Dual-mode enhancement of metallothionein protein with cell transduction and retention peptide fusion

Lim, K.S.,Lim, M.H.,Won, Y.W.,Kim, J.K.,Kang, Y.C.,Park, E.J.,Chae, J.W.,Kim, S.M.,Ryu, S.E.,Pak, Y.K.,Kim, Y.H. Elsevier Science Publishers 2013 Journal of controlled release Vol.171 No.2

Protein transduction domains (PTDs), also known as cell-penetrating peptides (CPPs), have been developed as effective systems for delivering bio-active cargos such as proteins, genes and particles. Further improvements on cell-specific targeting, intracellular organelle targeting and intracellular retention are still necessary to enhance the therapeutic effect of PTD fusion proteins. In order to enhance the cell transduction and retention of anti-oxidative metallothionein protein (MT), MT was recombinantly fused with transcriptional activator (Tat) with or without a short peptide (sMTS) derived from mitochondria malate dehydrogenase (mMDH). Cellular uptake and retention time of fusion protein were significantly increased in the H9c2 cell by sMTS. The Tat-sMTS-MT (TMM) fusion protein protected H9c2 cells more effectively against hypoxia, hyperglycemia and combination compared with Tat-MT (TM) by reducing intracellular ROS level. It maintained the normal blood glucose level over an extended period of time in a streptozotocin-induced diabetic mouse model. PTD-sMTS-MT fusion protein has a potential to be used as a therapeutic protein for the treatment or prevention of diabetes and diabetic complications.

Protective effects of protein transduction domain-metallothionein fusion proteins against hypoxia- and oxidative stress-induced apoptosis in an ischemia/reperfusion rat model

Lim, K.S.,Cha, M.J.,Kim, J.K.,Park, E.J.,Chae, J.W.,Rhim, T.,Hwang, K.C.,Kim, Y.H. Elsevier Science Publishers 2013 Journal of controlled release Vol.169 No.3

Ischemic heart diseases caused by insufficient oxygen supply to the cardiac muscle require pharmaceutical agents for the prevention of the progress and recurrence. Metallothionein (MT) has a potential as a protein therapeutic for the treatment of this disease due to its anti-oxidative effects under stressful conditions. In spite of its therapeutic potential, efficient delivery systems need to be developed to overcome limitations such as low transduction efficiency, instability and short half-life in the body. To enhance intra-cellular transduction efficiency, Tat sequence as a protein transduction domain (PTD) was fused with MT in a recombinant method. Anti-apoptotic and anti-oxidative effects of Tat-MT fusion protein were evaluated under hyperglycemia and hypoxia stress conditions in cultured H9c2 cells. Recovery of cardiac functions by anti-apoptotic and anti-fibrotic effects of Tat-MT was confirmed in an ischemia/reperfusion (I/R) rat myocardial infarction model. Tat-MT fusion protein effectively protected H9c2 cells under stressful conditions by reducing intracellular ROS production and inhibiting caspase-3 activation. Tat-MT fusion protein inhibited apoptosis, reduced fibrosis area and enhanced cardiac functions in I/R. Tat-MT fusion protein could be a promising therapeutic for the treatment of ischemic heart diseases.

