https://wiki.oroboros.at/index.php?title=Hoppel_2016b_Abstract_MitoFit_Science_Camp_2016&feed=atom&action=historyHoppel 2016b Abstract MitoFit Science Camp 2016 - Revision history2024-03-29T09:20:39ZRevision history for this page on the wikiMediaWiki 1.36.1https://wiki.oroboros.at/index.php?title=Hoppel_2016b_Abstract_MitoFit_Science_Camp_2016&diff=223710&oldid=prevGnaiger Erich at 16:44, 10 January 20222022-01-10T16:44:07Z<p></p>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>The goal of our study was to perform a quantitative and qualitative comparison of those two methods in muscle biopsy samples (''vastus lateralis''). The muscle sample from 40 patients was used for (1) isolation of mitochondria, and (2) muscle fibers permeabilization. Oxygen consumption was performed with the isolated mitochondria using a conventional oxygraph and standard protocols (performed by Mariana Rosca, M.D. [1]), while for the permeabilized fibers, high-resolution respirometry combined with a multiple substrate-uncoupler-inhibitor-titration (SUIT) protocol was used ([[Vazquez E|Edwin Vazquez]] [2]). Citrate synthase activity (Hiral Patel), a marker of mitochondrial content, was compared in homogenate of muscle and permeabilized fibers removed from the chamber after the estimation of OXPHOS. In a group identified as “pathological controls”, a quantitative comparison of the OXPHOS capacities obtained was performed between the two methods (studies done by [[Lemieux H|Helene Lemieux, Ph.D.]]). Oxygen flux in isolated mitochondria was expressed per g wet weight of muscle using the mitochondrial protein yield from the muscle, or per unit of citrate synthase activity. Furthermore, a factor was applied for the conversion of flux in isolated mitochondria measured at 30 °C to the corresponding flux in the permeabilized fibers measured at 37 °C [2]. Independent of how the data are expressed, the estimation of mitochondrial respiration for substrates entering at Complexes I, II, or IV were similar if not identical in both methods. The careful and quantitative evaluation of methods available for measurement of mitochondrial function is important for combining such results in a comparative data base [2].</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>The goal of our study was to perform a quantitative and qualitative comparison of those two methods in muscle biopsy samples (''vastus lateralis''). The muscle sample from 40 patients was used for (1) isolation of mitochondria, and (2) muscle fibers permeabilization. Oxygen consumption was performed with the isolated mitochondria using a conventional oxygraph and standard protocols (performed by Mariana Rosca, M.D. [1]), while for the permeabilized fibers, high-resolution respirometry combined with a multiple substrate-uncoupler-inhibitor-titration (SUIT) protocol was used ([[Vazquez E|Edwin Vazquez]] [2]). Citrate synthase activity (Hiral Patel), a marker of mitochondrial content, was compared in homogenate of muscle and permeabilized fibers removed from the chamber after the estimation of OXPHOS. In a group identified as “pathological controls”, a quantitative comparison of the OXPHOS capacities obtained was performed between the two methods (studies done by [[Lemieux H|Helene Lemieux, Ph.D.]]). Oxygen flux in isolated mitochondria was expressed per g wet weight of muscle using the mitochondrial protein yield from the muscle, or per unit of citrate synthase activity. Furthermore, a factor was applied for the conversion of flux in isolated mitochondria measured at 30 °C to the corresponding flux in the permeabilized fibers measured at 37 °C [2]. Independent of how the data are expressed, the estimation of mitochondrial respiration for substrates entering at Complexes I, II, or IV were similar if not identical in both methods. The careful and quantitative evaluation of methods available for measurement of mitochondrial function is important for combining such results in a comparative data base [2].</div></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>|mipnetlab=US OH Cleveland Hoppel CL<del style="font-weight: bold; text-decoration: none;">, AT Innsbruck Gnaiger E</del>, AT Innsbruck Oroboros</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>|mipnetlab=US OH Cleveland Hoppel CL, AT Innsbruck Oroboros</div></td></tr>
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<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">== Affiliations ==</ins></div></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">1-Center Mitochondr Diseases, Depts Pharmacology & Medicine, Case Western Reserve School Med, Cleveland, OH, USA. - charles.hoppel@case.edu</ins></div></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">2-Daniel Swarovski Research Laboratory, Dept General Transplant Surgery, Med Univ Innsbruck, Austria</ins></div></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">== References ==</ins></div></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">#Puchowicz MA, Varnes ME, Cohen BH, Friedman NR, Kerr DS, Hoppel CL (2004) Oxidative phosphorylation analysis: assessing the integrated functional activity of human skeletal muscle mitochondria-case studies. Mitochondrion 4:377-85. [[Puchowicz 2004 Mitochondrion|»Bioblast Link]]</ins></div></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">#Gnaiger E (2009) Capacity of oxidative phosphorylation in human skeletal muscle. New perspectives of mitochondrial physiology. Int J Biochem Cell Biol 41:1837-45. [[Gnaiger 2009 Int J Biochem Cell Biol|»Bioblast Link]]</ins></div></td></tr>
