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Chemical Analysis of Bioaerosols
Published in Christopher S. Cox, Christopher M. Wathes, Bioaerosols Handbook, 2020
Bioluminescence is caused by a sequence of reactions involving firefly luciferin, adenosine triphosphate, luciferase, magnesium and oxygen but the application of bioluminescence to the detection of microorganisms in the atmosphere is difficult because the luciferase becomes denaturated.33 On the other hand, the chemiluminescent techniques have been applied successfully in detecting bioaerosols.34,35 (see also Chapter 13) The bioaerosol is sampled by some suitable collection method (e.g., a wet cyclone air sampler) and the bacteria are collected into a solution. Iron porphyrins (e.g., haem) from bacteria catalyse the reaction between luminol and an oxidizing radical (e.g., perborate). Sodium hydroxide to causes bacterial lysis thereby releasing haem and is also necessary for the luminol reaction: Luminol+ROOHNaOHhematin→aminophthalate+ROH+N2+hν
Diazinon impairs bioenergetics and induces membrane permeability transition on mitochondria isolated from rat liver
Published in Journal of Toxicology and Environmental Health, Part A, 2020
Camila Araújo Miranda, Anilda Rufino de Jesus Santos Guimarães, Paulo Francisco Veiga Bizerra, Fábio Erminio Mingatto
ATP levels were determined in mitochondria energized with 5 mM glutamate plus 5 mM malate using the firefly luciferin-luciferase assay system (Lemasters and Hackenbrock 1976). The incubation medium consisted of 125 mM sucrose, 65 mM KCl, and 10 mM HEPES-KOH (pH 7.4), plus 0.5 mM EGTA and 10 mM K2HPO4, for a final volume of 1 ml. The reaction was started by the addition of mitochondria (1 mg protein), the medium was shaken, and after 10 min of incubation in the presence of DZN, the suspension was centrifuged at 9,000 × g for 5 min at 4°C, and the pellet was treated with 1 ml ice-cold 1 M HClO4. After centrifugation at 14,000 × g for 5 min at 4°C, 100 µl aliquots of the supernatants were neutralized with 5 M KOH, suspended in 100 mM TRIS-HCl (pH 7.8) (1 ml final volume), and centrifuged at 15,000 × g for 15 min. The supernatant was processed with a Sigma-Aldrich assay kit (Catalog Number FLAA) according to the manufacturer’s instructions and measured using a SIRIUS Luminometer (Berthold, Pforzheim, Germany).
Imidacloprid affects rat liver mitochondrial bioenergetics by inhibiting FoF1-ATP synthase activity
Published in Journal of Toxicology and Environmental Health, Part A, 2018
Paulo F. V. Bizerra, Anilda R. J. S. Guimarães, Marcos A. Maioli, Fábio E. Mingatto
ATP levels were determined in mitochondria in the presence of 5 mM glutamate plus 5 mM malate using the firefly luciferin-luciferase assay system (Lemasters and Hackenbrock 1976). The incubation medium contained 125 mM sucrose, 65 mM KCl, and 10 mM HEPES-KOH, pH 7.4, plus 0.5 mM EGTA and 10 mM K2HPO4 (1 ml final volume). The reaction was started by addition of mitochondria (1 mg protein), the medium was shaken, and after 10 min of incubation in the presence of IMD, the suspension was centrifuged at 9,000 × g for 5 min at 4°C, and pellet was treated with 1 ml ice-cold 1 M HClO4. After centrifugation at 14,000 × g for 5 min at 4°C, 100 µl aliquots of the supernatants were neutralized with 5 M KOH, suspended in 100 mM TRIS-HCl, pH 7.8 (1 ml final volume), and centrifuged at 15,000 × g for 15 min. The supernatant was processed with a Sigma-Aldrich assay kit (Catalog Number FLAA) according to the manufacturer’s instructions and measured using a SIRIUS Luminometer (Berthold, Pforzheim, Germany).