Immobilization of calmodulin to affi-gel 15 agarose for the purification of ins(1,4,5)P_3-kinase from rat brain

LEE, Kyeong Ai,SHIN, Yong Chul,LEE, Sang Yeol 慶尙大學校 기초과학연구소 1990 基礎科學硏究所報 Vol.6 No.-

각종 hormone, neurotransmitter, growth factor 등의 외부자극에 의하여 ins(1,4,5)P_3를 ins(1,3,4,5)P_4로 만드는 ins(1,4,5)P_3-kinase의 효소활성이 Ca^++과 calmodulin의 존재하에서 증가된다는 사실로부터, ins(1,4,5)P_3-kinase 효소분리에 calmodulin affi-gel agarose resin을 응용하였다. 소의 뇌로부터 순수분리된 calmodulin을 affi-gel 15 agarose에 고정화시켰다. 쥐의 뇌에서 0.1M Tris, pH 7.5 buffer로 추출한 homogenates로부터 green gel과 calmodulin affi-gel chromatography를 이용하여 ins(1,4,5)P_3-kinase를 분리한 결과 1320배 분리하여 0.792μ㏖/min./㎎ protein의 specific activity를 갖는 효소를 얻었으며, 이 분획을 SDS-전기 영동한 결과 ins(1,4,5)P_3-kinase의 분자량은 53,000인 것으로 추정되었다. Using the fact that ins(1,4,5)P_3-kinase which converts the ins(1,4,5)P_3 to ins(1,3,4,5) P_4 is activated by the Ca^++/calmodulin complex, it was tried to purify the ins(1,4,5) P_3-kinase from rat brain cytosol by the immobilized calmodulin affinity gel chromatography. The calmodulin which was originally discovered as an activator protein of cyclic nucleotide phosphodiesterase, was purified to complete homogeneity from bovine brain and immobilized to affi-gel 15 agarose gel matrix. The sequential chromatography of dye-matrix green gel and calmodulin affi-gel provided very good methods for removing huge amounts of impure proteins(1320 folds) in the purification of ins(1,4,5) P_3-kinase, resulting in a specific activity of 0.792 μ㏖e/min/㎎ protein. On the contrary to the other calmodulin-sensitive enzymes, i. e., phosphodiesterase, ATPase, adenylate cyclase, etc., which were eluted by the EGTA alone from the calmodulin gel column, the ins(1,4,5)P_3-kinase has a strong binding affinity not only to the calmodulin protein itself but also to the agarose gel matrix, so that it could be eluted by the 0.1M Tris-Cl pH 7.6 buffer containing both 1mM EGTA and 0.2M-KCL. SDS-PAGE of the enriched ins(1,4,5)P_3-kinase fraction obtained from the calmodulin affi-gel chromatography showed a few minor protein bands and an extensive major protein band with an apparent M.W of 53000.

