Immobilization of the Hyperthermophilic Archaeon Thermococcus onnurineus Using Amine-coated Silica Material for H₂ Production

배승섭,나정걸,이성목,강성균,이현숙,이정현,김태완,Bae, Seung Seob,Na, Jeong Geol,Lee, Sung-Mok,Kang, Sung Gyun,Lee, Hyun Sook,Lee, Jung-Hyun,Kim, Tae Wan The Korean Society for Microbiology and Biotechnol 2015 한국미생물·생명공학회지 Vol.43 No.3

초고온성 고세균 Thermococcus onnurineus NA1은 개미산, 일산화탄소, 또는 전분 등을 이용해서 수소를 생산하는 것으로 알려져 있다. 본 연구에서는 T. onnurineus NA1의 고정화 세포를 이용한 수소생산을 고찰하였다. 고정화 실험결과, T. onnurineus NA1은 표면에 아민기가 코팅된 규조토 담체에 정전기적 인력에 의해 효과적으로 고정화되었고, 1 g의 담체에 고정화 될 수 있는 최대 세포의 양은 71.7 mg-dcw로 확인되었다. 고정화 세포를 이용한 세 번의 반복회분식 배양을 통해 개미산으로부터 수소생산 특성을 고찰하였고, 그 결과 배양이 반복됨에 따라 고정화 세포 농도의 증가에 기인하여 초기수소생산속도가 2.3 에서 4.0 mmol l⁻¹ h⁻¹로 상당량 증가됨이 관찰되었다. 따라서, T. onnurineus NA1의 고정화세포 시스템은 수소생산을 위한 좋은 대안이 될 수 있을 것으로 사료된다. 본 연구는 초고온성 고세균의 고정화세포를 수소생산에 적용한 첫 번째 사례이다. Previously we reported that the hyperthermophilic archaeon, Thermococcus onnurineus NA1 is capable of producing hydrogen (H₂) from formate, CO or starch. In this study, we describe the immobilization of T. onnurineus NA1 as an alternative means of H₂ production. Amine-coated silica particles were effective in immobilizing T. onnurineus NA1 by electrostatic interaction, showing a maximum cell adsorption capacity of 71.7 mg-dried cells per g of particle. In three cycles of repeated-batch cultivation using sodium formate as the sole energy source, immobilized cells showed reproducible H₂ production with a considerable increase in the initial production rate from 2.3 to 4.0 mmol l⁻¹ h⁻¹, mainly due to the increase in the immobilized cell concentration as the batch culture was repeated. Thus, the immobilized-cell system of T. onnurineus NA1 was demonstrated to be feasible for H₂ production. This study is the first example of immobilized cells of hyperthermophilic archaea being used for the production of H₂.

임상 검체에서 분리된 nontyphoid Salmonella에서의 integron의 빈도와 특징

이정은,박수진,김성한,김미나,이남용,이복권,이상오,김양수,우준희,최상호 대한감염학회 2009 감염과 화학요법 Vol.41 No.3

Background : Nontyphoid Salmonella (NTS) is a leading cause of human food-borne enteritis, It has been known that integron, a naturally occurring gene capture and expression element, Plays an important role in the development and dissemination of multidrug-resistance. In this study, we investigated the prevalences and molecular characteristics of integrons in NTS clinical strains. Materials and Methods : Between 1995-96 and 2000-03, a total 261 NTS clinical strains comprising 39 serotypes were collected from clinical specimens. All strains were serotyped, and the MICs of ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, tetracycline, and trimethoprim were determined by agar dilution method. Integrons were detected by PCR amplification of integrase genes, and gene cassettes were determined by PCR and sequencing, Conjugation experiments were performed using E. coli J53 as a recipient. The clonal relationship was analyzed by pulsed-field gel electrophoresls (PFGE). Results : Of the 261 strains tested, class 1 integrons were present in 21 strains (8.0%). Class 2 and class 3 integrons were not found. The integron-positive rate was higher in S. Typhimurium (24.2% [8/33]) than in S. Enteritidis (2.0% [3/153]). Overall rates of antimicrobial resistance were higher in integron-positive strains. dhfr12-orfF-aadA2gene cassette was detected in 5 strains, aadA2in 4 strains, dhfr17-orfF-aadA5 in 2 strains, and addA1 in 1 strain. Ten integron-positive transconjugants were successfully selected. Among 8 integron-positive strains of S. Typhimurium, 7 had similar PFGE patterns. Conclusion: This study suggests that integrons are already playing a significant role in antimicrobial resistance in NTS. Continuous monitoring is needed to detect the emergence and spread of integron-mediated antimicrobial resistance.

