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Effects of exercise and respiration on hemodynamic efficiency in CFD simulations of the total cavopulmonary connection.

Congenital heart defects with a single functional ventricle, such as hypoplastic left heart syndrome and tricuspid atresia, require a staged surgical approach to separate the systemic and pulmonary circulations. Ultimately, the venous or pulmonary side of the heart is bypassed by directly connecting the vena cava to the pulmonary arteries with a modified t-shaped junction. The Fontan procedure (total cavopulmonary connection, TCPC) completes this process of separation. To date, computational fluid dynamics (CFD) simulations in this low pressure, passive flow, intrathoracic system have neglected the presumed important effects of respiration on physiology and higher "stress" states such as with exercise have never been considered. We hypothesize that incorporating effects of respiration and exercise would provide more realistic estimates of TCPC performance. Time-dependent, 3D blood flow simulations are performed by a custom finite element solver for two patient-specific Fontan models with a novel respiration model, developed to generate physiologic time-varying flow conditions. Blood flow features, pressure, and energy efficiency are analyzed at rest and with increasing flow rates to simulate exercise conditions. The simulations produce realistic pressure and flow data, comparable to that measured by catheterization and echocardiography, and demonstrate substantial increases in energy dissipation (i.e. decreased performance) with exercise and respiration due to increasing intensity of small scale vortices in the flow. As would be expected, these changes are highly dependent on patient-specific anatomy and Fontan geometry. We propose that respiration and exercise should be incorporated into TCPC CFD simulations to provide increasingly realistic evaluations of TCPC performance.

Blood Flow Velocity↗

Ergonomics assessment of selected dust respirators: Their use in the tropics.

The suitability and effectiveness of four different types of British made respirators were studied with respect to comfort, convenience and fit on wearers in Sri Lanka (a developing country). Objective and subjective assessments were made to evaluate the degree of discomfort and interferences to the use of senses. The study revealed that factors such as breathing resistance, work-rate and activity period affected the physiological responses. The weight of the respirator and the skin temperature had no direct relationship with the cardiovascular stress. Positive-pressure respirators that gave lower face temperatures than negative-pressure masks gave this type of respirator an additional advantage in hot environments. Respirators that restricted jaw movement affected the speech intelligibility of the wearer. Orinasal masks restricted vision more than the other types. The problem of fit was found negligible though head and face dimensions significantly differed between the British and the Sri Lankans. Subjective assessment correlated well with objective tests.

Journal Article↗

Rate control of phosphorylation-coupled respiration by rat liver mitochondria.

Liver mitochondria provided with an oxidizable substrate, ATP, oxygen, and an ADP-generating system (soluble F1-ATPase) were used to reevaluate the rate-controlling step(s) intrinsic to all of the processes of mitochondrial oxidative phosphorylation. The quantity termed "control strength" (C), previously defined as the fractional change in flux through a (system) induced by a fractional change in the concentration of an individual enzyme in the system, has been used to evaluate rate-influencing steps in this overall process by carefully defining the dimensions of the "system" under analysis. If the system is defined by a suspension of mitochondria provided with substrates, plus an extrinsic ADP-generating process (ATPase), the value of C of the latter for the overall process of phosphorylation-linked respiration is near 1.0 until the capacity of the mitochondria to phosphorylate ADP is approached, after which C for the soluble ATPase becomes zero as the maximum capacity for phosphorylation is attained. Carboxyatractyloside was found only marginally to inhibit respiration stimulated by ATPase, even when a large percentage of adenine nucleotide translocase molecules were immobilized. The relative lack of effect of carboxyatractyloside on phosphorylating respiration is explained by the readjustment of the concentration of one of the substrates (ADP) and an inhibitor (ATP), which results from inhibition of adenine nucleotide translocase. The residual blunted inhibition of respiration is explained by product inhibition of the ADP-regenerating ATPase, and not necessarily to any intrinsically mitochondrial intermediate process. The system being evaluated can be redefined to include only the processes intrinsic to mitochondria. This can be achieved by providing exactly comparable substrate concentrations to the mitochondria under comparable incubation conditions. Under these conditions, the adenine nucleotide translocase is the principal, if not the only, rate-controlling step in the overall process of oxidative phosphorylation until a new rate-limitation is attained (ATP synthesis). These data are consistent with the conclusion that, at intermediate rates of phosphorylation-coupled respiration, the extramitochondrial ATP/ADP ratio regulates this process through its kinetic effects on the catalytic properties of the adenine nucleotide translocase.

