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PubMed2005

A sulphite respiration system in the chemoheterotrophic human pathogen Campylobacter jejuni.

The ability to use sulphite as a respiratory electron donor is usually associated with free-living chemolithotrophic sulphur-oxidizing bacteria. However, this paper shows that the chemoheterotrophic human pathogen Campylobacter jejuni has the ability to respire sulphite, with oxygen uptake rates of 23 +/- 8 and 28 +/- 15 nmol O(2) min(-1) (mg cell protein)(-1) after the addition of 0.5 mM sodium sulphite or metabisulphite, respectively, to intact cells. The C. jejuni NCTC 11168 Cj0004c and Cj0005c genes encode a monohaem cytochrome c and molybdopterin oxidoreductase, respectively, homologous to the sulphite : cytochrome c oxidoreductase (SOR) of Starkeya novella. Western blots of C. jejuni periplasm probed with a SorA antibody demonstrated cross-reaction of a 45 kDa band, consistent with the size of Cj0005. The Cj0004c gene was inactivated by insertion of a kanamycin-resistance cassette. The resulting mutant showed wild-type rates of formate-dependent respiration but was unable to respire with sulphite or metabisulphite as electron donors. 2-Heptyl-4-hydroxyquinoline-N-oxide (HQNO), a cytochrome bc(1) complex inhibitor, did not affect sulphite respiration at concentrations up to 25 microM, whereas formate respiration (which occurs partly via a bc(1) dependent route) was inhibited 50%, thus suggesting that electrons from sulphite enter the respiratory chain after the bc(1) complex at the level of cytochrome c. Periplasmic extracts of wild-type C. jejuni 11168 showed a symmetrical absorption peak at 552 nm after the addition of sulphite, demonstrating the reduction of cytochrome c. No cytochrome c reduction was observed after addition of sulphite to periplasmic extracts of the Cj0004c mutant. A fractionation study confirmed that the majority of the SOR activity is located in the periplasm in C. jejuni, and this activity was partially purified by ion-exchange chromatography. The presence of a sulphite respiration system in C. jejuni is another example of the surprising diversity of the electron-transport chain in this small-genome pathogen. Sulphite respiration may be of importance for survival in environmental microaerobic niches and some foods, and may also provide a detoxification mechanism for this normally growth-inhibitory compound.

Amino Acid Sequence↗
PubMed1972

Effects of cytokinins on the respiration of soybean callus tissue.

A technique which incorporates a brief blending step to disperse callus tissue into small clumps of cells was developed, and the effects of cytokinins on respiration of soybean (Glycine max [L.] Merrill var. Acme) callus tissue prepared in this way were studied. Adenine alone did not affect respiration, but kinetin and zeatin showed effects correlating with their reported effects on growth of this tissue; after about 3 hours both hormones promoted respiration at concentrations which promote growth, while kinetin, but not zeatin, also exhibited inhibition at higher concentrations. Studies with 2,4-dinitrophenol led to the suggestion that although the respiration of this tissue is largely under the control of ATP levels, kinetin does not exert its control on respiration through effects on ATP levels or oxidative phosphorylation during the monitoring period. Further inhibitor and substrate studies provided evidence that the promotion of respiration by kinetin results from an increase in substrate entering the tricarboxylic acid cycle, perhaps by an effect on pyruvate metabolism.The inhibition of respiration by high concentrations of kinetin is partially due to effects on oxidative phosphorylation or ATP utilization, but 70% of the inhibition cannot be attributed to this.

Journal Article↗
PubMed1975

The effect of nucleotides and inhibitors on respiration in isolated wheat mitochondria.

