association of Animal Productio 2011 Animal Bioscience Vol.24 No.1
Methane is known to be one of the major greenhouse gases. On a global scale, livestock farming may contribute 18% of total greenhouse gas emissions. Though methane contribution is less than 2% of all the factors leading to global warming, it plays an important role because it is 21 times more effective than carbon dioxide. Methane emission is a direct result of the fermentation process performed by ruminal microorganisms and, in particular, the archael methanogens. Reducing methane emission would benefit both ruminant production and the environment. Methane generation can be reduced by electron-sink metabolic pathways to dispose of the reducing moieties. An alternative way for methane control in the rumen is to apply inhibitors against methanogens. Generating methane from manure has considerable merit because it appears to offer at least a partial solution to two pressing problems-environmental crisis and energy shortage. An obvious benefit from methane production is the energy value of the gas itself. Control of methane emission by rumen microbes in Korea has mainly been focused on application of various chemicals, such as BES and PMDI, that inhibit the growth and activity of methanogens in the rumen. Alternatives were to apply long-chain polyunsaturated fatty acids and oils with or without organic acids (malate and fumarate). The results for trials with methane reducing agents and the situation of biogas production industries and a typical biogas plant in Korea will be introduced here.
송기욱(K.W. Song),유근봉(K.B. You),현중섭(J.S. Hyun),장성호(S.H. Jang),하정수(J.S. Ha) 대한기계학회 2001 대한기계학회 춘추학술대회 Vol.2001 No.8
Main steam pipe in power plants is subjected to service conditions under which creep processes take place limiting the component's lifetime. To ensure a safe and economic operation it is necessary to get accurate information about the lifetime situation of single components as well as of the whole system. Careful evaluation is combined with FEM analysis, NDT, microstructure evaluation. Especially, 14MoV63 steel is used as material for main steam pipe for 30 years old power plants. In service inspections have shown an increasing number of cracks and and creep cavities beside stress concentration parts. A detailed analysis came to the conclusion that lifetime has been consumed to a high degree, 80%level.