Chinese hamster 난소세포에서 수용성 니켈에 의해 특징적으로 발현되는 유전인자들의 빠른 분석에 관한 연구

성근제,이상한,우기민,송호연,조만희 순천향의학연구소 2001 Journal of Soonchunhyang Medical Science Vol.7 No.2

니켈에 의한 암 발생기전 및 세포 독성기전을 이해하고 니켈에 의해 발현에 변화를 보이는 유전자들을 검출하기 위하여 니켈이 포함된 배지에서 배양된 Chinese hamster난소 세포부터 mRNA differential display 분석을 시행하였다. 본 연구에서는 동위원소나 DNA염기서열 분석용 젤을 사용하는 기존의 방법 대신에 훨씬 빠르고, 다루기 쉽고 안전성을 지닌 변형된 기법을 개발하여 적용하였다. 니켈에 의해 발현에 변화를 보이는 cDNA fragments들을 10% polyacrylamide mini젤상에서 분리 및 클로닝한 다음, DNA염기 서열 분석과 GenBank Blast search 프로그램을 이용하여 상동성 조사를 하였다. Reverse Northern blot을 이용하여 발현에 변화가 있는 cDNA들을 재확인한 결과, 검사된 19 cDNAs 들중 총 9종에서 발현의 변화를 보였으며, 이 중 3종은 니켈에 반응하여 발현에 증가를 보였고, 반면에 6종은 발현이 감소하였다.

정신과 입원환자의 서비스 만족척도의 개발

김철권,이지연,송영선,김규호,김경률,김제원,이동기,최병무 大韓神經精神醫學會 2006 신경정신의학 Vol.45 No.3

목적 본 연구의 목적은 국내 정신의료기관(정신병원, 종합병원 정신과)에서 간편하게 사용할 수 있는 신뢰도와 타당도가 검증된 입원환자의 서비스 만족척도를 개발하기 위한 것이다. 방법 여러 단계를 거쳐 국내 실정에 맞는 문항을 개발하였고,정신병원, 종합병원, 대학병원 정신과 병동에서 퇴원하는 348명의 환자들을 대상으로 신뢰도와 타당도를 조사하였다. 요인분석 결과 5개의 요인이 추출되었으며 전체 변량의 63.04%를 설명하였다. 내적 일관성에 대한 신뢰도 계수 (Crohnbach's α)는 0.95로 상당히 높은 수준이었고, 수정된 개별문항-총점 상관계수는 0.50부터 0.72까지의 범위에 속하였다. 요인분석을 통해 추출된 서비스 만족척도의 각 하위척도 점수와 서비스 만족척도에 첨부된 각 영역에 대해 전반적인 만족도를 묻는 세 개의 문항 점수 간의 상관성 역시 유의하게 높았다. 결과 진단명에 따른 서비스 만족척도의 전체 점수에서는 불안/신체형/강박장애 환자군과 기분장애 환자군이 각각 정신분열병 및 기타 정신병 환자군과 알코올중독 장애 환자군에 비하여 유의하게 높은 점수를 보였으며, 입원형태에 있어서는 자발적 입원군이 강제 입원군에 비하여 전체 만족점수가 유의하게 높았다. 그러나 성별, 월수입, 학력, 직업, 종교, 결혼상태 등의 변수에서는 서비스 만족척도의 전체 점수에서 유의한 차이가 발견되지 않았으며, 마찬가지로 연령, 입원일수, 입원횟수, 첫 발병나이, 유병기간 등의 임상적 변수에서도 서비스 만족척도의 전체 점수와 유의한 관계를 보이지 않았다. 서비스 만족척도의 전체점수에서 대학병원 환자군이 정신병원 환자군과 종합병원 환자군에 비하여 각각 유의하게 높았다. 결론 결론적으로 국내 정신의료기관에 입원한 환자들의 서비스 만족을 평가하기 위한 목적으로 개발된 본 척도는 높은 수준의 신뢰도와 타당도를 보였으며, 또 국내 정신의료 환경에 맞는 요인구조를 보였다. 따라서 정신의료 서비스에 대한 정신과 환자의 만족도에 대한 연구가 거의 없는 실정에서 본 척도의 개발은 향후 국내 정신의료의 질과 치료결과를 높이는 도구로 활용될 수 있을 것으로 기대된다. Objectives : To develop and test the validity and reliability of a brief self-completed questionnaire (Service Satisfaction Scale : SSS) for routinely assessing the quality of service in psychiatric ward inpatients. Methods : A 30-item multidimensional questionnaire was developed by several steps of face validity and content validity. The questionnaire was administered to inpatients (n=348) discharged from psychiatric hospitals, general hospitals, and University hospitals. Construct validity was supported by performing principal component analysis. Reliability was estimated by calculating internal consistency of Cronbach's alpha. Results : Factor analysis yielded five factors comprising staff attitude, treatment quality, ward environment, access/cost, and ward rule, which account for 63.04% of the common variance. The internal consistency of the scale was high (Cronbach's alpha=0.95). The concurrent validity was supported by the significant correlation of each of five factors with item that measured overall satisfaction of SSS. Patients with neurosis (anxiety disorder, somatoform disorder, obsessive compulsive disorder) and mood disorders were significantly satisfied than those with psychosis and alcoholic disorders. Patients who admitted voluntarily were more significantly satisfied than those who admitted involuntarily. Sociodemographic variables such as age, gender, marital status, monthly income, education level, employment status and religion were not significantly different at the total scores of SSS. Similarly, clinical characteristics such as age of onset, duration of illness, lengths of hospital stay and number of previous hospitalization did not associate significantly with the total scores of SSS. Patients discharged from university hospitals were significantly more satisfied than those of the general and psychiatric hospitals. Conclusion : SSS performed well in the validity and reliability, indicating that it can be a useful tool for measuring Satisfaction of psychiatric inpatients in Korea.

