G.I.S.H. 수술과 복식 전자궁적출술의 비교

지일운,허준용,서호석,박용균,조수용,주갑순,이규완,구병삼 대한산부인과내시경학회 1995 대한산부인과내시경학회잡지 Vol.7 No.1

protoplast-fusion에 依한 澱粉에서 Ethanol의 單段醱酵能 酵母 開發 : I. Characteristics of two yeast strains and conditions for the protoplast formation and reeneration as a preliminary step in interspecific protoplast-fusion I. Interspecific Protoplast-fusion 을 爲한 酵母菌林의 諸特性과 Protoplast 調製 및 Regeneration 條件

吳秉夏,黃殷成,李炯周,李啓瑚,朴官和,張海東,徐鉉昌 서울大學校農科大學 1984 서울대농학연구지 Vol.9 No.1

澱粉으로 부터의 alcohol 醱酵能을 增進시키기 爲하여 澱粉糖化性 菌株인 Saccharomyces diastaticus와 優秀한 alcohol 醱酵性 菌株인 Saccharomyces uvarum을 母菌株로 하여 이들간의 同屬異種間 原形質融合(interspecific protoplast fusion)을 通한 優秀한 澱粉醱酵 性 alcohol 生産性 菌株를 새로이 開發할 目的에서 다음과 같은 一漣의 實驗結果를 얻었다. S. diastaticus의 醱酵液과 S. diastaticus+S. uvarum 混合醱酵液의 風味特性등을 確認하였다. 風味成分 抽出은 methylene chloride와 diethylether를 가지고 neutral flavor fraction과 acidic flavor fraction으로 나누었고 gas chromatography를 通하여 同定 및 定量하였다. Neutral flavor fraction의 경우 S. diastaticus+S. uvarum 混合醱酵液이 S. diastaticus 醱酵液보다, ester成分中에서는 ethyl acetate와 ethyl undecanoate가 더 많았고, alcohol 成分中에서는 n-propanol과 n-butanol이 더 많았다. Acidic flavor fraction의 경우 C??~C?? fatty acid가 同定 및 定量되었는데 S. diastaticus+S. uvarum 混合醱酵液이 S. diastaticus 醱酵液보다 lauric acid, caprylic acid, capric acid 含量이 두드러지게 많았다. S. diastaticus의 glucoamylase 生産性, glucoamylase의 分離 精製, 酵素力價 그리고 酵素學的 特性에서 optimum pH는 5.0, optimum temperature는 55℃ 이었다. S. diastaticus와 S. uvarum을 母菌株로 이들 간의 protoplast fusion을 위한 基礎的인 硏究로서 두 菌株의 諸特性과 protoplast調製의 最適條件을 決定하고 protoplast의 regeneration 條件의 確立을 도모하였다.두 菌의 生育曲線에서 모두 培養開始 7~8 時間만에 對數期 中期에 到達되었으므로 protoplast 調製는 이 時期의 細胞를 쓰기로 하였다. Generation time은 S. diastaticus가 1.04, S. uvarum이 1.38 時間이었다. 細胞의 크기는 S. diastaticus 44.10?㎛³, S. uvarum 99.67㎛³로 S. uvarum이 2倍나 컸다. DNA 含量은 細胞 當 S. diastaticus 44.3fg, S. uvarum 37.6fg이었다. 30% glucose 및 soluble starch에 대한 두 菌株의 ethanol 醱酵能은 glucose에 對하여 S. uvarum 11.4%, S. diastaticus 8.9% 이었고 soluble starch에 對하여는 S. diastaticus 만이 6.9%이었다. 두 菌株는 generation time, 細胞크기 및 DNA 含量 等으로 보아 diploid strain임을 알 수 있었고, 融合株 選拔을 위한 marker 로는 Sacch. uvarum의 melibiose 資化能의 차이를 利用할 수 있음을 밝혔다. Protoplast의 調製에는 β-glucuronidase와 Zymoyase를 使用하였는데 두 酵素 反應最適條件은 β-glucuronidase는 pH 8.0에서 10% 濃度의 溶液으로, Zymolyase는 pH7.5에서 20㎛/ml의 濃度의 溶液으로 하여 모두 70分間 處理하는 것으로 決定하였으나 이 정도의 處理時間에서는 protoplast가 극히 不安定하게 되어 regeneration frequency가 떨어지는 것을 確認하였으며, 特히 Zymolyase 處理로 얻어진 protoplast의 regeneration率이 낮은 것은 Zymolyase中에 不純物로 微量 混在한 protease가 protoplast의 노출된 membrane-bound protein을 分解함으로써 protoplast를 破壞시키기 때문인 것으로 추측되었다. 