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<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del style="font-weight: bold; text-decoration: none;">== Affiliations ==</del></div></td><td colspan="2"></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del style="font-weight: bold; text-decoration: none;">1-Center Mitochondr Diseases, Depts Pharmacology & Medicine, Case Western Reserve School Med, Cleveland, OH, USA. - charles.hoppel@case.edu</del></div></td><td colspan="2"></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del style="font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2"></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del style="font-weight: bold; text-decoration: none;">2-Daniel Swarovski Research Laboratory, Dept General Transplant Surgery, Med Univ Innsbruck, Austria</del></div></td><td colspan="2"></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del style="font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2"></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del style="font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2"></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del style="font-weight: bold; text-decoration: none;">== References ==</del></div></td><td colspan="2"></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del style="font-weight: bold; text-decoration: none;">#Puchowicz MA, Varnes ME, Cohen BH, Friedman NR, Kerr DS, Hoppel CL (2004) Oxidative phosphorylation analysis: assessing the integrated functional activity of human skeletal muscle mitochondria-case studies. Mitochondrion 4:377-85. [[Puchowicz 2004 Mitochondrion|»Bioblast Link]]</del></div></td><td colspan="2"></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del style="font-weight: bold; text-decoration: none;">#Gnaiger E (2009) Capacity of oxidative phosphorylation in human skeletal muscle. New perspectives of mitochondrial physiology. Int J Biochem Cell Biol 41:1837-45. [[Gnaiger 2009 Int J Biochem Cell Biol|»Bioblast Link]]</del></div></td><td colspan="2"></td></tr>
</table>Gnaiger Erichhttps://wiki.oroboros.at/index.php?title=Hoppel_2016b_Abstract_MitoFit_Science_Camp_2016&diff=171293&oldid=prevBeno Marija at 13:32, 23 January 20192019-01-23T13:32:17Z<p></p>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>The goal of our study was to perform a quantitative and qualitative comparison of those two methods in muscle biopsy samples (''vastus lateralis''). The muscle sample from 40 patients was used for (1) isolation of mitochondria, and (2) muscle fibers permeabilization. Oxygen consumption was performed with the isolated mitochondria using a conventional oxygraph and standard protocols (performed by Mariana Rosca, M.D. [1]), while for the permeabilized fibers, high-resolution respirometry combined with a multiple substrate-uncoupler-inhibitor-titration (SUIT) protocol was used ([[Vazquez E|Edwin Vazquez]] [2]). Citrate synthase activity (Hiral Patel), a marker of mitochondrial content, was compared in homogenate of muscle and permeabilized fibers removed from the chamber after the estimation of OXPHOS. In a group identified as “pathological controls”, a quantitative comparison of the OXPHOS capacities obtained was performed between the two methods (studies done by [[Lemieux H|Helene Lemieux, Ph.D.]]). Oxygen flux in isolated mitochondria was expressed per g wet weight of muscle using the mitochondrial protein yield from the muscle, or per unit of citrate synthase activity. Furthermore, a factor was applied for the conversion of flux in isolated mitochondria measured at 30 °C to the corresponding flux in the permeabilized fibers measured at 37 °C [2]. Independent of how the data are expressed, the estimation of mitochondrial respiration for substrates entering at Complexes I, II, or IV were similar if not identical in both methods. The careful and quantitative evaluation of methods available for measurement of mitochondrial function is important for combining such results in a comparative data base [2].