A rationally designed small molecule for identifying an <i>in vivo</i> link between metal–amyloid-β complexes and the pathogenesis of Alzheimer's disease

Beck, Michael W.,Oh, Shin Bi,Kerr, Richard A.,Lee, Hyuck Jin,Kim, So Hee,Kim, Sujeong,Jang, Milim,Ruotolo, Brandon T.,Lee, Joo-Yong,Lim, Mi Hee Royal Society of Chemistry 2015 Chemical Science Vol.6 No.3

<▼1><P>An <I>in vivo</I> chemical tool designed to target metal–Aβ complexes and modulate their activity was applied to the 5XFAD mouse model of Alzheimer’s disease (AD) demonstrating the involvement of metal–Aβ in AD pathology.</P></▼1><▼2><P>Multiple factors, including amyloid-β (Aβ), metals, and reactive oxygen species (ROS), are involved in the development of Alzheimer's disease (AD). Metal ions can interact with Aβ species generating toxic oligomers and ROS <I>in vitro</I>; however, the involvement of metal–Aβ complexes in AD pathology <I>in vivo</I> remains unclear. To solve this uncertainty, we have developed a chemical tool (<B>L2-b</B>) that specifically targets metal–Aβ complexes and modulates their reactivity (<I>i.e.</I>, metal–Aβ aggregation, toxic oligomer formation, and ROS production). Through the studies presented herein, we demonstrate that <B>L2-b</B> is able to specifically interact with metal–Aβ complexes over metal-free Aβ analogues, redirect metal–Aβ aggregation into off-pathway, nontoxic less structured Aβ aggregates, and diminish metal–Aβ-induced ROS production, overall mitigating metal–Aβ-triggered toxicity, confirmed by multidisciplinary approaches. <B>L2-b</B> is also verified to enter the brain <I>in vivo</I> with relative metabolic stability. Most importantly, upon treatment of 5XFAD AD mice with <B>L2-b</B>, (i) metal–Aβ complexes are targeted and modulated in the brain; (ii) amyloid pathology is reduced; and (iii) cognition deficits are significantly improved. To the best of our knowledge, by employing an <I>in vivo</I> chemical tool specifically prepared for investigating metal–Aβ complexes, we report for the first time experimental evidence that metal–Aβ complexes are related directly to AD pathogenesis.</P></▼2>

Human coagulation factor VIII domain-specific recombinant polypeptide expression

Choi, Su Jin,Jang, Ki Jung,Lim, Jeong-A,Kim, Hye Sun Korean Society of Hematology; Korean Society of Bl 2015 Blood Research Vol.50 No.2

<P><B>Background</B></P><P>Hemophilia A is caused by heterogeneous mutations in <I>F8</I>. Coagulation factor VIII (FVIII), the product of <I>F8</I>, is composed of multiple domains designated A1-A2-B-A3-C1-C2. FVIII is known to interact with diverse proteins, and this characteristic may be important for hemostasis. However, little is known about domain-specific functions or their specific binding partners.</P><P><B>Methods</B></P><P>To determine <I>F8</I> domain-specific functions during blood coagulation, the FVIII domains A1, A2, A3, and C were cloned from Hep3B hepatocytes. Domain-specific recombinant polypeptides were glutathione S-transferase (GST)- or polyhistidine (His)-tagged, over-expressed in bacteria, and purified by specific affinity chromatography.</P><P><B>Results</B></P><P>Recombinant polypeptides of predicted sizes were obtained. The GST-tagged A2 polypeptide interacted with coagulation factor IX, which is known to bind the A2 domain of activated FVIII.</P><P><B>Conclusion</B></P><P>Recombinant, domain-specific polypeptides are useful tools to study the domain-specific functions of FVIII during the coagulation process, and they may be used for production of domain-specific antibodies.</P>

Studies on Simultaneous Determination of Chlorophyll a and b, Pheophorbide a, and $\beta-Carotene$ in Chlorella and Spirulina Products

이영자,김소희,김진숙,한정아,서해점,임효정,최수영,Lee Young Ja,Kim So Hee,Kim Jin-Sook,Han Jeong A,Seo Hae Jeom,Lim Hyo Jeong,Choi Soo Young The Korean Society of Food Hygiene and Safety 2005 한국식품위생안전성학회지 Vol.20 No.3