The effect of mono-, di-, and trinucleoside phosphates and respiratory inhibitors on respiration in winter wheat (Triticum aestivum L. cv. Rideau) mitochondria has been examined. When added during state 4 respiration, subsequent to addition of ADP, all of the dinucleotides stimulated oxidation and induced respiratory control with all substrates examined. Similar results were obtained with AMP, but other mononucleotides and all trinucleotides did not affect the rate of oxidation. Nucleoside diphosphates did not stimulate respiration when added prior to the addition of ADP, but subsequent addition of AMP, ADP, or ATP re-established coupled respiration in the presence of the dinucleotides.The duration of 2, 4-dinitrophenol stimulated respiration during oxidation of alpha-ketoglutarate was found to be dependent on the amount of AMP, ADP, or ATP added, either prior, or subsequent to, addition of the uncoupler. The addition of oligomycin during 2, 4-dinitrophenol stimulated respiration reestablished coupled respiration with low ADP/O ratios, when added after addition of ATP or conditions which allow formation of ATP from added ADP. The nucleoside diphosphates, other than ADP, did not stimulate oxidation of alpha-ketoglutarate in the presence of 2, 4-dinitrophenol until a small amount of adenine nucleotide was added to the system. The results suggest that dinucleotides other than ADP, are able to participate in the energy conversion processs of the mitochondria, probably via transphosphorylation reactions.

Journal Article↗
PubMed1977

Root and nodule respiration in relation to acetylene reduction in intact nodulated peas.

Inoculated pea plants (Pisum sativum L.) were grown with N-free nutrients in a controlled environment room and rates of respiratory CO(2) evolution and C(2)H(2) reduction by the intact nodulated roots were determined. Experiments followed changes related to diurnal cycles, light and dark treatments, partial defoliation, aging of plants and NH(4)NO(3) addition. In all experiments, changes in C(2)H(2) reduction were associated with parallel changes in the respiration rate, although in all but the defoliation experiment there was a basal level of respiration which was independent of the rate of C(2)H(2) reduction. In conditions which affected growth or plant size as well as C(2)H(2) reduction, respiration changed by an average of 0.42 mg CO(2) (mumol C(2)H(2) reduced)(-1). However, some treatments decreased C(2)H(2) reduction without greatly changing the growth and in these conditions respiration was decreased by an average of 0.27 mg CO(2) (mumol C(2)H(2) reduced)(-1). While this value may also include some respiration associated with other processes, it is proposed that it more closely estimates respiration directly associated with energy utilization for acetylene reduction; whereas the higher value includes respiration related to maintenance and growth processes as well.

Journal Article↗
PubMed1983

Appearance and Disappearance of Cyanide-Resistant Respiration in Vigna mungo Cotyledons during and following Germination of the Axis.

Mitochondrial preparations isolated from black gram (Vigna mungo L.) cotyledons exhibited cyanide-resistant respiration which was of mitochondrial origin. The appearance and the disappearance of this alternative respiration took place during and following imbibition. During the first 6 hours of imbibition, the respiration was completely inhibited by cyanide, but after this time the alternative respiration markedly developed, reaching a maximal cyanide-resistance 12 to 16 hours after the start of imbibition. Subsequently, the alternative respiration gradually disappeared. The actions of cycloheximide and chloramphenicol indicated that the appearance was dependent on cytoplasmic protein synthesis and that the disappearance depended on both cytoplasmic and mitochondrial protein synthesis. The alternative pathway contributed to state 4 respiration, but not to state 3 respiration, in mitochondria from 1-day-old cotyledons. On day 3, it contributed to neither state 3 nor state 4.

Journal Article↗
PubMed1984

Relationships between Respiration Rate and Adenylate and Carbohydrate Pools of the Soybean Fruit.