</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>The goal of our study was to perform a quantitative and qualitative comparison of those two methods in muscle biopsy samples (''vastus lateralis''). The muscle sample from 40 patients was used for (1) isolation of mitochondria, and (2) muscle fibers permeabilization. Oxygen consumption was performed with the isolated mitochondria using a conventional oxygraph and standard protocols (performed by Mariana Rosca, M.D. [1]), while for the permeabilized fibers, high-resolution respirometry combined with a multiple substrate-uncoupler-inhibitor-titration (SUIT) protocol was used ([[Vazquez E|Edwin Vazquez]] [2]). Citrate synthase activity (Hiral Patel), a marker of mitochondrial content, was compared in homogenate of muscle and permeabilized fibers removed from the chamber after the estimation of OXPHOS. In a group identified as “pathological controls”, a quantitative comparison of the OXPHOS capacities obtained was performed between the two methods (studies done by [[Lemieux H|Helene Lemieux, Ph.D.]]). Oxygen flux in isolated mitochondria was expressed per g wet weight of muscle using the mitochondrial protein yield from the muscle, or per unit of citrate synthase activity. Furthermore, a factor was applied for the conversion of flux in isolated mitochondria measured at 30 °C to the corresponding flux in the permeabilized fibers measured at 37 °C [2]. Independent of how the data are expressed, the estimation of mitochondrial respiration for substrates entering at Complexes I, II, or IV were similar if not identical in both methods. The careful and quantitative evaluation of methods available for measurement of mitochondrial function is important for combining such results in a comparative data base [2].</div></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>|mipnetlab=US OH Cleveland Hoppel CL, AT Innsbruck Gnaiger E, AT Innsbruck <del style="font-weight: bold; text-decoration: none;">OROBOROS</del></div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>|mipnetlab=US OH Cleveland Hoppel CL, AT Innsbruck Gnaiger E, AT Innsbruck <ins style="font-weight: bold; text-decoration: none;">Oroboros</ins></div></td></tr>
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</table>Beno Marijahttps://wiki.oroboros.at/index.php?title=Hoppel_2016b_Abstract_MitoFit_Science_Camp_2016&diff=123596&oldid=prevBeno Marija at 15:28, 7 November 20162016-11-07T15:28:23Z<p></p>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|preparations=Permeabilized cells, Homogenate, Isolated mitochondria</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|preparations=Permeabilized cells, Homogenate, Isolated mitochondria</div></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>|<del style="font-weight: bold; text-decoration: none;">substratestates</del>=<del style="font-weight: bold; text-decoration: none;">CI</del>, <del style="font-weight: bold; text-decoration: none;">CII</del>, CIV</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>|<ins style="font-weight: bold; text-decoration: none;">pathways</ins>=<ins style="font-weight: bold; text-decoration: none;">N</ins>, <ins style="font-weight: bold; text-decoration: none;">S</ins>, CIV</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|instruments=Oxygraph-2k</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|instruments=Oxygraph-2k</div></td></tr>
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</table>Beno Marijahttps://wiki.oroboros.at/index.php?title=Hoppel_2016b_Abstract_MitoFit_Science_Camp_2016&diff=115064&oldid=prevKandolf Georg at 07:07, 1 July 20162016-07-01T07:07:41Z<p></p>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|substratestates=CI, CII, CIV</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|substratestates=CI, CII, CIV</div></td></tr>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>== Affiliations ==</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>== Affiliations ==</div></td></tr>
</table>Kandolf Georghttps://wiki.oroboros.at/index.php?title=Hoppel_2016b_Abstract_MitoFit_Science_Camp_2016&diff=114795&oldid=prevKandolf Georg at 14:16, 27 June 20162016-06-27T14:16:17Z<p></p>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|abstract=We use the measurement of integrated mitochondrial function or oxidative phosphorylation (OXPHOS) to evaluate mitochondrial dysfunction in energy production as a diagnostic tool in skeletal muscle. During an one year period we used both isolated skeletal muscle mitochondria and permeabilized fibers from the same biopsy to evaluate OXPHOS capacity. </div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|abstract=We use the measurement of integrated mitochondrial function or oxidative phosphorylation (OXPHOS) to evaluate mitochondrial dysfunction in energy production as a diagnostic tool in skeletal muscle. During an one year period we used both isolated skeletal muscle mitochondria and permeabilized fibers from the same biopsy to evaluate OXPHOS capacity. </div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>The goal of our study was to perform a quantitative and qualitative comparison of those two methods in muscle biopsy samples (''vastus lateralis''). The muscle sample from 40 patients was used for (1) isolation of mitochondria, and (2) muscle fibers permeabilization. Oxygen consumption was performed with the isolated mitochondria using a conventional oxygraph and standard protocols (performed by Mariana Rosca, M.D. [1]), while for the permeabilized fibers, high-resolution respirometry combined with a multiple substrate-uncoupler-inhibitor-titration (SUIT) protocol was used ([[Vazquez E|Edwin Vazquez]] [2]). Citrate synthase activity (<del style="font-weight: