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      • SCOPUSSCIE

        Elucidation of ethanol tolerance mechanisms in <i>Saccharomyces cerevisiae</i> by global metabolite profiling

        Kim, Sooah,Kim, Jungyeon,Song, Ju Hwan,Jung, Young Hoon,Choi, Il‐,Sup,Choi, Wonja,Park, Yong‐,Cheol,Seo, Jin‐,Ho,Kim, Kyoung Heon WILEY‐VCH Verlag 2016 Biotechnology Journal Vol.11 No.9

        <P><B>Abstract</B></P><P>Ethanol, the major fermentation product of yeast, is a stress factor in yeast. We previously constructed an ethanol‐tolerant mutant yeast iETS3 by using the global transcriptional machinery engineering. However, the ethanol‐tolerance mechanism has not been systematically investigated. In this study, global metabolite profiling was carried out, mainly by gas chromatography/time‐of‐flight mass spectrometry (GC/TOF MS), to investigate the mechanisms of ethanol tolerance in iETS3. A total of 108 intracellular metabolites were identified by GC/TOF MS and high performance liquid chromatography, and these metabolites were mostly intermediates of the central carbon metabolism. The metabolite profiles of iETS3 and BY4741, cultured with or without ethanol, were significantly different based on principal component and hierarchical clustering analyses. Our metabolomic analyses identified the compositional changes in cell membranes and the activation of glutamate metabolism and the trehalose synthetic pathway as the possible mechanisms for the ethanol tolerance. These metabolic traits can be considered possible targets for further improvement of ethanol tolerance in the mutant. For example, the <I>KGD1</I> deletion mutant, with up‐regulated glutamate metabolism, showed increased tolerance to ethanol. This study has demonstrated that metabolomics can be a useful tool for strain improvement and phenotypic analysis of microorganisms under stress.</P>

      • KCI등재

        A Retrospective Clinical Evaluation of an Artificial Intelligence Screening Method for Early Detection of STEMI in the Emergency Department

        Kim Dongsung,Hwang Ji Eun,Cho Youngjin,Cho Hyoung-Won,Lee Wonjae,Lee Ji Hyun,Oh Il-Young,Baek Sumin,Lee Eunkyoung,Kim Joonghee 대한의학회 2022 Journal of Korean medical science Vol.37 No.10

        Background: Rapid revascularization is the key to better patient outcomes in ST-elevation myocardial infarction (STEMI). Direct activation of cardiac catheterization laboratory (CCL) using artificial intelligence (AI) interpretation of initial electrocardiography (ECG) might help reduce door-to-balloon (D2B) time. To prove that this approach is feasible and beneficial, we assessed the non-inferiority of such a process over conventional evaluation and estimated its clinical benefits, including a reduction in D2B time, medical cost, and 1-year mortality. Methods: This is a single-center retrospective study of emergency department (ED) patients suspected of having STEMI from January 2021 to June 2021. Quantitative ECG (QCG™), a comprehensive cardiovascular evaluation system, was used for screening. The non-inferiority of the AI-driven CCL activation over joint clinical evaluation by emergency physicians and cardiologists was tested using a 5% non-inferiority margin. Results: Eighty patients (STEMI, 54 patients [67.5%]) were analyzed. The area under the curve of QCG score was 0.947. Binned at 50 (binary QCG), the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were 98.1% (95% confidence interval [CI], 94.6%, 100.0%), 76.9% (95% CI, 60.7%, 93.1%), 89.8% (95% CI, 82.1%, 97.5%) and 95.2% (95% CI, 86.1%, 100.0%), respectively. The difference in sensitivity and specificity between binary QCG and the joint clinical decision was 3.7% (95% CI, −3.5%, 10.9%) and 19.2% (95% CI, −4.7%, 43.1%), respectively, confirming the non-inferiority. The estimated median reduction in D2B time, evaluation cost, and the relative risk of 1-year mortality were 11.0 minutes (interquartile range [IQR], 7.3–20.0 minutes), 26,902.2 KRW (22.78 USD) per STEMI patient, and 12.39% (IQR, 7.51–22.54%), respectively. Conclusion: AI-assisted CCL activation using initial ECG is feasible. If such a policy is implemented, it would be reasonable to expect some reduction in D2B time, medical cost, and 1-year mortality.

