Efficient Interleukin-21 Production by Optimization of Codon and Signal Peptide in Chinese Hamster Ovarian Cells

( Hee Jun Cho ),( Byung Moo Oh ),( Jong-tae Kim ),( Jeewon Lim ),( Sang Yoon Park ),( Yo Sep Hwang ),( Kyoung Eun Baek ),( Bo-yeon Kim ),( Inpyo Choi ),( Hee Gu Lee ) 한국미생물 · 생명공학회 2019 Journal of microbiology and biotechnology Vol.29 No.2

Interleukin-21 is a common γ-chain cytokine that controls the immune responses of B cells, T cells, and natural killer cells. Targeting IL-21 to strengthen the immune system is promising for the development of vaccines as well as anti-infection and anti-tumor therapies. However, the practical application of IL-21 is limited by the high production cost. In this study, we improved IL-21 production by codon optimization and selection of appropriate signal peptide in CHO-K1 cells. Codon-optimized or non-optimized human IL-21 was stably transfected into CHO-K1 cells. IL-21 expression was 10-fold higher for codon-optimized than non-optimized IL-21. We fused five different signal peptides to codon-optimized mature IL-21 and evaluated their effect on IL-21 production. The best result (a 3-fold increase) was obtained using a signal peptide derived from human azurocidin. Furthermore, codon-optimized IL-21 containing the azurocidin signal peptide promoted IFN-γ secretion and STAT3 phosphorylation in NK-92 cells similar to codon-optimized IL-21 containing original signal peptide. Collectively, these results indicate that codon optimization and azurocidin signal peptides provide an efficient approach for the high-level production of IL-21 as a biopharmaceutical.

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.

3T Multi Voxel Spectroscopy에서 SENSE와 NEX 변화에 따른 정상인 뇌 대사물질 변화 분석

성열훈,임재동,이재현,조성봉,우동철,최보영,Seong, Yeol-Hun,Rhim, Jae-Dong,Lee, Jae-Hyun,Cho, Sung-Bong,Woo, Dong-Chul,Choe, Bo-Young 한국의학물리학회 2008 의학물리 Vol.19 No.4