bold; text-decoration: none;">[[Patel H|</del>Hiral Patel<del style="font-weight: bold; text-decoration: none;">]]</del>), a marker of mitochondrial content, was compared in homogenate of muscle and permeabilized fibers removed from the chamber after the estimation of OXPHOS. In a group identified as “pathological controls”, a quantitative comparison of the OXPHOS capacities obtained was performed between the two methods (studies done by [[Lemieux H|Helene Lemieux, Ph.D.]]). Oxygen flux in isolated mitochondria was expressed per g wet weight of muscle using the mitochondrial protein yield from the muscle, or per unit of citrate synthase activity. Furthermore, a factor was applied for the conversion of flux in isolated mitochondria measured at 30 °C to the corresponding flux in the permeabilized fibers measured at 37 °C [2]. Independent of how the data are expressed, the estimation of mitochondrial respiration for substrates entering at Complexes I, II, or IV were similar if not identical in both methods. The careful and quantitative evaluation of methods available for measurement of mitochondrial function is important for combining such results in a comparative data base [2].</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>The goal of our study was to perform a quantitative and qualitative comparison of those two methods in muscle biopsy samples (''vastus lateralis''). The muscle sample from 40 patients was used for (1) isolation of mitochondria, and (2) muscle fibers permeabilization. Oxygen consumption was performed with the isolated mitochondria using a conventional oxygraph and standard protocols (performed by Mariana Rosca, M.D. [1]), while for the permeabilized fibers, high-resolution respirometry combined with a multiple substrate-uncoupler-inhibitor-titration (SUIT) protocol was used ([[Vazquez E|Edwin Vazquez]] [2]). Citrate synthase activity (Hiral Patel), a marker of mitochondrial content, was compared in homogenate of muscle and permeabilized fibers removed from the chamber after the estimation of OXPHOS. In a group identified as “pathological controls”, a quantitative comparison of the OXPHOS capacities obtained was performed between the two methods (studies done by [[Lemieux H|Helene Lemieux, Ph.D.]]). Oxygen flux in isolated mitochondria was expressed per g wet weight of muscle using the mitochondrial protein yield from the muscle, or per unit of citrate synthase activity. Furthermore, a factor was applied for the conversion of flux in isolated mitochondria measured at 30 °C to the corresponding flux in the permeabilized fibers measured at 37 °C [2]. Independent of how the data are expressed, the estimation of mitochondrial respiration for substrates entering at Complexes I, II, or IV were similar if not identical in both methods. The careful and quantitative evaluation of methods available for measurement of mitochondrial function is important for combining such results in a comparative data base [2].</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|mipnetlab=US OH Cleveland Hoppel CL, AT Innsbruck Gnaiger E, AT Innsbruck OROBOROS</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|mipnetlab=US OH Cleveland Hoppel CL, AT Innsbruck Gnaiger E, AT Innsbruck OROBOROS</div></td></tr>
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</table>Kandolf Georghttps://wiki.oroboros.at/index.php?title=Hoppel_2016b_Abstract_MitoFit_Science_Camp_2016&diff=114792&oldid=prevKandolf Georg at 14:15, 27 June 20162016-06-27T14:15:02Z<p></p>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|year=2016</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|year=2016</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|event=MitoFit Science Camp 2016 Kuehtai AT</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|event=MitoFit Science Camp 2016 Kuehtai AT</div></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>|abstract=We use the measurement of integrated mitochondrial function or oxidative phosphorylation (OXPHOS) to evaluate mitochondrial dysfunction in energy production as a diagnostic tool in skeletal muscle. During an <del style="font-weight: bold; text-decoration: none;">c. </del>one year period we used both isolated skeletal muscle mitochondria and permeabilized fibers from the same biopsy to evaluate OXPHOS capacity. </div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>|abstract=We use the measurement of integrated mitochondrial function or oxidative phosphorylation (OXPHOS) to evaluate mitochondrial dysfunction in energy production as a diagnostic tool in skeletal muscle. During an one year period we used both isolated skeletal muscle mitochondria and permeabilized fibers from the same biopsy to evaluate OXPHOS capacity. </div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>The goal of our study was to perform a quantitative and qualitative comparison of those two methods in muscle biopsy samples (''vastus lateralis''). The muscle sample from 40 patients was used for (1) isolation of mitochondria, and (2) muscle fibers permeabilization. Oxygen consumption was performed with the isolated