      • SCIESCOPUS

        CRISPR interference-guided balancing of a biosynthetic mevalonate pathway increases terpenoid production

        Kim, Seong Keun,Han, Gui Hwan,Seong, Wonjae,Kim, Haseong,Kim, Seon-Won,Lee, Dae-Hee,Lee, Seung-Goo Elsevier 2016 Metabolic engineering Vol.38 No.-

        <P><B>Abstract</B></P> <P>Methods for simple and efficient regulation of metabolic pathway genes are essential for maximizing product titers and conversion yields, and for minimizing the metabolic burden caused by heterologous expression of multiple genes often in the operon context. Clustered regularly interspaced short palindromic repeats (CRISPR) interference (CRISPRi) is emerging as a promising tool for transcriptional modulation. In this study, we developed a regulatable CRISPRi system for fine-tuning biosynthetic pathways and thus directing carbon flux toward target product synthesis. By exploiting engineered <I>Escherichia coli</I> harboring a biosynthetic mevalonate (MVA) pathway and plant-derived terpenoid synthases, the CRISPRi system successfully modulated the expression of all the MVA pathway genes in the context of operon and blocked the transcription of the acetoacetyl-CoA thiolase enzyme that catalyzes the first step in the MVA pathway. This CRISPRi-guided balancing of expression of MVA pathway genes led to enhanced production of (-)-α-bisabolol (C15) and lycopene (C40) and alleviation of cell growth inhibition that may be caused by expression of multiple enzymes or production of toxic intermediate metabolites in the MVA pathway. Coupling CRISPRi to cell growth by regulating an endogenous essential gene (<I>ispA</I>) increased isoprene (C5) production. The regulatable CRISPRi system proved to be a robust platform for systematic modulation of biosynthetic and endogenous gene expression, and can be used to tune biosynthetic metabolic pathways. Its application can enable the development of microbial ‘smart cell’ factories that can produce other industrially valuable products in the future.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A regulatable CRISPR interference (CRISPRi) system was developed. </LI> <LI> Developed CRISPRi system was successfully applied to terpenoid production. </LI> <LI> CRISPRi-guided balancing of MVA pathway greatly enhanced terpenoid production. </LI> <LI> Regulatable CRISPRi system can be a powerful tool for metabolic engineering. </LI> </UL> </P>

      • Comparison of Expert and Novice Pilot Situation Awareness during Landing

        Wonjae Lee,Chunghoon Kim,Sungho Kim 대한인간공학회 2017 대한인간공학회 학술대회논문집 Vol.2017 No.11

        Objective: This study was to compare and quantify the level of situation awareness (SA) between expert and novice pilots using eye tracking features. Background: Successful and safe landing requires a high SA of pilots. However, existing studies have examined simple SA status, which was lack of comprehensively understanding by linking SA model and quantitative features. Method: The experiments were conducted by five repeated landing for four expert and four novice pilots using flight simulator and eye tracking equipment. Three eye tracking features (visit frequency, dwell time ratio, scan path entropy) were calculated for reflecting three level of SA model (perception, comprehension, projection). The classification of SA status was performed by applying significant eye tracking features, developing binary logistic regression model, testing 5-fold cross validation. Results: Visit frequency of expert pilots was 138%, 47%, 85%, 67%, 117%, and 91% higher than novice pilots in RPM, VVI, altimeter, heading, airspeed, and attitude areas of interest (AOIs) respectively; while 50% and 33% lower in runway and outside AOIs respectively. Dwell time ratio of expert pilots was 38% and 42% higher than novice pilots in runway and outside AOIs respectively; while 62%, 62%, and 65% lower in altimeter, airspeed, and attitude AOIs respectively. Scan path entropy of expert pilots was 33% higher than novice pilots in outside AOI; while 29% lower in attitude AOI. Sensitivity of the classification model for SA high and low status was 73.7%.Conclusion: This study showed that the difference between SA level of expert and novice pilots could be identified by using novel eye tracking features and applied to develop classification model for SA high and low status. Application: The results of this study can be used as objective data of strategy establishment or training feedback for novice pilots.

