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.

Mannanase를 생산하는 Bacillus속 균의 RAPD-PCR 분석

윤기홍 우송대학교 2004 우송대학교 논문집 Vol.9 No.-

가정에서 제조된 된장에서 mannanase 생산성이 높은 균으로 분리된 9종의 균주는 모두 포자를 형성하며 그람양성 간균으로 Bacillus속에 속하는 것으로 추정된다. 이들 분리균간에 유전체의 유사성을 조사하기 위해 10 nucleotides로 구성된 oligonucleotides 6 종과 12 nucleotides로 구성된 oligonucleotides 2종을 primer로 사용하여 Random Amplified Polymorphic DNA-중합연쇄반응 (RAPD-PCR)의 조건을 검토한 결과 40℃보다는 30℃와 35℃에서 annealing 반응을 수행하는 것이 효율적이었다. RAPD-PCR에 의해 증폭된 DNA를 agarsoe gel 전기영동으로 분석한 결과 primer에 따라 증폭된 DNA의 양과 그 크기의 다양성이 달랐으며, 분리균에 따라서도 특이한 증폭 DNA가 관찰되었다. 균주에 따라 증폭된 DNA를 비교한 결과 된장에서 분리된 9 종의 균주는 Bacillus licheniformis와는 달랐으며, Bacillus subtilis와 유사하였다. 특히 분리균 중 5 종류는 B. subtilis와 유사도가 매우 높았으며, 분리균간에도 서로 유사도가 높은 균이 있었지만 동일하지는 않은 것으로 확인되었다. 따라서 RAPD-PCR 방법에 의해 균주간 유사성을 비교하는 것은 다양한 미생물 자원을 확보하는데 유용한 방법이라고 여겨진다. Nine bacterial strains producing the extracellular mannanase were isolated from Korean soybean paste. The isolates are regarded as Bacillus on the basis of their morphological properties, which are gram positive and rod in shape with spore. To examine the similarity between genomes of the isolates, Random Amplified Polymorphic DNA - Polymerase Chain Reaction (RAPD-PCR) was performed with 6 kinds of oligonucleotides consisting of 10 nucleotides and 2 kinds of oligonucleotides consisting of 12 nucleotides. As a results of RAPD-PCR at different annealing temperatures such as 30℃, 35℃ and 40℃, many different DNA fragments were generated at 30℃ and 35℃ more than 40℃. Relatedness of the nine isolates from soybean was determined by comparing the amplified DNA fragments between the isolate strains, Bacillus subtilis and B. licheniformis, expecting that the nine isolates are different from B. licheniformis and five of them are very similar to B. subtilis. It was also found that all isolates are different strains. Therefore, RAPD-PCR is regarded as a useful method for preparing the bank of the diverse microorganisms from nature.

대학생들의 안전교육 및 안전의식에 관한 연구

홍기헌 ; 윤석호 김천대학교 2014 김천대학교 논문집 Vol.35 No.-

The purpose of this study is to figure out current status of safety education until today and safety consciousness of university students. For this study, the 300 university students were taken among freshmen and sophomores in gimcheon university. Furthermore it is try to realize the necessity of additional safety education in universi-ty and the unification of emergency call numbers. The results are as follows: 1. It was showed that the most students prefer unified emergency call number not divided. 2. Marine accident safety education was lowered compared with other life safety ducations such as fire, earthquake, and traffic safety educations. 3. It was showed that many university students wish additional actual safety educa- tion in university as academic subjects. 4. University students consider that Sewol passenger ship accident was caused by eficient of vocation and safety frigidity of crew and company staffs. And in order to revent this kind of accident, crew vocation should be built up, students safety duca-tion should be strengthened and government agency should watch severely the hip remodelling