Efficacy of Long-Term Tenofovir-Based Therapy in Chronic Hepatitis B Patients with Previous Nucleos(t)ide Analogues Treatment Experience

( Na Eun Lee ),( Hong Seon Son ),( Sung Hoon Choi ),( Chang Hun Lee ),( Seung Young Seo ),( Seong Hun Kim ),( Sang Wook Kim ),( Seung Ok Lee ),( Soo Teik Lee ),( Dae Ghon Kim ),( In Hee Kim ) 대한간학회 2017 춘·추계 학술대회 (KASL) Vol.2017 No.1

Aims: Tenofovir disoproxil fumarate (TDF) is considered as the preferred treatment option for chronic hepatitis B (CHB) patients with treatment failure or resistance to prior nucleos(t)ide analogues (NAs) treatment. We investigated the efficacy of long-term TDF-based therapy in CHB patients with previous NAs-experience. Methods: This study included total 251 patients who had previous history of NAs therapy and were treated with TDF mono (n=173) or TDF combined with other NA (n=78) from August 2012 to March 2017. Virologic response (VR) was defined as undetectable serum HBV DNA by PCR (<20 IU/mL). Results: Mean age of patients was 49.3 years, median duration of TDF therapy was 27.2 months, 75.7% were HBeAg-positive, and median HBV DNA was 3.7 log10IU/mL. The cumulative rates of VR were 188/244 (77.0%), 180/211 (85.3%), and 146/161 (90.7%) at 12, 24, and 36 months, respectively. Multivariate analysis showed that body mass index (OR 0.77, 95% CI 0.61-0.95, p=0.0189) and duration of TDF therapy (OR 1.09, 95% CI 1.02-1.18, p=0.0221) was significantly associated with VR. TDF monotherapy, HBeAg-positivity, platelet count, serum albumin was associated with VR in the univariate analysis, but not significant in the multivariate analysis. In relation to renal safety, patients showed renal impairment (7, 3.0%), mild hypophosphatemia (15, 7.2%), severe hypophosphatemia (1, 0.4%). Conclusions: Long-term TDF-based therapy demonstrated highly effective in viral suppression and relatively favorable renal safety in CHB patients with previous NA-experience. The body mass index and duration of TDF therapy was independent factors associated with VR.

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.

위암과 식도암이 병발된 다발성 원발성 악성종양 1예

박사영,이나영,이효진,이선영,최진혁,이순남,심강섭,성순희,한운섭 梨花女子大學校 醫科大學 醫科學硏究所 1996 EMJ (Ewha medical journal) Vol.19 No.3

Multiple primary cancer means that more that two cancers occur independently in an individual. Recently, the incidence of multiple primary cancer has increased with lengthened survival, of cancer patients, development of new diagnostic technique and increased clinical evaluation. We report a patient who had adenocarcinoma of stomach combined with squamous cell carcinoma of esophagus simultaneously.

온도 증가에 따른 Na-A형 제올라이트의 비정질화 메커니즘 연구

김현나(Hyun Na Kim),이성근(Sung Keun Lee) 한국암석학회 2013 한국암석학회 학술발표회 논문집 Vol.2013 No.5

Original Articles : Effect of aldosterone on the amplification of oncolytic vaccinia virus in human cancer Lines

( Hyun Ju Lee ),( Ja Sung Rho ),( Shao Ran Gui ),( Mi Kyung Kim ),( Yu Kyoung Lee ),( Yeon Sook Lee ),( Jeong Eun Kim ),( Eu Na Cho ),( Mong Cho ),( Tae Ho Hwang ) 대한간학회 2011 Clinical and Molecular Hepatology(대한간학회지) Vol.17 No.3