Relationships between respiration rate and adenylate and carbohydrate pools of the soybean (Glycine max L. Merrill) fruit during rapid seed growth were evaluated. Plants at mid pod-fill were subjected to different concentrations of CO(2) to alter the amount of photosynthate produced and, thus, available to the fruit. Respiration rate of the intact fruits was measured, along with glucose, sucrose, and starch concentrations, adenylate energy charge (AEC), and total adenylate pool (SigmaAdN) in the pod wall, seed coat, and cotyledons. The concentration of sucrose remained relatively constant in the pod wall (1.0 milligram per 100 milligrams dry weight), seed coat (6.5 milligrams per 100 milligrams dry weight), and cotyledons (4.5 milligrams per 100 milligrams dry weight) at moderate and high respiration rates. Furthermore, AEC remained relatively constant in the pod wall (0.55), seed coat (0.24), and cotyledons (0.44) during changes in respiration rate. This suggests that the amount of assimilate transported to the fruit, and its flux through the sucrose pools of the fruit parts, were important in the regulation of the respiration rate of the fruit. The average SigmaAdN in the seed coat (1300 picomoles per milligram dry weight) was significantly greater than in the cotyledons (750 picomoles per milligram dry weight) and pod wall (300 picomoles per milligram dry weight). In addition, the SigmaAdN in the seed coat and cotyledons increased with increasing respiration rate of the fruit. The high SigmaAdN in the seed coat and its increase with increases in respiration rate of the fruit suggest that an energy-requiring process is involved in the movement of sucrose through the seed coat.

Journal Article↗
PubMed2005

Respiration metabolism of Group B Streptococcus is activated by environmental haem and quinone and contributes to virulence.

Group B Streptococcus (GBS) is a common constituent of the vaginal microflora, but its transmission to newborns can cause life-threatening sepsis, pneumonia and meningitis. Energy metabolism of this opportunist pathogen has been deduced to be strictly fermentative. We discovered that GBS undergoes respiration metabolism if its environment supplies two essential respiratory components: quinone and haem. Respiration metabolism led to significant changes in growth characteristics, including a doubling of biomass and an altered metabolite profile under the tested conditions. The GBS respiratory chain is inactivated by: (i) withdrawing haem and/or quinone, (ii) treating cultures with a respiration inhibitor or (iii) inactivating the cydA gene product, a subunit of cytochrome bd quinol oxidase, in all cases resulting in exclusively fermentative growth. cydA inactivation reduced GBS growth in human blood and strongly attenuated virulence in a neonatal rat sepsis model, suggesting that the animal host may supply the components that activate GBS respiration. These results suggest a role of respiration metabolism in GBS dissemination. Our findings show that environmental factors can increase the flexibility of GBS metabolism by activating a newly identified respiration chain. The need for two environmental factors may explain why GBS respiration metabolism was not found in previous studies.

Bacterial Proteins↗
PubMed2000

Regulation of mitochondrial respiration by oxygen and nitric oxide.

Although the regulation of mitochondrial respiration and energy production in mammalian tissues has been exhaustively studied and extensively reviewed, a clear understanding of the regulation of cellular respiration has not yet been achieved. In particular, the role of tissue pO2 as a factor regulating cellular respiration remains controversial. The concept of a complex and multisite regulation of cellular respiration and energy production signaled by cellular and intercellular messengers has evolved in the last few years and is still being researched. A recent concept that regulation of cellular respiration is regulated by ADP, O2 and NO preserves the notion that energy demands drive respiration but places the kinetic control of both respiration and energy supply in the availability of ADP to F1-ATPase and of O2 and NO to cytochrome oxidase. In addition, recent research indicates that NO participates in redox reactions in the mitochondrial matrix that regulate the intramitochondrial steady state concentration of NO itself and other reactive species such as superoxide radical (O2-) and peroxynitrite (ONOO-). In this way, NO acquires an essential role as a mitochondrial regulatory metabolite. No exhibits a rich biochemistry and a high reactivity and plays an important role as intercellular messenger in diverse physiological processes, such as regulation of blood flow, neurotransmission, platelet aggregation and immune cytotoxic response.

Electron Transport↗
PubMed1979

Respiration and viability of thermally injured Saccharomyces cerevisiae.

