Critical role of bile acid (BA) in the cellular entry and permissiveness of Hepatitis B virus in vitro

Hong‑Jai Lee,Bo‑Young Shin,Jae‑Seung Moon,Ailyn Fadriquela,Selikem Abla Nuwormegbe,Chun‑Chang Ho,Jin‑Su Shin,Jee‑Sang Yoon,Sang‑Kyou Lee,Soo‑Ki Kim 대한독성 유전단백체 학회 2020 Molecular & cellular toxicology Vol.16 No.3

Background Lack of universal replication system for hepatitis B virus with narrow host range and organ tropism has hampered to uncover the pathogenesis of HBV. Previously, we reported the essentiality of humoral milieu and its components toward HBV and hepatitis C virus survival/viability in vitro. Of these components, the precise role of enterohepatic humoral milieu such as bile acid (BA) on HBV cultivation in vitro and in vivo is unknown. Objective We explored whether BA, specifically taurochenodeoxycholic acid (tCDCA) would directly regulate the viral DNA and surface antigen expression of HBV in vitro, consequently rendering HBV to enter into human or murine immortalized hepatocytes, and non-hepatocytes. Result We found that higher concentration of taurochenodeoxycholic acid (tCDCA) is able to preserve the genomic stability of HBV in cell-free DMEM, showing higher the surface antigenicity than taurocholic acid (tCA). In line, we found that in vitro cell culture condition (100 μmol/L of tCDCA coupled with 1 × 108 g e/mL HBV) would be optimal for HBV entry into target cells. Using this, human (HepG2, Huh7), and rodent (Hepa1c1c7, H4-II-E) hepatoma cell lines were infected by HBV, as evidenced by the presence of HBV biomarkers (HBsAg, and HBV DNA in culture supernatant, as well as HBcAg in cell). Further, cellular entry test revealed that HBV is able to infect 12 different non-hepatic cell lines regardless of species, and organ/tissue, consequently reproducing progeny as confirmed by HBV biomarkers. Last, reinfection test showed that the progenies of HBV from immortalized HepG2, and Hepa1c1c7 cells are able to enter into each or vice versa naïve HepG2, and Hepa1c1c7 cells with or without BA. Conclusion This study demonstrates that enterohepatic humoral milieu such as BA, specifically tCDCA would directly regulate HBV DNA and its surface antigen expression in vitro, consequently rendering HBV to enter into human or murine immortalized hepatocytes, and non-hepatocytes. This is the first note to render HBV permissive to human or rodent hepatic and non-hepatic cells via sole manipulation of humoral milieu, thus establishing the platform for in vitro robust replication system of HBV.

The HBV DNA Amounts in Serum Have No relationship with ALT level and Hetergeneous Population Coexits in Chronic Hepatitis B Virus Infection

Chun, Yoon-Keun,Ha, Joo-hun,Hong-Jung-Woo,Oh, Soo-Myung,Kim, Sung-Soo 경희대학교 동서의학연구소 1999 INTERNATIONAL SYMPOSIUM ON EAST-WEST MEDICINE Vol.1999 No.1

Yoon-Keun Chun¹,Joohun Ha□Hong-Jung Woo□, Soo Myung Oh□,Sung Soo Kim□ ¹Department of Molecular Biology, College of Medicine,²Department of Surgery, college of Medicine,³Department of Internal Medicine, College of Oriental Medicine,and ⁴East-Weat Medical Reserch Institute,Kyung Hee University, Seoul, Korea. The HBV DNA Amounts in Serum Have No relationship with ALT level and Hetergeneous Population Coexits in Chronic Hepatitis B Virus Infection. Proceedings of International Symposium on East-West Medicine, Seoul. 212-230, 1999. -Hepatitis B is caused by hepadnavirus. Hepatitis B virus replicates through 3.5kb pregenomic RNA intermediate which is regulated by core promoter. Pathogenesis of hepatitis B virus has been bilieved the result of host immune response. But recently many studies have reported that high level of viral replication caused by mutation in core promoter might result in severs hepatitis. But these studies were performed in vitro, not in vivo. So there is yet debate about which factor, viral of host factor, is more important in pathogenesis of hepatitis B virus. So we measured real viral replication level in 204 chronic hepatitis B patients by quantifying HBV DNA from sera by our novel PCR-based more sensitive method, and compared these results with ALT level measured from same sera, which indicates liver cell damage. Surprisingly there are no significant correlation between HBV DNA quantity and ALT level. Then we cloned core promoter region. In SSCP, we found that many viral mutants coexist in one patient. Base on SSCP result, we chose main viral core promoter type in each patients, which is thought to determine overall viral replication level in this patient. Main type of core promoter region of each 41 patients were directly sequenced. And with these we measured promoter activity by luciferase assay system and compared promoter activity with on another. We found tha there were some differences in promoter activity according to core promoter sequences. And we constructed replication-competent viral constructs with core promoter from 41 patients and Transfected these into HepG2 cell and measured HBV DNA by southern blot. There were also differences in HBV DNA quantity according to core promoter sequences. On these all results we investigated correlation between the effect of HBV core promoter on viral replication in vitro and HBN DNA quantity, ALT level from sera of each patients. We found there is no significant correlation among them. As a result, we concluded that in determining severity chronic hepatitis B patients, host factors of each patient is more important rather than replicative activity of virus itself.