어린이집 보육교사의 휴게시간 실행에 대한 인식과 요구

이선애(Lee, Sun-Ai),강혜경(Kang, Hye-Kyeong) 한국영유아보육학회 2020 한국영유아보육학 Vol.0 No.122

본 연구는 어린이집 보육교사의 근무환경에 대한 인식과 근무환경을 개선하기 위해 휴게시간을 중심으로 알아보고자 한다. 연구대상은 경인지역의 설립유형별 보육교사로 각 4명씩 연령과 교육경력을 고려하여 24명을 선별하였다. 연구방법은 교사별 개인 인터뷰형식의 FGI질문법을 활용하였으며 질적인 분석을 위해 인터뷰 내용을 전사하여 범주화하였다. 연구결과, 보육교사는 휴게시간 시행을 정책적인 문제로 인식하였고 현장에서 휴게시간 시행은 보육교사의 직무 특성상 어려움이 있는 것으로 나타났다. 즉, 휴게시간 실행에 대한 보육교사인식은 현실적인 어려움, 가짜 휴게시간 환경조성, 업무의 과중, 원장과의 트러블 발생, 보육의 질 저하, 제도 자금 낭비로 나타났다. 개선방안은 조기 퇴근의 필요성, 쉼에 대한 인식의 개선, 보육교사에게 시간적 여유의 제공, 복지제도의 필요, 영 · 유아 안전보장대책의 필요, 인력 지원을 요구하였다. 장기적으로는 법적 보육시간의 수정이 필요하며 보육교사의 근무시간과 보육수요를 분석하여 2교대나 근무환경의 수정을 요구하였다. 결론적으로 보육교사의 근무환경개선은 대체교사활용과 초과근무수당의 지급으로 휴게시간 운영을 지원하고 장기적으로는 보육교사의 근무시간과 보육시설이용시간에 대한 보육지침의 수정이 필요하다. 또한, 정부는 보육수요자의 환경을 분석하여 다양한 보육서비스로 분류하고 보육현장의 근무 탄력제나 2교대 근무제를 수용하는 등 수요자와 공급자에 대한 맞춤형 정책과 인프라를 구성해야 할 것이다. We will execute an in-depth interview(called FGI) to the daycare center teachers. We can show how to improve the teachers’ rest period by researching on the actual conditions. It is for giving the basis to the policy improvement by suggesting the methods of making better working conditions for the teachers who can’t rest in field. The objects of study are 24 daycare center teachers from Seoul-Incheon area, 4 people each from the groups based on the age and experience. We interviewed each teacher in depth using FGI method, and drew the research question by categorizing the interview contents. It turned out that the daycare center teachers’ rest period policy has a problem and it can’t be solved in the field. Also the intensity of labor is high and rest time is lacked compared to the other occupations, because of the characteristic of children and service. Putting substitute teachers in field, flexible working system and overtime payment would be the solutions. And in the long term, the working hours and the environment should be changed by analyzing the demand in the field. The cognition of the daycare center teachers’ rest period has been known before, but it has been ignored because of the field conditions. For example the policy of rest time has been worked, but it didn’t work properly in reality. In other words, there was out of comprehension about the daycare occupation and its environment. Also the working hours are long and the intensity of the labor is high compared to other jobs. First of all, it is required to put the substitute teachers and give the overtime payment. In the long-term, we suggest an amendment of working hours and related policies. And the government should classify the daycare services into the groups and make flexible working system, and overall infrastructure.

직무 역량 강화를 위한 보육교사 교육프로그램이 사회적 자본과 교사효능감 향상에 미치는 효과 : 어린이집교사를 중심으로

이선애(Lee, Sun-Ai),강혜경(Kang, Hye-kyeong) 학습자중심교과교육학회 2021 학습자중심교과교육연구 Vol.21 No.23

목적 본 연구는 어린이집교사를 대상으로 교사교육프로그램을 적용해 사회적 자본과 교사효능감의 향상에 미치는 효과를 검증하였다. 방법 서울 및 수도권에 소재한 어린이집교사 13명을 대상으로 2020년 1월 2일부터 3월 6일까지 주2회 2시간씩 총 16회기의 교육프로그램을 적용하였다. 사회적 자본과 교사효능감의 효과 검증을 위해 교육프로그램 실시 전 사전 검사를, 교육프로그램이 완전히 종료된 후 사후 검사를 실시하였다. 통계패키지 SPSS 21.0을 이용해 윌콕슨의 대비된 쌍의 부호 순위 사전-사후 z검정을 실시하여 자료를 분석하였다. 결과 첫째, 교사교육프로그램 적용 전⋅후 어린이집교사의 사회적 자본은 유의미하게 변화되었다. 둘째, 교사교육프로그램 적용전⋅후 어린이집교사의 효능감은 유의미하게 향상되었다. 셋째, 교사효능감 수준에 따른 집단별 교사교육프로그램 적용 전⋅후, 교사효능감 수준이 높은 집단(N=7)에서만 사회적 자본이 유의미하게 향상되었다. 넷째, 교사교육프로그램 적용 전⋅후 사회적 자본 수준 집단별로 교사효능감의 변화는 유의미하지 않았다. 결론 이를 통해 교사교육프로그램이 보육 구성원의 사회적 관계의 중요성에 대한 어린이집교사의 내면을 긍정적으로 변화시키는 것과 더불어 교사의 효능감까지 향상시킬 수 있다는 시사점을 제시하였다. Objectives The purposes of this study were to specify variables related to convergence competency, and to analyze the structural relationships among them. The purposes of this study were to specify variables related to convergence competency, and to analyze the structural relationships among them. Methods From January 2 to March 6, 2020, a total of 16 educational programs were applied to 13 daycare center teachers located in Seoul and the metropolitan area twice a week. In order to verify the effectiveness of social capital and teacher efficacy, a pre-test was conducted before the education program was implemented, and a post-test was conducted after the education program was completely completed. Using the statistical package SPSS 21.0, data were analyzed by performing a pre-post z-test of Wilcoson s control code ranking pair. Results First, it was found that the social capital of daycare center teachers changed significantly before and after the application of the teacher education program. Second, before and after the application of the teacher education program, the work efficiency of daycare center teachers improved significantly. Third, social capital has improved significantly only in groups with high teacher efficacy (N=7) before and after applying teacher education programs for each group according to the level of teacher efficacy. Fourth, there was no significant change in the teacher effect by social capital level before and after the application of the teacher education program. Conclusions first, it was suggested that through this, the teacher education program can positively change the inside of daycare center teachers and increase the teacher s efficiency on the importance of the social relationship of childcare members.