mitochondria using a conventional oxygraph and standard protocols (performed by Mariana Rosca, M.D. [1]), while for the permeabilized fibers, high-resolution respirometry combined with a multiple substrate-uncoupler-inhibitor-titration (SUIT) protocol was used ([[Vazquez E|Edwin Vazquez]] [2]). Citrate synthase activity ([[Patel H|Hiral Patel]]), a marker of mitochondrial content, was compared in homogenate of muscle and permeabilized fibers removed from the chamber after the estimation of OXPHOS. In a group identified as “pathological controls”, a quantitative comparison of the OXPHOS capacities obtained was performed between the two methods (studies done by [[Lemieux H|Helene Lemieux, Ph.D.]]). Oxygen flux in isolated mitochondria was expressed per g wet weight of muscle using the mitochondrial protein yield from the muscle, or per unit of citrate synthase activity. Furthermore, a factor was applied for the conversion of flux in isolated mitochondria measured at 30 °C to the corresponding flux in the permeabilized fibers measured at 37 °C [2]. Independent of how the data are expressed, the estimation of mitochondrial respiration for substrates entering at Complexes I, II, or IV were similar if not identical in both methods. The careful and quantitative evaluation of methods available for measurement of mitochondrial function is important for combining such results in a comparative data base [2].</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>The goal of our study was to perform a quantitative and qualitative comparison of those two methods in muscle biopsy samples (''vastus lateralis''). The muscle sample from 40 patients was used for (1) isolation of mitochondria, and (2) muscle fibers permeabilization. Oxygen consumption was performed with the isolated mitochondria using a conventional oxygraph and standard protocols (performed by Mariana Rosca, M.D. [1]), while for the permeabilized fibers, high-resolution respirometry combined with a multiple substrate-uncoupler-inhibitor-titration (SUIT) protocol was used ([[Vazquez E|Edwin Vazquez]] [2]). Citrate synthase activity ([[Patel H|Hiral Patel]]), a marker of mitochondrial content, was compared in homogenate of muscle and permeabilized fibers removed from the chamber after the estimation of OXPHOS. In a group identified as “pathological controls”, a quantitative comparison of the OXPHOS capacities obtained was performed between the two methods (studies done by [[Lemieux H|Helene Lemieux, Ph.D.]]). Oxygen flux in isolated mitochondria was expressed per g wet weight of muscle using the mitochondrial protein yield from the muscle, or per unit of citrate synthase activity. Furthermore, a factor was applied for the conversion of flux in isolated mitochondria measured at 30 °C to the corresponding flux in the permeabilized fibers measured at 37 °C [2]. Independent of how the data are expressed, the estimation of mitochondrial respiration for substrates entering at Complexes I, II, or IV were similar if not identical in both methods. The careful and quantitative evaluation of methods available for measurement of mitochondrial function is important for combining such results in a comparative data base [2].</div></td></tr>
</table>Kandolf Georghttps://wiki.oroboros.at/index.php?title=Hoppel_2016b_Abstract_MitoFit_Science_Camp_2016&diff=114760&oldid=prevKandolf Georg at 07:55, 27 June 20162016-06-27T07:55:35Z<p></p>
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<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Revision as of 07:55, 27 June 2016</td>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>== References ==</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>== References ==</div></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>#Puchowicz MA, Varnes ME, Cohen BH, Friedman NR, Kerr DS, Hoppel CL (2004) Oxidative phosphorylation analysis: assessing the integrated functional activity of human skeletal muscle mitochondria-case studies. Mitochondrion 4:377-85.</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>#Puchowicz MA, Varnes ME, Cohen BH, Friedman NR, Kerr DS, Hoppel CL (2004) Oxidative phosphorylation analysis: assessing the integrated functional activity of human skeletal muscle mitochondria-case studies. Mitochondrion 4:377-85. <ins style="font-weight: bold; text-decoration: none;">[[Puchowicz 2004 Mitochondrion|»Bioblast Link]]</ins></div></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>#Gnaiger E (2009) Capacity of oxidative phosphorylation in human skeletal muscle. New perspectives of mitochondrial physiology. Int J Biochem Cell Biol 41:1837-45.</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>#Gnaiger E (2009) Capacity of oxidative phosphorylation in human skeletal muscle. New perspectives of mitochondrial physiology. Int J Biochem Cell Biol 41:1837-45. <ins style="font-weight: bold; text-decoration: none;">[[Gnaiger 2009 Int J Biochem Cell Biol|»Bioblast Link]]</ins></div></td></tr>
