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Sooah Jang,Hyunjeong Kim,Hyejin Kim,SuKyoung Lee,EunWoo Kim,KeeNam koong,Eosu Kim 대한신경정신의학회 2018 PSYCHIATRY INVESTIGATION Vol.15 No.2
Objective-Conventional methods for organotypic hippocampal tissue slice culture (OHSC) have shown several disadvantages or limitations regarding age of animals used, duration of culture and difficulty using neurodegenerative models. Therefore, we tried to establish OHSC from old 3xTg-Alzheimer’s disease (AD) mice for longer period (over 4 weeks) and to validate utility of this system as a valid platform for translational neuroscience of AD. Methods-OHSC was performed with old 3xTg-AD mice (12-14 months), old wild type mice (12-14 months) and young 3xTg-AD mice (2-4 months) using serum-free medium for 4 weeks. Hippocampal structure was evaluated by 4’, 6-diamidino-2-phenylindole (DAPI) intensity and neuronal metabolism was measured by Alamarblue assay. Pathologic characteristics of AD were also investigated; β-amyloid levels by ELISA, amyloid plaque deposition by Thioflavin-S staining, and glial activation by immunohistochemistry. Results-Following 4-week culture in serum-free media, hippocampal cells and layers were well preserved in cultured slices from old AD mice as was in those from young AD and old wild type mice. On the contrary, excessive regression of total visible cells was observed in conventional serum-containing medium regardless of genotype of mice. In parallel with this well preserved structure, major pathologic characteristics of AD were also well manifested in hippocampal slices from old AD mice. Conclusion-Our findings suggest that long-term OHSC from old 3xTg-AD mouse can serve as a promising ex vivo system for studies on pathophysiology of AD, especially with the minimum number of sacrifice of experimental animals.
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>
Evaluation and Optimization of Metabolome Sample Preparation Methods for Saccharomyces cerevisiae
Kim, Sooah,Lee, Do Yup,Wohlgemuth, Gert,Park, Hyong Seok,Fiehn, Oliver,Kim, Kyoung Heon American Chemical Society 2013 ANALYTICAL CHEMISTRY - Vol.85 No.4
<P>Metabolome sampling is one of the most important factors that determine the quality of metabolomics data. The main steps in metabolite sample preparation include quenching and metabolite extraction. Quenching with 60% (v/v) cold methanol at −40 °C has been most commonly used for Saccharomyces cerevisiae, and this method was recently modified as “leakage-free cold methanol quenching” using pure methanol at −40 °C. Boiling ethanol (75%, v/v) and cold pure methanol are the most widely used extraction solvents for S. cerevisiae. In the present study, metabolome sampling protocols, including the above methods, were evaluated by analyzing 110 identified intracellular metabolites of S. cerevisiae using gas chromatography/time-of-flight mass spectrometry. According to our results, fast filtration followed by washing with an appropriate volume of water can minimize the metabolite loss due to cell leakage as well as the contamination by extracellular metabolites. For metabolite extraction, acetonitrile/water mixture (1:1, v/v) at −20 °C was the most effective. These results imply that the systematic evaluation of existing methods and the development of customized methods for each microorganism are critical for metabolome sample preparation to facilitate the reliable and accurate analysis of metabolome.</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/ancham/2013/ancham.2013.85.issue-4/ac302881e/production/images/medium/ac-2012-02881e_0005.gif'></P><P><A href='http://pubs.acs.org/doi/suppl/10.1021/ac302881e'>ACS Electronic Supporting Info</A></P>
Kim, In Jung,Nam, Ki Hyun,Yun, Eun Ju,Kim, Sooah,Youn, Hak Jin,Lee, Hee Jin,Choi, In-Geol,Kim, Kyoung Heon Springer Berlin Heidelberg 2015 Applied microbiology and biotechnology Vol.99 No.20
<P>Auxiliary activity family 9 (AA9, formerly known as glycoside hydrolase family 61 or polysaccharide monooxygenase) is a group of fungal proteins that were recently found to have a significant synergism with cellulase in cellulose hydrolysis via the oxidative cleavage of glycosidic bonds of cellulose chains. In this study, we report the active expression of a recombinant fungal AA9 from <I>Chaetomium globosum</I> (<I>Cg</I>AA9) in a bacterial host, <I>Escherichia coli</I>, and the optimization of its synergistic activity in cellulose hydrolysis by using cellulase. The recombinant <I>Cg</I>AA9 (0.9 mg/g cellulose) exhibited 1.7-fold synergism in the hydrolysis of Avicel when incubated with 0.9 filter paper units of Celluclast 1.5 L/g cellulose. The first study of the active expression of AA9 using a bacterial host and its synergistic optimization could be useful for the industrial application of AA9 for the saccharification of lignocellulose.</P><P><B>Electronic supplementary material</B></P><P>The online version of this article (doi:10.1007/s00253-015-6592-3) contains supplementary material, which is available to authorized users.</P>