融合實驗에 利用할 수 있을 정도의 regeneration frequency를 얻기 위해서는 Zymolyase를 45分間 處理하여 얻은 protoplast를 1.5%의 polyvinylpyrrolicone이 加해진 OYPD培地에서 重層法으로 展開하여 regeneration시키는 것이 좋은 것으로 판명되었다. As preliminary steps of protoplast fusion between Saccharomyces diastaticus and S. uvarum to develop a fusant of higher ethanol production from starch, characteristics of the two presumptive parent strains, optimal conditions for protoplast preparation and conditions for highrer regeneration frequency were investigated. To determine flavor characteristics of the parent strains, neutral and acidic flavor fractions were extracted from liquids fermented by S. diastaticus and S. diastaticus + S. uvarum with methylene chloride and diethly ether. The liquid by the mixed culture produced more ethly acetate, ethyl undecanoate, n-propanol, n-butanol, lauric acid, caprylic acid and capric acid than that by S. diastaticus. Glucoamylase from S. diastaticus was purified and activity, productivity, and characteristics were determined. Optimum conditions for the enzyme were pH 5.0 and 55℃. The two strains reached logarithmic phase in 7-8h during growth and the generation time was 1.04 in S. diastaticus and 1.38 in S. uvarum. Cell size and DNA content per cell of S. diastaticus were 44.10㎛³and 44.3 fg, and for S. uvarum, 99.67㎛³and 37.6fg. Ethanol productivities of S. diastaticus were 8.9% from 30% glucose and 6.9% from 30% starch and 11.4% from glucose with S. uvarum. Through determination of generation time, cell size, and DNA content per cell, both strains appeared as diploids, and differences in assimilability of melibiose and soluble starch of the two strains were selected as markers to determine the fusant. The optimal condition for protoplast formation was treatment of both strains with 10% ß-glucuronidase at pH 8.0 or 20㎍/ml Zymolyase at pH 7.5 for 70 min. While the regeneration frequencies were very low at 70min exposure to Zymolyase because of the instability of protoplasts, the yeasts treated for 45min were better for regeneration. The regeneration frequencies were also enhanced by 3-6 times when the regeration was carried out with 1.5% polyvinylpyrrolidone which stabilized protoplasts.

