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Effect of exercise on hexokinase distribution and mitochondrial respiration in skeletal muscle.

Horses were subjected to treadmill running at 65% (submaximal) or 100% (maximal) VO2,max to examine the effects of exercise on subcellular distribution of hexokinase (HK) and on mitochondrial respiration. It is hypothesized that the fraction of HK bound to mitochondria will be reduced due to an elevation of glucose-6-phosphate (G-6-P) concentration in the exercising muscle and that such release of HK from mitochondria will depress oxidative phosphorylation. Changes in muscle G-6-P concentration, pH, subcellular HK distribution, mitochondrial respiration and other metabolites were determined in biopsy samples pre-exercise, immediately post-exercise and during the recovery phase. The fraction of HK associated with mitochondria decreased from 38% to 7% at the end of maximal exercise; exercise at VO2,max also reduced respiratory capacity of muscle homogenates by 20% and was associated with a fivefold increase in muscle [G-6-P], a potent agent known to dissociate HK from mitochondria. The HK distribution returned to normal within 60 min after exercise and the reassociation of the HK with mitochondria parallelled the removal of muscle G-6-P. No changes in muscle HK distribution and respiration were found following the submaximal exercise despite the fact that G-6-P was slightly elevated. Muscle concentrations of adenosine triphosphate, creatine phosphate and glycogen and pH dropped after exercise while lactate concentration increased. The amount of mitochondria-bound HK was also altered in vitro in a preparation of mitochondria isolated from rat skeletal muscle to examine the effect of the bound HK on mitochondrial respiration.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗

On the absence of correlation between cyanide-resistant respiration and cytochrome d content in bacteria.

The regularity of appearance of cyanide-resistant respiration and cytochrome d in various bacteria as well as the relationship between the degree of resistance of respiration to cyanide and cytochrome d content was studied. Bacteria able to synthesize cyanide-resistant respiration were shown to appear during transition of culture to the stationary phase of growth caused by the exhaustion of carbon source. No regulatory of appearance of cytochrome d was observed. There is no correlation between the degree of resistance to cyanide and cytochrome d content. It was concluded that the cyanide-resistant respiration of bacteria and eukaryotic microorganisms may be associated with the functioning of a non-cytochrome nature oxidase.

Bacteria↗

Role of cellular defense against hydrogen peroxide-induced inhibition of myocyte respiration.

Hydrogen peroxide (H2O2) serves as a precursor for highly reactive oxygen intermediates. However, the respiratory function of myocytes is relatively resistant to exogenously administered H2O2. In this study, we examined whether or not the reduction of cellular defense increases the toxicity of H2O2. Rat heart myocytes were isolated by collagenase digestion. Respiratory rates of myocytes, suspended in a medium containing sucrose, 3-N-morpholino-propanesulfonic acid, EGTA and bovine serum albumin, were determined polarographically in the presence of pyruvate and malate with or without 2,4-dinitrophenol (DNP). Mitochondrial membrane potentials were measured by using [3H]triphenylmethylphosphonium+. Cellular defense was attenuated by i) inhibiting the catalase activity by 3-amino-1,2,4-triazole (AT), ii) reducing the glutathione concentration by diethyl maleate (DEM) or ethacrinic acid (EA), and iii) permeabilizing the sarcolemmal membrane by saponin. The dose-response relationship between H2O2 (0.1-5 mM) and mitochondrial membrane potential was not greatly affected by these experimental conditions. Myocyte respiration was inhibited by 5 mM H2O2, particularly that measured in the presence of DNP (48% of control). DEM treatment did not significantly affect the respiratory inhibition by H2O2, whereas the degree of inhibition was somewhat greater following EA or AT treatment. By contrast, the sensitivity of cellular respiration to H2O2 was potentiated approximately two orders of magnitude by the permeabilization of sarcolemmal membrane; thus, 100 microM H2O2 inhibited both DNP-stimulated and unstimulated respiration to 17% and 35% of control, respectively. The results indicate that factors existing in the sarcolemma and/or in the cytosol, which become ineffective and/or are diluted, respectively, following permeabilization with saponin, are important cellular defense mechanisms in alleviating the toxic effect of exogenous H2O2 on the respiration of mitochondria in situ in myocytes.

Amitrole↗

The course of respiration and circulation in death due to typical hanging.