Adenosine Diphosphate↗

Respiration-dependent uptake and extrusion of Mg2+ by isolated heart mitochondria.

It has been known for some time that isolated heart mitochondria can both take up and extrude Mg2+ by respiration-dependent, uncoupler-sensitive processes. A re-examination of these reactions reveals that the respiration-dependent uptake of Mg2+ can be quite rapid and efficient and is apparently preceded by a passive binding to the inner membrane. The rate of Mg2+ uptake can exceed 30 ng ion/min/mg protein at an efficiency of about 1 ng ion Mg2+ accumulated per ng atom O2 consumed. Passive binding and respiration-dependent accumulation of Mg2+ are strongly inhibited by K+ and other monovalent cations and the uptake reaction is further decreased by the presence of ATP or ADP. Under conditions approaching those faced by mitochondria in situ (state 3 respiration in a KCl medium) the rate of Mg2+ uptake, as estimated from 28Mg2+ distribution, is no more than 0.25 ng ion/min/mg. When heart mitochondria are suspended in a Mg2+-free medium, a slow, respiration-dependent Mg2+ efflux is seen. This reaction is quite insensitive to external K+ and otherwise shows an inhibitor profile markedly different from that of the Mg2+ accumulation reaction. Neither the uptake nor the loss of Mg2+ is inhibited by ruthenium red or diltiazem. These reactions therefore appear unrelated to those involved in the uptake and release of Ca2+. It is concluded that heart mitochondria have separate pathways available for Mg2+ uptake and release.

Adenine Nucleotides↗

Characterization of the alpha-adrenergic stimulation of hepatic respiration.

The alpha-adrenergic agonist phenylephrine induces a biphasic stimulation of respiration in perfused isolated rat liver. The first phase, of rapid onset and short duration, is paralleled by increased glycogenolysis, glycolysis, and NAD redox potential. The second phase lasts for as long as the alpha-agonist is present and is accompanied by increased gluconeogenic flux. Only the second phase of sustained increased respiration is clearly dependent on extracellular Ca2+. In contrast, normal respiratory responses were obtained under Ca2+-loading conditions or in the presence of the Ca2+ ionophore A23187, indicating that the alpha-adrenergic action on respiration is not simply mediated by its ability to increase the cytosolic Ca2+ concentration. No stimulation of gluconeogenesis is observed in the absence of a sustained increase of respiration. However, it is not energy support that leads to the stimulation of glucose production. The adrenergic response is influenced by the nutritional status of the animal and the availability of oxidizable fuels. In livers from starved animals, the alpha-adrenergic respiratory response is abolished when long chain fatty acid oxidation is prevented by the addition of tetradecylglycidate. In the presence of pyruvate the respiratory response is partially restored. It is concluded that increased beta-oxidation is not mandatory for the alpha-adrenergic stimulation of respiration; however, maximal respiratory responses are obtained only when fatty acid oxidation is allowed to proceed. The latter finding appears to be the result of a limited flux through the tricarboxylic acid cycle when long chain fatty acid oxidation is impeded, secondary to a limiting acetyl CoA supply.

Animals↗

The respiratory chain of Paramecium tetraurelia in wild type and the mutant Cl1. II. Cyanide-insensitive respiration. Function and regulation.

1. The cyanide-insensitive respiration in Paramecium tetraurelia was found to be located in mitochondria. 2. Sensitivity of the mitochondrial respiration to cyanide depended on growth conditions. Under standard conditions of growth, 15--20% of respiration was insensitive to 1 mM cyanide. Full resistance to 1 mM cyanide was observed by growing cells in the presence of erythromycin (100--400 microgram/ml) 0.2 mM cyanide. The mitochondrial respiration of the mutant Cl1 harvested during the exponential phase of growth was largely insensitive to cyanide (more than 80%). 3. Pyruvate was oxidized at the same rate by wild type mitochondria and mitochondria of the mutant Cl1. In contrast, succinate oxidation was 2--3 times faster in mitochondria of the mutant Cl1 than in wild type mitochondria. 4. The cyanide-insensitive respiration was inhibited by 1 mM salicylhydroxamic acid to nearly 100%. Other efficient respiratory inhibitors included amytal and heptylhydroxyquinoline. Antimycin was not inhibitory even at concentrations as high as 5 microgram/mg protein, a finding consistent with the lack of antimycin binding sites.