Resting cells of Saccharomyces cerevisiae Y25 were heated at 56 degrees C for 0 to 2 min. Respiratory activity of the cells reflected the severity of the heat stress. The endogenous respiration was approximately 50 microliter of O2/mg per h for cells heated for 2 min at 56 degrees C as compared with 2 microliter of O2/mg per h for nonheated cells. There was a distinct decrease in respiration after 1 to 3 h, and after 20 h the respiration rate of heated cells was less than that of nonheated cells. Along with increased rates of endogenous respiration, respiratory quotients of cells were altered after heat stress. Addition of 2,4-dinitrophenol stimulated O2 (uptake) in nonheated cells but decreased O2 (uptake) of heated cells. Due to the high rate of endogenous respiration, addition of glucose resulted in no substantial change in the rate of respiration of heated cells. However, addition of glucose prolonged the presence of the high rates of respiration observed in heated cells.

Dinitrophenols↗
PubMed1994

Augmented rates of respiration and efficient nitrogen fixation at nanomolar concentrations of dissolved O2 in hyperinduced Azoarcus sp. strain BH72.

Azoarcus sp. strain BH72 is an aerobic diazotrophic bacterium that was originally found as an endophyte in Kallar grass. Anticipating that these bacteria are exposed to dissolved O2 concentrations (DOCs) in the nanomolar range during their life cycle, we studied the impact of increasing O2 deprivation on N2 fixation and respiration. Bacteria were grown in batch cultures, where they shifted into conditions of low pO2 upon depletion of O2 by respiration. During incubation, specific rates of respiration (qO2) and efficiencies of carbon source utilization for N2 reduction increased greatly, while the growth rate did not change significantly, a phenomenon that we called "hyperinduction." To evaluate this transition from high- to low-cost N2 fixation in terms of respiratory kinetics and nitrogenase activities at nanomolar DOC, bacteria which had shifted to different gas-phase pO2s in batch cultures were subjected to assays using leghemoglobin as the O2 carrier. As O2 deprivation in batch cultures proceeded, respiratory Km (O2) decreased and Vmax increased. Nitrogenase activity at nanomolar DOC increased to a specific rate of 180 nmol of C2H4 min-1 mg of protein-1 at 32 nM O2. Nitrogenase activity was proportional to respiration but not to DOC in the range of 12 to 86 nM O2. Respiration supported N2 fixation more efficiently at high than at low respiratory rates, the respiratory efficiency increasing from 0.14 to 0.47 mol of C2H4 mol of O2 consumed-1. We conclude that (i) during hyperinduction, strain BH72 used an increasing amount of energy generated by respiration for N2 fixation, and (ii) these bacteria have a high respiratory capacity, enabling them to develop ecological niches at very low pO2, in which they may respire actively and fix nitrogen efficiently at comparatively high rates.

Cell Division↗
PubMed1969

Effects of ultraviolet radiation on respiration and growth in radiation-resistant and radiation-sensitive strains of Escherichia coli B.

Ultraviolet (UV) irradiation at 254 nm causes different respiration and growth responses in log-phase cultures of Escherichia coli B/r and B(s-1). These differences are correlated with the ability and inability, respectively, of these bacterial strains to repair UV-induced lesions in deoxyribonucleic acid (DNA). After irradiation, B(s-1) cells (radiation-sensitive) exhibit uncoupling of growth and respiration; growth and synthesis cease, whereas respiration continues. B/r cells (radiation-resistant) grown on glycerol exhibit severe temporary inhibition of growth and respiration after UV, and the coupling of these two processes is maintained, except at a very high UV dose. Inhibition begins at about the time DNA synthesis resumes and continues for a period of time that is dependent upon dose. Glucose-grown cells do not exhibit severe respiratory, growth, and synthetic inhibitions; these processes remain coupled in the cells during the postirradiation period. Photoreactivation treatment delays uncoupling of growth and respiration in B(s-1) and prevents inhibition of respiration and growth in B/r. These results indicate that the postirradiation responses result from the presence of pyrimidine dimers in DNA. Ultraviolet irradiation of B/r and B(s-1) cells results in an accumulation of adenosine triphosphate by 30 min after UV. This accumulation decreases with time and does not appear to be related to the inhibition of respiration in glycerol-grown B/r cells. The results on B/r are interpreted in terms of a control mechanism for reestablishment of a balance among macromolecules in the irradiated cells so as to provide them with the potential to survive. The specific steps in such a reestablishment of balance appear to depend upon the substrate oxidized. In B(s-1) cells, which cannot repair UV-induced damage in DNA, some control mechanism that coordinates cellular processes may be inactivated.