자기공명분광법(magnetic resonance spectroscopy: MRS)은 인체내 대사물질을 정량분석하여 병변의 조기진단 및 정밀진단에 도움을 주고 있으며, 최근 임상에 이용되고 있는 자기공명분광법은 single voxel spectroscopy (SVS) 기법과 multi voxel spectroscopy (MVS) 기법이 있다. 본 연구에서는 SENSE와 NEX를 변화시킨 multi voxel spectroscopy (MVS)의 데이터와 기존 single voxel spectroscopy (SVS)의 데이터를 비교 분석하여, 각각의 데이터의 유의성 차이를 평가하고자 하였다. 정상 성인 지원자 13명(남자: 5명, 여자: 8명, 평균 41세, 표준편차 11.65세)을 대상으로 chemical shift image (CSI)를 이용한 MVS검사를 시행하였다. 장비는 3.0T Achieva Release Version 2.1 (Philips Medical System, Netherland)을 이용하였고, 8 channel head coil을 사용하여 brain thalamus 부위에서 CSI spectrum을 1 slice 획득하였다. Scan parameter로는 FOV (field of view): $230{\times}184mm^2$, TR (time to repetition): 2000 msec, TE (time to echo): 288 msec, matrix: $15{\times}12$, VOI(view of interest): $110{\times}110mm^2$, voxel size: $15{\times}15{\times}15mm^3$로 하였다. SENSE factor (S)와 NEX (N)는 S1*N1, S2*N1, S2*N2, S3*N2로 변화하여 스펙트럼을 획득하였고, 각 scan time은 5분 54초, 3분 32초, 6분 20초, 4분 20초였다. 얻은 모든 MRS 데이터는 jMRUI 3.0 Version 프로그램에서 분석하였고, SENSE factor와 NEX를 변화시켜 얻은 MVS data 그룹들이 정상 성인 뇌 대사물질의 변화에 영향을 주는지 검증하기 위해 그룹 간에 ANOVA분석을 실행하여 P 값이 0.05보다 크게 나오면 그룹들 사이에 유의한 차이가 없다고 분석하였다. NAA/Cr과 Cho/Cr의 상대적 비율은 MV와 SVS사이에서는 유의한 차이가 없었다. 즉, SENSE factor와 NEX를 변화시켜 얻은 MVS data에서 정상 성인 뇌조직의 대사물질의 변화를 관찰한 결과, S1*N1의 NAA/Cr은 $1.45{\pm}0.03$, Cho/Cr은 $0.88{\pm}0.03$이고, S2*N1의 NAA/Cr은 $1.44{\pm}0.03$, Cho/Cr은 $0.87{\pm}0.05$, S2*N2의 NAA/Cr은 $1.43{\pm}0.02$, Cho/Cr은 $0.87{\pm}0.04$이며, S3*N2의 NAA/Cr은 $1.45{\pm}0.03$, Cho/Cr은 $0.87{\pm}0.03$으로 나타났다(F-value : 1.37, D.F : 3, P-value : 0.262). 그러나 데이터의 질을 측정하기 위한 MVS 데이터의 NAA Peak line-width는 SVS 데이터의 NAA Peak line-width 보다 약 3배 정도 넓었다. 본 연구에서는 MVS에서 SENSE factor와 NEX 값을 다양하게 변화시킨 MVS의 데이터와 SVS의 데이터가 큰 차이가 없음을 확인하였다. 즉, 어는 특정 부위의 뇌 조직의 대사물질은 MVS와 SVS 기법 모두 큰 차이가 없음을 확인할 수 있었다. 그러므로 MVS는 SVS보다 광범한 부위를 짧은 시간 안에 검사할 수 있으므로 매우 유용한 방법이라고 사료된다. To evaluate the metabolic changes in normal adult brains due to alterations SENSE and NEX (number of excitation) by multi voxel MR Spectroscopy at 3.0 Tesla. The study group was composed of normal volunteers (5 men and 8 women) with a mean ($\pm$ standard deviation) age of 41 (${\pm}11.65$). Their ages ranged from 28 to 61 years. MR Spectroscopy was performed with a 3.0T Achieva Release Version 2.0 (Philips Medical System-Netherlands). The 8 channel head coil was employed for MRS acquisition. The 13 volunteers underwent multi voxel spectroscopy (MVS) and single voxel spectroscopy (SVS) on the thalamus area with normally gray matter. Spectral parameters were as follows: 15 mm of thickness; 230 mm of FOV (field of view); 2000 msecs of repetition time (TR); 288 msecs of echo time (TE); $110{\times}110$ mm of VOI (view of interest); $15{\times}15{\times}15$ mm of voxel size. Multi voxel spectral parameters were made using specially in alteration of SENSE factor (1~3) and 1~2 of NEX. All MRS data were processed by the jMRUI 3.0 Version. There was no significant difference in NAA/Cr and Cho/Cr ratio between MVS and SVS likewise the previous results by Ross and coworkers in 1994. In addition, despite the alterations of SENSE factor and NEX in MVS, the metabolite ratios were not changed (F-value : 1.37, D.F : 3, P-value : 0.262). However, line-width of NAA peak in MVS was 3 times bigger than that in SVS. In the present study, we demonstrated that the alterations of SENSE factor and NEX were not critically affective to the result of metabolic ratios in the normal brain tissue.

입원환자 간호관리료 차등제 도입이후 간호사 확보수준의 변화

조성현,전경자,김윤미,박보현 대한간호학회 간호행정학회 2008 간호행정학회지 Vol.14 No.2

Purpose: To examine the changes in nurse staffing in hospitals after implementing the policy of differentiating inpatient nursing fees by staffing grades. Method: The study sample included 43 tertiary hospitals, 185 general hospitals, and 282 non-general hospitals that were operating in both 1999 and 2008. Nurse staffing grade was categorized from Grade 1 (highest) to 6 (lowest) in 1999 or Grade 7 in 2008, based on the nurse-to-bed ratio. Results: Tertiary hospitals at Grade 3 and Grade 4 accounted for 49% and 35%, respectively, in 2008, whereas 63% were Grade 6 in 1999. General hospitals at Grade 6 decreased from 87% to 48%. In non-general hospitals, little change was found in the staffing distribution, in that 92% still remained in Grade 6 or 7 in 2008. Forty tertiary hospitals (93%) and 45% of general hospitals improved their staffing grades, while only 7% of non-general hospitals did. Greater likelihood of improvement in staffing grades was found in general hospitals located in metropolitan areas or having 250 or more beds. Conclusion: Elaboration of the financial incentive system is needed to increase the policy impact on staffing improvement.