Animals↗

Interrelation between mitochondrial respiration, substrate supply and redox ratio in perifused permeabilized rat hepatocytes.

A one-step perifusion technique is described for studying the regulation of energy metabolism in intact hepatocytes and in mitochondria of the same cells after their permeabilization by digitonin. Cell count and activities of glutamate dehydrogenase, the latter being used as an indicator of mitochondrial integrity, were found to be nearly unchanged after permeabilization and perifusion for at least 40 min at 37 degrees C. The residual activity of lactate dehydrogenase after permeabilized indicated that permeabilized cells were almost depleted of soluble cytosolic components. The composition of the perifusion medium was chosen so that various metabolic states could be adjusted of both intact and permeabilized hepatocytes without the need to change the perfusion medium. Oxidative phosphorylation of mitochondria within permeabilized hepatocytes remained intact throughout the perifusion as indicated by the response of respiration to the addition of ADP, carboxyatractyloside and uncoupler. The application of the perifusion technique allows us to sample indicator metabolites in the effluent medium like acetoacetate (AcAc) and 3-hydroxybutyrate (HB) for calculating the mitochondrial redox ratios and rates of ketogenesis. In the presence of octanoate and ADP, an improvement of substrate supply by glutamate and malate led to increases in the intramitochondrial HB/AcAc ratio and the respiration rate. Glutamate/malate concentrations of 1 mM resulted in maximal respiration rates, whereas concentrations of 5 mM further enhanced the HB/AcAc ratio. Mitochondria responded to increasing ATP/ADP ratios in the perifusion medium by decreased respiration rates at higher HB/AcAc ratios. By comparing respiration rates and redox ratios of mitochondria in permeabilized cells with those before permeabilization (gluconeogenic conditions of hepatocytes), it is concluded that in the intact cell oxidative phosphorylation is limited with respect to substrate supply as well as by the ATP demand.

3-Hydroxybutyric Acid↗

Cyanide resistant respiration is involved in temperature rise in ripening mangoes.

Thermogenesis is attributed to the cyanide resistant respiration in the inflorescence of Arum lilies. Although cyanide resistant respiration is ubiquitously operative in the plant systems, it has never been correlated with thermogenesis except for the above example. Internal temperature of the ripening mango increased from 29.0 degrees C to 38.9 degrees C during its ripening process. Concomitantly, it was coupled with the increase in total respiration and cyanide resistant respiration as well. Implication of cyanide resistant respiration in thermogenesis has been discussed in relation to the ripening fruit.

Carbohydrate Metabolism↗

Nippostrongylus brasiliensis and Ascaridia galli: mitochondrial respiration in free-living and parasitic stages.

Aerobic respiratory pathways have been delineated and respiratory efficiency has been assessed in mitochondria isolated from embryonated eggs, infective larvae, and adult Nippostrongylus brasiliensis and Ascaridia galli. Mitochondrial respiration in free-living stages of N. brasiliensis is mediated mainly by a mammalian-like antimycin A- and cyanide-sensitive pathway; specific respiratory activity is high and oxidative phosphorylation efficient. In mitochondria of adult N. brasiliensis, antimycin A- and cyanide-sensitive respiration is decreased relative to respiration though an alternative pathway, and specific respiratory activity and mitochondrial efficiency are lower. Respiration in mitochondria from embryonated eggs and tissues of adult A. galli is comparable, and apparently mediated by an antimycin A- and cyanide-insensitive alternative respiratory pathway; no evidence for the presence of a mammalian-like respiratory pathway in embryonated eggs of A. galli was found. The results of this study are compared to mitochondrial respiration in eggs, larvae, and adult body wall muscle of Ascaris suum.

Adenosine Diphosphate↗

Acetaminophen-induced inhibition of hepatic mitochondrial respiration in mice.