Adenosine Triphosphate↗
PubMed1992

Relation between exposure to respirable silica dust and silicosis in a tungsten mine in China.

To estimate the quantitative relation between exposure to respirable silica dust and risk of an attack of silicosis, 1151 workers exposed to silica dust and employed from 1958 to 1987 in a tungsten mine in China were investigated. The results showed that the ratio of respirable silica dust concentration to total silica dust concentration was 0.529. Then, the total silica dust concentration in historical surveillance and monitoring data was converted to respirable silica dust concentration. The free silica content in respirable dust determined by x ray diffraction averaged 24.7%. Multiple logistic regression was used for the dichotomous dependent variables (presence or absence of silicosis). The independent variables in the multiple logistic regression with presence of silicosis as the dependent variable were age when first exposed, tuberculosis (presence or absence), and cumulative exposure to respirable silica dust. The partial regression coefficient of individual cumulative exposure was estimated as 0.079. It implied a positive association between exposure to respirable silica dust and risk of an attack of silicosis. The exposure limit for respirable silica dust was estimated as 0.24 mg/m3 under given conditions.

Dust↗
PubMed1995

Enterocyte respiration rates in feline small intestine exposed to graded ischemia.

The purpose of our study was to investigate the changes in enterocyte cellular and mitochondrial respiration rates subsequent to ischemia of graded duration. The small intestine of anesthetized adult cats was assigned to one of five treatment regimens: control or ischemia of 15-, 30-, 60-, or 90-min duration. Cellular and mitochondrial respiration was measured using a Clark-type O2 electrode at 0 and 4 h postharvest. Ischemia of increasing duration caused a progressive decrease in cellular and mitochondrial respiration in enterocytes at 0 h postharvest. By 4 h postharvest, cellular and mitochondrial respiration rates for the 15-, 30-, and 60-min ischemic groups had recovered to near control levels, whereas the 90-min group showed minimal recovery. These data suggest that ischemia suppresses cellular and mitochondrial respiration of intestinal epithelial cells, the magnitude of which is related to the ischemic duration. The ischemia-induced suppression in cellular respiration primarily reflects a reduction in mitochondrial respiration.

Animals↗
PubMed2002

Increased concentrations of P(i) and lactic acid reduce creatine-stimulated respiration in muscle fibers.

We tested the hypothesis that the respiratory function of skeletal muscle mitochondria is impaired by lactic acidosis and elevated concentrations of P(i). The rate of respiration of chemically skinned fiber bundles from rat soleus muscle was measured at [P(i)] (brackets denote concentration) and pH values similar to those at rest (3 mM P(i), pH 7.0) and high-intensity exercise (20 mM P(i), pH 6.6). Respiration was measured in the absence of ADP and after sequential additions of 0.1 mM ADP, 20 mM creatine (Cr; V(Cr)), and 4 mM ADP. Respiration at 0.1 mM ADP increased after addition of Cr. However, V(Cr) was 23% lower (P < 0.05) during high-intensity conditions than during resting conditions. V(Cr) was also reduced when P(i) or H(+) was increased separately (P < 0.05). Respiration in the absence of ADP and after additions of 0.1 mM ADP and 4 mM ADP was not affected by changes in [P(i)] or [H(+)]. The response was similar, irrespective of when acidosis was induced (i.e., quiescent or actively respiring mitochondria). In conclusion, Cr-stimulated respiration is impaired by increases in [H(+)] and [P(i)] corresponding to those in exercising muscle. Although the reduced Cr-stimulated respiration could be compensated for by increased [ADP], this might have implications for intracellular homeostasis.