Experiments were carried out on 15 dogs to investigate the course of respiration and circulation during the agonal period of death due to typical hanging. Animals were asphyxiated by a method simulating typical hanging. Electrocardiogram (ECG), electroencephalogram (EEG), blood pressure (BP) in the femoral artery and intrathoracic pressure (ITP) were registered. In typical hanging, the course of respiration was characterized by shorter stages of dyspnoea and initial apnoea and a longer stage of the terminal respiration when compared with obstructive asphyxia. However, the whole time of the course of respiration and circulation in typical hanging was almost the same as that in obstructive asphyxia. The BP increased rapidly, was maintained during the dyspnoea stage, and then decreased gradually. EEG disappeared with, or a short time after, the end of the dyspnoea stage. The increased heart rate in the dyspnoea stage remained until circulatory collapse. The analysis of the ECG complexes revealed that the heart muscle adapted to oxygen deficiency during typical hanging.

Airway Obstruction↗

[Phosphate-hemoglobin interaction: concerning the respiration of adult man, human fetus, the llama and dromedary (author's transl)].

The perinatal control of the oxygen affinity of human blood and the high altitude respiration of the llama is discussed. Furthermore, the sequences of the beta-chains of the hemoglobin of man, llama, dromedary and the gamma-chains of human fetal hemoglobin are reported. Considering physical, physiological, and chemical data it is shown that the human fetal respiration and the high altitude respiration of llama are based on a similar molecular mechanism and are marked by a reduced mutual effect of the non alpha-chains to 2,3-bisphosphoglycerate. The reduced mutual effect is interpreted as a mutation and interruption of the phosphate contacts in gamma 143 Serin at the fetus and beta 2 Asparagin at the llama resulting in an increased oxygen affinity of the corresponding hemoglobins. The respiration of the fetus and that of the llama in the Andes is thereby molecularly interpreted.

2,3-Diphosphoglycerate↗

Glucose-dependent respiration in suspensions of rabbit cortical tubules.

The effects of glucose on cellular respiration were examined in suspensions of rabbit cortical tubules. When glucose was removed from the bathing fluid, oxygen consumption (QO2) decreased from 18.6 +/- 0.8 to 15.7 +/- 0.5 nmol O2/mg protein X min (P less than 0.01). The transported but nonmetabolized analogue of glucose, alpha-methyl-D-glucoside (alpha MG), was found to support QO2 to the same extent as glucose. These observations were also evident in the presence of butyrate, a readily oxidized substrate of the renal cortex. Additional studies with nystatin and ouabain indicated that glucose-related changes in QO2 were the result of changes in Na, K-ATPase associated respiration. The effect of glucose was localized to the luminal membrane since phlorizin (10(-5) M), a specific inhibitor of luminal glucose-sodium cotransport, also significantly reduced QO2 by 10 +/- 1%. Phlorizin inhibition of QO2 was also evident in the presence of alpha MG but was abolished when glucose was removed from the bathing medium. Finally, measurement of NADH fluorescence showed that addition of glucose (5 mM) to a tubule suspension causes an oxidation of NAD. These data are all consistent with glucose acting to increase respiration by stimulating sodium entry at the luminal membrane (via glucose-sodium cotransport) followed by increased sodium pump activity and its associated increase in mitochondrial respiration.

Animals↗

Inhibition of respiration and of respiratory development by different doses of chloramphenicol in frog embryos (Rana pipiens).

High doses of chloramphenicol (4.5 and 9 mM) directly inhibited respiration in conjunction with an unusual cleavage anomaly (blastomere refusion) in embryos of the frog, Rana pipiens. Lower doses (2.1-3 mM) did not affect respiration or cleavage and, when added later, did not directly affect respiration or development at any tested stage. Continuous incubation in the latter doses, however, did eventually, by day 6 or 7, produce a parallel retardation of embryonic development and of respiratory development relative to controls, suggestive of impaired mitochondrial biogenesis. A lower continuous dose (0.9 mM) affected neither respiration nor development at any stage.

Animals↗

Augmentation of Hypoxic Respiration after Brief Hyperoxia in the Anesthetized Cat: Putative Function of GABA(A) Neurotransmission.

