Effect of ER signal peptides (KDEL) on expression and function of monoclonal antibody in plant

Da-Young Park,Eun-Yi Oh,Zhe Lu,Yangkang So,Mi-Hyun Ahn,Mira Song,Kyung-Jin Lee,Doo-Byoung Oh,Youngkwan Kim,Kisung Ko 한국당과학회 2009 한국당과학회 학술대회 Vol.2009 No.1

Plant genetic engineering led to the production of plant-derived mAb (mAbP), which provides a safe and economically feasible alternative to the current animal expression systems. In this study, the expression level of mAbP SO57 between with and without Lys-Asp-Glu-Leu (KDEL) was compared in transgenic plant. PCR and PT-PCR analyse showed stable gene insertion transcription of heavy and light chain genes of mAbP SO57 with or without KDEL in plant, respectively. We validated expression of mAbP SO57 by western blot. Western blot showed the significantly higher expression level of mAbP SO57 with KDEL compared to without KDEL. Flow cytometry (FACS) analysis showed that the Fc domains of both purified mAbP and mammalian-derived mAb (mAbM) evidenced similar binding activity to the FcγRI receptor (CD64). High performance liquid chromatography (HPLC) analysed, glycosylation patterns of mAbP SO57 with or without KDEL. The mAbP SO57 with KDEL had glycan profile with both oligomannose type (47.6%) and golgi type (52.4%), while the mAbP SO57 without KDEL had only golgi type (100%) glycans. Neutralizing analysis with rabies virus CVS-11 showed the similar neutralizing activity between mAbP SO57 with and without KDEL. These results suggest the potential of mAbP SO57 for rabies immunotherapy, regardless of plant specific glycan structures.

Effect of ER signal peptides (KDEL) on expression and function of anti-virus monoclonal antibody in plant

Da-Young Park,Yangkang So,Kyung-Jin Lee,Zhe Lu,Eun-Yi Oh,Kwang-Wok Min,Seul-Ki Lee,Jung-Hwan Lee,Mi-Hyun Ahn,Mira Song,Doo-Byoung Oh,Youngkwan Kim,Kisung Ko 한국당과학회 2009 한국당과학회 학술대회 Vol.2009 No.1

Plant genetic engineering has led to the production of plant-derived mAb (mAbP), which provides a safe and economically feasible alternative to the current antibody expression systems. In this study, the expression levels of mAbP SO57 with or without ER-signal peptides (Lys-Asp-Gly-Leu;KDEL) in transgenic tobacco plants were analysed in transgenic plant. PCR and Reverse Transcription-PCR analyses showed existence of heavy and light chain genes of mAb with or without KEDL and their transcription in plant, respectively. Western blot showed that the expression levels of mAbP SO57 with KDEL were significantly higher than that without KDEL. Flow cytometry analysis showed that the Fc domains of both purified mAbP and mammalian-derived mAb have similar binding activity to the FcγRI receptor. High-performance liquid chromatography analysis showed that the mAbP SO57 with KDEL had glycan profile with both oligomannose and golgi type, whereas the mAbP SO57 without KDEL had only golgi type glycans. Neutralizing analysis with rabies virus CVS-11 showed the similar neutralizing activity between mAbP SO57 with and without KDEL. These results suggest that the potential of mAbP SO57 for rabies immunotherapy is regardless of plant specific glycan structures.

항암화학요법 관련 오심과 구토 완화를 위한 비약물요법에 관한 연구동향

소향숙,최자윤,조인숙,김영재,김지영,김애숙,김옥미,김춘심,김현오,설영애,안정옥,이애리,이영자 성인간호학회 2003 성인간호학회지 Vol.15 No.3

Purpose: Purposes of this study were to understand the current trends on complementary therapy in relieving chemotherapy-induced nausea and vomiting and to suggest the future research direction. Method: Subjects were selected on CINAHL, MEDLINE, Korean Academy Data Base from 1980 to 2001 which used nausea, vomiting, chemotherapy and complementary therapy as key words in experimental studies. Eight korean articles and twenty-one international articles were analyzed in terms of general characteristics, research methods, and types of complementary therapy. Data were analyzed by using descriptive statistical methods. Result: Since 2000, researchers have more actively used complementary therapy. In subject characteristics, mean age was 35.5 years old, 45% of the researchers were performed with high level of incidence of chemotherapy induced vomiting, 14% of them set limit of consecutive cycle during research, and 65% of them did not comment the selecting criteria of sample. About 60% of them were designed post-test only control group: 35% used INV by Rhodes, 31% used Likert scale, and 24% used VAS for dependent variable. Muscle relaxation therapy was mostly applied for relief of nausea and vomiting. Conclusion: Further studies will be needed to control extrinsic variables affecting nausea and vomiting in research design and to accumulate evidence with studies applying various complementary therapies.