Effects of pH on the Elaboration of Pullulan and the Morphology of Aureobasidium Pullulans

SHIN, Yong Chul,LEE, Kyeong Ai 慶尙大學校 기초과학연구소 1990 基礎科學硏究所報 Vol.6 No.-

The effects of pH on the cell growth, the elaboration of pullulan and the morphology of Aureobasidium pullulans IFO 4464 were examined. A. pullulans grew in yeast-like form at constant pH 7.5 and in mycelial form at constant pH 2.5. At the both pH conditions, the elaboration of pullulan was very low, about 6.0-6.5 g/l. The mixture of yeast-like form and mycelial form of cells was found at the constant pH 4.5, at which condition, the elaboration of pullulan was high, about 24.5 g/l. The pH shift experiments showed that the specific production rates of poullulan were 0.048(hr^-1) for the mycelial form and 0.058(hr^-1) for the yeast-like form, which indicated that the yeast-like form has the similar, only slightly higher, biosynthetic activity of pullulan to the mycelial form at pH 4.5 and the pH of culture broth is more important factor for the elaboration of pullulan than the morphology of A. pullulans.

Identification of Five Chitinase Isozymes from S. marcescens

Hwang, Jae Ryoung,Gal, Sang Wan,Lee, Kyeong Ai,Shin, Yong Chul,Cho, Moo Je,Lee, Sang Yeol 慶尙大學校 기초과학연구소 1991 基礎科學硏究所報 Vol.7 No.-

SDS 존재하에서 전기영동을 한 후, substrate activity staining을 한 결과, S. marcescens는 5가지의 chitinase isozyme을 생성함을 알아내었다. 이 효소는 inducible enzyme으로서, 30℃에서 7일간 진탕배양하였을 때 1.5% chitin을 첨가한 LB-배지에서 얻은 효소활성이 chitin을 첨가하지 않은 배지에서 얻은 것보다 약 7배의 효소활성의 증가를 보였다. Chitinase를 최대로 생성시키는 조건에서 S. marcescens를 배양한 후 regenerated chitin을 이용한 친화성 chromatography를 행하여, 이 배양상등액으로부터 5가지의 chitinase isozyme를 분리하였으며 이들의 분자량이 58, 47, 43, 37 및 21 kd임을 확인하였다. 충분히 세척한 chitin column으로부터 chitinase를 elution시키는 방법으로, pH3.3인 완충액을 사용하였을 때, 5가지 chitinases가 모두 용출되었으며, pH를 2.0인 완충액으로 바꾸어 용출시켰더니 5가지 중 분자량이 58, 47, 43 kd인 3가지 chitinase가 더 회수되었다. 또한 Chitinases 의 pH에 대한 안정성을 실험한 결과 S. marcescens가 분비하는 chitinases는 산성조건에서 매우 불안전한 chitinases와 비교적 안정한 chitinase의 두가지 group이 존재함을 알 수 있었다. S. marcescescens KCTC2172 produced five extracellular chitinases that were involved in chitin degradation. These enzymes were identified as chitinase isozymes by substrate activity staining in SDS-PAGE gel after removal of SDS, because chitinases were resistant to SDS and existed as monomeric proteins. Their apparent molecular weights obtained by SDS-PAGE were 58, 47, 43, 37, and 21kd. The chitinase activities were induced about seven folds by the addition of 1.5% chitin to LB-medium compared with those obtained without chitin, and the induction was dependent on the various forms of chitin. Chitinases were purified using the regenerated chitin column by successive pH changes of elution buffer. When the column was eluted with enough volume of pH 3.3 buffer, five chitinase isozymes were eluted. However, three of them whose molecular weights were 58,47 and 43 kd were further eluted by the pH changes of elution buffer from pH 3.3 to 2.0 Considering the pH dependent stability of chitinases, it was known that S. marcescens produced two groups of chitinase isozymes; One is resistant and the other is very sensitive to acidic conditions, at pH below 3.3.