In this study, we attempted to determine the role of GABA neurotransmission in augmentation of hypoxic respiration by antecedent hyperoxic breathing. The experiments were performed in anesthetized, paralyzed and vagotomized cats divided into control and bicuculline (a GABA(A) receptor blocker)-injected groups. The experimental protocol consisted of exposing the animals to successive hypoxic-hyperoxic-hypoxic conditions. Respiration was assessed using phrenic electroneurograms, from which the peak phrenic height, a surrogate of the tidal volume component, and respiratory rate were obtained, and their product, the respiratory minute output, was calculated. We found that prior hyperoxic ventilation increased the subsequent respiratory response to hypoxia by an average of 23.5%, compared with the preoxygen response. This increase was driven by volume respiration. The biphasic character of the hypoxic respiratory response, consisting of stimulatory and depressant phases, was sustained. Bicuculline abolished the augmentative effect on hypoxic respiration of prior hyperoxia, which suggests that oxygenation induces GABA(A)-mediated hyperexcitability of respiratory neurons, possibly by the liberation of reactive oxygen species. We concluded that GABA neurotransmission is pertinent to the effect of hyperoxia on hypoxic respiratory reactivity.

Animals↗

Cyanide-insensitive respiration in Schizophyllum commune.

Two mutants of the hymenomycete Schizophyllum commune, unable to use acetate as the sole carbon source for growth, were isolated. Growth of the mutants on a glucose minimal medium was only slightly inhibited by sodium azide. Genetic analysis revealed mutations in different chromosomal genes in the respective mutants. Both these mutants exhibited a high cyanide-insensitive endogenous respiration. The inhibition of the respiration by 8-hydroxyquinoline showed the mutants to respire predominantly by an alternative respiratory pathway observed in many fungal species, but not in the hymenomycetes so far. An enhanced cyanide-insensitive respiration was also found in a wild-type strain of Schizophyllum commune grown in the presence of sodium azide.

Acetates↗

Contribution of sediment respiration to summer CO2 emission from low productive boreal and subarctic lakes.

We measured sediment production of carbon dioxide (CO(2)) and methane (CH(4)) and the net flux of CO(2) across the surfaces of 15 boreal and subarctic lakes of different humic contents. Sediment respiration measurements were made in situ under ambient light conditions. The flux of CO(2) between sediment and water varied between an uptake of 53 and an efflux of 182 mg C m(-2) day(-1) from the sediments. The mean respiration rate for sediments in contact with the upper mixed layer (SedR) was positively correlated to dissolved organic carbon (DOC) concentration in the water (r(2) = 0.61). The net flux of CO(2) across the lake surface [net ecosystem exchange (NEE)] was also closely correlated to DOC concentration in the upper mixed layer (r(2) = 0.73). The respiration in the water column was generally 10-fold higher per unit lake area compared to sediment respiration. Lakes with DOC concentrations <5.6 mg L(-1) had net consumption of CO(2) in the sediments, which we ascribe to benthic primary production. Only lakes with very low DOC concentrations were net autotrophic (<2.6 mg L(-1)) due to the dominance of dissolved allochthonous organic carbon in the water as an energy source for aquatic organisms. In addition to previous findings of allochthonous organic matter as an important driver of heterotrophic metabolism in the water column of lakes, this study suggests that sediment metabolism is also highly dependent on allochthonous carbon sources.

Carbon↗

Temporal variation in delta(13)C of ecosystem respiration in the Pacific Northwest: links to moisture stress.

We measured seasonal and interannual variations in delta(13)C values within the carbon reservoirs (leaves and soil) and CO(2) fluxes (soil and ecosystem respired CO(2)) of an old growth coniferous forest in the Pacific Northwest USA with relation to local meteorological conditions. There were significant intra-annual and interannual differences in the carbon isotope ratios of CO(2) respired at both the ecosystem (delta(13)C(R)) and the soil levels (delta(13)C(R-soil)), but only limited variations in the carbon isotope ratios of carbon stocks. The delta(13)C(R) values varied by as much as 4.4 per thousand over a growing season, while delta(13)C(R-soil )values changed as much as 6.2 per thousand. The delta(13)C of soil organic carbon (delta(13)C(SOC)) and needle organic carbon (delta(13)C(P)) exhibited little or no significant changes over the course of this study. Carbon isotope discrimination within leaves (Delta(p)) showed systematic decreases with increased canopy height, but remained fairly constant throughout the year (Delta(p)=17.9 per thousand -19.2 per thousand at the top of the canopy, Delta(p)=19.6 per thousand -20.9 per thousand at mid-canopy, Delta(p)=23.3 per thousand -25.1 per thousand at the canopy base). The temporal variation in the delta(13)C of soil and ecosystem respired CO(2) was correlated ( r=0.93, P<0.001) with soil moisture levels, with dry summer months having the most (13)C-enriched values. The dynamic seasonal changes in delta(13)C of respired CO(2) are hypothesized to be the result of fast cycling of recently fixed carbon back to the atmosphere. One scaling consequence of the seasonal and interannual variations in delta(13)C(R) is that inversion-based carbon-cycle models dependent on observed atmospheric CO(2) concentration and isotope values may be improved by incorporating dynamic delta(13)C(R) values to interpret regional carbon sink strength.