당뇨병과 동반된 화농성 간농양의 임상상 및 예후인자

오은숙,강무일,이원영,오기원,임동준,이소영,이정민,고승현,김성래,안유배,손현식,윤건호,차봉연,이광우,손호영,강성구 대한당뇨병학회 2000 임상당뇨병 Vol.1 No.1

연구배경: 감염질환은 혈관질환과 함께 당뇨병의 중요한 사망원인의 하나이며, 혈당조절이 불량할수록 감염 빈도가 증가됨은 잘 알려져 있다. 간농양의 발생이 있어서도 당뇨병은 중요한 기저질환 중의 하나이다. 일반적으로 간농양의 가장 흔한 원인균은 Escherichi coli라고 알려져 있으나, 최근 당뇨병을 가진 간농양 환자에서 Klesiella pneumoniae의 검출이 증가되고 있다. 또 최근 연구에 의하면 K.pneumonie가 간농양의 가장 흔한 원인균이며, K. pneumoniae에 의한 간농양은 K.pneumoniae 이외의 균주에 의한 간농양보다 당뇨병과 밀접히 관련되어 있다고 보고된 바 있다. 이에 저자들은 간농양 환자를 대상으로 당뇨병환자의 빈도를 알아보고, 그 원인균 및 특징을 비당뇨병 환자들의 경우와 비교해 보고자 하였다. 방법: 1992년 1월부터 1999년 6월까지 방사선학적 검사 혹은 수술에 의해 간농양이 확인되거나, 경피적 천자나 수술을 통한 배농액 배양, 혹은 혈액 검사에서 원인균이 증명된 182명의 환자를 대상으로 후향적 임상고찰을 실시하였다. 결과: 화농성 농양을 가진 167명의 환자(남:94명, 여:73명)중 당뇨병을 가진 환자는 54명 (남:30명, 여:24명)으로 32.3%를 차지하였다. 환자의 평균나이는 당뇨군에서 62.1 ± 13.4세, 비당뇨군에서는 55.1 ±15.5세로 당뇨군에서 높았으며(p=0.0021), 남녀간의 평균나이 비교 시 두 군 모두 여성의 나이가 의미있게 높았다. 비당뇨군에 비하여 당뇨군의 경우 저알부민혈증의 빈도와 aspartate transaminase가 증가되어있었다. 원인균주는 K. pneumoniae (당뇨군54%, 비당뇨군 39.1%), E. coli(당뇨군 17.5%, 비당뇨군 18.8%), Pseudomonas aeruginosa, Streptococcus viridans, Enterococcus 순이었으며 두 군간의 의미는 차이는 없었으나 K. pneumoniae의 경우 다른 균종에 비해 단독감염의 빈도가 높았다(82.8%). 사망률은 당뇨군에서 16.7%, 비당뇨군에서 7.1%로 당뇨군에서 의미있게 높았으며(p=0.019), 당뇨군에서의 사망환자는 모두 패혈증이 동반되어 있었다. 당뇨환자의 사망군은 생존군에 비해 당뇨병 이환기간이 길고 혼합감염의 빈도가 높았다(p=0.046). 검사실 소견에서는 총 빌리루빈 및 알카리성 포스파타제의 증가가 관찰되어 있었다. 결론: E. coli가 간농양의 주된 원인균이라는 기존 보고와는 달리 당뇨군과 비당뇨군 모두 K. pneumoniae가 간농양의 주된 원인균이었다. 또한 당뇨 유병기간이 길고 당뇨조절이 불량한 환자에서 혼합감염이며 총 빌리루빈 및 알카리성 포스파타제의 증가등과 같은 위험인자가 존재할 경우 사망률이 증가될 수 있으므로 이들 군에서는 적극적인 치료가 필요할 것으로 사료된다. Background: Liver abscesses are commonly associated with underlying disease, particularly diabete mellitus. The number of the liver abscesses caused by Klebslella pneumoniae in diabetic patient has been increased in Korea nowadays. This study was conducted to clarify the clinical presentation and prognostic factors of pyogenic liver abscesses, especially in diabetic patients and to determine the proportion of K. pneunomiae as a pathogen in liver abscess in Korea. Methods: Medical records of 167 patients treated for pyogenic liver abscess from January, 1992 through June, 1999 were reviewed retrospectively in detail. Major Pathogenic organism, clinical manifestations, prognostic factors, the importance of diabetes mellilus as an underlying disease and its effect on clinical features and prognosis were analyzed. Results: Among 167 cases of pyogenic liver abscess, underlying diabetes mellitus was present in 32.3%. The mean age of patients was 62.7 13.4 years in diabetic liver abscess group and 55.1 15.5 years in non-diabetic liver abscess group. Most liver abscesses were cryptogenic in origin or secondary to the billary tree diseases. The clinical presentations among the two groups were not significantly different. When compared to patients without diabetes, patients with diabetes had significantly higher proportions of hypoalbuminemia and elevated aspartate transaminase level. The most common organism of the pyogenic abscess was K. pneumoniae in both groups. Percutaneous drainage of the abscess with appropriate antibiotics was the most commonly used therapeutic modality in liver abscess. The mortality rate of diabetic liver abscess is 16.1 % and 7.1% in nondiabetic liver abscess. Complications, especially septicemla, were found more frequently in patients with diabetes than in patients without diabetes (64.8% vs 28.3%, septicemia : 31.5% vs 9.7%). The factors influencing mortality in the diabetic liver abscess were disease duration, mixed infection, presence of septicemia, elevated bilirubin and alkaline phosphatase. Conclusions: In contrast to prior report that the E. coli was the most common pathogen in liver abscess, we found that K. pneumoniae was the most common organism cultured in liver abscess. Diabetic patients have more complications and higher mortality than patients without diabetes. Early detection and proper treatment are needed to improve the outcome for diabetic patient with liver abscess.