수아세포종의 예후에 영향을 미치는 요인 분석 : An Analysis of Factors Influencing on Its Prognosis

조병규,왕규창,김일한,이상일,심보성,최길수 대한신경외과학회 1988 Journal of Korean neurosurgical society Vol.17 No.5

Authors analyzed the results of treatment in 49 cases of medulloblastoma who had been operated upon at the Seoul National University Hospital from January 1972 to June 1987. There were 2 cases(4%) of postoperative mortality(death within postoperative one month) and 13 cases of late moratlity. The 3-year overall survival rate was 49.3% and 3-year disease-free survival rate was 48.5%. The risk period of recurrence was about 2 years after surgery. The 3-year survival rate was significantly better in "gross total resection" group, "radiation dose more than 50 Gy" group, and groupⅡ(23 cases operated after July 1982) (p<0.05). The rate of posterior fossa recurrence was lower in "gross total resection" group for the patients who received posteror fossa irradiation of more than 50 Gy.

TV 채널의 브랜드 이미지와 선호도 : 대구지역 공중파 TV를 중심으로

조성호,김보은 慶北大學敎 社會科學大學 2005 社會科學 Vol.17 No.-

이 연구는 대구 지역 세 공중파 TV 방송국의 이미지를 기능적 이미지와 상징적 이미지의 차원으로 조사하였고, 각 채널의 브랜드 이미지가 선호도에 어떤 영향을 미치는가를 분석하였다. 분석 결과, 첫째, 지역의 TV 채널은 기능적 이미지 -1)뉴스 프로그램에 대한 평가, 2)뉴스 이외의 지역 프로그램에 대한 평가, 3)방송사의 지역사회 공헌도, 4)프로그램 시청의 효용성- 에서 채널간 차이가 발견되었고, 이러한 차이는 대체적으로 대구KBS와 대구MBC, 대구KBS와 TBC 사이에서 크게 나타났다. 대구MBC는 뉴스와 프로그램을 잘 만드는 채널, TBC는 지역사회에 공헌하는 채널로 이미지가 형성되어 있는 반면, 대구KBS는 특정 부문에서 뛰어나다고 평가받는 이미지가 형성되어 있지 않았다. 둘째, 상징적 이미지인 채널 브랜드 개성에서도 채널간 차이가 발견되었다. 대구KBS가 신뢰할 만하고 전통적인 이미지, 대구MBC와 TBC는 진취적이고 성실한 이미지를 갖고 있는 것으로 나타났다. 셋째, 채널의 이미지와 선호도와의 관계에서는, 대구KBS는 ‘시청의 효용성’ 차원, 대구MBC는 ‘채널 브랜드 개성’ 차원, TBC는 ‘뉴스 이외의 지역 프로그램 평가’ 차원이 채널 선호도에 가장 영향을 미치는 이미지로 나타났다. 각 방송사별로 나름대로 우위에 있는 이미지를 살려 채널 브랜드 자산을 높이고, 자사 채널의 브랜드 이미지가 선호도와 연결되도록 하는 효과적인 마케팅 전략이 요구된다. This study examined channel images of Daegu local broadcasting stations. Further, it examined the relationship between TV channel brand images and channel preference. The symbolic image among channels showed that there were differences in channel brand personality. Symbolic image was divided into four factors such as 'progressiveness,' 'sincerity,' 'reliability,' 'tradition.' KBS was symbolized with reliability and tradition where as MBC and TBC with progressiveness and sincerity. The results of analysing how symbolic image(channel brand personality) have an effect on preference of each channel showed that progressiveness was the most effective variable for all channels. Reliability and tradition didn't have any direct effect on preference. It suggested that placing emphasis on progressiveness could lead to get more preference for local broadcasting. Finally the overall results suggested that stations have to provide useful informational programs with high quality in order to enhance the image of each channel. The effective channel image structure are closely related to channel preference.

소득작물 전후작지 적응 극단기성 벼 ‘중모 1032’

손영보,이종희,조준현,이지윤,오성환,송유천,박동수,박노봉,김상열,여운상,남민희,권영업 忠北大學校 農業科學硏究所 2015 農業科學硏究 Vol.31 No.2

