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The mechanism of calcium-dependent activation of respiration of liver mitochondria from hibernating ground squirrels, Citellus undulatus.

1. It is shown that Ca(2+)-dependent activation of respiration of liver mitochondria from hibernating ground squirrels is accompanied by mitochondrial swelling. 2. The swelling of mitochondria from hibernating ground squirrels, as well as the activation of mitochondrial respiration, is precluded by cyclosporin A, p-bromphenacylbromide and oligomycin. Carboxyatractiloside, on the contrary, under these conditions favors the swelling and the acceleration of respiration. 3. It was concluded that Ca(2+)-dependent activation of hibernating ground squirrel liver mitochondrial respiration resulted from the appearance of a non-specific permeability pathway and from swelling of mitochondria.

Animals↗

Inhibition of mitochondrial respiration by chalepin, a naturally occurring furocoumarin.

Polarographic measurements of the rates of oxygen consumption by isolated rat liver mitochondria respiring on pyruvate/malate in metabolic state 4 revealed that additions of micromolar amounts of chalepin, a naturally occurring furocoumarin resulted in significant decreases in respiratory rates. At 16 microM chalepin, respiration was inhibited by at least 40%. A maximum inhibition of 60% was obtained at 80 microM chalepin. Whereas 16 microM chalepin gave 76% reduction of respiratory control ratio, at most 80% reduction was obtained at greater than or equal to 80 microM chalepin. There was no significant effect on either metabolic state 4 respiration or respiratory control ratio when succinate was used as electron donor. A comparison with the effects of rotenone indicates that chalepin is probably only one-tenth as potent as this classical inhibitor of respiration. These results show that chalepin-like rotenone is an inhibitor of energy coupling site 1.

Animals↗

Copper-stimulated respiration in the unfertilized egg of the Eurasian perch, Perca fluviatilis (L).

The addition of Cu2+ (0.1-1.0 mM) to respiring, unfertilized eggs produced a marked stimulation in the rate of respiration whereas Zn2+ had no effect over the same concentration range. In the absence of Cu2+, temperature had little effect on unfertilized egg respiration but the Cu2+ stimulated respiratory rate showed the more normal response with a Q10 of 1.86 (10-20 degrees C). It appears that perch egg respiration is rate-limited by a physical event and it is suggested that Cu2+ may act by dissipating an oxygen permeability barrier located at the chorion.

Animals↗

The reduction of glutathione disulfide produced by t-butyl hydroperoxide in respiring mitochondria.

Factors affecting the reduction of GSSG by rat liver mitochondria after a t-butyl hydroperoxide-induced (t-BOOH) oxidative stress were studied. The amounts of ADP and mitochondrial protein were adjusted to consume less than 50% of the available oxygen during the 8-min experimental period. A 4-min treatment of mitochondria with 24 nmol t-BOOH/mg protein (60 microM) oxidized 91% of total glutathione. In the presence of glutamate/malate, succinate or ascorbate/N,N,N',N'-tetramethyl-p- phenylenediamine (TMPD) (state 4 respiration), 84, 84, and 28% of the GSSG formed during t-BOOH treatment was reduced after 4 min, respectively. A similar extent of reduction was seen during state 3 respiration (1.5 mM ADP) with glutamate/malate, but no reduction occurred during state 3 respiration with either succinate or ascorbate/TMPD. The succinate-supported reduction of GSSG was completely blocked by rotenone, antimycin A, carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP), or 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU). In contrast, oligomycin potentiated GSSG reduction using either glutamate/malate or succinate as substrates. Rotenone partially blocked glutamate/malate-supported GSSG reduction. NADPH levels were altered in direct proportion to the effects on GSSG reduction. The current data indicate that the reduction of GSSG in oxidatively-stressed isolated rat liver mitochondria occurs most efficiently during state 4 respiration and is independent of ATP synthesis. Both transhydrogenation and the transmembrane proton gradient appear to be important in NADPH regeneration and consequent GSSG reduction.

