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Property "Has-20abstract" (as page type) with input value "-5B-5BImage:MITOEAGLE-2Dlogo.jpg-7Cleft-7C100px-7Clink=http:-2F-2Fwww.mitoglobal.org-2Findex.php-2FMITOEAGLE-7CCOST-20Action-20MitoEAGLE-5D-5D-0AMitochondrial-20respiration-20is-20commonly-20studied-20using-20isolated-20mitochondria-20in-20skeletal-20muscle.-20Several-20data-20also-20showed-20that-20measuring-20respiration-20in-20permeabilized-20fibers-20(pf)-20that-20preserve-20mitochondrial-20morphology-20and-20intracellular-20interactions-20is-20of-20particular-20interest-20to-20get-20more-20information-20the-20regulation-20of-20this-20organelle-20activity-20in-20different-20knockout-20and-20transgenic-20mice-20models-20-5B1,2-5D.-0A-0ATo-20optimize-20the-20pf-20preparation-20from-20mouse-20-27-27Tibialis-20anterior-27-27-20in-20our-20lab,-20we-20used-20the-20Design-20of-20Experiments-20(DoE)-20methodology-20that-20evaluates-20the-20impact-20of-20several-20experimental-20conditions-20or-20factors,-20on-20the-20pf-20respiration-20parameters-20(Pyruvate-20Malate-20Succinate-20respiration-20(PMS-20leak)-20and-20respiratory-20control-20ratio-20(RCRPMS).-20This-20DoE-20uses-20the-20Hadamard-20matrix-20of-20order-208,-20corresponding-20to-20eight-20experiments-20with-204-20repetitions-20to-20evaluate-20the-20experiment-20variability.-20We-20measured-20the-20effect-20of-206-20factors-20such-20as-20fiber-20types-20(red-20vs-20white),-20manual-20teasing-20(gentle-20vs-20rough),-20sapogenin-20content-20of-20saponin-20(8-20-2D-2025-25-20vs-2020-20-2D-2035-25),-20saponin-20concentration-20for-20permeabilization-20(25-20vs-2050-20µg-2Fml),-20incubation-20time-20with-20saponin-20(10-20vs-2030-20mn)-20and-20muscle-20resting-20period-20in-20buffer-20before-20the-20teasing-20step-20(0h-20vs-206h).-0A-0AOur-20results-20showed-20the-20PMS-20leak-20respiration-20rate-20was-20the-20highest-20for-20pf-20that-20were-20prepared-20from-20red-20fibers-20with-20a-20rough-20teasing-20and-20sapogenin-20content-20of-2020-2D35-25.-20Muscle-20resting-20period-20did-20not-20significantly-20affect-20the-20respiration,-20with-20no-20deleterious-20effect-20of-20the-206h-20resting-20period.-20The-20two-20remaining-20factors-20did-20not-20impact-20PMS-20leak-20respiration-20rate-20(saponin-20concentration-20and-20permeabilization-20time).-20In-20addition,-20RCRPMS-20variability-20was-20the-20lowest-20using-20the-20optimized-20conditions-20determined-20for-20PMS-20leak-20respiration.-20-0A-0AUsing-20a-20DoE-20analysis,-20we-20were-20able-20to-20optimize-20pf-20assay-20conditions-20with-20a-20reduced-20number-20of-20experiments-20and-20animals,-20and-20rapidly-20obtain-20valuable-20data-20in-20accordance-20with-20ethical-20recommendations.-20The-20optimization-20of-20pf-20preparation-20by-20DoE-20will-20be-20pursued-20with-20two-20goals-20(i)-20studying-20the-20possible-20interactions-20existing-20between-20the-203-20factors-20related-20to-20saponin-20(sapogenin-20content,-20saponin-20concentration-20and-20incubation-20time),-20(ii)-20calculating-20the-20optimal-20sample-20size-20(n)-20needed-20to-20observe-20statistically-20significant-20differences-20between-20two-20animal-20groups." contains invalid characters or is incomplete and therefore can cause unexpected results during a query or annotation process.

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