인공신경망 모형을 이용한 울산공단지역 일 최고 SO2 농도 예측

이소영 ( So Young Lee ),김유근 ( Yoo Keun Kim ),오인보 ( In Bo Oh ),김정규 ( Jung Kyu Kim ) 한국환경과학회 2009 한국환경과학회지 Vol.18 No.2

Interacelluar reprogramming of expression, glycosylation, and function of plant-derived anti-virus therapeutic monoclonal antibody

Da-Young Park,Kyung-Jin Lee,Yangkang So,Zhe Lu,Seulki Lee,Jung-Hwan Lee,Seung Han Oh,Yeon Soo Han,Doo-Byoung Oh,Youngkwan Kim,Kisung Ko 한국당과학회 2010 한국당과학회 학술대회 Vol.2010 No.1

Plant genetic engineering has led to production of plant-derived mAb (mAbP), which can provides a safe and economically effective alternative to the conventional antibody expression methods. In this study, the expression levels of mAbP SO57 with or without ER-retention peptide extensions signal (Lys-Asp-Glu-Leu; KDEL) in transgenic tobacco plants were analysed in transgenic plant. The expression levels of mAbP SO57 with KDEL were significantly higher than that without KDEL regardless of transcription level. mAbP SO57 with KDEL localized surround to the nucleus suggesting that the mAbP with KDEL is localized in ER. The mAbP without KDEL and mAbH had mainly Golgi type glycans, whereas the ER-localized mAbP with KDEL had glycan profile with both oligomannose type (47.6%) and Golgi type (52.4%). The Fc domains of both purified mAbP (with and without KDEL) and human-derived mAb (mAbH) had similar bingding activity to the Fcγ RI receptor (CD64). Both mAbP (with and without KDEL) had a shorter half-life than mAbH. However, both mAbP with and without KDEL was as effective at neutralizing activity of the rabies virus CVS-11 as the mAbH. These results suggest that ER localization of recombinant mAbP by KDEL reprograms glycosylation and enhances production of the functional antivirus therapeutic antibody in the plant.

