Tissue-, substrate-, and site-specific characteristics of mitochondrial reactive oxygen species generation

Reactive oxygen species are a by-product of mitochondrial oxidative phosphorylation, derived from a small quantity of superoxide radicals generated during electron transport. We conducted a comprehensive and quantitative study of oxygen consumption, inner membrane potentials, and H(2)O(2) release in mitochondria isolated from rat brain, heart, kidney, liver, and skeletal muscle, using various respiratory substrates (alpha-ketoglutarate, glutamate, succinate, glycerol phosphate, and palmitoyl carnitine). The locations and properties of reactive oxygen species formation were determined using oxidative phosphorylation and the respiratory chain modulators oligomycin, rotenone, myxothiazol, and antimycin A and the Uncoupler CCCP. We found that in mitochondria isolated from most tissues incubated under physiologically relevant conditions, reactive oxygen release accounts for 0.1-0.2% of O(2) consumed. Our findings support an important participation of flavoenzymes and complex III and a substantial role for reverse electron transport to complex I as reactive oxygen species sources. Our results also indicate that succinate is an important substrate for isolated mitochondrial reactive oxygen production in brain, heart, kidney, and skeletal muscle, whereas fatty acids generate significant quantities of oxidants in kidney and liver. Finally, we found that increasing respiratory rates is an effective way to prevent mitochondrial oxidant release under many, but not all, conditions. Altogether, our data uncover and quantify many tissue-, substrate-, and site-specific characteristics of mitochondrial ROS release. (C) 2009 Elsevier Inc. All rights reserved.

Accuracy of six minute walk test, stair test and spirometry using maximal oxygen uptake as gold standard

OBJETIVO: Determinar a acurácia das variáveis: tempo de escada (tTE), potência de escada (PTE), teste de caminhada (TC6) e volume expiratório forçado (VEF1) utilizando o consumo máximo de oxigênio (VO2máx) como padrão-ouro. MÉTODOS: Os testes foram realizados em 51 pacientes. O VEF1 foi obtido através da espirometria. O TC6 foi realizado em corredor plano de 120m. O TE foi realizado em escada de 6 lances obtendo-se tTE e PTE. O VO2máx foi obtido por ergoespirometria, utilizando o protocolo de Balke. Foram calculados a correlação linear de Pearson (r) e os valores de p, entre VO2máx e variáveis. Para o cálculo da acurácia, foram obtidos os pontos de corte, através da curva característica operacional (ROC). A estatística Kappa (k) foi utilizada para cálculo da concordância. RESULTADOS: Obteve-se as acurácias: tTE - 86%, TC6 - 80%, PTE - 71%, VEF1(L) - 67%, VEF1% - 63%. Para o tTE e TC6 combinados em paralelo, obteve-se sensibilidade de 93,5% e em série, especificidade de 96,4%. CONCLUSÃO: O tTE foi a variável que apresentou a melhor acurácia. Quando combinados o tTE e TC6 podem ter especificidade e sensibilidade próxima de 100%. Estes testes deveriam ser mais usados rotineiramente, especialmente quando a ergoespirometria para a medida de VO2máx não é disponível.

Dihydrolipoyl dehydrogenase as a source of reactive oxygen species inhibited by caloric restriction and involved in Saccharomyces cerevisiae aging

Replicative life span in Saccharomyces cerevisiae is increased by glucose (G1c) limitation [ calorie restriction (CR)] and by augmented NAD(+). Increased survival promoted by CR was attributed previously to the NAD(+)-dependent histone deacetylase activity of sirtuin family protein Sir2p but not to changes in redox state. Here we show that strains defective in NAD(+) synthesis and salvage pathways (pnc1 Delta, npt1 Delta, and bna6 Delta) exhibit decreased oxygen consumption and increased mitochondrial H2O2 release, reversed over time by CR. These null mutant strains also present decreased chronological longevity in a manner rescued by CR. Furthermore, we observed that changes in mitochondrial H2O2 release alter cellular redox state, as attested by measurements of total, oxidized, and reduced glutathione. Surprisingly, our results indicate that matrix-soluble dihydrolipoyl-dehydrogenases are an important source of CR-preventable mitochondrial reactive oxygen species (ROS). Indeed, deletion of the LPD1 gene prevented oxidative stress in npt1 Delta and bna6 Delta mutants. Furthermore, pyruvate and alpha-ketoglutarate, substrates for dihydrolipoyl dehydrogenase-containing enzymes, promoted pronounced reactive oxygen release in permeabilized wild-type mitochondria. Altogether, these results substantiate the concept that mitochondrial ROS can be limited by caloric restriction and play an important role in S. cerevisiae senescence. Furthermore, these findings uncover dihydrolipoyl dehydrogenase as an important and novel source of ROS leading to life span limitation.