Mirror Syndrome Resulting from Metastatic Congenital Neuroblastoma to Placenta

( Sung Hyeon Park ),( Jung-man Namgoong ),( Kyeong Nam Ko ),( Chong Jai Kim ),( Pil-ryang Lee ),( Euiseok Jung ),( Byong Sop Lee ),( Ki-soo Kim ),( Ellen Ai-rhan Kim ) 대한주산의학회 2018 Perinatology Vol.29 No.4

Congenital neuroblastoma is a rare disease. Placental metastasis is extremely rare and poor prognosis has been reported in neonates. Mirror syndrome could occur in mother with placental metastasis with possibilities of hypertension and edema. We report a case of detection of left suprarenal mass in fetus at 31⁺⁵ weeks’ gestation. Mother presented with palpitation, edema, headache, and hypertension. Maternal 24 hours urine vanillylmandelic acid (VMA) and normetanephrine (NME) level at 34 weeks’ gestation were elevated. Consequently, emergent cesarean section was done. Based on abdominal ultrasonography and whole body magnetic resonance imaging, left adrenal tumor with liver metastasis was suspected. Neuroblastoma was confirmed by liver and placenta biopsy. Chemotherapy was started with Pediatric Oncology Group 9243 at day 7 and changed into Children's Oncology Group 3961 due to cholestasis and poor response during 2nd cycle. Plasma exchange was done for aggravated direct hyperbilirubinemia. The baby expired at 73 days due to multi-organ failure. Maternal symptoms were completely resolved in 2 weeks after delivery along with normalization of the elevated level of VMA and NME. We report a first case of mirror syndrome in Korean mother and fetus resulting from metastatic congenital neuroblastoma to placenta.

Serratia marcescens 로 부터 5 가지 Chitinase Isozymes 의 동정

황재령,갈상완,이경애,신용철,조무제,이상열 ( Jae Ryoung Hwang,Sang Wan Gal,Kyeong Ai Lee,Yong Chul Shin,Moo Je Cho,Sang Yeol Lee ) 생화학분자생물학회 1991 BMB Reports Vol.24 No.3

S. marcescens KCTC2172 produced five extracellular chitinases that were involved in chitin degradation. These enzymes were identified as chitinase isozymes by substrate activity staining in SDS-PAGE gel after removal of SDS, because chitinases were resistant to SDS and existed as monomeric proteins. Their apparent molecular weights obtained by SDS-PAGE were 58, 47, 43, 37, and 21 kd. The chitinase activities were induced about seven folds by the addition of 1.5% chitin to LB-medium compared with those obtained without chitin, and the induction was dependent on the various forms of chitin. Chitinases were purified using the regenerated chitin column by successive pH changes of elution buffer. When the column was eluted with enough volume of pH 3.3 buffer, five chitinase isozymes were eluted. However, three of them whose molecular weights were 58, 47 and 43 kd were further eluted by the pH changes of elution buffer from pH 3.3 to 2.0. Considering the pH dependent stability of chitinases, it was known that S. marcescens produced two groups of chitinase isozymes; One is resistant and the other is very sensitive to acidic conditions, at pH below 3.3.

Imaging of Activated Cortical Areas after Light and Electrical Stimulation of the Rabbit Retina: F-18 FDG PET-guided Brain Mapping

Se Joon Woo,Hum Chung,서종모,Jung Hyun Park,Su Jin Kim,Jae-Sung Lee,Yu-Kyeong Kim,Jing Ai Zhou,Eui Tae Kim,Sung June Kim 대한의용생체공학회 2012 Biomedical Engineering Letters (BMEL) Vol.2 No.2

Purpose Using 18F- fluorodeoxyglucose positron emission tomography (FDG PET), we aimed to assess the cortical areas activated by light and electrical stimuli in rabbits and to determine the nature of the electrically evoked visual perception by comparing the 2 activated cortical images. Methods Three adult white rabbits were used, and 5 18F -FDG PET scans were performed on 5 different days for each rabbit: a resting image, 2 light stimulation images, and 2electrical retinal stimulation images by suprachoroidal electrodes of the right and left eyes. A small-animal PET scanner (resolution: 1.18 to 1.45 mm) was used for image acquisition. Activated cortical areas were determined by voxel-wise statistical analysis of the difference between the stimulation and resting PET images, and three-dimensional brain imaging of functional activation was constructed. Results After visual and electrical stimulation of the rabbit retina, increased metabolism was found in 2 distinct cerebral cortical areas: (1) the ipsilateral frontal cortex (frontal eye field) and (2) the contralateral occipito-parietal cortex (visual cortex). Cortical areas activated after electrical retinal stimulation were nearly identical to the visually activated cortical areas. Conclusions Electrical stimulation of the rabbit retina using suprachoroidal electrodes induced a similar cortical activation response as that induced by light stimulation. Brain PET images obtained from stimulation of the retina with light and electric current increase our understanding of cortical perceptions by both stimuli and can be a useful methodology for retinal prosthesis research.