Research Articles : Physiological characteristics of the Large Copper butterfly, Lycaena dispar (Lepidoptera: Lycaenidae)

( Seong Hyun Kim ),( Seong Jin Hong ),( Young Bo Lee ),( Hae Chul Park ),( Yeon Ho Je1 ),( Nam Jung Kim ) 한국잠사학회 2011 International Journal of Industrial Entomology Vol.23 No.2

To establish an indoor-rearing system for the Large Copper butterfly, Lycaena dispar and the Small Copper butterfly, Lycaena phlaeas, the effect of temperature, photoperiod and host plants on larval development was investigated. The larvae of Lycaena dispar fed on Rumex crispus, whereas the larvae of Lycaena phlaeas ate both Rumex crispus and Rumex acetosa. The duration of the larval period of Lycaena dispar was 13.8 days on R. crispus and that, of Lycaena phlaeas was 15.9 days and 15.2 days on R. acetosa and R. crispus respectively. Laboratory experiments show that the Large Copper larvae are able to feed on other Rumex species without harming their overall survival and can utilize these alternative host plants at least as efficiently as their natural host plant. This result suggests that plant chemistry is not responsible for their lack of utilization of the alternative host plants in the wild. Host plant choice by ovipositing females was measured with the two alternative hosts. Lycaena dispar preferred R. crispus to R. acetosa, wheareas Lycaena phlaeas preferred R. acetosa to R. crispus. Temperature has been proposed as an important determinant of developmental rate, lifespan and mortality in invertebrates. As temperature increased, length of the developmental period gradually decreased. The developmental period of the Large Copper larvae was 11.0 days and 28.5 days at 30oC and 17.5oC, respectively.

Functional analysis and molecular characterization of two acetylcholinesterases from the German cockroach, Blattella germanica

Young Ho Kim,Jae Young Choi,Yeon Ho Je,Young Ho Koh,Si Hyeock Lee 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.10

Two acetylcholinesterases (AChEs; BgAChE1 and BgAChE2) from Blattella germanica were functionally expressed using the baculovirus system. Kinetic analysis demonstrated that BgAChE2 had higher catalytic efficiency but lower substrate specificity than BgAChE1. Except paraoxon, BgAChE1 was generally less sensitive to inhibitors than BgAChE2. Western blot analysis using anti-BgAChE antibodies revealed that BgAChE1 was far more abundant in all examined tissues compared to BgAChE2, which is only present in the central nervous system. Both BgAChEs existed in dimeric form, covalently connected via a disulfide bridge under native conditions. Most fractions of BgAChE1 had a glycophosphatidylinositol (GPI) anchor, but a small fraction comprised a collagenlike tail. BgAChE2 appeared to have a collagen-GPI-fused tail. Based on the kinetic and molecular properties, tissue distribution and abundance, BgAChE1 was confirmed to play a major role in postsynaptic transmission.