Higher respiratory activity decreases mitochondrial reactive oxygen release and increases life span in Saccharomyces cerevisiae

Increased replicative longevity in Saccharomyces cerevisiae because of calorie restriction has been linked to enhanced mitochondrial respiratory activity. Here we have further investigated how mitochondrial respiration affects yeast life span. We found that calorie restriction by growth in low glucose increased respiration but decreased mitochondrial reactive oxygen species production relative to oxygen consumption. Calorie restriction also enhanced chronological life span. The beneficial effects of calorie restriction on mitochondrial respiration, reactive oxygen species release, and replicative and chronological life span could be mimicked by uncoupling agents such as dinitrophenol. Conversely, chronological life span decreased in cells treated with antimycin (which strongly increases mitochondrial reactive oxygen species generation) or in yeast mutants null for mitochondrial superoxide dismutase (which removes superoxide radicals) and for RTG2 (which participates in retrograde feedback signaling between mitochondria and the nucleus). These results suggest that yeast aging is linked to changes in mitochondrial metabolism and oxidative stress and that mild mitochondrial uncoupling can increase both chronological and replicative life span.

Heat budgets, thermal structure and dissolved oxygen in Brazilian reservoirs

A comparison of the thermal regime and oxygen distribution patterns of some Brazilian reservoirs was made. A strong latitudinal dependence of surface temperature, annual mean and annual range was found. Except for Tucurui Reservoir, a reservoir of the Equatorial zone, the lowest surface temperature of the year in the other compared reservoirs was observed from June to August while the highest extended from October to February. The decreasing trend of annual mean temperatures and thermal ranges was due to the increasing seasonal variability of insolation with the latitude. A positive relationship between the increase on thermal surface-bottom differences and the depth of water column was found for reservoirs of similar latitudes. A long thermal stratification (around four months) was evidenced in the lacustrine zone of reservoirs with a residence time higher than 40 days. Low fluctuation (<2%) of the annual variability of heat contents was observed for the Tucurui Reservoir, while in das Garcas Reservoir a manmade lake located in the frontier between tropical and temperate regions, the annual coefficient of variation attained 13%. Concerning the heat budgets, the value for the Tucurui Reservoir was two times higher than in das Garcas Reservoir. Both the morphometric and climatological factors affected the heat contents of the two compared reservoirs. In deep eutrophic reservoirs, a significant reduction on the oxygen concentrations in the hypolimnetic zone was frequently observed. In some oligotrophic stratified reservoirs, a decrease on oxygen with depth occurred when the temperature of the hypolimnion was higher than 20 degrees C and caused a biochemical oxygen demand. In das Garcas Reservoir, the actual oxygen deficits ranged from 0.40 to 1.52 mg.O-2.cm(-2) and appear to be linked to oxygen consumption after the senescence of Microcystis aeruginosa populations in the spring. But, other factors such as the allochthonous loads of organic matter also had an important role on the oxygen balance of das Garcas Reservoir.

Estimation of the volumetric oxygen tranfer coefficient (KLa) from the gas balance and using a neural network technique

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Evaporative water loss and oxygen uptake in two casque-headed tree frogs, Aparasphenodon brunoi and Corythomantis greeningi (Anura, Hylidae)

Evaporative water loss (EWL) and oxygen uptake ((V) over dot o(2)) was measured in two species of tree frogs with cranial co-ossification, Aparasphenodon brunoi and Corythomantis greeningi. Both species use their head to seal the entrance of bromeliads, tree holes or rocky crevices used as shelters. EWL was significantly reduced in sheltered individuals of both species as compared with those exposed nude to desiccation. EWL per unit area through the head surface was significantly lower than the body skin for A. brunoi but not for C., greeningi. EWL per unit surface area through C. greeningi body skin was about 50% that of A. brunoi, indicating a less permeable skin in the former species. The relationship between cranial coossification and EWL is discussed. ((V) over dot o(2)) in A. brunoi was comparable with other anurans of similar size, whereas in C. greeningi, it was lower than predicted from body mass. Moreover, ((V) over dot o(2)), in C. greeningi showed less sensitivity to temperature increase than in A. brunoi. C. greeningi occurs in a drier environment than A. brunoi, and this appears to be reflected in their EWL and ((V) over dot o(2)) characteristics. (C) 1997 Elsevier B.V.