Structural basis for differential activities of enantiomeric PPARγ agonists: Binding of S35 to the alternate site

Jang, J.Y.,Koh, M.,Bae, H.,An, D.R.,Im, H.N.,Kim, H.S.,Yoon, J.Y.,Yoon, H.J.,Han, B.W.,Park, S.B.,Suh, S.W. Elsevier Science 2017 Biochimica et biophysica acta. Proteins and proteo Vol.1865 No.6

<P>Peroxisome proliferator-activated receptor gamma (PPAR gamma) is a member of the nuclear receptor superfamily. It functions as a ligand-activated transcription factor and plays important roles in the regulation of adipocyte differentiation, type 2 diabetes mellitus, and inflammation. Many PPAR gamma agonists bind to the canonical ligand-binding pocket near the activation function-2 (AF-2) helix (i.e., helix H12) of the ligand-binding domain (LBD). More recently, an alternate ligand-binding site was identified in PPAR gamma LBD; it is located beside the 52 loop between the helices H2 ' and H3. We reported previously that the chirality of two optimized enantiomeric PPAR gamma ligands (S35 and R35) differentiates their PPAR gamma transcriptional activity, binding affinity, and inhibitory activity toward Cdk5 (cyclin-dependent kinase 5)-mediated phosphorylation of PPAR gamma at Ser245 (in PPAR gamma 1 numbering; Ser273 in PPAR gamma 2 numbering). S35 is a PPAR gamma phosphorylation inhibitor with promising glucose uptake potential, whereas R35 behaves as a potent conventional PPAR gamma agonist. To provide a structural basis for understanding the differential activities of these enantiomeric ligands, we have determined crystal structures of the PPAR gamma LBD in complex with either S35 or R35. S35 and R35 bind to the PPAR gamma LBD in significantly different manners. The partial agonist S35 occupies the alternate site near the Omega loop, whereas the full agonist R35 binds entirely to the canonical LBP. Alternate site binding of S35 affects the PPAR gamma transactivation and the inhibitory effect on PPAR gamma Ser245 phosphorylation. This study provides a useful platform for the development of a new generation of PPAR gamma ligands as anti-diabetic drug candidates.</P>

Expression of potato S-adenosyl-l-methionine synthase (SbSAMS) gene altered developmental characteristics and stress responses in transgenic Arabidopsis plants

Kim, S.H.,Kim, S.H.,Palaniyandi, S.A.,Yang, S.H.,Suh, J.W. Gauthier-Villars ; Elsevier Science Ltd 2015 Vol. No.

S-adenosyl-l-methionine (SAM) synthase (SAMS) catalyze the biosynthesis of SAM, which is a precursor for ethylene and polyamines, and a methyl donor for a number of biomolecules. A full-length cDNA of SAMS from Solanum brevidens was expressed in Arabidopsis thaliana to study its physiological function. RT-PCR analysis showed that SbSAMS expression was enhanced significantly in S. brevidens leaves upon treatment with salt, mannitol, ethephon, IAA and ABA. The transgenic SbSAMS overexpression lines accumulated higher levels S-adenosyl homocysteine (SAHC) and ethylene concomitantly with increased SAM level. Expression levels of genes related to ethylene biosynthesis such as ACC synthase, but not polyamine biosynthesis genes were enhanced in SbSAMS overexpressing Arabidopsis lines. In addition, ABA responsive, wound and pathogen-inducible genes were upregulated in SbSAMS transgenic Arabidopsis plants. Transgenic Arabidopsis lines exhibited higher salt and drought stress tolerance compared to those of vector control. Based on these results we conclude that SbSAMS is expressed under abiotic stress to produce SAM as a broad-spectrum signal molecule to upregulate stress-related genes including ethylene and ABA biosynthetic pathway genes responsible for ABA, pathogen and wound responses.

A phase II study of everolimus (RAD001), an mTOR inhibitor plus CHOP for newly diagnosed peripheral T-cell lymphomas

Kim, S. J.,Shin, D.-Y.,Kim, J. S.,Yoon, D. H.,Lee, W. S.,Lee, H.,Do, Y. R.,Kang, H. J.,Eom, H. S.,Ko, Y. H.,Lee, S. H.,Yoo, H. Y.,Hong, M.,Suh, C.,Kim, W. S. Oxford University Press 2016 ANNALS OF ONCOLOGY Vol.27 No.4

<P>Everolimus plus CHOP is effective for newly diagnosed, chemotherapy-naive peripheral T-cell lymphoma patients with acceptable toxicity.Everolimus, an oral mTOR inhibitor, has single-agent activity against relapsed lymphomas. Thus, we carried out a phase II study of everolimus in combination with CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone) as a first-line treatment for patients with peripheral T-cell lymphoma (PTCL) based on our phase I study results. Participants (n = 30) received CHOP with 5 mg everolimus per day from day 1 to 14 every 21 days for a total of six cycles. The primary end point was the overall response rate (ORR), which included complete response (CR) and partial response (PR) to this regimen. Immunohistochemistry was used to evaluate the expression of phosphatase and tensin homology (PTEN) and phosphorylated S6 kinase (pS6K) as a response. The objective response rate was 90% with CR (n = 17) and PR (n = 10). The CR rate was different among subtypes; angioimmunoblastic T-cell lymphoma (AITL, n = 3) had a CR whereas PTCL-not-otherwise specified and ALK-negative anaplastic large-cell lymphoma (ALCL) patients showed 63% (12/19) and 29% (2/7) of CR rate, respectively. This difference in CR rate among subtypes was associated with PTEN loss because PTEN loss was not seen in AITL but 33% of ALCL patients. The most common toxicity was hematological, with 80% of patients experiencing at least one event of grade 3/4 neutropenia, and 60% of patients had grade 3/4 thrombocytopenia. The everolimus plus CHOP was effective for PTCL patients, and its efficacy might be related with the preservation of PTEN.</P>