蛋白資源으로서의 大豆 增産 및 利用性 向上에 관한 硏究

李弘석,潘性紈,柳順昊,蔡永岩,林善旭,李炯周,韓仁圭 서울大學校農科大學 1984 서울대농학연구지 Vol.9 No.1

1. 大豆의 需給에 관한 硏究. (1) 大豆는 直接 食用消費와 飼料로서 間接消費가 급격히 增加 하고 있다. (2) 1968∼1981년 사이에 大豆의 用途別 消費構造가 크게 變化되어 食用消費에서 飼 料用 消費로 移動하고 있는 것이 뚜렷하다. (3) 大豆 消費量을 推定한 결과 總消費量은 1981年의 ??에서 1993년에는 ??으로 약 2.4배가 될것으로 예상된다. (4) 消費를 충족시키기 위해서는 막대한 양의 輸入이 불가피하지만 國內生産의 增大 와 質的 向上에 관한 方案이 시급히 모색되어야 한다. 2. 良質高蛋白 多收性品種의 選拔 및 育成 (1) 良質高蛋白 多收性品種의 育成을 위하여 母本을 선정하고 人工交配를 實施하여 58組合 670粒의 交配種子를 얻었다. (2) 雜種世代로부터 41組合의 247 F₂個體, 32組合 203系統의 F₃系統, 10組合의 25F₄系統을 선발하였다. (3) 生産力 檢定豫備試驗에서 3系統이 그리고 本試驗에서 2系統이 극히 多收性으로 선발되어 有望視되며 다음 試驗에 供試될 豫定이다. 3. 耕種方法에 따른 大豆蛋白의 量的 및 質的 向上에 관한 硏究. (1) 大豆의 量的收量增加를 위해서는 適正量의 施肥가 要求된다. (2) 전반적으로 볼때 소석회나 요소 엽면 시비는 등숙, 분지수, 株當협수, 株당粒 數, 100粒重 및 收量에 영향을 주고 있지만 標準肥栽培로 충분한 것으로 판단된 다. (4) 含黃 아미노산인 methinie과 cystine의 함량증가에 유황은 效果가 없었다. 4. 大豆根瘤菌의 分離 및 選別 (1) 募集 分離된 菌은 培養的 特性 및 生理試驗에 의하여 大豆根瘤菌種임을 확인하 였다. (2) 大豆品種 東北太, 長葉콩 및 Wayne에서 分離한 것이 根瘤의 差生 및 colony 發 育相이 우수하였다. 5. 大豆의 品種과 施肥에 따른 蛋白質 및 아미노산의 造成變化 (1) 品種間 蛋白質 含量에 큰 차이를 나타내고 있었다. (2) 品種內에서 시비에 따른 蛋白質 含量에 차이가 없었다. (3) 含硫黃 아미노산의 함량이 蛋白質 100g당 5.1∼6.4g으로 상당히 높았으나 품종 간이나 시비 방법에 따른 차이는 크지 않았다. 6. 國內産 大豆의 Broiler에 대한 飼料的 價値에 關하나 硏究 (1) 증체량, 사료섭취량 및 사료효율은 공시품종간에 유의적인 차이를 나타내지 않 았으나 대두 품종 봉의의 증체량이 제일 높았다. (2) 공시 대두는 어분이나 casein에 비하여 높은 증체율을 보았다. (3) 공시품종간에 AME와 TME에는 유의적인 차이를 보이지 않았다. Soybean has been cultivated as main protein source for a long time and it is also an important oil crop. As our living standard is getting better, higher quality in soybean protein and larger quantity of soybean are demanded. Therefore this study was conducted to provide comprehensive understanding for better utilization and processing of soybean as well as improvement of soybean breeding lines for higher yield with better guality. The results are summarized as follows: 1. Study on the demand and supply relationslip of soybean. (1) Demand for soybean as direct daily use of food and feed grain has been rapidly increased for past 13 years. (2) Utilization pattern of soybean had been shifted from direct use of food to feed grain during 1968∼1981. (3) Consumption of soybean will be increased 2.4 fold from ?? in 1981 to ?? in 1993. (4) Huge amount of soyben sbould be imported to satisfy the demand. Therefore certain measures soybean production in land and improvement of soybean protein guality should be considered. 2. Study on the selection and breeding for high yielding varieties with high protein content and better quality of protein in soybean. (1) For the breeding of high yield and high protein content with better quality in soybean, artificial cross between selected parents were made and obtained 670 crossed seeds in 58 combinations. (2) Two hundred forty seven F₂plants of 41 combinations, 203 F₃lines of 32 combinations and 25 F₄lines of 10 combinations were selected from the hybrid population for next generation, respectivly. (3) The results of preliminary yield trial and yield trial, three ?? lines and two ?? lines were selected as the promising lines. These lines will be tested for further yield test and put in regional adaptive trials. 3. Study on the improvement of quantity and quality of soybean protein. (1) Proper fertilization levels must be studied for increase of soybean yield. (2) Although lime and leaf spray of urea influened on the maturity, branch numbers, pod numbers per plant and grain numbers per plant in general, however, these had no positive effects on yield increase under fertrlizer levels used in this experiment. (3) Sulfur from ammonium sulfate fertilizer was ineffective for increase sulfur containing amino acids such as methionine and cystine. 4. Isolation and selection of Rhizobium strains collectecd in soybean fields. (1) Selected strains were confirmed as soybean nodulation bacteria by their cultural characteristics and by physiological test. (2) Strains isolated from cv. Dongbuktae, Jangyeobkong and Wayne were good in nodulation and colony formation. 5. Study on the variation of protein and amino acid composition of protein due to varietal differences and fertilizations. (1) Protein content was varied with varieties. (2) Fertilizations used in this experiment did not affect on protein content. (3) Contents of sulfur bearing amino acids were relatively high in varieties used and were 5.1∼6.4 grams of amino acid per 100 grams of protein, but the differences among the varieties and different fertilizations were not significant. 6. Studies on the nutiritive values of locally produced soybean for the broiler chicken. (1) Although the weight gain of chicks, feed intake and feed efficiency wre not statistically different among three varieties used, however, weight gain of chicks fed with cv. Bongyi was the highest. (2) Chicken fed with soybean grew better than those fed with fish meal or casein. (3) There was no significant difference among soybean varieties used in apparent metabolizable energy and true metabolizable energy.