Animals↗

Novel inhibitors of the trypanosome alternative oxidase inhibit Trypanosoma brucei brucei growth and respiration.

African trypanosomiasis is a deadly disease for which few chemotherapeutic options are available. The causative agents, Trypanosoma brucei rhodesiense and T. b. gambiense, utilize a non-cytochrome, alternative oxidase (AOX) for their cellular respiration. The absence of this enzyme in mammalian cells makes it a logical target for therapeutic agents. We designed three novel compounds, ACB41, ACD15, and ACD16, and investigated their effects on trypanosome alternative oxidase (TAO) enzymatic activity, parasite respiration, and parasite growth in vitro. All three compounds contain a 2-hydroxybenzoic acid moiety, analogous to that present in SHAM, and a prenyl side chain similar to that found in ubiquinol. ACD15 and ACD16 are further differentiated by the presence of a solubility-enhancing carbohydrate moiety. Kinetic studies with purified TAO show that all three compounds competitively inhibit TAO, and two compounds, ACB41 and ACD15, have inhibition constants five- and three-fold more potent than SHAM, respectively. All three compounds inhibited the respiration and growth of continuously cultured T. b. brucei bloodstream cells in a dose-dependent manner. None of the compounds interfered with respiration of rat liver mitochondria, nor did they inhibit the growth of a continuously cultured mammalian cell line. Collectively, the results suggest we have identified a new class of compounds that are inhibitors of TAO, have trypanocidal properties in vitro, and warrant further investigation in vivo.

Animals↗

Test-retest reliability of heart rate variability and respiration rate at rest and during light physical activity in normal subjects.

BACKGROUND: A variable that remains stable over repeated measurements (in stable conditions) is ideal for tracking modifications of the clinical state. The aim of the present study is to examine test-retest reliability of time-domain heart rate variability and respiration rate measurements using a portable device on normal subjects during rest and light physical activity. METHODS: Twenty-six normal subjects [18 females and 8 males aged 28 +/- 6 years and 34 +/- 12 years (mean +/- SD), respectively] underwent two measurements for time-domain heart rate variability (SDNN and RMSSD) and respiration rate, with 7 days in between. Measurements took place under three conditions: lying down in a laboratory, cycling in a laboratory and sleeping in an ambulatory surrounding. Reliability was assessed statistically by calculating intra-class correlation coefficients (ICC). RESULTS: Reliability was found to be good to excellent for both time-domain heart rate variability (SDNN: ICC values between 0.74 and 0.85, RMSSD: ICC values between 0.75 and 0.98) and for respiration rate (ICC values between 0.77 and 0.96). CONCLUSIONS: Both time-domain heart rate variability and respiration rate can be reliably assessed. However, we advise reliability research in a clinical setting before using the device for tracking modifications in a clinical state.

Adult↗

Comparison of temperature effects on soil respiration and bacterial and fungal growth rates.

Temperature is an important factor regulating microbial activity and shaping the soil microbial community. Little is known, however, on how temperature affects the most important groups of the soil microorganisms, the bacteria and the fungi, in situ. We have therefore measured the instantaneous total activity (respiration rate), bacterial activity (growth rate as thymidine incorporation rate) and fungal activity (growth rate as acetate-in-ergosterol incorporation rate) in soil at different temperatures (0-45 degrees C). Two soils were compared: one was an agricultural soil low in organic matter and with high pH, and the other was a forest humus soil with high organic matter content and low pH. Fungal and bacterial growth rates had optimum temperatures around 25-30 degrees C, while at higher temperatures lower values were found. This decrease was more drastic for fungi than for bacteria, resulting in an increase in the ratio of bacterial to fungal growth rate at higher temperatures. A tendency towards the opposite effect was observed at low temperatures, indicating that fungi were more adapted to low-temperature conditions than bacteria. The temperature dependence of all three activities was well modelled by the square root (Ratkowsky) model below the optimum temperature for fungal and bacterial growth. The respiration rate increased over almost the whole temperature range, showing the highest value at around 45 degrees C. Thus, at temperatures above 30 degrees C there was an uncoupling between the instantaneous respiration rate and bacterial and fungal activity. At these high temperatures, the respiration rate closely followed the Arrhenius temperature relationship.