서울의 Penicillinase Producing Neisseria gonorrhoeae 발생빈도(1998)

김재홍,김준호,반재용,이정우,황성주,정준규,정성태,강진문,조흔정,홍창의,정혜신,이한승,김이선,이봉길,이종호,선영우,한기덕,윤성필,이성훈,안종성,박석범,문승현,조항래,김형섭,류지호,황재영,박준홍,손상욱 한양대학교 의과대학 2001 한양의대 학술지 Vol.21 No.1

In recent years, gonorrhea has been pandemic and remains one of the most common STDs in the world, especially in developing countries. For the detection of a more effective therapeutic regimen and assessing the prevalence of Penicillinase Producing Neisseria gonorrhoeae(PPNG), we have been trying to study the patients who have visited the Venereal Disease Clinic of Choong-Ku Public Health Center in Seoul since 1980 by menas of the chromogenic cephalosporin method. In 1998, 93 strians of N. genorrhoeae were isolated, among which 60(64.5%) were PPNG. The prevalence of PPNG in Seoul, which had been decreased to 39% in 1996 after a peak of 74.3% in 1993, is increased to 64.5% in 1998.

서울의 Penicillinase Producing Neisseria Gonorrhoeae 발생빈도(1996)

김재홍,황동규,전재홍,김윤석,김중환,김용준,이창균,임동진,김현수,조창근,김경문,박상훈,전우형,김희성,이호정,차명수,김갑형,김형석,김석우,황지환,박병순,권오상,이민수,송기훈,성소영,이인섭,부태성 대한화학요법학회 1999 대한화학요법학회지 Vol.17 No.2

Background : In recent years, gonorrhea has been panedemic and remains one of the most commom STDs in the world, especially in developing countries. Objective & Methods: For the detection of a more effective therapeutic regimen and assessing the prevalence of PPNG, we have been trying to study the patients who have visited the VD Clinic of Choong-Ku Public Health Center in Seoul since 1980 by means of the chromogenic cephalosporin method. Results: In 1996, 139 strains of N. gonorrhoeae were isolated, among which 53(39.0%) were PPNG. Conclusion: Our results suggests that after a peak of 74.3% in 1993, the prevalence of PPNG in Seoul is gradually declining.

정신분열병에 대한 리스페리돈의 효과 및 안정성

이민수,김용구,김영훈,연병길,오병훈,윤도준,윤진상,이철,정희연,강병조,김광수,김동언,김명정,김상훈,김희철,나철,노승호,민경준,박기창,박두병,백기청,백인호,손봉기,손진욱,양병환,양창국,우행원,이정호,이종범,이홍식,임기영,전태연,정영조,정영철,정인과,정인원,지익성,채정호,한상익,한선호,한진희,서광윤 大韓神經精神醫學會 1998 신경정신의학 Vol.37 No.1