Protective Effects of Portulaca oleracea L. Extract against Matrix Metalloproteinase Production and Reactive Oxygen Species Generation Induced by Ultraviolet B Radiation in Human Keratinocytes

Jung Hwan Oh(오정환),Fatih Karadeniz(파티 카라데니즈),Jung Im Lee(이정임),So Young Park(박소영),Youngwan Seo(서영완),Chang-Suk Kong(공창숙) 한국생명과학회 2018 생명과학회지 Vol.28 No.8

쇠비름(Portulaca oleracea.L)은 쇠비름과에 속하는 한해살이풀로서 리놀렌산과 같은 불포화지방산, 페놀성 화합물, 플라보노이드, 비타민 C, 미네랄 함량이 높은 것으로 보고되어 있다. 본 연구에서는 쇠비름 추출물을 이용하여 UVB를 조사한 인간각질형성세포에서 광노화 억제능을 확인하였다. Matrix metalloproteinases는 세포의 기질을 분해하는 효소로 MMP-1는 collagenase, MMP-2와 MMP-9는 gelatinases로 피부 진피층을 구성하는 type Ⅰ collagen을 분해시키는데 영향을 미친다. UVB를 조사한 인간각질형성세포에서 쇠비름 추출물을 처리했을 때 MMP-1, -2, -9의 발현이 감소하였으며, type Ⅰ procollagen의 발현은 증가하는 것으로 나타났다. 또한 쇠비름 추출물을 처리한 군에서 UV에 의한 ROS 생성이 감소하였는데 이는 Nrf-2의 활성화를 통한 항산화 인자 SOD-1과 OH-1의 발현 증가로 인해 세포내 ROS 생성이 감소한 것으로 사료된다. 따라서 본 연구 결과를 통해 쇠비름 추출물이 UVB를 조사한 인간각질형성세포에서 MMP 인자 및 항산화 인자의 발현 조절을 통해 광노화로부터의 세포 보호능을 가지는 것을 확인하였으며 나아가 화장품 소재로서의 개발 가능성을 확인하였다. Portulaca oleracea L. is an edible plant widely consumed in daily diet throughout Europe, Asia and America. In this study, protective effects of P. oleracea L. extracts against oxidative stress and matrix metalloproteinase (MMP) activity induced by ultraviolet B (UVB) radiation were investigated using HaCaT immortal human keratinocytes. In this context, the mRNA and protein productions of MMPs (MMP-1, -2, and -9) and type I procollagen, which are major markers of photoaging induced by UVB radiation in HaCaT keratinocytes, were evaluated. Furthermore, UVB-induced reactive oxygen species (ROS) generation and mRNA and protein expression levels of superoxide dismutase-1 (SOD-1), oxygenase-1 (OH-1), and nuclear factor-erythroid 2-related factor-2 (Nrf-2), all of which are associated with the antioxidant balance, were investigated. As shown by the results, UVB radiation induced ROS formation and led to increased production of MMPs and decreased collagen production in human keratinocytes, which resulted in skin photoaging or photodamage. The treatment with P. oleracea L. extracts downregulated MMP (MMP-1, -2, and -9) production and upregulated type I procollagen expression in UVB-induced HaCaT cells. Furthermore, treatment with the extracts decreased UVB-induced ROS generation and increased the expression of antioxidant enzymes, such as SOD-1 and OH-1, through the Nrf-2 pathway. Taken together, these results suggest that P. oleracea L. extracts could be a potential cosmeceutical agent for the prevention of skin photoaging or photodamage.

초·중·고등학교 학생, 학부모, 일반교사의 보건교육 요구

윤순녕,김영임,최정명,조희순,김영희,박영남,오경순,이분옥,조선녀,조소영,한선희,하영미 韓國學校保健學會 2004 韓國學校保健學會誌 Vol.17 No.2

Purpose : The purpose of this study was to identify needs of health education of the students, parents, general teachers, and health teacher of the elementary, middle, and high schools. Method : 279 schools of elementary, middle, and high school were sampled nonproportionally during the period from June 28 to July 26, 2004 . The data were collected by the questionnaire from 1,888 students, 1,876 parents, 1,695 general teachers, 279 health teachers. A frequency, % and χ2-test were used to analysis by SAS program. Result : 1. It showed that 98.1% of elementary school students, 92.3% of middle school students, 89.6% of high school students answered that they need health education. The most of students, parents, and general teachers had high interests in health education. 2. In the case of students, main causes of needs of health education was 'to prevent diseases or accidents''. But parents answered that it was 'to build up a healthier behavior'. General teachers answered that it was 'more effective systematic and continuous health education'. 3. Over 80% of students, parents, and general teachers about the question of who qualified person is to teach health education as a regular class responded that health teacher is available. Especially 93% of elementary school students answered like that. 4. The most of students, parents, and general teachers answered that health education in the classroom is favorable 1 to 2 hours per week. Conclusion : As results of the study , the majority of students, parents, general teachers agreed needs of health education. Therefore, on the basis of this study, systematic and continuous health education is necessary. Additionally the establishment of independent health subject is required certainly.