Acetates↗

The impact of respiration on left atrial and pulmonary venous anatomy: implications for image-guided intervention.

BACKGROUND: Image-guided intervention using pre-acquired CT/MR 3-dimensional images is an emerging strategy for atrial fibrillation (AF) ablation but may be limited by its use of static images to depict dynamic physiology. The effect of biologic factors such as respiration on the left atrial-pulmonary venous (LA-PV) anatomy is not well understood but is likely to have important implications. Conventional CT/MR imaging is performed during an inspiratory breath-hold, while electroanatomical mapping (EAM) during "quiet" breathing approximates an expiratory breath-hold. This study examined the effects of respiration on LA-PV anatomy and the error introduced by respiration on the integration of EAM with 3D MR imaging. METHODS: Pre-procedural MRI angiography was performed at both end-expiration (EXP) and end-inspiration (INSP) in 20 patients undergoing AF catheter ablation. 3D INSP and EXP surface reconstructions of the LA-PVs were compared. In selected pts, EAM data acquired during the ablation procedure (n=7) were integrated with the 3D MRI datasets. RESULTS: Qualitative assessment of the INSP and EXP 3D images revealed splaying of the PVs and reduction in PV caliber of the right-sided PVs during held inspiration. After aligning these two datasets, the average surface-to-surface distance calculated by region ranged from 1.99mm (right middle PV) to 3.79mm (left superior PV). Registration of the EAM to the MRI models was better for the EXP dataset (2.30+/-0.73mm) than the INSP dataset (3.03+/-0.57mm; p=0.004). CONCLUSION: There are significant changes in LA-PV anatomy with respiration. MR images acquired during standard held inspiration may introduce unnecessary errors in registration during image-guided intervention.

Aged↗

Changes in lead availability affect bacterial community structure but not basal respiration in a microcosm study with forest soils.

This study investigates the effects of Pb during time on the bacterial communities of forest soils using water-extractable Pb concentrations in the soil solution as predictors of Pb bioavailability. In a microcosm experiment we applied increasing concentrations of Pb(NO(3))(2) solutions (0.5, 2, 8, 32 mM) to 5 forest soils of pH<5 and to a calcareous soil of pH>6.5. Sampling of the microcosms was performed after 3, 30 and 90 days of incubation. Community analysis included basal respiration rates and changes in the structure of the bacterial communities through T-RFLP fingerprinting. We also investigated functional stability in terms of resistance, expressed as the effects on basal respiration after 3 days of incubation, and of resilience, expressed as the recovery of bacterial community structure and of respiration rates after 90 days of incubation. Water-extractable Pb increased with time in most of the soils, in parallel with an increase of water-extractable dissolved organic carbon (DOC). The increased concentrations slightly affected bacterial community structure, although OTU (operational taxonomic unit) richness was not significantly reduced with Pb concentrations in any of the soils. The highest Pb treatment (32 mM) caused significant effects on basal respiration in some of the acidic soils, but no clear trend was observed in relation to increased Pb bioavailability with time. Resistance to Pb additions was evident in five of the six soils, but only two showed resilience after 90 days. This is the first study showing the effects of time on Pb bioavailability in soils and on the resulting reactions of the soil microbial communities.

DNA, Bacterial↗

Alterations in respiration rate of isolated rainbow trout hepatocytes exposed to the P-glycoprotein substrate rhodamine 123.