Carbon Dioxide↗

Tree-girdling to separate root and heterotrophic respiration in two Eucalyptus stands in Brazil.

The release of carbon as CO2 from belowground processes accounts for about 70% of total ecosystem respiration. Insights about factors controlling soil CO2 efflux are constrained by the challenge of apportioning sources of CO2 between autotrophic tree roots (and mycorrhizal fungi) and heterotrophic microorganisms. In some temperate conifer forests, the reduction in soil CO2 efflux after girdling (phloem removal) has been used to separate these sources. Girdling stops the flow of carbohydrates to the belowground portion of the ecosystem, which should slow respiration by roots and mycorrhizae while heterotrophic respiration should remain constant or be enhanced by the decomposition of newly dead roots. Therefore, the reduction in CO2 efflux after girdling should be a conservative estimate of the belowground flux of C from trees. We tested this approach in two tropical Eucalyptus plantations. Tree canopies remained intact for more than 3 months after girdling, showing no reduction in light interception. The reduction in soil CO2 efflux averaged 16-24% for the 3-month period after girdling. The reduction in CO2 efflux was similar for plots with one half of the trees girdled and those with all of the trees girdled. Girdling did not reduce live fine root biomass for at least 5 months after treatment, indicating that large reserves of carbohydrates in the root systems of Eucalyptus trees maintained the roots and root respiration. Our results suggest that the girdling approach is unlikely to provide useful insights into the contribution of tree roots and heterotrophs to soil CO2 efflux in this type of forest ecosystem.

Brazil↗

An experimental comparison of respiration measuring techniques in fermenters and shake flasks: exhaust gas analyzer vs. RAMOS device vs. respirometer.

Respiration measurement is applied as a universal tool to determine the activity of biological systems. The measurement techniques are difficult to compare, due to the vast variety of devices and analytical procedures commonly in use. They are used in fields as different as microbiology, gene engineering, toxicology, and industrial process monitoring to observe the physiological activity of living systems in environments as diverse as fermenters, shake flasks, lakes and sewage plants. A method is introduced to determine accuracy, quantitation limit, range and precision of different respiration measurement devices. Corynebacterium glutamicum cultures were used to compare an exhaust gas analyzer (EGA), a RAMOS device (respiration measurement in shake flasks) and a respirometer. With all measuring devices it was possible to determine the general culture characteristics. The EGA and the RAMOS device produced almost identical results. The scatter of the respirometer was noticeably higher. The EGA is the technique of choice, if the reaction volume is high or a short reaction time is required. The possibility to monitor cultures simultaneously makes the RAMOS device an indispensable tool for media and strain development. If online monitoring is not compulsive, the respiration of the investigated microbial system extremely low, or the sample size small, a respirometer is recommended.

Bacteriological Techniques↗

Heart and respiration rate changes in the neonate during electroencephalographic seizure.

An investigation of changes in the neonatal electrocardiogram (ECG) and respiration signals from labelled seizure data for five neonatal patient records is reported. A decrease during seizure of 5.70% in the mean RR-interval was found. The mean respiration rate per epoch was found to decrease by a mean 18.44% during seizure. No significant change in the RR-interval standard deviation or in the mean respiration amplitude during seizure was observed. These results raise the possibility of using ECG and respiration based methods instead of existing electrcoenceplogram (EEG) based methods, or in concert with EEG-based neonatal seizure detection methods to improve on previously reported seizure detection methods.

Analysis of Variance↗

The effect of orthodontic force application on the pulpal tissue respiration rate in the human premolar.