      • KCI등재

        중년 대상자의 유료노인요양시설에 대한 요구 조사

        김옥수,김소선,김경옥,김영애,김희승,박정숙,최원자 노인간호학회 2006 노인간호학회지 Vol.8 No.1

        Purpose: The purpose of this study was to investigate the needs for paying nursing homes for the middle-aged. Method: The subjects consisted of 365 people aged from 40 to 59 residing in Seoul. The questionnaire was composed of the general characteristics of the subjects, recognition and the intention of utilizing paying nursing homes, expenses of paying nursing homes, the operator of paying nursing homes, environment of paying nursing homes, and the service of paying nursing homes. Result: The frequency of the subjects with intentions of utilizing nursing homes was 48.2%. Gerontological nurse practitioners were preferred as the operators of nursing homes by 72.9% of the subjects. The majority of the subjects hoped to receive medical and nursing service in paying nursing homes. Conclusion: The results indicate that there are needs for high quality, low expense paying nursing homes. Therefore, it is necessary to reflect the needs of the subjects when designing the facility.

      • SCISCIESCOPUS

        Synthesis and antiviral evaluation of novel 4'-trifluoromethylated 5'-deoxyapiosyl nucleoside phosphonic acids.

        Kim, Seyeon,Kim, Eunae,Lee, Wonjae,Hee Hong, Joon Marcel Dekker 2014 NUCLEOSIDES NUCLEOTIDES AND NUCLEIC ACIDS Vol.33 No.12

        <P>On the basis of the discovery that the threosyl nucleoside phosphonate PMDTA is a potent anti-HIV compound, we synthesized several 4'-trifluoromethyl-5'-deoxyapiosyl nucleoside phosphonic acids and evaluated their anti-HIV activity. An efficient synthetic route was optimized, starting from an α-trifluoromethyl-α,β-unsaturated ester. Glycosylation of the purine nucleosidic bases with a glycosyl donor yielded modified nucleoside intermediates, which were then phosphonated and hydrolyzed to provide the targeted nucleoside analogs. Once synthesized, the anti-HIV and cytotoxic activities of each analog were evaluated. None of the analogs showed significant anti-HIV activity at concentrations up to 100?μM.</P>

      • KCI등재

        Extensive Genomic Rearrangement of Catalase-Less Cyanobloom-Forming Microcystis aeruginosa in Freshwater Ecosystems

        Kim Minkyung,Jung Jaejoon,Kim Wonjae,Park Yerim,Jeon Che Ok,Park Woojun 한국미생물학회 2024 The journal of microbiology Vol.62 No.11

        Many of the world’s freshwater ecosystems suffer from cyanobacteria-mediated blooms and their toxins. However, a mechanistic understanding of why and how Microcystis aeruginosa dominates over other freshwater cyanobacteria during warmer summers is lacking. This paper utilizes comparative genomics with other cyanobacteria and literature reviews to predict the gene functions and genomic architectures of M. aeruginosa based on complete genomes. The primary aim is to understand this species' survival and competitive strategies in warmer freshwater environments. M. aeruginosa strains exhibiting a high proportion of insertion sequences (~ 11%) possess genomic structures with low synteny across different strains. This indicates the occurrence of extensive genomic rearrangements and the presence of many possible diverse genotypes that result in greater population heterogeneities than those in other cyanobacteria in order to increase survivability during rapidly changing and threatening environmental challenges. Catalase-less M. aeruginosa strains are even vulnerable to low light intensity in freshwater environments with strong ultraviolet radiation. However, they can continuously grow with the help of various defense genes (e.g., egtBD, cruA, and mysABCD) and associated bacteria. The strong defense strategies against biological threats (e.g., antagonistic bacteria, protozoa, and cyanophages) are attributed to dense exopolysaccharide (EPS)- mediated aggregate formation with efficient buoyancy and the secondary metabolites of M. aeruginosa cells. Our review with extensive genome analysis suggests that the ecological vulnerability of M. aeruginosa cells can be overcome by diverse genotypes, secondary defense metabolites, reinforced EPS, and associated bacteria.

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