A genetic variation in microRNA target site of <i>KRT81</i> gene is associated with survival in early-stage non-small-cell lung cancer

Lee, S. Y.,Choi, J. E.,Jeon, H. S.,Hong, M. J.,Choi, Y. Y.,Kang, H. G.,Yoo, S. S.,Lee, E. B.,Jeong, J. Y.,Lee, W. K.,Lee, J.,Cha, S. I.,Kim, C. H.,Kim, Y. T.,Jheon, S.,Son, J. W.,Park, J. Y. Oxford University Press 2015 ANNALS OF ONCOLOGY Vol.26 No.6

<P>In this study, <I>KRT81</I> rs3660G>C was associated with survival of patients with NSCLC after surgical resection. Mechanistic study suggested that the G-to-C change caused reduced binding efficiency of miRNA, leading to decreased translational repression, thereby increased <I>KRT81</I> expression. The <I>KRT81</I> rs3660G>C may be a useful prognostic biomarker in early-stage NSCLC patients.</P><P><B>Background</B></P><P>MicroRNAs (miRNAs) have a key role in carcinogenesis through negative regulation of their target genes. Therefore, genetic variations in miRNAs or their target sites may affect miRNA–mRNA interactions, thereby result in altered expression of target genes. This study was conducted to investigate the associations between single-nucleotide polymorphisms (SNP) located in the miRNA target sites (poly-miRTSs) and survival of patients with early-stage non-small-cell lung cancer (NSCLC).</P><P><B>Methods</B></P><P>Using public SNP database and miRNA target sites prediction program, 354 poly-miRTSs were selected for genotyping. Among these, 154 SNPs applicable to Sequenom's MassARRAY platform were investigated in 357 patients. A replication study was carried out on an independent patient population (<I>n</I> = 479). <I>Renilla</I> luciferase assay and reverse transcription-polymerase chain reaction were conducted to examine functional relevance of potentially functional poly-miRTSs.</P><P><B>Results</B></P><P>Of the 154 SNPs analyzed in a discovery set, 14 SNPs were significantly associated with survival outcomes. Among these, <I>KRT81</I> rs3660G>C was found to be associated with survival outcomes in the validation cohort. In the combined analysis, patients with the rs3660 GC + CC genotype had a significantly better overall survival compared with those with GG genotype [adjusted hazard ratio (aHR) for OS, 0.65; 95% confidence interval (CI) 0.50–0.85; <I>P</I> = 0.001]. An increased expression of the reporter gene for the C allele of rs3660 compared with the G allele was observed by luciferase assay. Consistently, the C allele was associated with higher relative expression level of <I>KRT81</I> in tumor tissues.</P><P><B>Conclusion</B></P><P>The rs3660G>C affects KRT81 expression and thus influences survival in early-stage NSCLC. The analysis of the rs3660G>C polymorphism may be useful to identify patients at high risk of a poor disease outcome.</P>

Reactivities of organic isothiocyanates and thiocyanates toward dialkyl bis(phosphine) complexes of palladium(II) and platinum(II)

Lee, S.G.,Choi, K.Y.,Kim, Y.J.,Park, S.,Lee, S.W. Pergamon Press 2015 Polyhedron Vol.85 No.-