</table>Kandolf Georghttps://wiki.oroboros.at/index.php?title=Hoppel_2016b_Abstract_MitoFit_Science_Camp_2016&diff=114753&oldid=prevKandolf Georg at 07:46, 27 June 20162016-06-27T07:46:09Z<p></p>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>The goal of our study was to perform a quantitative and qualitative comparison of those two methods in muscle biopsy samples (''vastus lateralis''). The muscle sample from 40 patients was used for (1) isolation of mitochondria, and (2) muscle fibers permeabilization. Oxygen consumption was performed with the isolated mitochondria using a conventional oxygraph and standard protocols (performed by Mariana Rosca, M.D. [1]), while for the permeabilized fibers, high-resolution respirometry combined with a multiple substrate-uncoupler-inhibitor-titration (SUIT) protocol was used ([[Vazquez E|Edwin Vazquez]] [2]). Citrate synthase activity ([[Patel H|Hiral Patel]]), a marker of mitochondrial content, was compared in homogenate of muscle and permeabilized fibers removed from the chamber after the estimation of OXPHOS. In a group identified as “pathological controls”, a quantitative comparison of the OXPHOS capacities obtained was performed between the two methods (studies done by [[Lemieux H|Helene Lemieux, Ph.D.]]). Oxygen flux in isolated mitochondria was expressed per g wet weight of muscle using the mitochondrial protein yield from the muscle, or per unit of citrate synthase activity. Furthermore, a factor was applied for the conversion of flux in isolated mitochondria measured at 30 °C to the corresponding flux in the permeabilized fibers measured at 37 °C [2]. Independent of how the data are expressed, the estimation of mitochondrial respiration for substrates entering at Complexes I, II, or IV were similar if not identical in both methods. The careful and quantitative evaluation of methods available for measurement of mitochondrial function is important for combining such results in a comparative data base [2].</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>The goal of our study was to perform a quantitative and qualitative comparison of those two methods in muscle biopsy samples (''vastus lateralis''). The muscle sample from 40 patients was used for (1) isolation of mitochondria, and (2) muscle fibers permeabilization. Oxygen consumption was performed with the isolated mitochondria using a conventional oxygraph and standard protocols (performed by Mariana Rosca, M.D. [1]), while for the permeabilized fibers, high-resolution respirometry combined with a multiple substrate-uncoupler-inhibitor-titration (SUIT) protocol was used ([[Vazquez E|Edwin Vazquez]] [2]). Citrate synthase activity ([[Patel H|Hiral Patel]]), a marker of mitochondrial content, was compared in homogenate of muscle and permeabilized fibers removed from the chamber after the estimation of OXPHOS. In a group identified as “pathological controls”, a quantitative comparison of the OXPHOS capacities obtained was performed between the two methods (studies done by [[Lemieux H|Helene Lemieux, Ph.D.]]). Oxygen flux in isolated mitochondria was expressed per g wet weight of muscle using the mitochondrial protein yield from the muscle, or per unit of citrate synthase activity. Furthermore, a factor was applied for the conversion of flux in isolated mitochondria measured at 30 °C to the corresponding flux in the permeabilized fibers measured at 37 °C [2]. Independent of how the data are expressed, the estimation of mitochondrial respiration for substrates entering at Complexes I, II, or IV were similar if not identical in both methods. The careful and quantitative evaluation of methods available for measurement of mitochondrial function is important for combining such results in a comparative data base [2].</div></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|mipnetlab=US OH Cleveland Hoppel CL, AT Innsbruck Gnaiger E, AT Innsbruck OROBOROS</ins></div></td></tr>
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</table>Kandolf Georghttps://wiki.oroboros.at/index.php?title=Hoppel_2016b_Abstract_MitoFit_Science_Camp_2016&diff=114752&oldid=prevKandolf Georg at 07:45, 27 June 20162016-06-27T07:45:24Z<p></p>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>{{Abstract</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>{{Abstract</div></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>|title=Comparison of permeabilized skeletal muscle fibers or isolated mitochondria for the detection of oxidative phosphorylation defects</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>|title=Comparison of permeabilized skeletal muscle fibers or isolated mitochondria for the detection of oxidative phosphorylation defects<ins style="font-weight: bold; text-decoration: none;">.