ONTOGENIC VARIATION OF OXYGEN-UPTAKE IN THE PITVIPER BOTHROPS MOOJENI (SERPENTES, VIPERIDAE)

The scaling of oxygen uptake was measured along the ontogeny, in the neotropical pitviper Bothrops moojeni. Allometric relationship between oxygen uptake and body mass, quantified for juveniles, sub-adults and adults, showed the same mass coefficient and exponent. The uniformity of mass constants along ontogeny suggests that B. moojeni is energetically homomorphic. Variation in mass seem to be the sole determinant of oxygen uptake, and structural modifications have no effect on the metabolic rate. Applications of the homomorphism principle to assess variations in mass coefficient and exponent for intraspecific analysis of metabolism in reptiles are discussed. B. moojeni had an oxygen consumption in the range reported for viperids, but lower than that for colubrid snakes of similar size. Possible causative reasons for this pattern is discussed.

Weekend ethanol consumption and high-sucrose diet: resveratrol effects on energy expenditure, substrate oxidation, lipid profile, oxidative stress and hepatic energy metabolism

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Anaerobic contribution during maximal anaerobic running test: correlation with maximal accumulated oxygen deficit

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Is coronary sinus blood oxygen tension behavior determined by myocardial oxygen tension variation during cardiac reperfusion?

The relationship between coronary sinus blood oxygen tension (CSPO 2) and myocardial oxygen tension (MPO 2) variations during cardiac ischemia and reperfusion was studied in anesthetized open-chest dogs. Oxygen tension was measured by a polarographic method. Ischemia resulted in a slightly decreased CSPO 2 and a more pronounced reduction of MPO 2. After reperfusion the CSPO 2 rose rapidly and transiently before it returned gradually to the control level. By contrast, during the recovery period, the MPO 2 increased slowly, with recovery occurring long after the peak of CSPO 2. These data suggest that during the reperfusion phase, the CSPO 2 variation is probably due to opening of the myocardial arteriovenous shunts instead of an increase of flow through the myocardial capillary bed.

Oxygen uptake kinetics and time to exhaustion in cycling and running: A comparison between trained and untrained subjects

The objective of the present study was to compare pulmonary gas exchange kinetics (VO 2 kinetics) and time to exhaustion (Tlim) between trained and untrained individuals during severe exercise performed on a cycle ergometer and treadmill. Eleven untrained males in running (UR) and cycling (UC), nine endurance cyclists (EC), and seven endurance runners (ER) were submitted to the following tests on separate days: (i) incremental test for determination of maximal oxygen uptake (VO 2max) and the intensity associated with the achievement of VO 2max (IVO 2max) on a mechanical braked cycle ergometer (EC and UC) and on a treadmill (ER and UR); (ii) all-out exercise bout performed at IVO 2max to determine the time to exhaustion at IVO 2max (Tlim) and the time constant of oxygen uptake kinetics (τ). The τ was significantly faster in trained group, both in cycling (EC = 28.2 ± 4.7 s; UC = 63.8 ± 25.0 s) and in running (ER = 28.5 ± 8.5 s; UR = 59.3 ± 12.0 s). Tlim of untrained was significantly lower in cycling (EC = 384.4 ± 66.6 s vs. UC; 311.1 ± 105.7 s) and higher in running (ER = 309.2 ± 176.6 s vs. UR = 439.8 ± 104.2 s). We conclude that the VO 2 kinetic response at the onset of severe exercise, carried out at the same relative intensity is sensitive to endurance training, irrespective of the exercise type. The endurance training seems to differently influence Tlim during exercise at IVO 2max in running and cycling. © 2003 Taylor & Francis Ltd.

Two levels of the inspired oxygen fraction in propofol-anesthetized dogs with high intracranial pressure: cardiopulmonary function

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A technique for the determination of the available oxygen in living carp (Cyprinus carpio) muscle

1. 1. A polarographic method for the measurement of the available oxygen in the muscle of living carp by the use of a platinum microelectrode is proposed. 2. 2. The oxygen and the reference electrodes were assembled in a single insertion piece which was implanted in the muscle of a living carp maintained in a special experimental chamber. 3. 3. Curves for normal oxygen levels corresponding to air-saturated water, as well as to a carbogene-saturated water, were obtained. 4. 4. The method can be considered adequate for the measurement of tissue oxygen in living fishes. © 1984.