Current Status and Future Prospects of Microbial Insecticide

Yeon Ho Je 한국응용곤충학회 2015 한국응용곤충학회 학술대회논문집 Vol.2015 No.10

During the last century, the overuse of synthetic chemical insecticides led to problems associated with natural environment such as harmful effects on nontarget organisms and development of resistance. Therefore, the application of insect pathogenic microorganisms including Bacillus thuringiensis (Bt), baculoviruses, and entomopathogenic fungi has been an environmentally favorable methods for insect control. However, their use has been limited by several factors such as narrow host spectrum and slow speed of insecticidal activities. A number of approaches have been taken to overcome these defects of microbial insecticides. A novel recombinant baculovirus, NeuroBactrus, was constructed to develop an improved baculovirus insecticide with additional beneficial properties, such as a higher insecticidal activity and improved recovery, compared to wild-type baculovirus. For the construction of the NeuroBactrus, the Bt cry1-5 crystal protein gene was introduced into the Autographa californica nucleopolyhedrovirus (AcMNPV) genome by fusion of polyhedrin-cry1-5-polyhedrin under the control of the polyhedrin promoter. In the opposite direction, an insect-specific neurotoxin gene, AaIT, from Androctonus australis was introduced under the control of an early promoter from Cotesia plutellae bracovirus by fusion of a partial fragment of orf603. The Polyhedrin-Cry1-5-Polyhedrin fusion protein expressed by the NeuroBactrus was not only occluded into the polyhedra, but also activated by treatment with trypsin, resulting in an approximately 65-kDa active toxin. The NeuroBactrus showed a high level of insecticidal activity against Plutella xylostella larvae and a significant reduction in the median lethal time (LT50) against Spodoptera exigua larvae compared to those of wild-type AcMNPV. Expression of the foreign proteins (Bt toxin and AaIT) was continuously reduced during the serial passage of the NeuroBactrus. These results suggested that the NeuroBactrus could be recovered to wild-type AcMNPV through serial passaging. The cry gene from Bt, encoding the Cry protein, has been recently introduced into crops to generate transgenic plants that are resistant to pest insects. Through the 3D structure prediction and accompanying mutagenesis study for the Mod-Cry1Ac, 7 and 16 amino acid residues from domain I and II, respectively, responsible for its insecticidal activity against larvae of S. exigua and Ostrinia furnacalis were identified. We used site-directed mutagenesis to improve the insecticidal activity of Mod-Cry1Ac, resulted 34 mutant cry genes. These mutant cry genes were expressed, as a polyhedrin fusion form, using a baculovirus expression system. The expressed proteins were 95 kDa and SDS-PAGE analysis of the recombinant polyhedra revealed that expressed Cry proteins was occluded into polyhedra and activated stably to 65 kDa by trypsin. When the insecticidal activities of these mutant Cry proteins against to larvae of P. xylostella, S. exigua and O. furnacalis were assayed, they showed higher or similar insecticidal activity compared to those of Cry1Ac and Cry1C. Especially, Mutant-N16 is considered to have the potential for the efficacious biological insecticide since it showed the highest insecticidal activity.

Expression of a Recombinant Bacillus thuringiensis $\delta$-Endotoxin Fused with Enhanced Green Fluorescent Protein in Escherichia coli

Je, Yeon-Ho,Roh, Jong-Yul,Li, Ming-Shun,Chang, Jin-Hee,Shim, Hee-Jin,Jin, Byung-Rae,Boo, Kyung-Saeng Korean Society of Sericultural Science 2004 International Journal of Industrial Entomology Vol.8 No.2

The expression of a fusion protein comprised of the B. thuringiensis crystal protein, Cry1Ac, and enhanced green fluorescent protein (EGFP) in Escherichia coli XLl-blue was examined. Three recombinant plasmids were transformed into E. coli XL1-blue and named as ProAc/Ec, MuEGFP/Ec and ProMu-EGFP/Ec, respectively. All transformants were observed by light and fluorescence microscopy at mid-log phase. The expression in E. coli transformants, ProMu-EGFP/Ec and MuEGFP/Ec, exhibited bright enough fluorescence to be observed. Furthermore, ProMu-EGFP/Ec produced fluorescent inclusions, which may have been recombinant crystals between EGFP and Cry1Ac while MuEGFP/Ec expressed soluble EGFP in cell. In SDS-PAGE, ProAc/Ec had 130 kDa crystal protein band and MuEGFP/Ec had thick 27 kDa EGFP band. However, ProMu-EGFP/Ec had about 150 kDa fusion protein band. Accordingly, these results indicated that a fusion protein between the B. thuringiensis crystal protein and a foreign protein under the lacZ promoter was successfully expressed as granular structure in E. coli. It is suggested that the E. coli expression system by N-terminal fusion of B. thuringiensis crystal protein may be useful as excellent means for fusion expression and characterization of B. thuringiensis fusion crystal protein.