Crystal structure of Rv2258c from Mycobacterium tuberculosis H37Rv, an S-adenosyl-l-methionine-dependent methyltransferase

Im, H.N.,Kim, H.S.,An, D.R.,Jang, J.Y.,Kim, J.,Yoon, H.J.,Yang, J.K.,Suh, S.W. Academic Press 2016 Journal of structural biology Vol.193 No.3

<P>The Mycobacterium tuberculosis Rv2258c protein is an S-adenosyl-L-methionine (SAM)-dependent methyltransferase (MTase). Here, we have determined its crystal structure in three forms: a ligand-unbound form, a binary complex with sinefungin (SFG), and a binary complex with S-adenosyl-L-homocysteine (SAH). The monomer structure of Rv2258c consists of two domains which are linked by a long alpha-helix. The N-terminal domain is essential for dimerization and the C-terminal domain has the Class I MTase fold. Rv2258c forms a homodimer in the crystal, with the N-terminal domains facing each other. It also exists as a homodimer in solution. A DALI structural similarity search with Rv2258c reveals that the overall structure of Rv2258c is very similar to small-molecule SAM -dependent MTases. Rv2258c interacts with the bound SFG (or SAH) in an extended conformation maintained by a network of hydrogen bonds and stacking interactions. Rv2258c has a relatively large hydrophobic cavity for binding of the methyl-accepting substrate, suggesting that bulky nonpolar molecules with aromatic rings might be targeted for methylation by Rv2258c in M. tuberculosis. However, the ligand-binding specificity and the biological role of Rv2258c remain to be elucidated due to high variability of the amino acid residues defining the substrate-binding site. (C) 2016 Elsevier Inc. All rights reserved.</P>

The effects of mesenchymal stem cells injected via different routes on modified IL-12-mediated antitumor activity

Seo, S H,Kim, K S,Park, S H,Suh, Y S,Kim, S J,Jeun, S-S,Sung, Y C Nature Publishing Group 2011 Gene Therapy Vol.18 No.5

<P>Owing to its tumor tropism and prolonged transgene expression, mesenchymal stem cell (MSC) has been considered as an ideal delivery vehicle for cancer gene therapies or therapeutic vaccines. In this study, we demonstrated that intratumoral (i.t.) injection of MSCs expressing modified interleukin-12 (MSCs/IL-12M) exhibited stronger tumor-specific T-cell responses and antitumor effects as well as more sustained expressions of IL-12 and interferon (IFN)-γ in both sera and tumor sites than did IL-12M-expressing adenovirus (rAd/IL-12M) in mice bearing both solid and metastatic tumors. Subcutaneous (s.c.) injection of MSCs/IL-12M at contralateral site of tumor exhibited similar levels of serum IL-12 and IFN-γ as i.t. injection, but much weaker antitumor effects in both B16F10 melanoma and TC-1 cervical cancer models than i.t. injection. Although intravenous (i.v.) injection elicited earlier peak serum levels of cytokines, it induced weaker tumor-specific T-cell responses and antitumor effects than i.t. injection, indicating that serum cytokine levels are not surrogate indicators of antitumor effects. Taken together, these results indicated that MSC is more efficient than adenovirus as a cytokine gene delivery vehicle and that i.t. injection of MSCs/IL-12M is the best approach to induce strong tumor-specific T-cell responses that correlate with anti-metastatic effects as well as inhibition of solid tumor growth, although MSCs themselves have an ability to migrate into the tumor site. In addition, MSCs/IL-12M embedded in Matrigel (MSCs/IL-12M/Matrigel) exhibited significant antitumor effects even in immunodeficient mice such as SCID and BNX mice lacking T, B and natural killer (NK) cells, but not in IFN-γ knockout mice. Our findings provide an optimal approach for designing an efficient clinical protocol of MSC-based cytokine gene therapy to induce strong tumor-specific T-cell responses and therapeutic anticancer efficacy.</P>

