Abstract
This study investigated the feasibility of using a phosphorescence oxygen analyser to measure cellular respiration (mitochondrial O2 consumption) in foreskin samples and their fibroblast-rich cultures. Foreskin specimens from normal infants were collected immediately after circumcision and processed for measuring cellular respiration and for culture. Cellular mitochondrial O2 consumption was determined as a function of time from the phosphorescence decay of the Pd (II) meso-tetra-(4-sulfonatophenyl)-tetrabenzoporphyrin. In sealed vials containing a foreskin specimen and glucose, O2 concentration decreased linearly with time, confirming the zero-order kinetics of O2 consumption by cytochrome oxidase. Cyanide inhibited O2 consumption, confirming that the oxidation occurred mainly in the mitochondrial respiratory chain. The rate of foreskin respiration (mean ± SD) was 0.074 ± 0.02 μM O2 min(-1) mg(-1) (n = 23). The corresponding rate for fibroblast-rich cultures was 9.84 ± 2.43 μM O2 min(-1) per 10(7) cells (n = 15). Fibroblast respiration was significantly lower in a male infant with dihydrolipoamide dehydrogenase gene mutations, but normalised with the addition of thiamine or carnitine. The foreskin and its fibroblast-rich culture are suitable for assessment of cellular respiration. However, the clinical utility of foreskin specimens to detect disorders of impaired cellular bioenergetics requires further investigation.
Highlights
Advances in Knowledge - This study demonstrates the feasibility of using foreskin samples to measure cellular respiration
Application to Patient Care - Foreskin specimens and their fibroblast-rich cultures can be used to screen for disorders of impaired cellular bioenergetics. - This study shows the potential use of fibroblast respiration to predict responses to therapeutic interventions
The reference values for lymphocyte respiration are shown in Table 1.10 The rate of respiration in foreskin samples that was determined within one hour of circumcision was 0.076 ± 0.01
Summary
Advances in Knowledge - This study demonstrates the feasibility of using foreskin samples to measure cellular respiration (cellular mitochondrial oxygen consumption). Application to Patient Care - Foreskin specimens and their fibroblast-rich cultures can be used to screen for disorders of impaired cellular bioenergetics. Al-Jasmi et al described the use of a phosphorescence oxygen analyser to measure lymphocyte respiration in patients.[10] Their method was based on previously published principles.[11] The lymphocytes from patients were shown to be suitable for the screening of certain mitochondrial disorders. The primary aim of this study was to investigate the use of the phosphorescence oxygen analyser to measure cellular respiration (mitochondrial O2 consumption) in foreskin specimens and their fibroblast-rich cultures. The secondary aim was to utilise the foreskin to screen for metabolic disorders that impair cellular respiration (mitochondrial O2 consumption and accompanying adenosine triphosphate [ATP] synthesis). The hypothesis was that the foreskin and its fibroblast-rich culture can be utilised for assessment of cellular metabolic fuels and their energy conversion processes
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