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랫드에서 l-muscone의 급성독성 및 아급성독성시험 연구
오승민,연제덕,남혜운,박대규,조명행,정규혁 성균관대학교 약학연구소 1998 成均藥硏論文集 Vol.10 No.1
l-Muscone is synthesized for use as substitutive material of musk which is the active ingredient of woohwangchungsimwon. The objective of this investigation was to evaluate the acute and subacute toxicity of l-muscone in rats. In oral acute toxicity test, SPF Sprague-Dawley male and female rats were gavaged with l-muscone of two doses(0, 5.0g/㎏). No dead animal and abnormal autopsy findings were found in control and treated group. Body weights were slightly decreased in both sexes of rats treated with 5.0g/㎏. Therefore, oral LD_50 of l-muscone was consider to be higher than 5.0g/㎏ in male and female rats. In intraperitoneal acute toxicity test, rats were injected intraperitoneally with dosages of 0, 1,000, 1,316, 1,732, 2,279 and 3,000 ㎎/㎏. Decreased body weights and motor activities were observed at high dose group. Intraperitoneal LD_50 of l-muscone were 1,920 ㎎/㎏ in male and female rats. In the subacute study. l-muscone was administrated orally to both sexes of rats for 4 weeks as several doses(0, 10, 100 and 1,000 ㎎/㎏). There were neither dead animals nor significant changes of body weights during the experimental period. In addition, no differences were found between control and treated groups in clinical signs, urinalysis, hematology, serum biochemical analysis, and other findings. Above data suggest that no observed adverse effect level of l-muscone in rats might be over 1,000 ㎎/㎏/day in this study.
오승민,남혜윤,김준수,연제덕,신대희,이진영,박대규,조명행,정규혁 성균관대학교 약학연구소 1998 成均藥硏論文集 Vol.10 No.1
The acute and subacute toxicity of New Woohwangchungsimwon(NWCH) which was used l-muscone as substitutive material of musk were investigated in S.D. rats. In intraperitoneal acute toxicity test, rats(Sprague-Dawley, SPF) were injected intraperitoneally with dosages of 0, 540, 750, 1,070, 1,500 and 3,000 ㎎/㎏. Body weights were significantly decreased at 540 ㎎/㎏ dose group in both sexes and abnormal autopsy findings were founded in both sexes at all treated groups. Intraperitoneal LD_50 of NWCH was 812.3 ㎎/㎏ in male and 872.3 ㎎/㎏ in female rats. In the subacute toxicity study, NWCH was administrated orally to both sexes of rats for 4 weeks as several doses(0, 320, 800 and 2,000 ㎎/㎏). There were neither dead animals nor significant changes of body weights during the experimental period. In addition, no differences were found between control and treated groups in clinical signs, urinalysis, hematology, serum biochemical analysis, and other findings. Above data strongly suggest that no observed adverse effect level of NWCH might be over 2,000 ㎎/㎏/day in this study.
Structural insight into the heme-based redox sensing by DosS from Mycobacterium tuberculosis.