Morphological changes are observed in mitochondria early in the course of acetaminophen (APAP) hepatotoxicity. In order to determine if functional deficits also occur, this study examined the effect of APAP, in vivo and in vitro, on mitochondrial respiration in fasted, male CD-1 mice (3-4 months old). After a hepatotoxic dose of APAP (600 mg/kg, po), when glutamate was used as the respiratory substrate, state 3 respiration (ADP-stimulated) was inhibited and this was reflected in a decreased respiratory control ratio (RCR). In contrast, when succinate was the respiratory substrate, the decreased RCR was reflective of an increase in state 4 (resting) respiration. There was no detectable effect after a nonhepatotoxic dose of APAP (300 mg/kg, po). These APAP-induced respiratory effects and hepatotoxicity were prevented by piperonyl butoxide pretreatment, and were absent in 1- and 2-month-old mice, which are resistant to APAP-induced damage. Since the APAP-induced inhibition of mitochondrial respiration, in vivo, correlated with age-related and piperonyl butoxide-dependent differences in toxicity, the data suggest that the in vivo effects result, at least in part, from a mixed-function oxidase generated metabolite. In vitro, both state 3 and state 4 respiration, as well as the RCR, were inhibited by APAP in a concentration-dependent manner with glutamate as substrate. However, no effects were observed with succinate as substrate, thereby contrasting with results obtained following in vivo exposure. Therefore the in vitro effects of APAP are different from those observed in vivo and may result from a direct insult of the parent compound. These studies suggest that early alterations in mitochondrial function may be mechanistically important in APAP hepatotoxicity.

Acetaminophen↗

Respiration and energy conservation in the filarial worm Litomosoides carinii.

The average rate of endogenous respiration of intact Litomosoides carinii was 2.24 muatom O min-1 g-1 worm wet wt. No significant difference was observed in respiration capacities between male and female worms. Rates of oxygen uptake decreased progressively during disruption and fractionation of the parasite tissue and very few respiration capabilities remained in the mitochondrial fraction. Added substrates increased the respiratory rates of the intact filariid and cell-free extracts by a factor of 1.4 to 2.3, depending on the tissue system and substrate species used. Rotenone and cyanide strongly inhibited respiration in all incubations, whereas antimycin A, in most cases, suppressed oxygen consumption only partially. ATP conservation in cell-free extracts of L. carinii, as determined by the incorporation of 32Pi into the organic phosphate fraction, was twice as high in the presence of air as under an atmosphere of nitrogen. Anaerobically, rates of phosphorylation in these extracts were similar to the amounts of lactate. Phosphorylation in mitochondria isolated from the filarial worm was supported by malate, succinate, pyruvate and TMPD/ascorbate, whereas L-glutamate and beta-hydroxybutyrate exhibited only little or no effect, respectively. P/O ratios for pyruvate-supported oxidative phosphorylation were found to approach a value of 3. Electron transport inhibitors, oligomycin and 2,4-dinitrophenol strongly inhibited substrate-dependent mitochondrial phosphorylation. The data of the present investigation, together with other recent findings made by the same authors, have provided evidence that in L. carinii mitochondria a mammalian-type of respiratory system capable of carrying out oxidative phosphorylation is functional. It seems likely that this respiration-dependent chemical energy, proceeding in addition to that generated through fermentation processes, may be vital for muscular contraction and survival of this filarial parasite.

Adenosine Triphosphate↗

Sulfate reduction and oxic respiration in marine sediments: implications for organic carbon preservation in euxinic environments.

Compilations have been made of sulfate reduction rates and oxic respiration rates over the entire range of marine sedimentation rates, and sedimentary environments, including several euxinic sites. These data show, consistent with the findings of Jorgensen (1982, Nature, 296, 643-645), that sulfate reduction and oxic respiration oxidize equal amounts of organic carbon in nearshore sediments. As sedimentation rates decrease, oxic respiration, becomes progressively more important, and in deep-sea sediments 100-1000 times more organic carbon is oxidized by oxic respiration than by sulfate reduction. By contrast, nearly as much organic carbon is oxidized by sulfate reduction in euxinic sediments as is oxidized by the sum of sulfate reduction and oxic respiration in normal marine sediments of similar deposition rate. This observation appears at odds with the enhanced preservation of organic carbon observed in euxinic sediments. However, only small reductions in (depth-integrated) organic carbon decomposition rates (compared to normal marine) are required to give both high organic carbon concentrations and enhanced carbon preservation in euxinic sediments. Lower rates of organic carbon decomposition (if only by subtle amounts) are explained by the diminished ability of anaerobic bacteria to oxidize the full suite of sedimentary organic compounds.