곤충세포주에서 Autographa californica 핵다각체병 바이러스의 다각체 단백질과 Bacillus thuringiensis cryIA(c) 내독소 단백질의 융합 단백질 발현

Yeon Ho JE,Byung Rae JIN,Hyun Woo PARK,Jong Yul ROW,Jin Hee CHANG,Soo Dong WOO,Seok Kwon KANG 한국응용곤충학회 1997 한국응용곤충학회지 Vol.36 No.4

남부지방에서 분리한 곤충병원성곰팡이 Beauveria bassiana GY1-17균주를 이용하여 산림해충인 오리나무잎벌레(Agelastica coerulea), 밤나무혹나방(Meganola melancholica), 회양목명나방(Glyphodes perspectalis), 잔디해충인 등얼룩풍뎅이(Blitopertha orientalis), 채소해충인 배추좀나방(Plutella xylostella) 및 거세미나방(Agrotis segetum)의 생물적 방제 가능성을 알아보기 위하여 실험한 결과, 오리나무잎벌레와 배추좀나방유충은 7.0~2.0 conidia/ml 농도에서 처리 7일과 5일후 100%의 치사율을 나타내었다. 밤나무혹나방유충은 0.03875~3.1 conidia/ml 처리에서 66.7~100%의 높은 치사율을 보였으나 회양목명나방유충은 2.0~2.0conidia/ml 처리에서도 전혀 치사되지 않았다. 등얼룩풍뎅이유충은 3.7 conidia/ml 농도에서 46.7%의 치사율을 보였고, 거세미나방유충은 2.5 conidia/ml 농도에서 63.3% 치사율을 나타내었다. Biological control of forest insect pests, Agelastica coeruleci, Meganola n~elancholia,a nd Glyphodes perspectalis, turfgrass insect pest, Blitoperfhu orientr~lis,v egetable insect pests, Plutella xylostella and Agrotis segetun~ with entomopathogenic fungus, Beciuveria hassinna GY 1 - 17 isolated from rice paddy of Yangsan in the southern part of Korea were investigated. Mortalities of A. coeruleu and P. ~ylo.~rellluar vae were 100% at the rate of 7.0 and 2.0 x 107conidia/ml after 7 and 5 days and those of M. melancholia were 66.7 - 100% at the rate of 0.03875-3.1 X 107conidia/ml. However, G. perspectulis was not affected at the rate of 2.0 x lo7 to X I04conidia/ml. And mortalites of B. orientcilis and A. segetum larvae were 46.7% at 3.7 x 107conidia/ml and 63.3% at 2.5 X 107conidia/ml.

Generation of Baculovirus Expression Vector Using Detective Autographa California Nuclear Polyhedrosis Virus Genome Maintained in Escherichia coli for $Occ^{+}$ Virus Production

Je, Yeon-Ho,Chang, Jin-Hee,Roh, Jong-Yul,Jin, Byung-Rae Korean Society of Sericultural Science 2001 International Journal of Industrial Entomology Vol.2 No.2

We have generated a novel baculovirus genome which can be maintained in Escherichia coli that facilitates the rapid and efficient generation of recombinant baculovirus expression vectors. To make $Occ^{+}$ recombinant expression vectors, polyhedrin gene under the control of p10 promoter was inserted to bAcGOZA and this genome was designated bApGOZA. As in bAcGOZA, bApGOZA lacks a portion of the essential ORF1629 gene, but includes a mini-F replicon and selectable kanamycin-resistance marker, This occasion-producing activity of bApGOZA can be used very conveniently for its oral infectivity to insect larvae in mass production of foreign protein and insecticides.

Construction of a Novel Recombinant Baculovirus Producing Polyhedra with a Bacillus thuringiensis Cry1Ac Crystal Protein

Je, Yeon-Ho,Jin, Byung-Rae,Roh, Jong-Yul,Chang, Jin-Hee,Kang, Seok-Kwon The Korean Society of Virology 1999 Journal of Bacteriology and Virology Vol.29 No.3

We have now constructed a novel recombinant baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV) producing polyhedra with Bacillus thuringiensis (Bt) CryIAc crystal protein. The recombinant polyhedra produced by the recombinant baculovirus, Btrus, in insect cells was characterized. The recombinant baculovirus has two independent transcription units in opposite orientations with two promoters, p10 or polyhedrin gene promoter each initiating transcription of either native polyhedrin or fusion protein with polyhedrin and Bt Cry1Ac crystal protein. Surprisingly, this recombinant baculovirus stably produced recombinant polyhedra which were nearly similar to those of wild-type AcNPV. The immunogold staining experiment showed that the recombinant polyhedra were assembled with polyhedrin and Bt Cry1Ac crystal protein, and contained virus particles. Insecticidal toxicity of recombinant polyhedra of Btrus to the fall webworm, Hyphantria cunea, was strikingly improved in comparison with the wild-type AcNPV.

Genetic Subdivision of the Firefly, Luciola lateralis (Coleoptera: Lampyridae), in Korea Detected by Mitochondrial COI Gene Sequences

Je Yeon-Ho,Chang Jin-Hee,Kim Mi-Hyang,Roh Jong-Yul,Jin Byung-Rae,O'Reilly David R,Boo Kyung-Saeng 한국응용곤충학회 2001 한국응용곤충학회 학술대회논문집 Vol.2001 No.1