역상 컬럼을 이용하여 건강기능식품 중 클로렐라 및 스피루리나제품에 함유되어 있는 엽록소 a, b,페오포르바이드 a 및 $\beta$-카로틴의 HPLC동시분석법을 확립하였으며, 첨가농도 $50\;\mug/ml$에서 엽록소 a, b, 페오포르바이드 a 및 $\beta$-카로틴에 대한 회수율시험결과, 각각 2.8, 6.0, 10.6 및 $10.4\%$의 상대표준편차와 70.3, 71.6, 60.1 및 $90.5\%$의 회수율을 각각 나타냈다. 이때 검출한계는 $0.1\sim1.0\;\mug/ml$, 정량한계는 $0.2\sim2.0\;\mug/ml$이었으며 검량선 상관계수도 0.995 이상의 직선성을 보여주었다. 국내유통 클로렐라 및 스피루리나제품에 대한 엽록소 a, b,페오포르바이드 a및 $\beta$-카로틴의 함유량을 분석한 결과 엽록소 a $121.g\sim543$, 엽록소 b $0.6\sim160.0$, 페오포르바이드 a 및 P-카로틴 $383.6\sim1713.7mg/ml$ 수준으로 나타났다. 엽록소 b의 함유량은 클로렐라제품에서 평균 374.0 mg/100 g 으로 스피루리나제품의 평균 10.5 mg/100 g 보다 30배 이상 함유하고 있는 것으로 확인되었다. 그러나 $\beta$-카로틴의 함유량은 스피루리나제품이 평균 1335.4 mg/100 g 로 클로렐라제품의 평균 495.0 mg/100 g 보다 평균 함유량에서 2.7배 높은 것으로 나타났다. 국내 건강기능식품공전 중 클로렐라 및 스피루리나제품의 엽록소 a b, 및 페오포르바이드 항목의 규격검사를 본 연구의 동시분석법으로 개정함으로써 각 성분 함량의 정량, 분석시간의 단축 및 비용절감 둥 시험방법을 크게 개선할 수 있을 것으로 기대된다. A simple and sensitive analysis method based on reverse phase (RP) HPLC with UV detector was developed for simultaneous determination of chlorophyll a and b, pheophorbide a and $\beta-Carotene$ in Chlorella and Spirulina products. For added concentration $(50\;\mug/ml)$ of chlorophyll a and b, pheophorbide a and $\beta-Carotene$, recoveries of those were 70.3, 71.6, 60.1 and $90.5\%$, respectively, with relative standard deviations of 2.8,6.0, 10.6 and $10.4\%$. Limit of detection and quantification had ranges of $0.1\sim1.0\;\mug/ml$ and $0.2\sim2.0\;\mug/ml$, respectively. Calibration curve was linear with correlation coefficient of 0.995 for chlorophyll a and b, pheophorbide a and $\beta-Carotene$. Results of simultaneous determination in Chlorella and Spirulina products were showed ranges of $121.g\sim543.0\;\mug/ml$ for chlorophyll a,$0.6\sim160.0\;\mug/ml$ for chlorophyll b, $19.2\sim60.3\;\mug/ml$ for pheophorbide a and $383.6\sim1713.7\;\mug/ml$ for $\beta-Carotene$, respectively. Chlorophyll b contents in Chlorella products were detected above 30 times level to those in Spirulina products. $\beta-Carotene$ contents in Spirulina products were detected 2.7 times level to those in Chlorella products.

Human coagulation factor VIII domain-specific recombinant polypeptide expression

Su Jin Choi,Ki Jung Jang,Jeong-A Lim,Hye Sun Kim 대한혈액학회 2015 Blood Research Vol.50 No.2

BackgroundHemophilia A is caused by heterogeneous mutations in F8. Coagulation factor VIII (FVIII),the product of F8, is composed of multiple domains designated A1-A2-B-A3-C1-C2. FVIIIis known to interact with diverse proteins, and this characteristic may be important forhemostasis. However, little is known about domain-specific functions or their specificbinding partners. MethodsTo determine F8 domain-specific functions during blood coagulation, the FVIII domainsA1, A2, A3, and C were cloned from Hep3B hepatocytes. Domain-specific recombinantpolypeptides were glutathione S-transferase (GST)- or polyhistidine (His)-tagged,over-expressed in bacteria, and purified by specific affinity chromatography. ResultsRecombinant polypeptides of predicted sizes were obtained. The GST-tagged A2 polypeptideinteracted with coagulation factor IX, which is known to bind the A2 domain of activatedFVIII. ConclusionRecombinant, domain-specific polypeptides are useful tools to study the domain-specificfunctions of FVIII during the coagulation process, and they may be used for productionof domain-specific antibodies.