건국대학교 의과대학의 통합강의 모델에 관한 연구

조성일,최진영,김보경 건국대학교 의과학연구소 1999 건국의과학학술지 Vol.9 No.-

To develop a proper model of block lecture as an educational program, we surveyed the educational environments and problems in Konkuk University Medical College(KUMC). The major findings were as follows; 1. There is no significant differences in ratio of lecture/laboratory hour between KUMC and average of medical colleges in Korea. 2. The total lecture time is higher(138%) in KUMC than the minimal lecture time(1204 hrs) for basic medical science in the guideline of the Korean Medical Association(KMA). 3. The real lecture hours was 2,184 hrs for 4 years that is less than the time(2786 hrs) reported to Council of Korean Medical Dean. 4. The time estimated for block lecture in all course at KUMC is 1428.5 hrs. 5. A number of specialists(professors) for each course is not sufficient and many courses have to depend on lecture rather than laboratory or clinical bedside working. 6. The professors are separated at three distant sites resulting in inefficient structure. 7. The educational space in the two hospitals is not sufficient for block lecture program. 8. In KUMC at present, 9 courses can be performed with block lecture program, those are Diagnostics, Medical Ethics, Forensic Medicine, Introduction to Biomedicine, Legal Medicine, Emergency Medicine, Community Medicine, Psychiatry, and Pediatrics. And 20 courses need the further investigation to develop block lecture program. More intensive study is needed for 6 courses. From these results, although KUMC needs more space and specialists and needs effective concentration of manpower to perform block lecture, 35 block lectures can be introduced after investigation. And it is suggested that introducing the block lecture system in whole curriculum is not possible at present status of KUMC.

비특징 기반의 차량 번호판 인식

조보호,정성환 國立 昌原大學校 精報通信硏究所 1998 精報通信論文集 Vol.2 No.-

In this paper, we propose a nonfeature-based method to recognize vehicle plate characters. Previous feature-based requires complex preprocessing on a vehicle plate area. The problems of these approaches are the plate binarization process which causes distorted characters and difficult feature vector extraction. In the proposed method, we simplify the previous complex preprocessing. We also use the location information of characters in the vehicle plate area and the color intensity difference between the character region and the background region in the vehicle plate area. Our method then directly recognizes the extracted character region by using ART2 neural network. We perform experiments with extracted 77 vehicle image under various lighting condition. In the experimental results, our proposed method is not sensitive to the lighting condition. It also shows about 92%, 0.1 sec in terms of the total recognition rate and the recognizing time, respectively.

방사선 조사된 참깨에서 hydrocarbon류 검지를 위한 초임계 유체 추출 공정의 이용

조성길,서상철,이보경,홍주헌,최용희 한국산업식품공학회 2003 산업 식품공학 Vol.7 No.4

초임계 유체 추출 공정을 이용하여 참깨에서 hydrocarbon류를 검출하여 참깨의 방사선 조사 여부 검지 전처리 기술에 활용하고자 하였다. 참깨를 2 kGy의 감마선을 조사하여 hexane으로 지방을 추출한 용매 추출과 초임계 유체 추출을 이용하여 추출한 지방을 Florisil coulumn으로 분리한 후 hydrocarbon 류를 GC-MS로 분석하였다. 용매 추출에서와 같이 초임계 유체 추출 공정을 이용하여 검출된 hydrocarbon 류은 동일하게 검지되었으며 그 marker 로써 활용 가능한 hydrocarbon 류인 13:3, 17:2, 16:2, 17:1은 비조사 참깨에서는 검출되지 않아TEk. 보조 용매르 사용한 초임계 추출 공정에서 각 보조용매에서 추출된 지방량은 용매 추출보다 많았으며 보조용매를 사용하여 검출된 hydrocarbon 류의 검출량은 methanol을 제외하고 전반적으로 많이 검출되었다. 특히 acetone의 경우 marker로 사용가능한 hydrocarbon류는 용매 추출에서보다 3~4배 정도, CO₂만을 사용함 초임계 유체 추출 공정에서 보다 거의 2배의 검출 특성을 보여주었다. 이러한 결과는 초임계 유체 추출 공정이 방사선 조사 여부 검지 기술에 있어서 기존의 용매 추출에 비해 지방 추출시 용매 소모량의 감소, hydrocarbon 류의 검출량 증가 등의 이유로 hydrocarbon 류의 검지 기술에 있어 전처리 기술로 활용 가능하리라 사료된다. Radiation-induced hydrocarbons from the irradiated sesame were investigated in this study to compare the chemical properties of irradiated and non-irradiated foodstuff. The detective volatile hydrocarbons for the irradiated sesame were (C14:0), 1-tetradence (C14:1), n-pentadecane (C15:0), n-hexadecane (C16:0), 1-hexadecane (C16:1), n-heptadecane (C17:0), 8-heptadecene (C17:0), 1,7-hexadecadiene (C16:2) 1,7,10-hexadecadiene (C16:3) 6,9-heptadecadiene (C17:2). The characteristic of extracts was also investigated at the change of pressure and temperature. And then it was found that the increase of the pressure and the decrease of the temperature in supercritical fluid extraction have made possible much more rapid extraction of volatile hydrocarbons. The amounts of volatile hydrocabons for irradiated and non-irradiated foodstuff were definitely differentiate. It could be concluded that the supercritical fluid extraction was possible for detective extraction method irradiated foods.