Metabolomic elucidation of recovery of <i>Melissa officinalis</i> from UV-B irradiation stress
Kim, Sooah,Lee, Hojoung,Kim, Kyoung Heon Elsevier 2018 INDUSTRIAL CROPS AND PRODUCTS Vol.121 No.-
<P><B>Abstract</B></P> <P>UV irradiation is a major stress and leads to the accumulation of secondary metabolites in plants as a protective mechanism. The altered metabolism caused by the stress will eventually return to basal conditions, however, the recovery mechanism after UV irradiation stress remains unknown. To understand how plant metabolism recovers following UV irradiation stress, global metabolite profiling of <I>Melissa officinalis</I> (lemon balm) was performed using gas chromatography/mass spectrometry (GC/MS). Principal component and hierarchical clustering analyses showed the significant discrimination of metabolite profiles between the control (non-irradiated), UV-irradiated <I>M. officinalis</I>, and <I>M. officinalis</I> allowed to recover from the UV stress. The glycolysis and phenylpropanoid pathway rapidly reverted to their original states. In contrast, the TCA cycle and amino acid biosynthesis returned slowly to their original states. This study determined that the metabolism and metabolite levels recover their original conditions after the removal of UV irradiation, and that the recovery time of each metabolic pathway differs.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Metabolite profiles reverted to their original states during recovery. </LI> <LI> The glycolysis and phenylpropanoid pathway recovered rapidly after UV stress. </LI> <LI> The TCA cycle and amino acid biosynthesis recovered slowly after UV stress. </LI> <LI> Metabolomics is a powerful tool to investigate metabolic recovery from stress. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>
Sooah Jang,In-Young Kim,Sun-Woo Choi,Anna Lee,Ju-Yeal Lee,Hyunkyung Shin,Junwoo Lee,Mikyeong Lee,Kyoung-Ryul Lee,Saeeun Jung,Jin Sun Ryu,Jihee Oh,Manjae Kwon,Joohan Kim,Ryunsup Ahn,Young-Chul Jung,Jeo 대한신경정신의학회 2024 PSYCHIATRY INVESTIGATION Vol.21 No.3
Objective This study evaluated the clinical effectiveness of Minds.NAVI, a depression screening kit combining psychometric measures and stress hormone biomarkers, in a prospective clinical trial. The objective was to assess its potential as a depression screening tool and investigate the associations between psychological assessments, salivary hormone staging, and depression severity.Methods Thirty-five participants with major depressive disorder and 12 healthy controls (HCs) were included. The Minds.NAVI software, utilizing the PROtective and Vulnerable factors battEry Test (PROVE) and salivary cortisol/dehydroepiandrosterone (DHEA) analysis, was employed. The PROVE test is a comprehensive self-report questionnaire that assesses depressive symptoms, suicide risk, attachment style, adverse childhood experiences, mentalization capacity, and resilience. In addition, salivary cortisol and DHEA levels were measured to evaluate the functional stage of the hypothalamic–pituitary–adrenal (HPA) axis.Results Minds.NAVI exhibited 100% sensitivity, 91.7% specificity, and 97.9% accuracy in distinguishing depression from HCs within an exploratory small group. Salivary stress hormone phases showed changes with depression stage (p=0.030), and the proportion of patients with “adrenal exhaustion stage” was higher in the moderate/severe depression group (p=0.038). Protective/vulnerable factors differed significantly between controls and depressed groups (p<0.001). Cortisol awakening response inversely correlated with depressive symptom severity (r=-0.31, p=0.034).Conclusion This study suggested possible clinical effectiveness of Minds.NAVI, a depression screening tool that integrates psychometric measures and stress hormone biomarkers. The findings support the potential association between depression, chronic stress, and HPA axis hyporesponsiveness.