Difference between revisions of "Pecina 2014 Biochim Biophys Acta Clinical"

Latest revision as of 17:21, 4 June 2017

Pecina P, Houstkova H, Mracek T, Pecinova A, Nuskova H.Tesarova M, Hansikova H, Janota J, Zemanc J, Houstek J (2014) Noninvasive diagnostics of mitochondrial disorders in isolated lymphocytes with high resolution respirometry. Biochim Biophys Acta Clinical 2:62–71.

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Pecina P, Houstkova H, Mracek T, Pecinova A, Nuskova H.Tesarova M, Hansikova H, Janota J, Zemanc J, Houstek J (2014) BBA Clinical

Abstract: Mitochondrial diseases belong to the most severe inherited metabolic disorders affecting pediatric population. Despite detailed knowledge of mtDNA mutations and progress in identification of affected nuclear genes, diagnostics of a substantial part of mitochondrial diseases relies on clinical symptoms and biochemical data from muscle biopsies and cultured fibroblasts.

To investigate manifestation of oxidative phosphorylation defects in isolated lymphocytes, digitoninpermeabilized cells from48 children were analyzed by high resolution respirometry, cytofluorometric detection of mitochondrial membrane potential and immunodetection of respiratory chain proteins with SDS and Blue Native electrophoreses.

Evaluation of individual respiratory complex activities, ATP synthesis, kinetic parameters of mitochondrial respiratory chain and the content and subunit composition of respiratory chain complexes enabled detection of inborn defects of respiratory Complexes I, IV and V within 2 days. Low respiration with NADH-dependent substrates and increased respiration with glycerol-3-phosphate revealed Complex I defects; changes in p50 for oxygen and elevated uncoupling control ratio pointed to complex IV deficiency due to SURF1 or SCO2 mutation; high oligomycin sensitivity of state 3-ADP respiration, upregulated mitochondrial membrane potential and low content of Complex V were found in lymphocytes with ATP synthase deficiency due to TMEM70 mutations.

Based on our results, we propose the best biochemical parameters predictive for defects of respiratory Complexes I, IV andVmanifesting in peripheral blood lymphocytes.

The noninvasiveness, reliability and speed of an approach utilizing novel biochemical criteria demonstrate the high potential of isolated lymphocytes for diagnostics of oxidative phosphorylation disorders in pediatric patients. • Keywords: Lymphocytes, Respirometry, Oxidative phosphorylation, Mitochondrial diseases, Diagnostics

• O2k-Network Lab: CZ Prague Houstek J

Labels: MiParea: Respiration, Instruments;methods, Patients

Organism: Human Tissue;cell: Blood cells, Lymphocyte Preparation: Permeabilized cells

Regulation: Inhibitor Coupling state: LEAK, OXPHOS Pathway: N, Gp, CIV, Other combinations, ROX HRR: Oxygraph-2k