Bacteria, Anaerobic↗

Increased respiration in skeletal muscle mitochondria from cold-acclimated ducklings: uncoupling effects of free fatty acids.

Intermyofibrillar mitochondria from skeletal muscle (m. gastrocnemius) and liver mitochondria were isolated from cold-acclimated (4 degrees C) or control (30 degrees C) 4-week old ducklings. The respiratory rate of isolated mitochondria, with Na-succinate as substrate, was followed polarographically at 25 degrees C in order to determine the basal respiratory rate, the rate of respiration in the presence of free fatty acids (FFA) (Na-palmitate), and the fully uncoupled rate, after addition of FCCP. The basal respiration (which in liver mitochondria was unaffected by acclimation to cold) was higher (+53%) in intermyofibrillar mitochondria from cold-acclimated ducklings than from controls, and the maximal FCCP-stimulated respiration was also increased (+98%) by acclimation to cold. FFA-stimulated respiration increased as a function of FFA concentration in both types of mitochondria. The increase in respiration due to FFA was about double in intermyofibrillar mitochondria from cold-acclimated ducklings than that of controls, but in liver mitochondria there was no increase due to cold. The membrane potential was estimated by the dye safranine in the absence or in the presence of FFA in the incubation medium. There were no significant differences in the basal membrane potential in the two groups and the addition of FFA led to the same depolarization in both groups. The significance of these alterations for acclimation to cold is discussed.

Acclimatization↗

Support of respiration and citrulline synthesis by isolated hepatic mitochondria from Squalus acanthias by acyl-CoAs and other nitrogen-donating substrates.

The effects of different substrates supporting respiration and glutamine-dependent citrulline synthesis from ornithine, ammonia, and bicarbonate by isolated hepatic mitochondria from Squalus acanthias (spiny dogfish) were determined. Highest rates of respiration were achieved with succinate, palmitoyl-CoA, and beta-hydroxybutyrate as oxidizable substrates. All acyl-CoAs tested (from C-2 to C-22) supported carnitine-dependent respiration at a substantial rate. Short-chain fatty acids did not support respiration. Ammonia required for citrulline synthesis could be formed from glutamate, or from leucine plus alpha-ketoglutarate which gives rise to glutamate by transamination, as the result of glutamate dehydrogenase activity, but the reaction was inhibited by succinate or other oxidizable substrates. Alanine or ornithine could not be substituted for leucine, suggesting that leucine may specifically activate glutamate dehydrogenase. Glutamate required for citrulline synthesis could be formed from alpha-ketoglutarate and ammonia as the result of glutamate dehydrogenase activity if succinate was present. Transamination of alpha-ketoglutarate with ornithine present in the reaction mixtures provided glutamate at a rapid rate whether or not succinate was present. These results are consistent with the view that hepatic dogfish mitochondria efficiently utilize acyl-CoAs derived from triglyceride stores in the liver to support respiration, glutamine-dependent citrulline synthesis from ammonia, and formation of ketone bodies as a major fuel for muscle.

Acyl Coenzyme A↗

Regulation of the rate of respiration and oxidative phosphorylation in liver mitochondria from hibernating ground squirrels, Citellus undulatus.

1. The rates of oxidation of various substrates (beta-hydroxybutyrate, succinate, ascorbate + TMPD) and the rate of ATP synthesis in liver mitochondria from active and hibernating ground squirrels were measured. 2. It was shown that the rate of mitochondrial respiration is significantly lower in hibernating animals than in active animals. 3. The degree of inhibition of mitochondrial respiration in hibernating ground squirrels was found to correlate with the length of the respiratory chain fragment involved in the oxidation of a given substrate. 4. The inhibition of mitochondrial respiration in hibernating animals was accompanied by a decrease in the rate of ATP synthesis. 5. The activity of phospholipase A2 in liver mitochondria from hibernating ground squirrels was found to be decreased. The activation of phospholipase A2 by Ca2+ ions eliminated the inhibition of respiration almost completely. 6. It was assumed that the inhibition of mitochondrial respiration during hibernation is (a) related to the suppression of phospholipase A2 activity and (b) caused by the reduced rates of electron transport through the respiratory chain and/or of substrate transport across the mitochondrial membrane.