소아의 급성 인두염에서 연쇄구균 신속항원검사의 유용성

임기현 ( Ki Hyun Lim ),최우진 ( Woo Jin Choi ),김미진 ( Mi Jin Kim ),김영호 ( Young Ho Kim ),정지아 ( Ji A Jung ),양승 ( Seung Yang ),황일태 ( Il Tae Hwang ),이혜란 ( Hae Ran Lee ) 대한소아알레르기호흡기학회(구 대한소아알레르기 및 호흡기학회) 2006 소아알레르기 및 호흡기학회지 Vol.16 No.1

목 적 : 최근 소아 급성 인두염에서 부적절한 항생제 투여가 문제시 되고 있으나 증상만으로 항생제 사용 여부를 결정하기는 어려우며 인두배양검사가 확진 방법이다. 그러나 검사에 시간이 요구되므로 미국소아과학회에서는 배양검사와 신속항원검사를 시행하여 신속항원검사가 음성이면 항생제 사용을 보류하도록 권하고 있다. 저자들은 이와 같은 지침을 국내에서도 적용할 수 있는지 알아보고자 연쇄구균 신속항원검사의 유용성을 조사하였다. 방 법 : 2004년 9월부터 2005년 8월까지 한림대학교 강동성심병원 소아과를 방문하여 급성 인두염으로 진단받은 70명을 대상으로 하였다. 편도 삼출, 발열, 피부 발진 등의 임상적 특징과 내원 전 항생제 투여 여부를 조사하고, CBC, ASO, CRP 등의 혈액 검사와 인두에서 연쇄구균 신속항원검사와 배양검사를 시행하였다. 결 과 : 1) 인두배양검사에서 A군 연쇄구균이 분리된 양성례는 14례(20.0%)이었으며, 연쇄구균 신속항원검사는 22례(31.4%)에서 양성이었다. 인두배양검사에 대한 신속항원검사의 민감도는 78.6 %, 특이도는 80.4%이었다. 양성예측치는 50.0%, 음성예측치는 93.8%이었다. 2) 발열 유무에 따라 인두배양검사와 신속항원검사에는 차이가 없었다. 3) 편도삼출의 여부에 따라 인두배양검사 양성률에 차이가 없었으며, 신속항원검사는 오히려 편도삼출이 없는 경우에서 양성이 더 많았다. 4) 성홍열양 발진이 있는 경우에 인두배양검사와 신속항원검사에서 양성이 많았다. 5) 인두배양검사 양성은 항생제를 이전에 투여한 경우에 35례 중 3례(8.57%) 이전에 항생제를 투여하지 않은 경우의 35례 중 11례(31.43%)로 낮아 항생제 사용이 인두배양검사에 영향을 주었다. 이전에 인두배양검사는 양서d음성이어도 신속항원검사는 양성인 경향이 있었다. 6) 인두배양검사와 신속항원검사에 따라 혈액의 백혈구 수, ASO, CRP 값은 각각 통계학적인 차이가 없었다. 결 론 : 소아 급성 인두염에서 신속항원검사는 연쇄구균 감염의 진단에 유용한 도구로서 항생제의 조기 투여 여부를 결정하는 데 도움이 되리라 사료된다. Purpose : The purpose of this study was to evaluate the utility of rapid antigen detection test for group A streptococci in children with acute pharyngitis. Methods : Seventy children with acute pharyngitis enrolled the study, who visited the Department of Pediatrics, Kangdong Sacred Heart Hospital, Hallym Medical Center from September 2004 to August 2005. The presence of clinical features such as tonsillar exudates, fever, scarletiniform rash and previous antibiotic use were evaluated and throat swab culture, rapid antigen detection test for group A Streptococci, CBC, ASO, and CRP were performed. Results : Among 70 children with acute pharyngitis, the prevalence of group A beta-hemolytic Streptococcus by throat swab culture and rapid antigen detection test was 14(20.0 %) and 22(31.4%), respectively. The sensitivity of the rapid antigen detection test was 78.6 %, specificity was 80.4%, the positive predictive value was 50.0% and the negative predictive value was 93.8%. The positivity of the throat swab culture and rapid antigen detection test showed no signigicant differences according to the presence of the clinical symptoms except scarletiniform rash. Laboratory data such as CBC, ASO or CRP proved not valuable to predict the positivity of the throat swab culture and/or rapid antigen detection test for group A Streptococcus. There was a tendency of negative throat swab culture and positive rapid antigen detection test in cases of previous antibiotic use. Conclusion : The rapid antigen detection test can be a useful diagnostic tool for early detection of the group A Streptococcus in children with acute pharyngitis and can be helpful for the clinician to reduce the inappropriate use of antibiotics. [Pediatr Allergy Respir Dis (Korea) 2006;16:57-65]