Room-temperature reactions of trans-[PdEt<SUB>2</SUB>L<SUB>2</SUB>] (L=PMe<SUB>3</SUB>, PEt<SUB>3</SUB>, PMe<SUB>2</SUB>Ph) with organic isothiocyanates [R-NCS; R=benzyl; CH(CH<SUB>3</SUB>)Ph, R-(-) and S-(+); indanyl, S-(+)] afforded the S,S-coordinated Pd(II) complexes [Pd(S<SUB>2</SUB>C?N-R)L<SUB>2</SUB>] containing a dithiocarbonimidato (S<SUB>2</SUB>C?N-R) group. Similar reactions involving allyl isothiocyanates produced the cationic η<SUP>3</SUP>-allyl Pd complex [Pd(η<SUP>3</SUP>-allyl)(PMe<SUB>3</SUB>)<SUB>2</SUB>]<SUP>+</SUP>(NCS)<SUP>-</SUP>. When [Pd(S<SUB>2</SUB>C?N-R)(PMe<SUB>3</SUB>)<SUB>2</SUB>] was treated with 1equiv of a chelating phosphine [L-L=depe (1,2-bis(diethylphosphino)ethane) and dmpe (1,2-bis(dimethylphosphino)ethane)], the corresponding complexes [Pd(S<SUB>2</SUB>C?N-R)(L-L)] were produced. Reactions of trans-[PdEt<SUB>2</SUB>L<SUB>2</SUB>] (L=PMe<SUB>3</SUB>, PMe<SUB>2</SUB>Ph) with organic thiocyanates (R-SCN; R=benzyl, Et) resulted in the formation of [Pd(CN)<SUB>2</SUB>L<SUB>2</SUB>] and an organic disulfide by S-C bond cleavage of R-SCN. However, similar reactions of the dimethyl analogs, trans-[PdMe<SUB>2</SUB>L<SUB>2</SUB>] (L=PMe<SUB>3</SUB>, PEt<SUB>3</SUB>), with benzyl thiocyanate afforded different products, [Pd(NCS)<SUB>2</SUB>L<SUB>2</SUB>] or [PdMe(NCS)L<SUB>2</SUB>]. Treating [Pt(styrene)(PMe<SUB>3</SUB>)<SUB>2</SUB>] with benzyl isothiocyanate gave the S-coordinated dithiocarbonimidato Pt(II) complex, [Pt(S<SUB>2</SUB>C?N-R)(Me<SUB>3</SUB>P)<SUB>2</SUB>] (R=benzyl). In contrast, cis-[PtEt<SUB>2</SUB>(PMe<SUB>3</SUB>)<SUB>2</SUB>] reacted with the isothiocyanate to afford the trialkyl Pt(IV) complex [PtEt<SUB>2</SUB>(SCN)(CH<SUB>2</SUB>Ph)(PMe<SUB>3</SUB>)<SUB>2</SUB>].

콩 탈곡기를 활용한 들깨 탈곡 기계화 연구

김형곤 ( H. G. Kim ),전현종 ( H. J. Jun ),김진구 ( J. G. Kim ),유승화 ( S. H. Yu ),김영근 ( Y. K. Kim ),강태경 ( T. G. Kang ),최일수 ( I. S. Choi ),이상희 ( S. H. Lee ),최용 ( Y. Choi ) 한국농업기계학회 2017 한국농업기계학회 학술발표논문집 Vol.22 No.2

들깨 생산을 위한 농작업에서 경운· 정지, 피복, 방제 등은 기계화 되었으나 파종, 정식, 예취· 탈곡은 기계화가 미흡한 실정이다. 본 연구에서는 기계화가 미흡한 들깨의 탈곡작업 기계화 촉진 및 범용화를 통한 들깨 기계화기술을 향상 하고자 수행하였다. 첫 번째 수행내용은 기존 콩 탈곡기 송풍 팬에 의해 손실되는 들깨를 줄이고자 송풍 팬 속도(450~1350 RPM)에 따른 풍구속도(m/s) 및 손실량(g)을 측정하기 위해 탈곡된 들깨 500 g을 탈곡기에 투입하여 실험하였다. 두 번째 수행내용은 투입된 작물의 줄기가 탈곡 시 배출이 용이하도록 탈곡기 덮개 안쪽에 가이드를 설치하여 탈곡에 미치는 영향을 실험하였다. 탈곡시험은 가이드를 설치하지 않은 경우와 설치한 경우로 분리하여 비교시험을 하였다. 가이드는 급동 회전방향과 10°, 20°, 30°를 유지하면서 배출구까지 도달하도록 설치하였고, 각도에 따른 잔물배출시간(s), 잔물배출무게(g) 및 잔물 파쇄 길이(cm)을 측정하였다. 시험결과 들깨의 손실률이 발생되는 구간은 1050 RPM, 풍구속도 5.27 m/s였으며 송풍 팬 회전속도가 증가할수록 손실률은 급격하게 증가하였다. 이는 실험 탈곡기 구조의 들깨 종말속도가 대략 5m/s로 송풍팬 회전속도를 1000 RPM, 풍구속도를 5m/s 이하로 설정해야 할 것으로 판단된다. 가이드 각도에 따른 잔물배출시간은 각도가 클수록 잔물배출시간이 단축되었고 특히 10°(18.90 s)는 무 설치(29.23 s) 보다 약 10초 단축되어 가이드 설치가 잔물배출에 효과적으로 나타났다. 잔물배출무게는 가이드 각도에 영향을 받지 않은 반면 잔물 파쇄 길이는 가이드 각도가 클수록 길게 나타났다. 결과적으로 실험 콩 탈곡기 적용 들깨 탈곡 시 5 m/s 이하의 풍속에서 잔물배출시간과 파쇄정도에 따른 들깨 손실 등을 고려한 복합적인 연구가 필요할 것으로 판단되었다.