Adenosine Triphosphate↗

Sedimenticola selenatireducens, gen. nov., sp. nov., an anaerobic selenate-respiring bacterium isolated from estuarine sediment.

The respiration of selenate, as a terminal electron acceptor has been known for over a decade, but the microorganisms involved in this respiration are largely unknown. Here we characterize a novel selenate-respiring bacterium, strain AK4OH1, isolated from an estuarine sediment enrichment culture. Strain AK4OH1 has the unique capability to oxidize aromatic acids, such as benzoate, 4-hydroxybenzoate and 3-hydroxybenzoate, coupled to selenate respiration. This novel respiratory coupling has not been described before. Reduction of selenate is followed by stoichiometric accumulation of selenite. The strain grows in agar shake tubes forming bright red colonies due to precipitation of elemental selenium. Strain AK4OH1 is a strictly anaerobic bacterium, which can also respire nitrate and nitrite via denitrification. Analysis of the 16S rRNA gene sequence shows that this strain clusters with another selenate-reducing bacterium and a (per) chlorate reducing bacterium, within the Gammaproteobacteria, along with symbionts of bivalves and tubeworms. Based on its unique physiological capabilities and its 16S rRNA gene sequence phylogeny, we classify this strain AK4OH1 as a new genus and species with the proposed name Sedimenticola selenatireducens.

Base Sequence↗

Kinetics of the inhibition of mitochondrial respiration by NO.

The kinetics of the inhibition of mitochondrial respiration by NO was examined in isolated mitochondria (here obtained from rat brown adipose tissue). The Ki of NO for the inhibition was approximately 27 nM; the IC50 of NO increased in proportion to the square of an increase in O2 tension. The Km of O2 for respiration was approximately 16 microM; in the presence of NO, the dependence of respiration on O2 tension had a Hill coefficient of approximately 2. The unusual kinetics is probably related to the ability of cytochrome c oxidase to use 2 NO or 1 O2 as electron acceptor. The interaction between NO and O2 in the control of respiration could be described by the formula VO2(O2, NO) = VO2max x ([O2]2/((16 microM x (1 + [NO]/27 nM))2 + [O2]2)). Thus, the kinetics is such that respiration in the presence of physiological levels of NO is very sensitive to decreasing O2 tension.

Adipose Tissue, Brown↗

Effects of temperature and dietary sucrose concentration on respiration in the silverleaf whitefly, Bemisia argentifolii.

A system consisting of a flow-through chamber connected to a commercial infrared gas analysis system was developed to measure homopteran respiration during feeding. Using this system, respiration rates of 202 and 206 µmol CO(2) h(-1) g(-1) (4.96 and 5.04 ml CO(2) h(-1) g(-1)) were determined for whiteflies and cotton aphids, respectively, at 25 degrees C on diets containing 15% sucrose. These rates were considerably higher than those of other stationary insects, indicating that whiteflies and aphids maintain a relatively high metabolic rate when feeding. Whitefly respiration increased with temperature from 25 to 46 degrees C with a Q(10) of about 2 on diets containing 10, 15 and 20% sucrose, but less than 2 on diets containing 2.5 and 5% sucrose. Respiration rates were similar on the diets containing >10% sucrose, but were generally lower on the diets containing <10% sucrose. Respiration rates decreased upon extended exposure to 47 degrees C; the rate of decrease was inversely related to the dietary sucrose concentration up to 15%. The results indicate that whiteflies require a sucrose concentration of between 5 and 10% (i.e. 0.15 and 0.3 M) for maximum rates of metabolism while feeding. Higher concentrations of sucrose in the diet delayed high-temperature mortality, possibly a reflection of the high sucrose requirement for sorbitol synthesis in whiteflies.

Journal Article↗