Design, Synthesis and Binding Affinity of 3´-Fluoro Analogues of Cl-IB-MECA as Adenosine A₃ Receptor Ligands

Lim, Moo Hong,Kim, Hea Ok,Moon, Hyung Ryoung,Lee, Seung Jin,Chun, Moon Woo,Gao, Zhan-Guo,Melman, Neli,Jacobson, Kenneth A.,Kim, Joong Hyup,Jeong, Lak Shin 梨花女子大學校 藥學硏究所 2003 藥學硏究論文集 Vol.- No.12

Several 3'-fluoro analogues, la, lb, and lc of selective and potent adenosine A₃ receptor agonist, Cl-IB-MECA were synthesized from D-xylose via highly regioselective opening of lyxo-epoxides, 8a and 8b with fluoride anion. Compared to the high binding affinity of CI-IB-MECA to the A₃ adenosine receptor, the corresponding 3'-fluoro derivative showed remarkably decreased binding affinity, indicating that 3'-hydroxyl group acts as hydrogen bonding acceptor, not hydrogen bonding donor like fluorineatom in binding to the A₃ adenosine receptor.

Upregulation of NAD(P)H:quinone oxidoreductase by radiation potentiates the effect of bioreductive beta-lapachone on cancer cells.

Choi, Eun K,Terai, Kaoru,Ji, In-Mi,Kook, Yeon H,Park, Kyung H,Oh, Eun T,Griffin, Robert J,Lim, Byung U,Kim, Jin-Seok,Lee, Doo S,Boothman, David A,Loren, Melissa,Song, Chang W,Park, Heon Joo Stockton Press 2007 Neoplasia Vol.9 No.8

<P>We found that beta-lapachone (beta-lap), a novel bioreductive drug, caused rapid apoptosis and clonogenic cell death in A549 human lung epithelial cancer cells in vitro in a dose-dependent manner. The clonogenic cell death caused by beta-lap could be significantly inhibited by dicoumarol, an inhibitor of NAD(P)H:quinone oxido-reductase (NQO1), and also by siRNA for NQO1, demonstrating that NQO1-induced bioreduction of beta-lap is an essential step in beta-lap-induced cell death. Irradiation of A549 cells with 4 Gy caused a long-lasting upregulation of NQO1, thereby increasing NQO1-mediated beta-lap-induced cell deaths. Although the direct cause of beta-lap-induced apoptosis is not yet clear, beta-lap treatment reduced the expression of p53 and NF-kappaB, whereas it increased cytochrome C release, caspase-3 activity, and gammaH2AX foci formation. Importantly, beta-lap treatment immediately after irradiation enhanced radiation-induced cell death, indicating that beta-lap sensitizes cancer cells to radiation, in addition to directly killing some of the cells. The growth of A549 tumors induced in immunocompromised mice could be markedly suppressed by local radiation therapy when followed by beta-lap treatment. This is the first study to demonstrate that combined radiotherapy and beta-lap treatment can have a significant effect on human tumor xenografts.</P>

Intestinal Nematodes from Small Mammals Captured near the Demilitarized Zone, Gyeonggi Province, Republic of Korea

Deok-Gyu Kim,Jae-Hwan Park,Jae-Lip Kim,Bong-Kwang Jung,Sarah Jiyoun Jeon,Hyemi Lim,Mi Youn Lee,Eun-Hee Shin,Terry A. Klein,Heung-Chul Kim,Sung-Tae Chong,Jin-Won Song,Luck-Ju Baek,Jong-Yil Chai 대한기생충학열대의학회 2015 The Korean Journal of Parasitology Vol.53 No.1