Cho, Ha Yeon,Cho, Hyo Je,Kim, Young Min,Oh, Jeong Il,Kang, Beom Sik American Society for Biochemistry and Molecular Bi 2009 The Journal of biological chemistry Vol.284 No.19
<P>Mycobacterium tuberculosis is thought to undergo transformation into its non-replicating persistence state under the influence of hypoxia or nitric oxide (NO). This transformation is thought to be mediated via two sensor histidine kinases, DosS and DosT, each of which contains two GAF domains that are responsible for detecting oxygen tension. In this study we determined the crystal structures of the first GAF domain (GAF-A) of DosS, which shows an interaction with a heme. A b-type heme was embedded in a hydrophobic cavity of the GAF-A domain and was roughly perpendicular to the beta-sheet of the GAF domain. The heme iron was liganded by His-149 at the proximal heme axial position. The iron, in the oxidized form, was six-coordinated with a water molecule at the distal position. Upon reduction, the iron, in ferrous form, was five-coordinated, and when the GAF domain was exposed to atmospheric O(2), the ferrous form was oxidized to generate the Met form rather than a ferrous O(2)-bound form. Because the heme is isolated inside the GAF domain, its accessibility is restricted. However, a defined hydrogen bond network found at the heme site could accelerate the electron transferability and would explain why DosS was unable to bind O(2). Flavin nucleotides were shown to reduce the heme iron of DosS while NADH was unable to do so. These results suggest that DosS is a redox sensor and detects hypoxic conditions by its reduction.</P>
Yeon-Je Cho,Yun-Hee Choi,Byung-Loc Kim,Min-Hee Han,Hak-Sung Lee,Yong-Gi Jeong 한국피부과학연구원 2021 아시안뷰티화장품학술지 Vol.19 No.4
목적: 본 연구는 전남산 황칠, 천일염, 시금치 및 다시마 등 추출물의 최적 배합비율(optimal mixing ratio, OMR)을 선정하고, 이 를 이용하여 탈모증상 완화에 도움을 줄 수 있는 기능성 화장품 개발을 위해 수행되었다. 방법: 본 연구팀은 세포독성과 세포증식율 시험을 통해 OMR을 선정하였고, 최종 선정된 OMR에 대하여 항염증 및 항균활성 효과를 확인하였다. 결과: 세포독성은 OMR이 0.1:1:5:1 일 때 낮은 반면, 세포증식율은 높게 나타났으며, 항염증은 IL-6 및 iNOS의 발현을 효과적으로 억제하였다. 또한 항균활 성은 녹농균과 황색포도상구균에서 효과를 보였다. 결론: 본 연구는 탈모증상 완화에 도움을 줄 수 있는 기능성 화장품 개발을 위한 황칠 및 천일염 등에 대한 OMR (0.1:1:5:1)을 선정하였다. 최종 선정한 OMR은 낮은 세포독성과 높은 세포증식율, IL-6 및 iNOS 의 발현 억제 및 항균활성 효과가 확인되었다. 따라서 향후 탈모증상 완화에 도움을 줄 수 있는 기능성 화장품으로서 활용 가능성이 기대된다. Purpose: This study was conducted to select the optimal mixing ratio (OMR) of extracts including Dendropanax , sea salt, and others from Jeollanam-do and to develop functional cosmetics that can help alleviate hair loss symptoms. Methods: Our research team determined the OMR through cytotoxicity and cell proliferation tests, and confirmed the anti-inflammatory and anti-microbial effects of the final selected OMR. Results: The cytotoxicity was low when the OMR was 0.1:1:5:1, but cell proliferation was high, and anti-inflammatory activity effectively inhibited the expression of IL–6 and iNOS. The anti-microbial activity also had an effect on Pseudomonas aeruginosa and Staphylococcus aureus. Conclusion: This study selected OMR (1:0.1:5:1) to develop functional cosmetics that can help alleviate hair loss symptoms. The final selection of OMR confirmed low cytotoxicity, high cell proliferation, inhibition of expression of IL–6 and iNOS, and anti-microbial activity. Therefore, it is expected to serve as a functional cosmetic that can help alleviate hair loss symptoms in the future.