Reducing intracellular xenobiotic concentration is an important defence strategy used by cells challenged with foreign chemicals. One mechanism used to achieve this goal is via the use of P-glycoproteins (P-gps), ATP-dependent transporters that mediate the removal of hydrophobic compounds from cells. The energetic costs of this mechanism are unknown, therefore, the activity and respiratory costs associated with the P-gp-mediated efflux of rhodamine 123 (R123) was measured in isolated rainbow trout hepatocytes. The accumulation of R123 was rapid and concentration-dependent. Initial accumulation rates were 1.79+/-0.41, 7.29+/-1.06 and 15.30+/-1.74ngR123/min/10(6)cells when exposed to 1, 5 and 10 microM R123, respectively. Efflux was measured in cells 'pre-loaded' with R123 at each concentration, resulting in initial efflux rates of 0.77+/-0.12, 2.02+/-0.35 and 3.51+/-0.84ngR123/min/10(6)cells, respectively. The baseline oxygen consumption rate of hepatocytes was 33.21+/-1.09 ng O2/min/10(6)cells. Respiration rates were significantly higher in cells exposed to 5 and 10 microM R123 (39.08+/-0.80 and 41.72+/-0.61ng O2/min/10(6)cells), representing increases over basal rates of 18.5 and 25.7%, respectively. Measurements of isolated mitochondrial respiration established that changes in hepatocyte oxygen consumption were not through the direct effects of R123 on mitochondria. The P-gp inhibitor, XR9576 significantly inhibited R123 efflux from cells with a concomitant return of respiration rates to baseline values. This study demonstrates that increased P-gp transport of xenobiotics can significantly raise cellular respiration rates and may result in higher energy costs for organisms living in P-gp-substrate contaminated environments.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Collection of three bacterial aerosols by respirator and surgical mask filters under varying conditions of flow and relative humidity.

A variety of respirator filters and surgical masks were challenged with three aerosolized bacteria: Mycobacterium abscessus (M.a.) (a rod), Staphylococcus epidermidis (S.e.) (a sphere), and Bacillus subtilis (B.s.) (a rod). Tests were conducted at two flow rates (45 and 85 l./min) and two humidity levels (30 and 70%). Aerosols were measured with a total-particle, direct-reading, spectrometer and a viable particle cascade impactor. Measurements up- and downstream of the filter or mask were used in determining aerosol penetration; respirator or surgical mask fit was not evaluated. Bioaerosol penetration measured with two aerosol sampling instruments was found to correlate. Additionally, bioaerosol test parameters were evaluated with respect to their effect on penetration. Increasing flow resulted in increased penetration of all organisms while an increase in relative humidity did not exert a consistent effect on all organisms. Of the respirators approved by the National Institute for Occupational Safety and Health (NIOSH), filter efficiency was as expected with dust/mist respirators having the lowest and HEPA filters the highest efficiency. Surgical masks were the least efficient of all filters tested; these are not certified by NIOSH. Bioaerosol penetration was compared to that of a polystyrene latex sphere (PSL) aerosol. Penetration of the test aerosols was predicted on the basis of particle aerodynamic diameter and was expected to be in this order: PSL > M.a. > S.e. = B.s. The PSL aerosol was the most penetrating, as predicted. However, results showed that B.s. was more penetrating than S.e. The aerodynamic diameter may not be the best parameter for predicting aerosol penetration of non-spherical particles in these filters.

Aerosols↗

Estimating factors to convert Chinese 'Total Dust' measurements to ACGIH respirable concentrations in metal mines and pottery industries.

Historical data on the dust exposures of Chinese workers in metal mines (iron/copper, tin, tungsten) and pottery industries are being used in an ongoing joint Chinese/United States epidemiological study to investigate the exposure-response relationship for the development of silicosis, lung cancer, and other diseases. The historical data include 'total dust' concentrations determined by a Chinese method. Information about particle size distribution and the chemical and mineralogical content of airborne particles is generally not available. In addition, the historical Chinese sampling strategy is different from a typical American eight-hour time-weighted average (TWA) sampling strategy, because the Chinese samples were collected for approximately 15 minutes during production so the sample could be compared to their maximum allowable concentration (MAC) standard. Therefore, in order to assess American respirable dust exposure standards in light of the Chinese experience, factors are needed to convert historical Chinese total dust concentrations to respirable dust concentrations. As a part of the joint study to estimate the conversion factors, airborne dust samples were collected in 20 metal mines and 9 pottery factories in China during 1988 and 1989 using three different samplers: 10mm nylon cyclones, multi-stage 'cassette' impactors, and the traditional Chinese total dust samplers. More than 100 samples were collected and analysed for each of the three samplers. The study yielded two different estimates of the conversion factor from the Chinese total dust concentrations (measured during production processes) to respirable dust concentrations. The multivariate analysis of variance (MANOVA) reveals that, with a fixed sampling/analysis method, conversion factors were not statistically different among the different job titles within each industry. It also indicates that conversion factors among the industries were not statistically different. However, the two estimates consistently showed that conversion factors were the lowest in the pottery industry. Average conversion factors were then calculated for each of the estimates across the industries studied. A pooled mean conversion factor, 0.25+/-0.04, was then derived for all the job titles and industries. Respirable dust levels were estimated from the historical 'total dust' concentrations collected between 1952 and 1992 by adopting the American standard.