Antimicrobial activity and synergism of Sami-Hyanglyun-Hwan with ciprofloxacin against methicillin-resistant Staphylococcus aureus

Choi, J.G.,Choi, J.Y.,Mun, S.H.,Kang, O.H.,Bharaj, P.,Shin, D.W.,Chong, M.S.,Kwon, D.Y. Elsevier 2015 Asian Pacific journal of tropical medicine Vol.8 No.7

Objective: To investigate the antibacterial activity of SHH extracted with either water or ethanol against methicillin-resistant Staphylococcus aureus (MRSA) and combinatory antimicrobial effect with ciprofloxacin (CIP) by time kill assay and checkerboard dilution test. Methods: The antibacterial activity determined by broth dilution method indicated that the antibacterial activity of Sami-Hyanglyun-Hwan (SHH) water extract (SHHW) and SHH ethanol extract (SHHE) ranged from 250 to 2000 μg/mL and 125 to 1000 μg/mL against MRSA, respectively. Results: In the checkerboard method, the combinations of SHHE with CIP had a partial synergistic or synergistic effect against MRSA. The time-kill curves showed that a combined SHHE and CIP treatment reduced the bacterial counts dramatically after 24 h. Conclusions: The present study demonstrates the therapeutic ability of SHHE against MRSA infections.

In vitro antibacterial activity and major bioactive components of Cinnamomum verum essential oils against cariogenic bacteria, Streptococcus mutans and Streptococcus sobrinus

Choi, O.,Cho, S.K.,Kim, J.,Park, C.G.,Kim, J. China Humanity Technology Publishing House 2016 Asian Pacific journal of tropical biomedicine Vol.6 No.4

Objective: To evaluate the antibacterial activity of Cinnamomum verum (C. verum) from 32 different essential oils against cariogenic bacteria, Streptococcus mutans (S. mutans) and Streptococcus sobrinus (S. sobrinus). Methods: The antibacterial activities of each essential oil were individually investigated against S. mutans and S. sobrinus. The essential oil of C. verum was selected for further evaluation against S. mutans and S. sobrinus. Gas chromatography mass spectrometry was used to determine the major constituents of C. verum essential oil. In addition, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration of the most effective constituent was investigated. Results: The essential oil from C. verum exhibited the greatest antibacterial activity. Gas chromatography mass spectrometry analysis revealed that the major components of C. verum essential oil were cinnamaldehyde (56.3%), cinnamyl acetate (7.1%) and β-phellandrene (6.3%). The MIC of cinnamaldehyde was measured using broth dilution assays. The MIC of cinnamaldehyde was 0.02% (v/v) against both bacterial strains tested. The minimum bactericidal concentration of cinnamaldehyde against S. mutans and S. sobrinus were 0.2% and 0.1% (v/v), respectively. Conclusions: The essential oil of C. verum and its major component cinnamaldehyde possessed considerable in vitro antibacterial activities against cariogenic bacteria, S. mutans and S. sobrinus strains. These results showed that the essential oil of C. verum and its bioactive component, cinnamaldehyde, have potential for application as natural agents for the prevention and treatment of dental caries.

Anti-inflammatory and anti-proliferative effects of Rhus verniciflua Stokes in RAW264.7 cells

Choi, H.-S.,Seo, H.S.,Kim, S.R.,Choi, Y.K.,Jang, B.-H.,Shin, Y.-C.,Ko, S.-G. SPANDIDOS PUBLICATIONS 2014 MOLECULAR MEDICINE REPORTS Vol.9 No.1

Inflammatory response is a major defense mechanism against pathogens and chemical or mechanical injury. Rhus verniciflua Stokes (RVS) has traditionally been used as an ingredient in East Asian medicine for the treatment of gastritis, stomach cancer and atherosclerosis. The aim of the current study was to analyze the effect of RVS on LPS-induced inflammatory responses in the RAW264.7 mouse macrophage cell line. RAW264.7 cells were treated with various concentrations of RVS and LPS at specific time points. WST assay, trypan blue assay and quantification of activated cells revealed that RVS suppressed cell proliferation in a dose-dependent manner. RVS induced G1 cell cycle arrest, suppressed iNOS and COX-2 mRNA expression induced by LPS and decreased intracellular ROS levels induced by LPS. In addition, RVS induced PARP and caspase-3 cleavage suggesting that RVS causes cell death. Results of the present study indicate that RVS may be advantageous in treating inflammatory disease.