</ins></div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|authors=Hoppel CL, Gnaiger E</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|authors=Hoppel CL, Gnaiger E</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|year=2016</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|year=2016</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|event=MitoFit Science Camp 2016 Kuehtai AT</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|event=MitoFit Science Camp 2016 Kuehtai AT</div></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>|abstract=We use the measurement of integrated mitochondrial function or oxidative phosphorylation (OXPHOS) to evaluate mitochondrial dysfunction in energy production as a diagnostic tool in skeletal muscle. During an c. one year period we used both isolated skeletal muscle mitochondria and permeabilized fibers from the same biopsy to evaluate OXPHOS capacity. <del style="font-weight: bold; text-decoration: none;">The goal of our study was to perform a quantitative and qualitative comparison of those two methods in muscle biopsy samples (vastus lateralis). The muscle sample from 40 patients was used for (1) isolation of mitochondria, and (2) muscle fibers permeabilization. Oxygen consumption was performed with the isolated mitochondria using a conventional oxygraph and standard protocols (performed by Mariana Rosca, M.D. [1]), while for the permeabilized fibers, high-resolution respirometry combined with a multiple substrate-uncoupler-inhibitor-titration (SUIT) protocol was used (Edwin Vazquez [2]). Citrate synthase activity (Hiral Patel), a marker of mitochondrial content, was compared in homogenate of muscle and permeabilized fibers removed from the chamber after the estimation of OXPHOS. In a group identified as “pathological controls”, a quantitative comparison of the OXPHOS capacities obtained was performed between the two methods (studies done by Helene Lemieux, Ph.D.). Oxygen flux in isolated mitochondria was expressed per g wet weight of muscle using the mitochondrial protein yield from the muscle, or per unit of citrate synthase activity. Furthermore, a factor was applied for the conversion of flux in isolated mitochondria measured at 30 °C to the corresponding flux in the permeabilized fibers measured at 37 °C [2]. Independent of how the data are expressed, the estimation of mitochondrial respiration for substrates entering at Complexes I, II, or IV were similar if not identical in both methods. The careful and quantitative evaluation of methods available for measurement of mitochondrial function is important for combining such results in a comparative data base [2].</del></div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>|abstract=We use the measurement of integrated mitochondrial function or oxidative phosphorylation (OXPHOS) to evaluate mitochondrial dysfunction in energy production as a diagnostic tool in skeletal muscle. During an c. one year period we used both isolated skeletal muscle mitochondria and permeabilized fibers from the same biopsy to evaluate OXPHOS capacity. </div></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div> </div></td><td colspan="2"></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">The goal of our study was to perform a quantitative and qualitative comparison of those two methods in muscle biopsy samples (''vastus lateralis''). The muscle sample from 40 patients was used for (1) isolation of mitochondria, and (2) muscle fibers permeabilization. Oxygen consumption was performed with the isolated mitochondria using a conventional oxygraph and standard protocols (performed by Mariana Rosca, M.D. [1]), while for the permeabilized fibers, high-resolution respirometry combined with a multiple substrate-uncoupler-inhibitor-titration (SUIT) protocol was used ([[Vazquez E|Edwin Vazquez]] [2]). Citrate synthase activity ([[Patel H|Hiral Patel]]), a marker of mitochondrial content, was compared in homogenate of muscle and permeabilized fibers removed from the chamber after the estimation of OXPHOS. In a group identified as “pathological controls”, a quantitative comparison of the OXPHOS capacities obtained was performed between the two methods (studies done by [[Lemieux H|Helene Lemieux, Ph.D.]]). Oxygen flux in isolated mitochondria was expressed per g wet weight of muscle using the mitochondrial protein yield from the muscle, or per unit of citrate synthase activity. Furthermore, a factor was applied for the conversion of flux in isolated mitochondria measured at 30 °C to the corresponding flux in the permeabilized fibers measured at 37 °C [2]. Independent of how the data are expressed, the estimation of mitochondrial respiration for substrates entering at Complexes I, II, or IV were similar if not identical in both methods. The careful and quantitative evaluation of methods available for measurement of mitochondrial function is important for combining such results in a comparative data base [2].