Trigonopsis variabilis의 D-amino acid oxidase 정제

윤기홍 우송대학교 1999 우송대학교 논문집 Vol.4 No.-

배지에 methionine을 0.2% 이상 첨가하였을 때 Trigonopsis variabilis 배양균체내 D-amino acid oxidase (D-AAO)의 비활성도는 증가하였으나 균의 성장도는 매우 갑소되었다. 배양 후 배지의 pH가 급격히 감소하였으며 이에 따라 균의 성장이 억제되는 것으로 확인 되었는데 배양액의 pH를 유지할 경우 D-AAO의 생산성을 높일 수 있을 것으로 판단된다. T. variabilis 균체파쇄액으로 부터 DEAE-Sepharose와 Mone Q column을 사용항여 D-AAO의 비활성이 7.23 U/mg protein이 되도록 정제하였는데 정제과정 중 균체파쇄액에 미약하게 존재하는 catalase가 제거되었다. 정제된 D-AAO를 이용하여 cephalosporin C (CPC)의 탈아미노 반응을 수행한 결과 반응액 중에 catalase를 첨가하였을 때는 CPC가 ketoadipyl 7-aminocephalosporanic acid (7-ACA)로 전환되었으나 catalase를 첨가하지 않았을 때는 반응산물의 대부분이 glutaryl 7-ACA로 전환되고 미량의 ketoadipyl 7-ACA와 미지의 물질이 생겨나는 것으로 확인되었다. Specific activity of D-amino acid oxidase (D-AAO) was estimated to be increased in Trigonopsis variabilis cells grown on the broth supplemented with methionine over the concentration of 0.2%, but the cell growth was severely inhibited by the methionine, Final pH of the medium was found to decrease dramatically after the cell growth, suggesting that the dectrased pH caused to inhibit the growth of T. variabilis cell. In order to imprpve the D-AAO productivity of T. variabilis it is required to maintain the medium pH during cell growth. D-AAO was purified from cell-free extract of T. variabilis by DEAE-Sepharose and Mono Q culumn chromatography with specitic acitivity of 7.23 U/mg protein. Catalase activity was not detected in the purified enzyme. When the T. variabilis D-AAO reacted cephalosporin C (CPC) as a substrate with catalase or not, CPC was mainly converted to ketoadipyl 7-aminocephalosporanic acid (7-ACA) in the presence of catalse. In the absence of catalase, the reactants were, however, identified to be glutaryl 7-ACA with trace amount of ketoadipyl 7-ACA and unkwon compounds.

대두 및 대두발효식품의 항돌연변이성

윤기도,권동진,홍석산,김수일,정건섭 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.4

To investigate the inhibitory effect of soybean and Korean traditional fermented soybean products on the chemically induced mutagenesis, we extracted soybean, Kanjang, Doenjang, Kochujang, and Chonkukjang with water, methanol and hexane. Inhibitory effect of extracts was assayed by the SOS chromotest using Escherichia coli PQ37 as a test strain. 4-nitroquinoline-1-oxide(4NQO), N-methyl-N'-nitro-N-nitrosoguanidine(MNNG), and aflatoxin B_1(AFB_1) were used as mutagens. Methanol extracts showed relatively higher inhibitory effect than water and hexane extracts. Methanol extracts of soybea, Doenjang, Kochujang, and Chongkukjang showed inhibitory effect of 68.4, 96.3, 17.5, and 100.9%, against MNNG, and 28.6, 109.1, 41.3, and 101.8% against AFB_1., respectively. Doenjang methanol extract showed inhibitory effect of 51.0, 96.3, and 109.1% against 4NQO, MNNG, and AFB_1. Inhibitory effect of heat-treated Doenjang and Chongkukjang methanol extracts on the mutagenicity of MNNG and AFB_1 was remained over 95% of the inhibitory effect of heat-untreated extracts, demonstrating the heat stability of the potent antimutagenic activity.