Crystal structures of LacD from Staphylococcus aureus and LacD.1 from Streptococcus pyogenes: Insights into substrate specificity and virulence gene regulation

Lee, S.J.,Kim, H.S.,Kim, D.J.,Yoon, H.J.,Kim, K.H.,Yoon, J.Y.,Suh, S.W. North-Holland Pub ; Elsevier Science Ltd 2011 FEBS letters Vol.585 No.2

Staphylococcus aureus LacD, a Class I tagatose-1,6-bisphosphate (TBP) aldolase, shows broadened substrate specificity by catalyzing the cleavage of 1,6-bisphosphate derivatives of d-tagatose, d-fructose, d-sorbose, and d-psicose. LacD.1 and LacD.2 are two closely-related Class I TBP aldolases in Streptococcus pyogenes. Here we have determined the crystal structures of S. aureus LacD and S. pyogenes LacD.1. Monomers of both enzymes are folded into a (β/α)<SUB>8</SUB> barrel and two monomers associate tightly to form a dimer in the crystals. The structures suggest that the residues E189 and S300 of rabbit muscle Class I fructose-1,6-bisphosphate (FBP) aldolase are important for substrate specificity. When we mutated the corresponding residues of S. aureus LacD, the mutants (L165E, L275S, and L165E/L275S) showed enhanced substrate specificity toward FBP. Structured summary: lacDbinds to lacD by X-ray crystallography(View interaction) lacD1binds to lacD1 by X-ray crystallography(View interaction)

Synthesis and biological evaluation of C-ring truncated deguelin derivatives as heat shock protein 90 (HSP90) inhibitors

Kim, H.S.,Hong, M.,Ann, J.,Yoon, S.,Nguyen, C.T.,Lee, S.C.,Lee, H.Y.,Suh, Y.G.,Seo, J.H.,Choi, H.,Kim, J.Y.,Kim, K.W.,Kim, J.,Kim, Y.M.,Park, S.J.,Park, H.J.,Lee, J. Elsevier/Pergamon 2016 Bioorganic & medicinal chemistry Vol.24 No.22

Based on the lead compound L-80 (compound 2), a potent heat shock protein 90 (HSP90) inhibitor, a series of C-ring truncated deguelin analogs were designed, synthesized and evaluated for Hypoxia Inducible Factor-1α (HIF-1α) inhibition as a primary screening method. Their structure-activity relationship was investigated in a systematic manner by varying the A/B ring, linker and D/E ring, respectively. Among the synthesized inhibitors, compound 5 exhibited potent HIF-1α inhibition in a dose-dependent manner and significant antitumor activity in human non-small cell lung carcinoma (H1299), with better activities than L-80. It also inhibited in vitro hypoxia-mediated angiogenic processes in human retinal microvascular endothelial cells (HRMEC). The docking study of 5 showed a similar binding mode as L-80: it occupied the C-terminal ATP-binding pocket of HSP90, indicating that the anticancer and antiangiogenic activities of 5 were derived from HIF-1α destabilization by inhibiting the C-terminal ATP-binding site of hHSP90.

한국인 Long Term Non Progressors로부터 분리된 HIV-1 env 및 nef 유전자와 이들의 면역 유전학적 특성 분석에 의한 질병진전 요인 규명

이주실,강춘,김성순,박근용,고운영,기미경,최병선,남정구,서순덕,이성래,구본기,박혜경,도경미,김태규,최희백,조현일,김요숙,김지영,이해정,안수진 국립보건연구원 1999 국립보건원보 Vol.36 No.-