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Preparation of Dopamine Transporter-specific Antibodies Using Molecular Cloned Genes
Lee, Shee Yong,Im, Suhn Young,Kim, Kyeong Man 전남대학교 약품개발연구소 1999 약품개발연구지 Vol.8 No.1
Dopamine transporter (DAT) plays the most important role in terminating the actions of dopamines released into the synaptic deft DAT is also the target of various psychotropic drugs such as cocaine and amphetamine. In this study we prepared DAT specific antibodies using the 2nd extracellular loop of rat DAT as an antigen. The 2nd extracellular loop of the rat DAT was expressed in bacteria as a fusion protein with glutathione-S-transferase, and injected into rabbits to raise antibodies. Produced antibodies dearly recognized the rat DAT in ELISA, immunoblotting, and immunoprecipitation. As expected from the high sequence homology between the rat and human DAT, the antibodies raised for the rat DAT cross-reacted with the human DAT in the immunoblotting. Considering die specificity for DAT with wide range of applications such as ELISA, immunoblotting, and immunoprecipitation, these antibodies would be valuable tool for understanding the pharmacological actions of dopamine transporter and drug addiction.
Preparation of Dopamine Transporter-specific Antibodies Using Molecular Cloned Genes
Lee, Shee-Yong,Im, Suhn-Young,Kim, Kyeong-Man The Pharmaceutical Society of Korea 1999 Archives of Pharmacal Research Vol.22 No.3
Dopamine transporter (DAT) plays the most important role in terminating the actions of dopamines released into the synaptic cleft. DAT is also the target of various psychotropic drugs such as cocaine and amphetamine. In this study were prepared DAT-specific antibodies using the 2nd extracellular loop of rat DAT as an antigen. The 2nd extracellular loop of the rat DAT was expressed in bacterial as a fusion protein with glutathione-S-transferase, and injected ito rabbits to raise antibodies. Produced antibodies clearly recognized the rat DAT in ELISA, immunoblotting, and immumoprecipitation. As expected from the high sequence homology between the rat and human DAT, the antibodies raised for the rat DAT cross-reacted with the human DAT in the immunoblotting. Considering the specificity for DAT with wide range of applications such as ELISA, immunoblotting, and immunoprecipitation, these antibodies would be valuable tool for understanding the pharmacological actions of dopamine transporter and drug addition.
Cho, Dong-Im,Koo, Na-Youn,Chung, Woon-Jae,Kim, Tae-Sung,Ryu, Shi Yong,Im, Suhn Young,Kim, Kyeong Man 전남대학교 약품개발연구소 2002 약품개발연구지 Vol.11 No.-
NF-_κB that plays an important role in iNOS expression is one of the targets of various potential anti-inflammatory agents including resveratrol. Resveratol contains a structural similarity with estrogen, and there has been speculation about resveratrol as estrogen agonist. In this study, the mechanism and structural requirements of resveratrol and related hydroxystilbenes for the inhibition of LPS-induced nitric oxide production were studied in macrophage cells (RAW 264.7 and J774) by comparing its effect on LPS-induced NF-_κB translocation and nitric oxide production, and by considering the possibility of involvement of an estrogen receptor. LPS-induced nitric oxide production was inhibited only when cells were treated with resveratol prior to stimulation with LPS, suggesting that resveratol does not affect the enzyme itself, A higher concentration of resveratol than needed for the inhibition of nitric oxide production was required for the inhibition of NF-_κB mobilization or iNOS expression. Estrogen and diethylstilbesterol, an estrogen agonist, caused only weak inhibition of nitric oxide production, and the effects of resveratol were not noticeably blocked by ICI-182780, an estrogen antagonist. Structure-activity analysis of resveratol and nine hydroxystilbenes suggests that the structural balance between oxygen functional groups on the bezene rings is important for their activity. Our results suggest that resveratol might act on other cellular targets as well as NF-_κB at the initial stage of gene expression. Unique structural features of hydroxystilbenes are needed for suppression of nitric oxide production and it is unlikely that estrogen receptor is involved in it.