연구목적 : 본 시험의 목적은 임상시험 시작전에 연구자들을 대상으로 PANSS Workshop을 통하여 PANSS, ESRS에 대한 국내에서의 표준화 작업을 구축하고 새로운 정신병 치료제인 리스페리돈의 효과와 안정성을 재확인하여 리스페리돈 사용에 대한 적정화를 이루는데 있다. 연구방법 : 1996년 4월부터 1996년 9월까지 국내 39개 대학병원 정신과에 입원중인 혹은 증상이 악화되어 입원하는 정신분열병 환자 377명을 대상으로 다시설 개방 연구를 시행하였다. 1주일간의 약물 배설기간을 가진후, 리스페리돈을 8주간 투여하였고, 기준점, 1주, 2주, 4주, 그리고 8주후에 평가되었다. 용량은 제1일에는 리스페리돈 1mg씩 1일 2회, 제2일에는 2mg씩 1일 2회, 제3∼7일에는 3mg씩 1일 2회 투여하였다. 이후 환자의 임상상태에 따라 임의로 증량할 수 있으며, 최대 일일 16mg을 초과하지 않도록 하였다. 추체외로 증상을 조절하기 위한 투약을 허용하였다. 임상증상 및 부작용의 평가는 PANSS(Positive and Negative Syndrome Scale), CGI(Clinical Global Impression) 그리고 ESRS(Extrapyramidal Symptom Rating Scale)을 사용하였다. 연구결과 : 377명중 343명(91%)이 8주간의 연구를 완결하였다. 치료 종결시점인 8주후 PANSS 총점수가 20% 이상 호전된 경우를 약물 반응군으로 정의할때, 약물반응군은 81.3%였다. 리스페리돈에 반응하는 예측인자로는 발병연령, 이전의 입원 횟수, 유병기간이 관련 있었다. 리스페리돈은 1주후부터 PANSS양성, 음성, 및 일반정신병리 점수상에 유의한 호전을 보여 효과가 빨랐다. CGI의 경우도 기준점에 비해 1주후부터 유의한 감소를 나타내었다. ESRS의 경우, 파킨슨 평가점수는 기준점과 비교해 투여 1주, 2주, 4주후 유의하게 증가되었다가 8주후 기준점과 차이가 없었다. Dystonia 평가점수는 1주후만 유의한 증가를 보였으며, dyskinesia 평가점수는 유의한 차이가 없었다. 혈압, 맥박수의 생명징후 및 일반 혈액학 검사, 생화학적 검사, 심전도 검사에서 유의한 변화는 없었다. 결 론 : 이상의 다시설 개방 임상 연구를 통해 리스페리돈은 정신분열병 환자에서 양성증상뿐만 아니라 음성증상 및 전반적인 증상에도 효과적인 것으로 사료된다. 보다 명확한 평가를 위해서는 다른 항정신병약물과의 이중맹검 연구가 필요할 것으로 생각되며, 또한 장기적 치료에 대한 평가도 함께 이루어져야 하겠다. Objective : The purpose of this study was to investigate the efficacy and safety of risperidone in the treatment of Korean schizophrenic patients. Method : This multicenter open study included 377 schizophrenic patients drawn from 39 university hospitals. After a wash-out period of 1 week, the schizophrenic patients were treated with risperidone for 8 weeks and evaluated at 5 points ; at baseline, and 1, 2, 4 and 8 weeks of treatment. The dose was increased from 2mg/day(1mg twice daily) to 6mg/day(3mg twice daily) during the first week and adjusted to a maximum of 16mg/day over the next 7 weeks according to the patient's clinical response. Medication to control extrapyramidal symptoms was permitted. The psychiatric and neurological status of the patients was assessed by PANSS, CGI, and ESRS scales. Results : 343(91%) of 377 patients completed the 8-week trial period. Clinical improvement, as defined by a 20% or more reduction in total PANSS score at end point, was shown by 81.3% of patients. The predictors of response to risperidone were associated older age, shorter duration of illness, fewer previous hospitalization. Risperidone had rapid onset of action ; a significant decrease of the total PANSS and three PANSS factor(positive, negative, general), and CGI was already noticed at the end of first week. For the ESRS, parkinsonism rating scores were significantly increased until week 4 comparing with baseline. Dystonia rating scores were significantly increased until week 1, and dyskinesia rating scores were not significantly changed during the study. Laboratory parameters including vital sign, EKG, hematological, and biochemical values showed no significant changes during the trial. Conclusions : This study suggests that risperidone is generally safe and effective against both the positive and negative symptoms in our group of patients.