Air Pollutants, Occupational↗

Regulation of mitochondrial respiration by nitric oxide inhibition of cytochrome c oxidase.

Nitric oxide (NO) and its derivatives inhibit mitochondrial respiration by a variety of means. Nanomolar concentrations of NO immediately, specifically and reversibly inhibit cytochrome oxidase in competition with oxygen, in isolated cytochrome oxidase, mitochondria, nerve terminals, cultured cells and tissues. Higher concentrations of NO and its derivatives (peroxynitrite, nitrogen dioxide or nitrosothiols) can cause irreversible inhibition of the respiratory chain, uncoupling, permeability transition, and/or cell death. Isolated mitochondria, cultured cells, isolated tissues and animals in vivo display respiratory inhibition by endogenously produced NO from constitutive isoforms of NO synthase (NOS), which may be largely mediated by NO inhibition of cytochrome oxidase. Cultured cells expressing the inducible isoform of NOS (iNOS) can acutely and reversibly inhibit their own cellular respiration and that of co-incubated cells due to NO inhibition of cytochrome oxidase, but after longer-term incubation result in irreversible inhibition of cellular respiration due to NO or its derivatives. Thus the NO inhibition of cytochrome oxidase may be involved in the physiological and/or pathological regulation of respiration rate, and its affinity for oxygen.

Animals↗

Effects of size and light on respiration and activity of walleye pollock (Theragra chalcogramma) larvae.

The respiration rate and swimming activity of walleye pollock (Theragra chalcogramma) larvae were measured in the laboratory to determine how these were affected by body size (measured as dry weight), and amount of light. Size influenced respiration rates, but not activity. Activity increased with increased light, and as walleye pollock larvae developed, light had an increasingly important effect on respiration rate. For older larvae, light is an important factor affecting respiration rate and this may be due to an increased sensitivity to light. Thus, in addition to size, light plays an important role in the energetics of walleye pollock larvae.

Journal Article↗

Surviving the flood: plastron respiration in the non-tracheate arthropod Phrynus marginemaculatus (Amblypygi: Arachnida).

Specimens of Phrynus marginemaculatus can remain responsive when submerged in water for more than 24 hours. Behavioral data indicate that P. marginemaculatus utilizes dissolved oxygen from the surrounding water. Scanning electron miscroscopy and light microscope sections show cuticular modifications for plastron respiration. All previous examples of plastron respiration have involved animals with tracheal systems, but amblypygids respire through the use of two pairs of book lungs. This study provides the first example of plastron respiration not only in the order Amblypygi, but also, in any non-tracheate arthropod.

Journal Article↗

Mitochondrial respiration following acute hypoxia in the perfused rat heart.