고온성 Bacillus sp.의 Xylanases 특성 비교

윤기홍 우송대학교 1997 우송대학교 논문집 Vol.2 No.-

토양에서 분리된 cellulase-free xylanase 생산균인 고온성 Bacillus sp. KK-1의 배양상등액으로 부터 내열성 xylanase S를 정제하였다. Bacillus sp. KK-1의 xylanase 유전자를 확보하기 위해 총 염색체 DNA를 이용하여 xylanase 유전자를 대장균에 크로닝한 결과 xylanase 활성을 보이는 30개 대장균 형질전환주를 얻어 이들이 생산하는 xylanases의 열안정성을 조사한 결과 모두 xylanase S보다 낮았다. 또한 형질전환주에서 분리된 plasmids를 여러가지 제한효소로 절단하여 분석한 결과 동일한 부분의 DNA조각을 공유하고 있었다. 이로써 이들 형질전환주는 동일한 xylanase를 생산하고 있다고 판단되며 이를 xylanase Y라 명명하고 대장균 형질전환주 균체로 부터 xylanase Y를 정제하였다. 정제된 xylanase S와 xylanase Y의 성질을 비교 분석하였는데 이들은 전혀 다른 성질을 보였으므로 Bacillus sp. KK-1이 최소한 두 종류의 xylanases를 생산하는 것을 알 수 있다. 특히 xylanase S는 열안정성이 매우 높아 65C에서도 약 10시간 방치하여도 효소 활성이 75% 이상 유지 되었으나 xylanase Y는 50C에서 30분간 방치하여도 활성을 약 25% 정도 상실하였다. Xylanase Y가 열안정성이 xylanase S에 비해 매우 낮으므로 Bacillus sp. KK-1를 50C에서 약 16시간 동안 배양하여 얻은 배양상등액에서는 xylanase S와는 달리 xylanase Y의 활성이 거의 상실되어 정제될 수 없었던 것으로 여겨진다. A thermostable xylanase, designated xylanase S, was purified from the culture supernatant of Bacillus sp. KK-1 which was isolated from natural soil. In order to obtain Bacillus sp. KK-1 xylanase gene. xylanase gene was cloned in Escherichia coli from Bacillus sp. KK-1. From approximately 20,000 transformants. 30 E, coli colonies showing xylanase activity was obtained. It was found that almost all xylanases from the E, coil clones was less stable than xylanase S at 60℃. Issert DNAs of recombinant plasmids, isolated from E. coil clones, shared the common DNA fragment of Bacillus sp. KK-1, suggesting that E coli clones produced an identical xylanase. Xylanase, named xylanse Y, was therefore purified from cell free extract of Em coli, The physical and chemical properties fo purified xylanase Y were identified to be different from those of xylanase S, resulting that Bacillus sp. KK-1 produced at least two xylanses. Especcially ,xylanase S retained over 75% io its maximum activity at 65℃ for 10 h. But , xylanase Y lost 25% of its activity at 50℃ within 30 min. From this result. it was supposed that xylanase Y could not purified from the culture supernatant of Banillus sp. KK-1 growing for 16 h at 50℃ because of inactivation of the xylanase Y unlike xylanase S.

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.

내열성 α-Amylase 생산균의 탐색과 분리균 α-Amylases의 특성

윤기홍 우송대학교 1998 우송대학교 논문집 Vol.3 No.-

토양으로 부터 고온성 미생물을 1,286 주 분리하고 이들의 수용성 전분 분해능을 조사하여 α-amylase를 생산하는 균주를 402 주 확보하였다. 이들 중 불용성 전분이 가수분해능이 우수한 α-amylase 생산균 65 주를 각각 액체배양하여 배양상등액을 이용하여 전분분해능을 검토하여 전분분해능이 우수한 14주를 선발하였다. Denaturing-polyacrylamidegel에서 α-amylase 활성염색을 수행하여 4 주가 α-amylase 생산능이 우수한 것으로 나타났으며 이들 중 고온에서 효소활성과 열안정성이 높은 것이 4 주로 확인되었다. 특히 열안정성이 가장 뛰어난 α-amylase를 생산하는 분리균 16-7을 동정한 결과 Bacillus licheniformis로 판명되었다. 분리균 No. 16-7이 생산하는 α-amylase를 정제하여 효소 특성을 분석한 결과 N-말단의 아미노산 잔기배열과 분자량이 B. licheniformis의 효소와 동일하였으나 열안정성과 최적반응온도는 더 높았다. Among the thermophilic bacteria of 1,286 isolated from the natural soil. 402 a-amylase-producing strains were selected based on their liquefying activity of the soluble starch. The 65 isolates were further screened to produce the a-amylase hydrolyzing efficiently the insoluble starch from 402 isolates. They were grown in the complex broth, and then the culture supernatants were inverstigated for degradastion of the starch, resulting that the 14 isolates showed high activities for starch hydrolysis. Activity staining of the crude enzymes from the 14 isolates was done on the denaturing-polyacrylamide gel. From the result. the 4 isolates were found to produce the thermostable a-amylases with high activities. Especially, isolate 16-7 produced the a-amylase showing the highest thermostability. It was identified as Bacillus Iicheniformis. The a-amylase was purified and analyzed to have the N-terrminal amino acid sequences identical to that of B. Iicheniformis a-amylase. But, the 16-7 a-amylase was more thermostable than B. Iichniformis a-amylase.