발목 삠을 경험한 20대 성인에게 적용한 종아리뼈 재위치 테이핑이 균형과 심리적 수준에 미치는 영향
임진구 ( Jin-gu Im ),김가은 ( Ka-eun Kim ),김주원 ( Joo-won Kim ),염민우 ( Min-woo Yum ),위정은 ( Jeong-eun Wi ),장윤정 ( Yoon-jeong Jang ),최용준 ( Yong-jun Choi ),김선엽 ( Suhn-yeop Kim ) 대전대학교 기초과학연구소 2023 혜화보건의료학회지 Vol.1 No.1
Background: Many people suffer from an ankle sprain, which is damaged by a sudden external rotation of the ankle. Ankle sprain causes the distal fibula to be positioned anteriorly and ankle instability develops after an injury. Purpose: This study aims to investigate the effects of fibular repositioning taping (FRT) on the static, dynamic balance and perception level of stability, confidence, and reassurance of the ankle in subjects who have experienced ankle sprain (AS) and subjects who have never experienced it (NAS).Methods: In this study, 54 young adults (male: 30, female: 24) participated and were divided into AS group (n=24) and the NAS group (n=30). Subjects were tested in the single-leg stance test for static balance, the star excursion balance test, and the dynamic leap balance test for dynamic balance in a randomized trial. After the FRT was applied, three tests were performed once again and a perception level test was performed last. The static balance test was measured by using Wii balance board. Result: The results of this study showed that there was not a significant difference between the two subject groups. However, after FRT was applied, two subject groups showed significant increases in static and dynamic balance regardless of experienced an ankle sprain or not. In the perception level test, two subject groups showed increased stability, confidence, and reassurance after applying the FRT. Conclusions: The FRT is effective for improving static balance, dynamic balance, perception level.
Choi, Hong-Seok,Cho, Dong-Im,Choi, Hoo-Kyun,Im, Suhn-Yong,Ryu, Shi-Yong,Kim , Kyeong-Man The Pharmaceutical Society of Korea 2004 Archives of Pharmacal Research Vol.27 No.12
The effects of four tanshinones isolated from Tanshen (the root of Salvia miltiorrhiza Bunge, Labiatae) were tested for their inhibition of nitric oxide production in macrophage cells, and the underlying molecular mechanisms studied. Of the four tanshinones used, 15, 16-dihydrotanshinone- I, tanshinone-IIA and cryptotanshinone, but not tanshinone I, demonstrated significant inhibition of the LPS-induced nitric oxide production in RAW 264.7 cells, with calculated $IC_{50}$ values of 5, 8, and 1.5 ${\mu}M$ , respectively. Tanshinones exerted inhibitory activities on the LPS-induced nitric oxide production only when applied concurrently with LPS, and tanshinone- IIA and cryptotanshinone were found to inhibit LPS-induced NF-$_KB$ mobilization and extracellular- regulated kinase (ERK) activation, respectively. These results suggest that tanshinones inhibit LPS-induced nitric oxide generation by interfering with the initial stage of LPS-induced expression of certain genes. NF-$_KB$ and ERK could be the molecular targets for tanshinones for the inhibition of LPS-induced nitric oxide production in macrophage cells.
인체 인터루킨 - 2 - ( IL - 2 ) Peptide 의 단일클론 항체 생산
이영익,김영훈,임선영,함경수,염현정,한문희 ( Young Ik Lee,Young Hoon Kim,Suhn Yong Im,Hyeon Jeong Yeom,Kyong So Hahm,Moon Hi Han ) 생화학분자생물학회 1991 BMB Reports Vol.24 No.2
Monoclonal antibody against Interleukin-2 peptide was generated. One hundred four hybridomas which were survived in HAT media produced anti-IL2 antibody. Among 104 hybridomas, eleven clones has been identified to produce IgG1 type monoclonal antibody which has kappa light chain after ELISA subtyping analysis. After purification of IgG1 monoclonal antibody, heavy and light chain of immunoglobulin has been identified whose molecular weights are 55 kD and 25 kD, respectively. These IL-2 antibodies were confirmed using immunoblotting techniques. IgGl type IL-2 monoclonal antibody were bound to 133 amino acids IL-2 antigen whose M.W. is 15 kD. These monoclonal antibodies were used for purifying IL-2 peptides. IL-2 ELISA systems were generated using immuno-affinity purified IL-2 as antigens.