A total of 1,708 small mammals (1,617 rodents and 91 soricomorphs), including Apodemus agrarius (n = 1,400), Microtus fortis (167), Crocidura lasiura (91), Mus musculus (32), Myodes (= Eothenomys) regulus (9), Micromys minutus (6), and Tscherskia (= Cricetulus) triton (3), were live-trapped at US/Republic of Korea (ROK) military training sites near the demilitarized zone (DMZ) of Paju, Pocheon, and Yeoncheon, Gyeonggi Province from December 2004 to December 2009. Small mammals were examined for their intestinal nematodes by necropsy. A total of 1,617 rodents (100%) and 91 (100%) soricomorphs were infected with at least 1 nematode species, including Nippostrongylus brasiliensis, Heligmosomoides polygyrus, Syphacia obvelata, Heterakis spumosa, Protospirura muris, Capillaria spp., Trichuris muris, Rictularia affinis, and an unidentified species. N. brasiliensis was the most common species infecting small mammals (1,060; 62.1%) followed by H. polygyrus (617; 36.1%), S. obvelata (370; 21.7%), H. spumosa (314; 18.4%), P. muris (123; 7.2%), and Capillaria spp. (59; 3.5%). Low infection rates (0.1-0.8%) were observed for T. muris, R. affinis, and an unidentified species. The number of recovered worms was highest for N. brasiliensis (21,623 worms; mean 20.4 worms/infected specimen) followed by S. obvelata (9,235; 25.0 worms), H. polygyrus (4,122; 6.7 worms), and H. spumosa (1,160; 3.7 worms). A. agrarius demonstrated the highest prevalence for N. brasiliensis (70.9%), followed by M. minutus (50.0%), T. triton (33.3%), M. fortis (28.1%), M. musculus (15.6%), C. lasiura (13.2%), and M. regulus (0%). This is the first report of nematode infections in small mammals captured near the DMZ in ROK.

Inappropriate Gonadotropin Secretion in Polycystic Ovary Syndrome: The Relationship with Clinical, Hormonal and Metabolic Characteristics

( A Ra Shim ),( Yu Im Hwang ),( Kyung Jin Lim ),( Young Mi Choi ),( Young Eun Jeon ),( Seok Kyo Seo ),( Si Hyun Cho ),( Young Sik Choi ),( Byung Seok Lee ) 대한산부인과학회 2011 Obstetrics & Gynecology Science Vol.54 No.11

Objective To evaluate the clinical, hormonal and metabolic characteristics according to luteinizing hormone (LH)/follicle stimulating hormone (FSH) ratio in women with polycystic ovary syndrome (PCOS). Methods A total of 225 women with PCOS were included in this study. They were divided into two groups according to LH/FSH ratio; group A (LH/FSH < 2, n=160) and group B (LH/FSH > 2, n=65). We compared clinical, hormonal and metabolic characteristics including age at menarche, body mass index (BMI), blood pressures, ovarian volume, and serum androgen levels between the two groups. Serum glucose and insulin levels, fasting glucose/insulin ratio, homeostatic model assessment of insulin resistance (HOMA-IR), and lipid profi les were also compared between the two groups. Pearson correlation coeffi cients were used to evaluate correlation between the LH/FSH ratio and various parameters. Results Age of menarche in the group B was signifi cantly later than that in the group A. BMI, waist-hip ratio and blood pressures were signifi cantly lower in the group B compared to those of the group A. Ovarian volume and serum levels of LH, estradiol, total testosterone, sex-hormone binding globulin, 17-hydroxyprogesterone (OHP), and high density lipoprotein were signifi cantly higher in the group B. Postprandial 2 hours glucose, insulin and HOMA-IR were signifi cantly higher in the group A. After adjustment of BMI, the LH/FSH ratio was signifi cantly positively correlated with age at menarche, ovarian volume, total testosterone levels, and 17-OHP levels. Conclusion The inappropriate gonadotropin secretion may be negatively correlated with BMI, and positively with age of menarche and increased ovarian androgen production irrespective of BMI in women with PCOS. However, it may not be associated with metabolic characteristics.

3-D 동작 분석 장치를 이용한 한국성인(20대, 30대)의 동작 범위에 관한 연구

임현균,김진호,박수찬,류신아,이영신 한국운동역학회 1998 한국운동역학회지 Vol.8 No.2

Devices, such as a goniometer, have been used to measure a human being's active range of motion, but interferences between subjects and testers have affected the result precision. In this study, 56 dynamic motion ranges were measured using a 3-D motion analysis system which is quite accurate (2mm error for 1m). The 3-D motion analysis system consists of VICON 140, data acquisition system, and data analysis program (KRISSMAS). 80 Korean subjects (40 males, 28±6.1 aged and 40 females, 28.4±6.9 aged) participated. Subjects were divided into 4 groups by Ro¨hrer's index based on National Anthropometric Survey of Korea 1997. Basic statistical analysis were performed using SPSS (ver. 8.0) programs after refining data. Various comparisons were performed for the right and left side, male and female, age groups (20's, 30's. and 40's-60's) using previous studies. The results show that female subjects have more flexible joints in the neck, hip, ankle, and wrist, while male subjects have more flexibility in their waist. But most of the significances were not high (>0.05). The prediction that the right side of Korean bodies would be mort flexible was not a good hypothesis. And the joints flexibilities are not correlated with Ro¨hrer's Index.