Fatty acid biosynthesis and lipogenic enzyme activities in subcutaneous adipose tissue of feedlot steers fed supplementary palm oil or soybean oil

Choi, S. H.,Gang, G. O.,Sawyer, J. E.,Johnson, B. J.,Kim, K. H.,Choi, C. W.,Smith, S. B. American Society of Animal Science 2013 Journal of animal science Vol.91 No.5

<P>We hypothesized that supplementing finishing diets with palm oil would promote adipocyte differentiation in subcutaneous adipose tissue of feedlot steers, and that soybean oil supplementation would depress adipocyte differentiation. Twenty-eight Angus steers were assigned randomly to 3 groups of 9 or 10 steers and fed a basal diet without additional fat (control), with 3% palm oil (rich in palmitic acid), or with 3% soybean oil (rich in polyunsaturated fatty acids), for 10 wk, top-dressed daily. Palm oil had no effect (<I>P</I> > 0.05) on ADG, food intake, or G:F, whereas soybean oil depressed ADG (<I>P</I> = 0.02), food intake (<I>P</I> = 0.04), and G:F (<I>P</I> = 0.05). Marbling scores tended (<I>P</I> = 0.09) to be greater in palm oil-fed steers (Modest<SUP>09</SUP>) than in soybean oil-fed steers (Small<SUP>55</SUP>). Subcutaneous adipocyte mean volume was greater in palm oil-fed steers (515.9 pL) than in soybean-supplemented cattle (395.6 pL; <I>P</I> = 0.01). Similarly, glucose and acetate incorporation into total lipids in vitro was greater in subcutaneous adipose tissue of palm oil-fed steers (119.9 and 242.8 nmol·3h<SUP>–1</SUP>·10<SUP>5</SUP> cells, respectively) than adipose tissue of soybean oil-fed steers in (48.9 and 95.8 nmol·3h<SUP>–1</SUP>·10<SUP>5</SUP> cells, respectively). Glucose-6-phosphate dehydrogenase and NADP-malate dehydrogenase activities were greater (<I>P</I> ≤ 0.05) in subcutaneous adipose tissue of palm oil-fed steers than in adipose tissue of control steers. Palm oil did not increase palmitic acid or decrease oleic acid in subcutaneous adipose tissue or LM, but decreased (<I>P</I> ≤ 0.05) myristoleic, palmitoleic, and <I>cis</I>-vaccenic acid in adipose tissue, indicating a depression in stearoyl-coenzyme A desaturase activity. Soybean oil increased the proportion of α-linolenic acid in adipose tissue and muscle and increased linoleic acid and 18:1<I>trans</I>-10 in muscle. We conclude that palm oil supplementation promoted lipid synthesis in adipose tissue without depressing feed efficiency or increasing the palmitic acid content of beef.</P>

Clinical and histopathological study of Charcot-Marie-Tooth neuropathy with a novel S90W mutation in BSCL2.

Choi, B-O,Park, M-H,Chung, K W,Woo, H-M,Koo, H,Chung, H-K,Choi, K-G,Park, K D,Lee, H J,Hyun, Y S,Koo, S K Oxford University Press 2013 Neurogenetics Vol.14 No.1

<P>The objective of the study was to investigate the disease-causing mutation in an autosomal dominant Charcot-Marie-Tooth disease type 2 family and examine the clinical and histopathological evaluation. We enrolled a family of Korean origin with axonal Charcot-Marie-Tooth disease neuropathy (FC305; 13 males, six females) and applied genome-wide linkage analysis. Whole exome sequencing was performed for two patients. In addition, sural nerve biopsies were obtained from two patients. Through whole exome sequencing, we identified an average of 20,336 coding variants from two patients. We also found evidence of linkage mapped to chromosome 11p11-11q13.3 (LOD score of 3.6). Among these variants in the linkage region, we detected a novel p.S90W mutation in the Berardinelli-Seip congenital lipodystrophy 2 (BSCL2) gene, after filtering 31 Korean control exomes. Our p.S90W patients had frequent sensory disturbances, pyramidal tract signs, and predominant right thenar muscle atrophy in comparison with reported p.S90L patients. The phenotypic spectra were wide and demonstrated intrafamilial variability. Two patients with different clinical features underwent sural nerve biopsies; the myelinated fiber densities were increased slightly in both patients, which differed from two previous case reports of BSCL2 mutations (p.S90L and p.N88S). This report expands the variability of the clinical spectrum associated with the BSCL2 gene and describes the first family with the p.S90W mutation.</P>

Pinoresinol-4,4'-di-O-β-d-glucoside from Valeriana officinalis root stimulates calcium mobilization and chemotactic migration of mouse embryo fibroblasts