Inhibition of interleukin-12 production in lipopolysaccharide-activated macrophages by curcumin
Kang, Bok Yun,Chung, Su Wol,Chung, Woon Jae,Im, Suhn Young,Hwang, Seung Yong,Kim, Tae Sung 전남대학교 약품개발연구소 1999 약품개발연구지 Vol.8 No.1
Pharmacological control of interleukin-12 production may be a key therapeutic strategy for modulating immunological diseases dominated by type-1 cytokine responses. In this study we investigated the effects of curcumin (1,7-bis[4-hydroxy-3-methoxyphenyl]-1,6-heptadiene-3,5-dione) on the production of interleukin-12 from mouse macrophages stimulated with lipopolysaccharide. Curcumin potently inhibited the production of interleukin-12 in a dose-dependent manner. The effect of curcumin on interleukin-12 p40 promoter activation was analyzed by transfecting RAW264.7 monocytic cells with p40 promoter/reporter constructs. The repressive effect mapped to a region in the p40 promoter containing a binding site for nuclear factor κB (p40-κB). Futhermore, activation of macrophages by lipopolysaccharide resulted in markedly enhanced binding activity to the κB site, which significantly upon addition of curcumin. These results suggest that curcumin-induced inhibition of interleukin-12 production in macrophages may explain some of the biological effects of curcumin including its anti-inflammatory activity.
Choi, Il-Whan,Sun-Kim, Il-Whan,Kim, Young-Suk,Ko, Hyun-Mi,Im, Suhn-Young,Kim, Jae-Hong,You, Hye-Jin,Lee, Yong-Chul,Lee, Jae-Hoon,Park, Young-Min,Lee, Hern-Ku Elsevier 2005 The Journal of allergy and clinical immunology Vol.116 No.3
<P><B>Background</B></P><P>Late-phase airway hyperresponsiveness (AHR) in asthma is considered the event leading to persistent inflammation in the lungs, but the molecular mechanisms involved in this process are poorly understood.</P><P><B>Objective</B></P><P>To examine the role of TNF-α in the development of a late AHR and airway inflammation in asthma.</P><P><B>Methods</B></P><P>We established a murine model of asthma with not only biphasic AHR to methacholine but also airway eosinophilia. The effect of TNF-α blockade was determined by using anti–TNF-α antibody and TNF-α knockout mice. Cytosolic phospholipase A<SUB>2</SUB> (cPLA<SUB>2</SUB>) mRNA expression and activity were assessed by using RT-PCR and 1-stearoyl-2-[1-<SUP>14</SUP>C] arachidonyl-sn-glycero-3-phosphocholine as the substrate, respectively.</P><P><B>Results</B></P><P>TNF-α blockade resulted in significant inhibition of the late AHR without affecting the early AHR, and reduction in airway eosinophilia and inflammation. cPLA<SUB>2</SUB> activity was increased in asthmatic lungs in a TNF-α–dependent way, and cPLA<SUB>2</SUB> inhibitor blocked late AHR and airway eosinophilia. TNF-α also stimulated the synthesis of cPLA<SUB>2</SUB> metabolites such as leukotriene B<SUB>4</SUB> and platelet-activating factor in the airway. Specific inhibitors of cPLA<SUB>2</SUB> metabolites inhibited the late AHR and airway eosinophilia.</P><P><B>Conclusions</B></P><P>TNF-α is the proximal key cytokine capable of developing late-phase AHR and subsequent airway inflammation through expression/activation of cPLA<SUB>2</SUB>.</P>