Blood cell preparation

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 {{Publication
-|title=Pecina P, Houstkova H, Mracek T, Pecinova A, Nuskova H.Tesarova M, Hansikova H, Janota J, Zemanc J, Houstek J (2014) Noninvasive diagnostics of mitochondrial disorders in isolated lymphocytes with high resolution respirometry. BBA Clinical  2: 62–71.
+|title=Pecina P, Houstkova H, Mracek T, Pecinova A, Nuskova H.Tesarova M, Hansikova H, Janota J, Zemanc J, Houstek J (2014) Noninvasive diagnostics of mitochondrial disorders in isolated lymphocytes with high resolution respirometry. Biochim Biophys Acta Clinical 2:62–71.
+|info=[http://www.sciencedirect.com/science/article/pii/S2214647414000208 open access]
 |authors=Pecina P, Houstkova H, Mracek T, Pecinova A, Nuskova H.Tesarova M, Hansikova H, Janota J, Zemanc J, Houstek J
 |year=2014
 |journal=BBA Clinical
-|abstract=Mitochondrial diseases belong to the most severe inherited metabolic disorders affecting pediatric
+|abstract=Mitochondrial diseases belong to the most severe inherited metabolic disorders affecting pediatric population. Despite detailed knowledge of mtDNA mutations and progress in identification of affected nuclear genes, diagnostics of a substantial part of mitochondrial diseases relies on clinical symptoms and biochemical data from muscle biopsies and cultured fibroblasts.
-population. Despite detailed knowledge of mtDNA mutations and progress in identification of affected nuclear
-genes, diagnostics of a substantial part of mitochondrial diseases relies on clinical symptoms and biochemical
-data from muscle biopsies and cultured fibroblasts.
-To investigate manifestation of oxidative phosphorylation defects in isolated lymphocytes, digitoninpermeabilized
+To investigate manifestation of oxidative phosphorylation defects in isolated lymphocytes, digitoninpermeabilized cells from48 children were analyzed by high resolution respirometry, cytofluorometric detection of mitochondrial membrane potential and immunodetection of respiratory chain proteins with SDS and Blue Native electrophoreses.
-cells from48 children were analyzed by high resolution respirometry, cytofluorometric detection
-of mitochondrial membrane potential and immunodetection of respiratory chain proteins with SDS and Blue
-Native electrophoreses.
-Evaluation of individual respiratory complex activities, ATP synthesis, kinetic parameters of mitochondrial
+Evaluation of individual respiratory complex activities, ATP synthesis, kinetic parameters of mitochondrial respiratory chain and the content and subunit composition of respiratory chain complexes enabled detection of inborn defects of respiratory Complexes I, IV and V within 2 days. Low respiration with NADH-dependent substrates and increased respiration with glycerol-3-phosphate revealed Complex I defects; changes in p50 for oxygen and elevated uncoupling control ratio pointed to complex IV deficiency due to SURF1 or SCO2 mutation; high oligomycin sensitivity of state 3-ADP respiration, upregulated mitochondrial membrane potential and low content of Complex V were found in lymphocytes with ATP synthase deficiency due to TMEM70 mutations.
-respiratory chain and the content and subunit composition of respiratory chain complexes enabled detection of
-inborn defects of respiratory complexes I, IV and V within 2 days. Low respiration with NADH-dependent
-substrates and increased respiration with glycerol-3-phosphate revealed complex I defects; changes in p50 for
-oxygen and elevated uncoupling control ratio pointed to complex IV deficiency due to SURF1 or SCO2 mutation;
-high oligomycin sensitivity of state 3-ADP respiration, upregulated mitochondrial membrane potential and low
-content of complex V were found in lymphocytes with ATP synthase deficiency due to TMEM70 mutations.
-Based on our results, we propose the best biochemical parameters predictive for defects of respiratory
+Based on our results, we propose the best biochemical parameters predictive for defects of respiratory Complexes I, IV andVmanifesting in peripheral blood lymphocytes.
-complexes I, IV andVmanifesting in peripheral blood lymphocytes.
-The noninvasiveness, reliability and speed of an approach utilizing novel biochemical criteria
+The noninvasiveness, reliability and speed of an approach utilizing novel biochemical criteria demonstrate the high potential of isolated lymphocytes for diagnostics of oxidative phosphorylation disorders in pediatric patients.
-demonstrate the high potential of isolated lymphocytes for diagnostics of oxidative phosphorylation disorders in
-pediatric patients.
 |keywords=Lymphocytes, Respirometry, Oxidative phosphorylation, Mitochondrial diseases, Diagnostics
 |mipnetlab=CZ Prague Houstek J
 }}
 {{Labeling
-|area=Respiration, Patients
+|area=Respiration, Instruments;methods, Patients
 |organism=Human
-|tissues=Blood cells
+|tissues=Blood cells, Lymphocyte
-|model cell lines=Lymphocyte
 |preparations=Permeabilized cells
-|injuries=Mitochondrial Disease; Degenerative Disease and Defect
+|topics=Inhibitor
 |couplingstates=LEAK, OXPHOS
-|substratestates=CI, GpDH, CIV, ROX
+|pathways=N, Gp, CIV, Other combinations, ROX
 |instruments=Oxygraph-2k
-|additional=Labels
+|additional=Blood cell preparation
 }}