Cho, Eun Sook,Jin, Byung Rae,Sohn, Hung Dae,Choi, Kwang Ho,Kim, Soung Ryul,Kang, Seok Woo,Yun, Eun Young,Kim, Sang Hyun,Kim, Keun Young,Je, Yeon Ho,Kang, Seok Kwon 한국잠사학회 1998 한국잠사곤충학회지 Vol.40 No.2
To construct transfurmed Bm5 cells, Autographa californica nuclear polyhedrosis virus(AcNPV) IE1 gene, an immediate early viral gene was firstly used in this study. AcNPV IE1 gene, which shares on 95.3% nucleotide sequence homology with Bombyx mori nuclear polyhedrosis virus (BmNPV) IE1 gene, was isolated and cloned into pBluescript. Neomycin gene from pKO-neo was inserted under the control of the IE1 promoter to yield pAcIE1-neo. The plasmid pAcIE1-neo was transfected into Bm5 or Sf9 cells, and neomycin-resistant cells were selected in TC100 medium containing 10% fetal bovine serum (FBS) and 1 mg/ml G4l8 for two weeks. Individual clones were picked and each was amplified for further characterization. The genomic DNA from neomycin-resistant cells was isolated and characterized by PCR using AcNPV IEI gene-specific primers and by Southern blot analysis using neomycin gene probe. We concluded that AcNPV IE1 gene was functional in B. mori-derived Bm5 cells as well as Spodoptera fugjprrda-derived Sf9 cells to produce stably-transformed insect cells
Cho, Ha Yeon,Cho, Hyo Je,Kim, Myung Hee,Kang, Beom Sik Elsevier 2011 FEBS letters Vol.585 No.12
<P><B>Highlights</B></P><P>► A channel to the heme in DosS can be blocked by E87. ► DosT has an open channel to heme due to G85. ► DosS mutants with an open channel bind oxygen when they are exposed to air. ► DosT mutant is oxidized similarly to DosS without formation of an oxy-ferrous form.</P> <P><B>Abstract</B></P><P>Two sensor kinases, DosS and DosT, are responsible for recognition of hypoxia in <I>Mycobacterium tuberculosis</I>. Both proteins are structurally similar to each other, but DosS is a redox sensor while DosT binds oxygen. The primary difference between the two proteins is the channel to the heme present in their GAF domains. DosS has a channel that is blocked by E87 while DosT has an open channel. Absorption spectra of DosS mutants with an open channel show that they bind oxygen as DosT does when they are exposed to air, while DosT G85E mutant is oxidized similarly to DosS without formation of an oxy-ferrous form. This suggests that oxygen accessibility to heme is the primary factor governing the oxygen-binding properties of these proteins.</P>
Cho, Ha Yeon,Maeng, Seo Jin,Cho, Hyo Je,Choi, Yoon Seo,Chung, Jeong Min,Lee, Sangmin,Kim, Hoi Kyoung,Kim, Jong Hyun,Eom, Chi-Yong,Kim, Yeon-Gil,Guo, Min,Jung, Hyun Suk,Kang, Beom Sik,Kim, Sunghoon American Society for Biochemistry and Molecular Bi 2015 The Journal of biological chemistry Vol.290 No.49
<P>Many multicomponent protein complexes mediating diverse cellular processes are assembled through scaffolds with specialized protein interaction modules. The multi-tRNA synthetase complex (MSC), consisting of nine different aminoacyl-tRNA synthetases and three non-enzymatic factors (AIMP1–3), serves as a hub for many signaling pathways in addition to its role in protein synthesis. However, the assembly process and structural arrangement of the MSC components are not well understood. Here we show the heterotetrameric complex structure of the glutathione transferase (GST) domains shared among the four MSC components, methionyl-tRNA synthetase (MRS), glutaminyl-prolyl-tRNA synthetase (EPRS), AIMP2 and AIMP3. The MRS-AIMP3 and EPRS-AIMP2 using interface 1 are bridged via interface 2 of AIMP3 and EPRS to generate a unique linear complex of MRS-AIMP3:EPRS-AIMP2 at the molar ratio of (1:1):(1:1). Interestingly, the affinity at interface 2 of AIMP3:EPRS can be varied depending on the occupancy of interface 1, suggesting the dynamic nature of the linear GST tetramer. The four components are optimally arranged for maximal accommodation of additional domains and proteins. These characteristics suggest the GST tetramer as a unique and dynamic structural platform from which the MSC components are assembled. Considering prevalence of the GST-like domains, this tetramer can also provide a tool for the communication of the MSC with other GST-containing cellular factors.</P>