젖소 말초혈액 림프구로부터 소백혈병 바이러스 배양 및 전자현미경적 관찰

윤순식,박중원,변재원,강문일,유한상,한홍율 韓國電子顯微鏡學會 2005 Applied microscopy Vol.35 No.1

국내 젖소의 54.2%가 BLV에 감염되어 있지만 현재까지 국내에서는 소백혈병 바이러스 (BLV) 입자를 확인한 연구 보고가 없기 때문에 BLV 항체 양성 소의 말초혈액림프구를 배양, BLV를 발현시켜 전자현미경으로 바이러스 입자를 검출하였고 배양조건에 따른 바이러스 발현율 및 발현 시간을 비교하였다. 검사 결과 전형적인 C-형 바이러스를 확인할 수 있었으며 BLV 단크론 항체를 이용한 면역염색결과 BLV 항원 양성으로 확인되었다. BLV 는 대부분 세포 외부에 분포하고 있었으며 세포질 막에서 생성, 발아되어 나오는 것도 관찰되었다. 전체 바이러스의 크기는 90~100 nm였으며 nucleocapsid는 40~60nm였다. 소태아혈청 (FBS)과 T- 및 B-림프구 분열촉진물질(mitogen)을 각각 첨가하여 배양한 결과 두 군 모두에서 BLV 발현이 확인되었다. Lipopolysaccharide 첨가군은 배양 12시간, Conconavalin A 첨가군은 배양 24시간에 각각 림프구의 10%에서 바이러스가 관찰되었다. 또한 FBS만 첨가한 군과 FBS와 mitogen을 모두 첨가하지 않은 군에서도 관찰되었으나 바이러스의 수는 적었다. 본 연구에서 확립된 BLV 배양 기법을 활용하면 BLV에 감염된 소 중 바이러스를 발현하는 소, 즉 전파능이 있는 개체를 찾아내어 우선적으로 도태할 수 있기 때문에 BLV 감염으로 인한 피해를 막는데 효과적으로 이용될 수 있을 것으로 사료된다. Many studies have been performed on the bovine leukemia virus (BLV) since bovine leukosis had been reported in 1968 in Korea. However, there was no report on the ultrastructural examination of BLV. An attempt to detect C-type viral particles in the cultured peripheral blood lymphocytes of Holstein-Friesian dairy cattle, was made to determine whether in vitro viral expression might be used as a reliable method to identify the cow which is likely to transmit BLV. In transmission electron microscopic (TEM) examination, the virus particles were found predominantly outside of the lymphocytes even though a few particles were also observed within the membrane bound cytoplasmic vacuoles. All of them were C-type particles consisting of a central, electron-dense core separated by a clear area from a limiting envelope with a unit membrane structure. Virus particles were easily detected in the lymphocyte which was cultured with medium supplemented with either T-lymphocyte mitogen (conconavalin A) or B-lymphocyte mitogen (lipopolysaccharide). Identical viral particles, although fewer, were also consistently present in the lymphocytes cultured with medium which was containing foetal bovine serum (FBS) only and which was containing neither FBS or mitogen. By contrast, no virus particle was detected in extensive examination of lymphocytes before culture. In conclusion, the BLV cultivation and detection methods established in this study could be used as a tool to identify and eliminate the cattle which can transmit the BLV.

親環境農業을 위한 有機農業의 作付體系 確立에 관한 硏究

홍성찬,강희경,윤영상 공주대학교 자원과학연구소 2000 資源科學硏究論文集 Vol.8 No.-

This study was carried out to produce safe agricultural product and to improve the soil environment of crop cultivation using the reasonable cropping system in organic farming to the case study of organic farmhouse in Chungnam Hongdong area for sustainable agriculture. In the ratio of farming forms in these organic farmhouses, paddy field-upland field-greenhouse form, paddy field-upland field form, crop-livestock form and only greenhouse form were 23.8%, 23.8%, 23.8% and 19.0% respectively. In the greenhouse area of organic farmhouse, the 50% of total greenhouse number were 600~1,000 pyung(200~330a) and the 40% of total were 1,000~1,500 pyung(330~500a). In the area of upland field, the 80% of total upland field number were 500~1,000 pyung(167~330a). In the cropping systems of surveyed farmhouses, the number of crop-combination patterns were twenty-seven and the 96.3% of those was combined with vegetable. Vegetable-vegetable pattern, cereal crops-vegetable pattern and root and tuber crop-vegetable pattern were 44.4%, 18.5% and 14.8% respectively. Leguminous crop combined pattern was 7.4%, but patterns combined with barley or wheat did not exist. Among the vegetable combined patterns, leaf vegetables combined pattern, fruit vegetables combined pattern and root vegetables combined pattern were 66.7%, 31.0% and 22.2% respectively. Among the vegetable-vegetable patterns, the portion of fruit vegetables-leaf vegetables pattern was the highest and that of root and tuber vegetables-root and tuber vegetables pattern was the lowest. Among the twenty-seven crop-combination patterns, unreasonable crop rotation pattern was 50% over and especially the almost crop-combination pattern of greenhouse was unreasonable to crop rotation pattern. The farmhouse that control the amount of compost application by means of periodical soil analysis did not exist in this area.