Mitochondria isolated from tissues of hypoxic animals have increased respiratory capacity (State 3 respiration) when assayed in vitro at ambient oxygen tensions. The present study utilized the isolated perfused rat heart to determine whether or not this change could be produced in the absence of the neural and hormonal changes that accompany hypoxia in vivo. Following 10-min single pass retrograde perfusion with normoxic Krebs-Henseleit buffer (PO2 greater than or equal to 600 mmHg), perfusion was continued for up to 15 min with either normoxic or hypoxic buffer (PO2 less than or equal to 150 mmHg). After 10 min of hypoxic perfusion State 3 respiration of the mitochondria from the hypoxic hearts was 13 to 15% higher (P less than or equal to 0.05) than that of normoxic hearts when assayed with either glutamate/malate or succinate as substrate but was unchanged when TMPD and ascorbate was the substrate. Succinate-supported State 4 respiration of the hypoxic mitochondria also showed a small (10%) but significant (P less than or equal to 0.05) increase. These changes were not abolished by preperfusing the heart with propranolol (10(-7), 10(-6), or 10(-5) M) indicating that the response was not attributable to release of local stores of catecholamines. Respiratory control and ADP/O ratios as well as contents of cytochrome c and aa3 of the mitochondria from the hypoxic hearts were similar to those of normoxic hearts indicating that the mitochondria remained intact and tightly coupled. We concluded that the hypoxia-induced increase in mitochondrial State 3 respiration, while independent of neural and hormonal influences from the body requires an intracellular event, since they cannot be reproduced by subjecting isolated mitochondria to hypoxia in vitro.

Adenosine Diphosphate↗

Concentrations of the major birch tree allergen Bet v 1 in pollen and respirable fine particles in the atmosphere.

BACKGROUND: Birch tree pollen allergens are an important cause of early spring hay fever and allergic asthma. Pollen counts provide a guide for individuals with birch pollen allergy. However, birch pollen, because of its size, has a low probability of entering the lower airways to trigger asthma. Yet birch pollen allergens are known to be associated with respirable particles present in the atmosphere. OBJECTIVE: We sought to determine the concentration of major allergen Bet v 1 in birch pollen and respirable particles in the atmosphere during the birch pollen season. METHODS: We used a two-site monoclonal antibody-based assay (ELISA) to quantitate Bet v 1 in pollen extracts and high-volume air sampler filters collecting particles larger and smaller than 7.2 microm. RESULTS: Bet v 1 (0.006 ng) is detectable per birch pollen grain, of which 0.004 ng is present in aqueous extracts (13.9% of soluble proteins). Atmospheric Bet v 1 concentrations are correlated with birch pollen counts. Heavy rainfall tended to wash out pollen and particles, indicated by a mean daily Bet v 1 concentration of 0.12 ng/m3 (20 pollen equivalents), but light rainfall produced a dramatic increase in allergen-loaded respirable particles with Bet v 1 concentrations of 1.2 ng/m3 (200 pollen equivalents). CONCLUSION: These results highlight the different environmental risk factors for hay fever and allergic asthma in patients sensitized to Bet v 1. Light rainfall causes an increase in respirable particles; hence, this is an important risk factor for asthma.

Air Pollution↗

Monitoring microbial biomass and respiration in different soils from the Czech Republic--a summary of results.

The microbial biomass (Cbio), respiration (basal respiration (BR) and potential respiration (PR)), and derived indices for 520 independent soil samples of 117 different soils from the Czech Republic were statistically analysed. The broad range of soil samples allowed the stepwise breakdown of the database into six reasonable categories of soil: arable soils, loamy grassland soils, sandy grassland soils with weak organic matter content, sandy grassland soils with moderate organic matter content, forest soils with moderate organic matter content, and forest organic soils with rich organic matter content. Because soil microbiology lacks benchmarking values, the ranges of the microbial characteristics for these categories were stated and are presented here. The separation into soil groups narrowed the ranges enough to be useful for comparative purposes. The groups displayed significant differences in basal microbial parameters. The lowest microbial biomass was found in arable soils and grassland sandy soils with weak organic matter content. The highest microbial biomass was shown by loamy grassland soils and organic forest soils. Respiration displayed similar results to the microbial biomass. The derived indices revealed less significant differences confirming their inner-standard nature. The relationships between the soil contamination and microbial parameters were not explored because of the confounding effect of soil organic matter. However, it was not shown by the category of grassland sandy soils with weak organic matter content suggesting they could be especially suitable for the biomonitoring of harmful effects of chemicals on soil microorganisms.

Biomass↗