Protease 생산성이 향상된 Bacillus natto 돌연변이주의 개발

윤기홍 우송대학교 2000 우송대학교 논문집 Vol.5 No.-

청국장 발효균인 Bacillus natto의 protease 생산성이 향상된 변이주를 개발하기 위해 N-methyl-N'-nitro-N-nitrosoguanidine (NTG)를 사용하여 돌연변이를 유발하였다. 돌연변이는 B. natto의 사멸률이 90%가 되도록 중기대수기의 B. natto 배양액에 NTG를 100㎍/㎖ 첨가하여 10 분간 수행하였다. Protease 생산성이 우수한 변이주는 2% skim milk와 2% tween 20을 첨가한 LB 평판배지에서 형성된 콜로니 주변의 skim milk 분해환의 크기와 선명도를 관찰함으로써 선발하였다. 이로써 변이주 M8을 최종적으로 선발하였는데 M8은 모균주에 비해 액체배지에서 최대 성장정도가 약간 저하되었지만, protease 생산성은 8.3 배 증가한 것으로 나타났으며 최대 효소생산성은 46.4 U/㎖ 이었다. 또한 protease는 중기대수기를 지나면서 다량 생성되기 시작하였으며 정지기에 도달하여 그 생산성이 최대에 이르는 것으로 나타났다. 본 실험을 통해 개발된 변이주 M8은 protease 생산성이 향상되었으므로 콩 발효시 단백질의 분해력이 우수할 것으로 예상되며 청국장 발효균으로서의 가치가 높다고 판단된다. Bacillus natto cells, natto-fermenting food microorganism, were mutagenized by N-methly-N'-nitro-N-nitrosoguanidine (NTG) for increasing its protease productivity. Mutagenesis was performed by treating the B. natto cells with 100㎍/㎖ of NTG for 10 min, which caused approx. ninety percent of the cells to die. Mutant strains were isolated by scoring the size and transparency of halo formed around the colonies grown on LB a gar plates containing skim milk (2%) and tweeny 20 (2%). A mutant strain of protease hyper-producer, designated as M8, was finally obtained, and its growth and protease production were investigated. In flask cultures, growth pattern of M8 was similar to that of parent strain, though maximum growth level of M8 decreased little less. Nevertheless, the maximum protease production of mutant M8 was 46.4 U/㎖ corresponding to 8.3-fold increase over the parent strain. It was additionally found that the protease production of mutant M8 was done after mid-logarismic growth phase and reached maximum level at stationary phase. Increasement of its protease productivity is useful for the efficient hydrolysis of soybean protein, expecting that mutant M8 is a promising strain for natto.

국내 토착 담수어류의 장내미생물 분리와 특성

윤기홍 우송대학교 2003 우송대학교 논문집 Vol.8 No.-

자연에서 포획된 담수어 5종과 해삼 1종 및 양식어 4종으로부터 장내 미생물을 분리한 결과 양식어 보다는 자연에서 포획된 수계 생물에서 분리균의 다양성이 높은 것으로 나타났다. 분리균 총 52종의 형태적·생리적 특성을 조사한 결과 담수어에서 분리된 균은 그람 양성균과 음성균이 고르게 분포되어 있으나, 해수 생물에서 분리된 균은 모두 그람 양성균으로 확인되었다. 또한 분리균의 가수분해 효소 생산을 조사한 결과 담수어에서 유래된 미생물은 amylase, cellulase, xylanase, protease, esterase, gelatinase의 활성을 보이는 것이 다수 존재한 반면에 해수 생물에서 분리된 미생물은 대부분 이들 가수분해 효소를 생성하지 못하였다. 분리균 중 다양한 가수분해 효소를 생산하는 8-6번 균주를 배양한 결과 매우 빠른 성장을 보였으며, 말기 대수기부터 효소 생산이 이루어지는 것으로 확인되었다. Intestinal microorganims were isolated from ten kinds of aquatics. It was found that diversity of the bacterial isolates was higher in aquatics catched from nature than aquaculture. As a result of gram staining, gram-negative and gram-positive bacteria were distributed in the bacterial isolates from freshwater fishes, while almost all of bacterial isolates from marine aquatics was gram-positive. Many bacterial isolates from freshwater fishes showed hydrolytic activities on macromolecules such as starch, carboxymethyl cellulose, skim milk, tributyrin, gelatin and oat spelt xylan, indicating that they produced α-amylase, cellulase, protease, esterase, gelatinase and xylanase. However, few bacterial isolates from marine aquatics showed hydrolytic activity on the macromolecules except tributyrin. It was identified that a bacterial isolate 8-6 producing various hydrolytic enzymes grew fast and produced the amylase, cellulase, and xylanase at the late-logarithmic growth stage.