실리카흄을 흔입한 고강도 콘크리트 개발

홍창우,김태경,김경진,김성환,김남윤,심도식,이정호,윤청호,백민경,원치문,박제선,이주형,정경일 강원대학교 석재복합신소재제품연구센터 1996 석재연 논문집 Vol.1 No.-

콘크리트 구조물이 대형화됨에 따라 설계단면이 증대되어 상대적으로 많은 경제적 손실을 부담하게 된다. 따라서 단면을 감소시키면서도 소요의 하중에 안전한 구조물을 건설하기 위해서는 우선적으로 구조물 건설에 기초가 되는 고품질, 고내구성의 고강도 콘크리트 개발이 절실히 요구된다 본 연구에서는 일정 시멘트비 및 혼화재 비율하에서 소요의 워커빌리티가 확보되는 고강도 콘크리트를 시간과 온도의 변화에 따라 증기 양생하여, 압축, 인장, 휨강도 뿐만 아니라 파괴특성을 실험적으로 연구하였다. 시료 제작시 시멘트 입자 사이의 공극 및 불연속 영역을 충전하여 고밀도화하기 위한 콘크리트용 혼화재로 시멘트 비표면적이 상당히 작은 초미립 분말인 실리카흄을 이용하였다. 또한 AE감수제 및 고성능 유동화제를 사용하여 혼화재의 첨가에 의하여 발생될 수 있는 워커빌리티의 감소를 방지하였다. 실험결과에 의하면 일정 양생 지속 시간하에서 온도의 증가에 따라 콘크리트의 압축, 인장 및 휨강도가 전반적으로 증가하였다. 동일하게 일정 온도하에서 양생 지속시간이 커짐에 따라 강도들이 증가됨을 알 수 있었다. 따라서 상대적으로 높은 온도와 긴 지속 양생 조건하에서 파괴에 대한 저항력이 크게 나타났다. The compressible, tensible, and flexibleresistance of the high strength concrete is analyzed by the experimentation in the present study. For the test, we cure several samples with the silica fume as a mixture being become dense the spaces between the particle of cement under the variation of both the temperature and the curing-interval. Then, the superplasticizer and the ezcon are also used to satisfy the required workability for construction. The compressible, tensible, and flexible resistances to a stress are increased as increasing the temperature and the time interval for the curing. Therefore it is concluded that the overal fractural and mechanical properties is improved by mixing the silica fume into the cement.

타이어 설계인자를 고려한 자동차의 조종 및 안정 특성에 관한 연구

윤성운,홍석주,최규남 朝鮮大學校 機械技術硏究所 2004 機械技術硏究 Vol.7 No.2

As the technique of automobile industry is being advanced, the advancement of performance for vehicle is being required. In order to achieve this purpose, the study on the handling and stability which are produced by moving vehicle is carried out actively. Ability to predict vehicle performance using computer simulation is essential to reducing the development time for tire companies as well as car makers. From the previous research. we showed that handling simulation tool developed for J-turn and lane change maneuvers on ADAMS could be used in predicting the performance difference among tires with different specifications in vehicle mild handling behaviors. Through this study, some meaningful correlations were achieved. Also barriers to direct application of the simulation technique to tire development process and several technical problems that should be resolved were iden tified. In this results, ability to predict vehicle performance using computer simulation is essential to reducing the development time for tire companies as well as car makers.

응급의료 정보전달매체로서의 구급활동일지의 유용성

윤영철,안성훈,이경호,전형대,김원율,조영식,성백문,김홍용,김경환,구본관 대한응급의학회 1998 대한응급의학회지 Vol.9 No.2

Regulation for new standardized rescue record form of the 119 EMS system was established by law from February 1, 1988. We have recently evaluated the effectiveness of the record form. Records of 192 patients who visited the Emergency department of Sangggye Paik Hospital, Inje University by the 119 EMS system from February 1 to February 28, 1998 were analyzed statistically according to the contents of the record form. In conclusion, the new standardized rescue record form of the 119 EMS system was effective in giving information of patients' history and analysing of the 119 EMS system. But, Detailed modification of the record form about disease entities is needed for proper initial evaluation of emergent patient.