Background/Aims: JX-594 is an oncolytic virus derived from the Wyeth vaccinia strain that causes replication-dependent cytolysis and antitumor immunity. Starting with a cross-examination of clinical-trial samples from advanced hepatocellular carcinoma patients having high levels of aldosterone and virus amplification in JX-594 treatment, we investigated the association between virus amplification and aldosterone in human cancer cell lines. Methods: Cell proliferation was determined by a cell-counting-kit-based colorimetric assay, and vaccinia virus quantitation was performed by quantitative polymerase chain reaction (qPCR) and a viral plaque assay. Also, the intracellular pH was measured using a pH-sensitive dye. Results: Simultaneous treatment with JX-594 and aldosterone significantly increased viral replication in A2780, PC-3, and HepG2 cell lines, but not in U2OS cell lines. Furthermore, the aldosterone treatment time altered the JX-594 replication according to the cell line. The JX-594 replication peaked after 48 and 24 hours of treatment in PC-3 and HepG2 cells, respectively. qPCR showed that JX-594 entry across the plasma membrane was increased, however, the changes are not significant by the treatment. This was inhibited by treatment with spironolactone (an aldosterone-receptor inhibitor). JX-594 entry was significantly decreased by treatment with EIPA [5-(N-ethyl-N-isopropyl)amiloride; a Na+/H+-exchange inhibitor], but aldosterone significantly restored JX-594 entry even in the presence of EIPA. Intracellular alkalization was observed after aldosterone treatment but was acidified by EIPA treatment. Conclusions: Aldosterone stimulates JX-594 amplification via increased virus entry by affecting the H+ gradient. (Korean J Hepatol 2011;17:213-219)

Comparison of Virologic Response and Renal Safety of Long-Term Antiviral Therapy with Tenofovir and Entecavir in Naive Patient with Chronic Hepatitis B

( Hong Seon Son ),( Sung Hoon Choi ),( Na Eun Lee ),( Chang Hun Lee ),( Seung Young Seo ),( Seong Hun Kim ),( Sang Wook Kim ),( Seung Ok Lee ),( Soo Teik Lee ),( Dae Ghon Kim ),( In Hee Kim ) 대한간학회 2017 춘·추계 학술대회 (KASL) Vol.2017 No.1

Aims: Tenofovir (TDF) entecavir (ETV) are considered as the preferred treatment options for treatment-naïve chronic hepatitis B (CHB) patients. We compared the virologic response and renal safety of long-term TDF versus ETV therapy in naïve CHB patients. Methods: This retrospective study included total 432 patients who were treated with TDF (n=205) or ETV (n=227) from August 2012 to March 2017. Virologic response (VR) was defined as undetectable serum HBV DNA by PCR (<20 IU/mL). Results: Mean age of patients, sex, baseline serum levels of AST, ALT, creatinine, and HBV DNA were not significantly different between TDF and ETV groups. The cumulative rates of VR between TDF and ETV groups were 71.6% vs. 61.8% (p=0.477), 88.1% vs. 79.6% (p=0.058), and 84.8% vs. 83.7% (p=0.966) at 1, 2, and 3 years, respectively. The cumulative rates of genotypic resistance between TDF and ETV groups were 0% vs. 0%, 0% vs. 1.5% (p=0.404), and 0% vs. 2.2% (p=0.447) at 1, 2, and 3 years, respectively. Incidences of renal impairment and hypophosphatemia during treatment up to 3 years were not significantly different between two groups. Multivariate analysis showed that HBeAg-positivity (OR 0.27, 95% CI 0.12-0.58, p=0.0012) and serum HBV DNA at 1 year 2000 IU/mL (OR 0.09, 95% CI 0.03-0.25, p<0.0001) were significantly associated with VR. Conclusions: Long-term TDF and ETV treatments appear to have similar virologic response and renal safety in naïve CHB patient. However, long-term ETV therapy might to be associated with genotypic resistance in 2.2% up to 3 years, while none of patients on TDF therapy did.

Expression of the Na⁺-HCO₃⁻cotransporter and its role in pH_iregulation in guinea pig salivary glands

Li, Jingchao,Koo, Na-Youn,Cho, Ik-Hyun,Kwon, Tae-Hwan,Choi, Se-Young,Lee, Sung J.,Oh, Seog B.,Kim, Joong-Soo,Park, Kyungpyo American Physiological Society 2006 American journal of physiology, Gastrointestinal a Vol.291 No.6

<P>Patterns of salivary HCO3<SUP>−</SUP>secretion vary and depend on species and gland types. However, the identities of the transporters involved in HCO3<SUP>−</SUP>transport and the underlying mechanism of intracellular pH (pHi) regulation in salivary glands still remain unclear. In this study, we examined the expression of the Na<SUP>+</SUP>-HCO3<SUP>−</SUP>cotransporter (NBC) and its role in pHiregulation in guinea pig salivary glands, which can serve as an experimental model to study HCO3<SUP>−</SUP>transport in human salivary glands. RT-PCR, immunohistochemistry, and pHimeasurements from BCECF-AM-loaded cells were performed. The amiloride-sensitive Na<SUP>+</SUP>/H<SUP>+</SUP>exchanger (NHE) played a putative role in pHiregulation in salivary acinar cells and also appeared to be involved in regulation in salivary ducts. In addition to NHE, NBC also played a role in pHiregulation in both acini and ducts. In the parotid gland, NBC1 was functionally expressed in the basolateral membrane (BLM) of acinar cells and the luminal membrane (LM) of ducts. In the submandibular gland, NBC1 was expressed only in the BLM of ducts. NBC1 expressed in these two types of salivary glands takes up HCO3<SUP>−</SUP>and is involved in pHiregulation. Although NBC3 immunoreactivity was also detected in submandibular gland acinar cells and in the ducts of both glands, it is unlikely that NBC3 plays any role in pHiregulation. We conclude that NBC1 is functionally expressed and plays a role in pHiregulation in guinea pig salivary glands but that its localization and role are different depending on the type of salivary glands.</P>

