"2014-03-21+01:00"^^ . "mitochondrial diseases" . . "To adapt and optimize modern biochemical and functional methods for analysis of mitochondrial energetic metabolism in blood lymphocytes: (i) To use highly sensitive oxygraphy of permeabilized lymphocytes, luciferase analysis of ATP production, spectrophotometric analysis of enzyme activities, analysis of mitochondrial membrane potential by cytofluorometric probe TMRM and by TPP+ electrode for analysis of energetic function of mitochondria. (ii) To apply BN-PAGE, SDS-PAGE and their combinations together with western blot detection using specific antibodies to structural subunits of respiratory complexes for protein analysis of lymphocyte mitochondria. (iii) To use fluorescent probes AMPLEX UltraRed and DCFDA for ROS production analysis. (iv) To analyze expression of selected genes by RT-PCR. To apply this methodical approach in diagnostics of mitochondrial diseases caused by yet unknown genetic defects, and in monitoring the course of the disease and effect of therapy."@en . . "NT12370" . "Vyu\u017Eit\u00ED lymfocyt\u00E1rn\u00EDch mitochondri\u00ED pro detekci a prospektivn\u00ED anal\u00FDzu mitochondri\u00E1ln\u00EDch chorob" . " energy provision" . "0"^^ . . "Adaptovat a optimalizovat modern\u00ED biochemick\u00E9 a funk\u010Dn\u00ED metody pro anal\u00FDzu mitochondri\u00E1ln\u00EDho energetick\u00E9ho metabolizmu v krevn\u00EDch lymfocytech: (i) Pro anal\u00FDzy energetick\u00E9 funkce mitochondri\u00ED vyu\u017E\u00EDt vysoce citlivou oxygrafii izolovan\u00FDch a permeabilizovan\u00FDch lymfocyt\u016F, lucifer\u00E1zov\u00E9 m\u011B\u0159en\u00ED tvorby ATP, spektrofotometrick\u00E9 m\u011B\u0159en\u00ED enzymov\u00FDch aktivit, m\u011B\u0159en\u00ED energizace mitochondri\u00E1ln\u00ED membr\u00E1ny pomoc\u00ED cytofluorometrick\u00E9 sondy TMRM a TPP+ elektrody. (ii) Pro proteinov\u00E9 anal\u00FDzy vyu\u017E\u00EDt BN-PAGE, SDS-PAGE a jejich kombinace a western blot detekci specifick\u00FDmi protil\u00E1tkami proti strukturn\u00EDm podjednotk\u00E1m respira\u010Dn\u00EDch komplex\u016F. Pro anal\u00FDzu ROS vyu\u017Eit sondy AMPLEX UltraRed a DCFDA. (iii) Pro anal\u00FDzu zm\u011Bn exprese vybran\u00FDch gen\u016F vyu\u017E\u00EDt RT-PCR Tento metodick\u00FD p\u0159\u00EDstup aplikovat p\u0159i diagnostice mitochondri\u00E1ln\u00EDch onemocn\u011Bn\u00ED na podklad\u011B nezn\u00E1m\u00FDch genetick\u00FDch defekt\u016F a p\u0159i sledov\u00E1n\u00ED v\u00FDvoje mitochondri\u00E1ln\u00EDch onemocn\u011Bn\u00ED a vlivu terapie." . . "2015-01-22+01:00"^^ . . . "mitochondrial diseases; oxidative phosphorylation; energy provision; lymphocytes"@en . . . . "1"^^ . "http://www.isvav.cz/projectDetail.do?rowId=NT12370"^^ . . . "2"^^ . . "3"^^ . . "2015-12-31+01:00"^^ . "3"^^ . . . "2011-06-01+02:00"^^ . "Use of lymphocyte mitochondria for detection and prospective analysis of mitochondrial diseases."@en . . " oxidative phosphorylation" . .