This study investigates the effect of an orthodontic force on pulpal respiration in the human premolar. The participants in this study required the removal of four first premolars for orthodontic treatment. After written consent was obtained, these teeth were used in the following manner. Randomly, the premolars on one side of the mouth were designated as experimental and the premolars on the opposing side served as controls. The four teeth were removed following a 3-day application of an orthodontic force. The pulp tissue was then extirpated and used for the investigation. A radioactively labeled carbon dioxide production system was used to evaluate the effect on pulpal respiration of the orthodontic force. It is thought that this method has been proved to provide a viable and sensitive biochemical analysis of tissue respiration on very small samples over short observation periods. The data were collected and prepared for statistical analysis. This investigation demonstrated that the pulp tissue respiration in seventeen subjects was depressed an average of 27% as a result of orthodontic force application. A positive correlation between the age of the participant and the amount of tissue respiratory depression was also demonstrated. It was therefore concluded that orthodontic forces of very short duration do cause biochemical and biologic pulpal tissue alterations and that orthodontic forces may be less biologically safe as the age of the patient increases.

Adolescent↗

A practical mechanical respirator, 1929: the "iron lung".

No satisfactory mechanical respirator existed before 1929, when Philip Drinker and Louis Shaw described an apparatus of their own design. This machine was in the form of a cylindrical tank enclosing the patient's body and chest, leaving the head outside the chamber under atmospheric pressure. Air pumps, later a bellows, raised and lowered pressure within the tank to assume the entire work of breathing. Popularly named the iron lung, the Drinker respirator supported thousands of patients afflicted with respiratory paralysis during the polio era. It was being superseded by positive-pressure airway ventilators just as the polio era came to a close. Today the Drinker respirator has disappeared virtually without a trace. Although its disadvantage was its cumbersome size, we must concede that it supported patients over the long term with fewer complications than do the respirators of today.

Disease Outbreaks↗

In vitro effects of acetaminophen metabolites and analogs on the respiration of mouse liver mitochondria.

Acetaminophen, an analgesic and antipyretic, is toxic in overdose to liver and kidney. The effects on mitochondrial respiration of acetaminophen, its less toxic analog, 3-hydroxyacetanilide, and metabolites which arise from these compounds have been investigated. The parent compounds inhibited NADH-linked respiration reversibly, whereas the metabolites inhibit all mitochondrial respiration, apparently in the Complex III region of the respiratory chain. The quinone derivatives, 4-acetamido-o-benzoquinone and 2-acetamido-p-benzoquinone, are the best inhibitors, with the onset of inhibition dependent on active respiration, suggesting interaction of these compounds with oxidized components of the electron transport chain.

Acetaminophen↗

Cytochrome c dependent, antimycin-A resistant respiration in mitochondria from potato tuber (Solanum tuberosum L.). Influence of wounding and storage time on outer membrane NADH-cytochrome-c-reductase.

Cytochrome c has two stimulatory effects on respiration of mitochondria especially those from wounded potato tuber. In the first place a stimulation of succinate- and NADH-consuming, antimycin-A-sensitive respiration, which reaches a maximal value at low cytochrome c concentrations, has been found. In the second place, at higher concentrations of cytochrome c a stimulation of NADH-consuming respiration occurs, which is antimycin-A-resistant, but KCN-sensitive. This antimycin-A-resistant, NADH-consuming respiration is absent, when no cytochrome c is added to the reaction medium. It is insensitive to metal chelators, to which the antimycin-A-and KCN-resistant plant mitochondrial alternative oxidase is sensitive. By measurements of NADH-cytochrome c reductase activities a corresponding antimycin-A-resistant NADH-cytochrome c reductase has been found, which is insensitive to osmotic shock treatment. A localization of this antimycin-A-resistant electron transport with NADH as the electron donor in the outer mitochondrial membrane is likely. In the mitochondrial preparations cytochrome c might stimulate by acting as an electron-carrier between the outer membrane reductase and the inner membrane cytochrome oxidase. A big increase of the outer membrane mediated electron transport in the mitochondria has been observed after wounding of potato tuber tissue. The ability of the tissue to produce this electron transport pathway after wounding disappeared after prolonged storage of the tubers. A possible function of this electron transport pathway in fatty acid desaturation during the wound-reaction is suggested.

Antimycin A↗