</ins></div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>}}</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>}}</div></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>{{</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>{{<ins style="font-weight: bold; text-decoration: none;">Labeling</ins></div></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|area=Respiration, Patients</ins></div></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|organism=Human</ins></div></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|tissues=Skeletal muscle</ins></div></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|preparations=Permeabilized cells, Homogenate, Isolated mitochondria</ins></div></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|substratestates=CI, CII, CIV</ins></div></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|instruments=Oxygraph-2k</ins></div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>}}</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>}}</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>== Affiliations ==</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>== Affiliations ==</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>1-Center Mitochondr Diseases, Depts Pharmacology & Medicine, Case Western Reserve School Med, Cleveland, OH, USA. - charles.hoppel@case.edu</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>1-Center Mitochondr Diseases, Depts Pharmacology & Medicine, Case Western Reserve School Med, Cleveland, OH, USA. - charles.hoppel@case.edu</div></td></tr>
<tr><td colspan="2"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>2-Daniel Swarovski Research Laboratory, Dept General Transplant Surgery, Med Univ Innsbruck, Austria</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>2-Daniel Swarovski Research Laboratory, Dept General Transplant Surgery, Med Univ Innsbruck, Austria</div></td></tr>
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</table>Kandolf Georghttps://wiki.oroboros.at/index.php?title=Hoppel_2016b_Abstract_MitoFit_Science_Camp_2016&diff=114751&oldid=prevKandolf Georg: Created page with "{{Abstract |title=Comparison of permeabilized skeletal muscle fibers or isolated mitochondria for the detection of oxidative phosphorylation defects |authors=Hoppel CL, Gnaiger E..."2016-06-27T07:27:15Z<p>Created page with "{{Abstract |title=Comparison of permeabilized skeletal muscle fibers or isolated mitochondria for the detection of oxidative phosphorylation defects |authors=Hoppel CL, Gnaiger E..."</p>
<p><b>New page</b></p><div>{{Abstract<br />
|title=Comparison of permeabilized skeletal muscle fibers or isolated mitochondria for the detection of oxidative phosphorylation defects<br />
|authors=Hoppel CL, Gnaiger E<br />
|year=2016<br />
|event=MitoFit Science Camp 2016 Kuehtai AT<br />
|abstract=We use the measurement of integrated mitochondrial function or oxidative phosphorylation (OXPHOS) to evaluate mitochondrial dysfunction in energy production as a diagnostic tool in skeletal muscle. During an c. one year period we used both isolated skeletal muscle mitochondria and permeabilized fibers from the same biopsy to evaluate OXPHOS capacity. The goal of our study was to perform a quantitative and qualitative comparison of those two methods in muscle biopsy samples (vastus lateralis). The muscle sample from 40 patients was used for (1) isolation of mitochondria, and (2) muscle fibers permeabilization. Oxygen consumption was performed with the isolated mitochondria using a conventional oxygraph and standard protocols (performed by Mariana Rosca, M.D. [1]), while for the permeabilized fibers, high-resolution respirometry combined with a multiple substrate-uncoupler-inhibitor-titration (SUIT) protocol was used (Edwin Vazquez [2]). Citrate synthase activity (Hiral Patel), a marker of mitochondrial content, was compared in homogenate of muscle and permeabilized fibers removed from the chamber after the estimation of OXPHOS. In a group identified as “pathological controls”, a quantitative comparison of the OXPHOS capacities obtained was performed between the two methods (studies done by Helene Lemieux, Ph.D.). Oxygen flux in isolated mitochondria was expressed per g wet weight of muscle using the mitochondrial protein yield from the muscle, or per unit of citrate synthase activity. Furthermore, a factor was applied for the conversion of flux in isolated mitochondria measured at 30 °C to the corresponding flux in the permeabilized fibers measured at 37 °C [2]. Independent of how the data are expressed, the estimation of mitochondrial respiration for substrates entering at Complexes I, II, or IV were similar if not identical in both methods. The careful and quantitative evaluation of methods available for measurement of mitochondrial function is important for combining such results in a comparative data base [2].<br />
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== Affiliations ==<br />
1-Center Mitochondr Diseases, Depts Pharmacology & Medicine, Case Western Reserve School Med, Cleveland, OH, USA. - charles.hoppel@case.edu<br />
2-Daniel Swarovski Research Laboratory, Dept General Transplant Surgery, Med Univ Innsbruck, Austria<br />
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== References ==<br />
#Puchowicz MA, Varnes ME, Cohen BH, Friedman NR, Kerr DS, Hoppel CL (2004) Oxidative phosphorylation analysis: assessing the integrated functional activity of human skeletal muscle mitochondria-case studies. Mitochondrion 4:377-85.<br />
#Gnaiger E (2009) Capacity of oxidative phosphorylation in human skeletal muscle. New perspectives of mitochondrial physiology. Int J Biochem Cell Biol 41:1837-45.</div>Kandolf Georg