초·중·고등학교 학생·학부모·일반교사의 보건교육 영역별 요구도와 보건교육

윤순녕,김영임,최정명,조희순,김영희,박영남,오경순,이분옥,조선녀,조소영,한선희,하영미 韓國學校保健學會 2005 韓國學校保健學會誌 Vol.18 No.1

Purpose: The purpose of this study was to identify the needs of health education in students, their parents and teachers in the elementary, middle and high schools and the current situation of health education class. Method: The subjects of this study were a total of 9450 persons including students, their parents and teachers from 279 schools throughout the country. They were selected through convenient sampling. Data were analyzed through x^(2)­test and ANOVA. Result: Students, their parents and teachers replied that 18 dimensions of health education class (DHEC) are necessary. The four DHEC - healthier life style, sex education, mental health and safety education - showed high educational needs in students, their parents, and teacher. High school students had higher educational need of 'symptom management for daily living' than elementary and middle school students. Students, their parents and teachers in elementary school had higher educational needs of 17 DHEC that those in middle and high school. The percentages of schools with health education class taught by health teachers were 99.2%, 75.5% and 66.0% respectively in elementary, middle and high schools. Health education was given mainly using physical education classes at elementary schools, and creative class hours at middle and high schools. In general, health education took 1-3 hours per week at elementary schools, and less than an hour at middle and high schools. Conclusion: Therefore, based on the results, systematic health education class should begin from elementary school to meet the need of health education in students, their parents and teachers, and further study should be made on the number of hours required and the amount of contents of 18 DHEC.

남자대학생의 흡연태도와 행동에 관한 연구

강경진,김정미,민혜영,안소진,오경숙,은경희,이현,이희선,홍주미,김수지 이화여자대학교 간호과학대학 2003 이화간호학회지 Vol.- No.37

There have been so many researches about problems of cigarettes and relation of smoking and heath since 1950's and generally known that smoking is one of the worst factor of increasing disease and mortality. No matter what such many proofs about various problems of smoking have been researched, smoking rate of current male collage students has not been decreased. Besides, most of the researches about cigarette use and its addiction have been focused on the adolescent or adult women smokers or adult male, not male collage student. Therefore, the subject of this study is about smoking attitude and smoking behavior of male collage students by general features. Smoking attitude was analyzed by modifying 'Guidelines for the content of tobacco smoking surveys of the general population'(WHO, 1982). Two hundred of male collage students participated to the distributed questionnaire for one week in May. The smoking attitude was categorized into duration of smoking and number of cigarette per day(CPD), and nicotine dependency, investigated by Fagerstrom Tolerance Questionnaire(FTQ) (Fagerstorm & Schneider, 1989). All statistics were performed by procedure of SAS according to number, percentage, mean, standard deviation, and ANOVA. The results are as follows: 1. Smoking attitude level was mean 25.73±5.15 marks, ranging from 13 to 39 marks of the total 40 marks. 2. Duration of cigarettes, number of cigarette per day, and nicotine dependency were mean 5.14±2.84 years, 12.88±6.85 pieces, and 4.78±1.69, respectively. 3. Smoking attitude showed significance according to general features such as marital state and allowance, but the significance according to marital state is not valuable because the numerical difference is so big between married one and single that they cannot be compared with each other. 4. Duration of smoking was significant according to general features such as age and allowance. 5. Smoking amount showed significance the factors of age only. 6. ll1ere was no significance in nicotine dependency according to general features. As a result, smoking attitude of male collage student in this study shows moderate level. Smoking attitude and duration is significant according to allowance, and smoking amount is significant according to age. Nicotine dependency according to general features is not significant but comparing with the preceding study about adult male, male collage students in this study are respectively more nicotine-dependent than adult male smokers. In conclusion, the right allowance usage strategies and effective heath education focused on the young are needed. In addition, helpful programs and information spreading strategies are necessary for establishing higher level of smoking attitude. At last, systemic regulations in daily living for ""anti-nico-dependent"" and perspective reforming of male university student are needed for reducing nicotine dependency.