녹동균 세포외막 단백질 백신 CFC-1-101의 안정성 및 면역원성 검토 : 임상 제 Ⅰ/Ⅱa상 시험

장인진,김익상,유경상,임동석,김형기,신상구,장우현,박완제,이나경,정상보,안동호,조양제,안보영,이윤하,김영지,남성우,김현수 대한감염학회 1998 감염 Vol.30 No.3

목적 : 제일제당에서는 녹농균의 세포외막 단밸질을 유효성분으로 하는 백신인 CFC-101을 개발하였으며, 동물시험에서 이 백신의 안전성과 유효성을 입증하였다. 본 연구에서는 이 녹농균 백신의 인체에 대한 안전성과 면역원성을 평가하는 동시에 인체 접종시의 최적 투여 용량을 결정하기 위하여 제 I/Ⅱa상 임상시험을 수행하였다. 방법 : 건강한 성인 남자를 피험자로 선별하여 각 용량군에 백신투여자 6명, 위약투여자 2명을 배정하였다. 백신 투여군은 0.25mg, 0.5mg 또는 1.0mg 용량의 녹농균 백신을 7일 간격으로 3회에 걸쳐 근육주사 하였으며, 위약 투여군에게는 세포외막 단백질을 제외한 동일한 성분을 투여하였다. 백신접종 후 국소적 또는 전신적인 반응의 발생여부를 관찰하고, 혈액시료를 체취하여 백신의 역가와 유효성을 검정하였다. 결과 : 녹농균 백신 CFC-101은 모든 접종자에서 양호한 내약성을 보였다. 또한 0.5mg 과 1.0mg 백신 투여군에서는 100%의 항체양전율을 나타내었다. 생성된 항체는 녹농균 세포외막단백질에 특이성을 보였고, 녹농균 감염에 대해 방어효능이 있었다. 결론 : 이와같은 결과로부터 이 녹농균 백신은 인체에 안전하게 투여할 수 있으며, 높은 항체 생성능으로 감염방어 효능을 보이고 0.5mg과 1.0mg이 최적용량인 것으로 판단되었다. Background : We developed a Pseudomonas aeruginosa outer membrane protein(OMP) vaccine CFC-101, and the prophylactic efficacy of which has been demonstrated in animal models. In order to evaluate the safety and immunogenicity of the P. aeruginosa vaccine, we carried out a phase I/Ⅱa clinical trial in healthy male volunteers. Methods : Groups of eight volunteers, including two placebo subjects, were vaccinated intramuscularly with three doses of 0.25, 0.5 or 1.0 mg of the vaccine at one week intervals. Sings of systemic and local reactions observed after vaccination were recorded for each vaccinee for 5 days. Physical examinations were performed on days 0, 1, 7, 8, 14, 15, 21, and 42, and clinical laboratory tests were done on days 0, 3, and 21. Blood samples for assay of serum antibody levels were obtained up to 42 days after the first vaccination. Results : The vaccine was generally well tolerated by all vaccinees, showing no significant side effects. In the three dosage groups, all vaccinees, except one receiving the 0.25 mg dose, showed significant elevation in serum IgG antibody titers against the vaccine proteins, indicating 100% seroconversion in 0.5 and 1.0 mg groups. The human antibodies induced by the vaccine were specific for P. aeruginosa OMPs, as confirmed by western blot analysis and immunoprecipitation assays. The capacity of the human antisera to enhance opsonophagocytic killing activity by polymorphonuclear leukocytes and to confer protection against P. aeruginosa infections indicates that the antibodies elicited by the vaccine have protective efficacy. Conclusion : We conclude that the P. aeruginosa OMP vaccine is safe and effective for human use and its optimal dose to be 0.5 or 1.0 mg.