Do, K.H.,Choi, Y.W.,Kim, E.K.,Yun, S.J.,Kim, M.S.,Lee, S.Y.,Ha, J.M.,Kim, J.H.,Kim, C.D.,Son, B.G.,Kang, J.S.,Khan, I.A.,Bae, S.S. G. Fischer 2009 Phytomedicine Vol.16 No.6

Lignans are major constituents of plant extracts and have important pharmacological effects on mammalian cells. Here we showed that pinoresinol-4,4'-di-O-β-d-glucoside (PDG) from Valeriana officinalis induced calcium mobilization and cell migration through the activation of lysophosphatidic acid (LPA) receptor subtypes. Stimulation of mouse embryo fibroblast (MEF) cells with 10μM PDG resulted in strong stimulation of MEF cell migration and the EC<SUB>50</SUB> was about 2μM. Pretreatment with pertussis toxin (PTX), an inhibitor of G<SUB>i</SUB> protein, completely blocked PDG-induced cell migration demonstrating that PDG evokes MEF cell migration through the activation of the G<SUB>i</SUB>-coupled receptor. Furthermore, pretreatment of MEF cells with Ki16425 (10μM), which is a selective antagonist for LPA<SUB>1</SUB> and LPA<SUB>3</SUB> receptors, completely blocked PDG-induced cell migration. Likewise, PDG strongly induced calcium mobilization, which was also blocked by Ki16425 in a dose-dependent manner. Prior occupation of the LPA receptor with LPA itself completely blocked PDG-induced calcium mobilization. Finally, PDG-induced MEF cell migration was attenuated by pretreatment with a phosphatidylinositol 3-kinase (PI3K) inhibitor such as LY294002. Cells lacking downstream mediator of PI3K such as Akt1 and Akt2 (DKO cells) showed loss of PDG-induced migration. Re-expression of Akt1 (but not Akt2) completely restored PDG-induced DKO cell migration. Given these results, we conclude that PDG is a strong inducer of cell migration. We suggest that the pharmacological action of PDG may occur through the activation of an LPA receptor whereby activation of PI3K/Akt signaling pathway mediates PDG-induced MEF cell migration.

Five-year clinical outcomes in patients with significant coronary artery spasm: A propensity score-matched analysis

Choi, B.G.,Park, S.H.,Rha, S.W.,Park, J.Y.,Choi, S.Y.,Park, Y.,Xu, S.,Ngow, H.A.,Ali, J.,Li, H.,Kim, J.B.,Lee, S.,Na, J.O.,Choi, C.U.,Lim, H.E.,Kim, J.W.,Kim, E.J.,Park, C.G.,Seo, H.S.,Oh, D.J. Elsevier/North-Holland Biomedical Press 2015 INTERNATIONAL JOURNAL OF CARDIOLOGY Vol.184 No.-

Background: Coronary artery spasm (CAS) is known to be a risk factor of acute coronary syndrome and angina pectoris. However, there is no currently available data with larger study population regarding long-term clinical outcomes of CAS in real world clinical practice. Objectives: We evaluated the prevalence of CAS and the impact of CAS on 5-year clinical outcomes in a series of Asian CAS patients documented by intracoronary acetylcholine (Ach) provocation test. Methods: A total of 1413 consecutive patients without significant coronary artery disease (CAD) who underwent Ach provocation test between Nov. 2004 and Oct. 2008 were enrolled. Significant CAS was defined as >70% of narrowing by incremental intracoronary injection of 20, 50 and 100μg. Patients were divided into two groups based on the presence of significant CAS (the non-CAS group: n=640, the CAS group; n=773). To adjust potential confounders, a propensity score matched (PSM) analysis was performed using the logistic regression model. Results: A total of 54.7% (773/1413) patients were diagnosed as CAS documented by Ach provocation test. After PSM analysis, 2 propensity-matched groups (451 pairs, n=902, C-statistic=0.677) were generated. Despite of similar incidence of individual hard endpoints including mortality, myocardial infarction and revascularization, the CAS group showed the higher trend of recurrent angina requiring follow up angiography than the non-CAS group up to 5years (HR; 1.56, 95% C.I.; 0.99-2.46, p=0.054). Conclusions: The prevalence of CAS was 54.7%. Although the cumulative incidence of recurrent angina requiring follow up coronary angiography seems to be increased up to 5years in CAS patients, CAS patients was not associated with major individual and composite clinical outcomes such as mortality, MI, PCI, CVA with optimal medical therapy as compared with patients without CAS.