Enhanced Lysosomal Activity Is Involved in Bax Inhibitor-1-induced Regulation of the Endoplasmic Reticulum (ER) Stress Response and Cell Death against ER Stress. INVOLVEMENT OF VACUOLAR H⁺-ATPASE (V-ATPASE)

Lee, G.-H.,Kim, D.-S.,Kim, H.-T.,Lee, J.-W.,Chung, C.-H.,Ahn, T.,Lim, J.M.,Kim, I.-K.,Chae, H.-J.,Kim, H.-R. The American Society for Biochemistry and Molecula 2011 The Journal of biological chemistry Vol.286 No.28

Comparison of dual antiplatelet therapy prescribed as one-pill versus two-pill regimen

Lim, W.-H.,Chae, I.-H.,Yoon, C.-H.,Choi, D.-J.,Lim, S. W.,Park, W. J.,Doh, J.-H.,Kim, S.-H.,Kim, M.-A.,Lee, S.-H. F K SCHATTAUER VERLAGSGESELLSCHAFT MBH 2016 Thrombosis and haemostasis Vol.116 No.1

Development of An Intelligent Laboratory Information System for Health Promotion

Chae, Y. M.,Lim, H. S.,Lee, J. H.,Bae, J. H.,Bae, M. Y.,Cho, K. W. 한국전문가시스템학회 1998 학술대회 Vol.2 No.1

This study aimed to develop an Intelligent Laboratory Information System (ILIS) for the community health promotion center in Kwachun city to help process an increasing number of laboratory test data in an efficient manner and to support the clinical decision-making of public health doctors. An ILIS consists of two modules: result interpretation and therapy advisor. An object-oriented rule editor, the Intelligent Rule Element (IRE), was used to develop the system. A sample of 188 cases was used for validation of the system. The cases were put through the system to determine whether they were in agreement with the physician's judgement. Overall, the system correctly predicted 89% of the cases. The integration of an ILIS with the health promotion information system will automate the entire laboratory test process from order entry to diagnosis and therapy advice, improving efficiency of the process and reducing workloads of health workers. Implications for future improvement and the role of an ILIS were also discussed.

Fabrication and magnetic properties of Fe nanostructures in anodic alumina membrane

Lim, J.-H.,Chae, W.-S.,Lee, H.-O.,Malkinski, L.,Min, S.-G.,Wiley, J. B.,Jun, J.-H.,Lee, S.-H.,Jung, J.-S. American Institute of Physics 2010 JOURNAL OF APPLIED PHYSICS - Vol.107 No.9

Usefulness of serum bilirubin levels as a biomarker for long-term clinical outcomes after percutaneous coronary intervention

Kim, H. W.,Choi, D. H.,Lim, L.,Lee, Y. M.,Kang, J. T.,Chae, S. S.,Ki, Y. J.,Song, H.,Koh, Y. Y. Springer Science + Business Media 2015 Heart and vessels Vol.30 No.6

Cell cloning-on-the-spot by using an attachable silicone cylinder

Park, H.B.,Son, W.,Chae, D.H.,Lee, J.,Kim, I.W.,Yang, W.,Sung, J.K.,Lim, K.,Lee, J.H.,Kim, K.H.,Park, J.I. Academic Press 2016 Biochemical and biophysical research communication Vol.474 No.4

Cell cloning is a laboratory routine to isolate and keep particular properties of cultured cells. Transfected or other genetically modified cells can be selected by the traditional microbiological cloning. In addition, common laboratory cell lines are prone to genotypic drift during their continual culture, so that supplementary cloning steps are often required to maintain correct lineage phenotypes. Here, we designed a silicone-made attachable cloning cylinder, which facilitated an easy and bona fide cloning of interested cells. This silicone cylinder was easy to make, showed competent stickiness to laboratory plastics including culture dishes, and hence enabled secure isolation and culture for days of selected single cells, especially, on the spots of preceding cell-plating dishes under microscopic examination of visible cellular phenotypes. We tested the silicone cylinder in the monoclonal subcloning from a heterogeneous population of a breast cancer cell line, MDA-MB-231, and readily established independent MDA-MB-231 subclones showing different sublineage phenotypes.

Shini-San Inhibits Mast Cell-Dependent Immediate-Type Allergic Reactions

Kim, H.M.,Lee, Y.H.,Chae, H.J.,Kim, H.R.,Baek, S.H.,Lim, K.S.,Hwang, C.Y. The Society of Korean Medicine Ophthalmology 2000 한방안이비인후피부과학회지 Vol.13 No.2

Shini-San has been used for treatment of allergic disease in Korea. However, its effect in experimental models remains unknown. The mast cell plays a pivotal role in initiating al1ergic response by secreting intracytoplasmic granular mediators such as histamine. The present report describes an inhibitory effect of Shini-San on mast cell-mediated immediate-type al1ergic reactions. Topical application of compound 48/80 can induce an ear swelling response in normal ($WBB6F_1-+/+$) mice but not in congenic mast cell-deficient $WBB6F_1-W/W^v$ mice. Shini-San inhibited concentration dependent mast cell-dependent ear swelling response induced by compound 48/80 in normal mice. Shini-San inhibited concentration-dependent passive cutaneous anaphylaxis induced by anti-dinitrophenyl (DNP) immunoglobulin E (IgE) in rats by topical application. Shini-San also inhibited in concentration-dependent fashion the histamine release from the rat peritoneal mast cells by compound 48/80 or anti-DNP IgE. Moreover, Shini-San had a significant inhibitory effect on compound 48/80-induced systemic anaphylactic reaction. These results indicate that Shini-San inhibits immediate-type allergic reactions by inhibition of mast cell degranulation in vivo and in vitro.