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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>
Christoph Niessen,Ernst Michael Jung,Walter A. Wohlgemuth,Benedikt Trabold,Michael Haimerl,Andreas Schreyer,Christian Stroszczynski,Philipp Wiggermann 대한영상의학회 2013 Korean Journal of Radiology Vol.14 No.5
We report in a 65-year-old man hepatocellular carcinoma adjacent to a transjugular intrahepatic portosystemic shunt stent-graft which was successfully treated with irreversible electroporation (IRE). IRE is a new non-thermal tissue ablation technique which uses electrical pulses to induce cell necrosis by irreversible membrane poration. IRE proved to be more advantageous in the ablation of perivascular tumor with little injury to the surrounding structures.
Mitochondrial iron accumulation with age and functional consequences
Seo, Arnold ,Y. ,Xu, Jinze ,Servais, Stephane ,Hofer, Tim ,Marzetti, Emanuele ,Wohlgemuth, Stephanie ,E. ,Knutson, Mitchell ,D. ,Chung, Hae ,Young ,Leeu Blackwell Publishing Ltd 2008 Aging Cell Vol.7 No.5
<P>Summary</P><P>During the aging process, an accumulation of non-heme iron disrupts cellular homeostasis and contributes to the mitochondrial dysfunction typical of various neuromuscular degenerative diseases. Few studies have investigated the effects of iron accumulation on mitochondrial integrity and function in skeletal muscle and liver tissue. Thus, we isolated liver mitochondria (LM), as well as quadriceps-derived subsarcolemmal mitochondria (SSM) and interfibrillar mitochondria (IFM), from male Fischer 344× Brown Norway rats at 8, 18, 29 and 37 months of age. Non-heme iron content in SSM, IFM and LM was significantly higher with age, reaching a maximum at 37 months of age. The mitochondrial permeability transition pore (mPTP) was more susceptible to the opening in aged mitochondria containing high levels of iron (i.e. SSM and LM) compared to IFM. Furthermore, mitochondrial RNA oxidation increased significantly with age in SSM and LM, but not in IFM. Levels of mitochondrial RNA oxidation in SSM and LM correlated positively with levels of mitochondrial iron, whereas a significant negative correlation was observed between the maximum Ca<SUP>2+</SUP> amounts needed to induce mPTP opening and iron contents in SSM, IFM and LM. Overall, our data suggest that age-dependent accumulation of mitochondrial iron may increase mitochondrial dysfunction and oxidative damage, thereby enhancing the susceptibility to apoptosis.</P>