Acidosis↗
PubMed1996

Role of nitric oxide and its interaction with superoxide in the suppression of cardiac muscle mitochondrial respiration. Involvement in response to hypoxia/reoxygenation.

BACKGROUND: Nitric oxide (NO); superoxide anion (O2.d-); the reaction product of NO with O2.d-, peroxynitrite (ONOO-); and ischemia/reperfusion have all been reported to inhibit respiration in isolated mitochondria. However, the specific species involved in the inhibition of respiration in intact tissues are poorly understood. METHODS AND RESULTS: O2 consumption in isolated cardiac muscle from bovine calf hearts was quantified by use of a Clark-type electrode. Exogenous and endogenous sources of NO, from S-nitroso-N-acetylpenicillamine (SNAP) and bradykinin or carbachol, reversibly inhibited respiration, whereas the O2.- releasing agent, pyrogallol (PG), inhibited respiration in a manner that was only partially reversed when examined 15 minutes after the removal of PG. The generation of ONOO- with SNAP + PG caused a potentiation of the O2(-)-elicited inhibition of respiration when examined 15 minutes after the removal of the ONOO- generating system. Tiron (a scavenger of O2.-) did not alter the actions of SNAP, but it attenuated the direct inhibitory effects of PG +/- SNAP and essentially eliminated the suppression of respiration observed 15 minutes after removal of the O2.- or ONOO- generating system. Urate (a scavenger of ONOO-) antagonized only the actions of PG + SNAP. After exposure of muscle slices to a model of hypoxia (15 minutes) and reoxygenation (10 minutes), respiratory inhibition was observed. This reoxygenation-induced inhibition was potentiated by L-arginine, the substrate for NO biosynthesis, and was markedly blocked by nitro-L-arginine (an NO synthase inhibitor), Tiron, or urate. CONCLUSIONS: The potentially physiological reversible regulation of respiration in cardiac muscle by NO is converted to an effect that does not show rapid reversibility under conditions in which ONOO- forms, and this could contribute to cardiac dysfunction in situations such as hypoxia/reoxygenation.

Animals↗
PubMed1995

Mitochondrial respiration scavenges extramitochondrial superoxide anion via a nonenzymatic mechanism.

We determined that mitochondrial respiration reduced cytosolic oxidant stress in vivo and scavenged extramitochondrial superoxide anion (O2-.) in vitro. First, Saccharomyces cerevisiae deficient in both the cytosolic antioxidant cupro-zinc superoxide dismutase (Cu,Zn-SOD) and electron transport (Rho0 state) grew poorly (P < 0.05) in 21% O2 compared with parent yeast and yeast deficient only in electron transport or Cu,Zn-SOD, whereas anaerobic growth was the same (P > 0.05) in all yeast. Second, isolated yeast and mammalian mitochondria scavenged extramitochondrial O2-. generated by xanthine/xanthine oxidase. Yeast mitochondria scavenged 42% more (P < 0.05) extramitochondrial O2-. during pyruvate/malate-induced respiration than in the resting state. Addition of either antimycin (respiratory chain inhibitor) or FCCP (respiratory chain uncoupler) prevented increased O2-. scavenging. Mitochondria isolated from yeast deficient in the mitochondrial manganous superoxide dismutase (Mn-SOD) increased (P < 0.05) O2-. scavenging 56% during respiration. This apparent SOD activity, expressed in units of SOD activity per milligram of mitochondrial protein, was the same (9 +/- 0.6 vs. 10 +/- 1.0; P = 0.43) as the O2-. scavenging of mitochondria with Mn-SOD, suggesting that respiration-dependent mitochondrial O2-. scavenging was nonenzymatic. Finally, isolated rat liver and lung mitochondria also increased (P < 0.05) O2-. scavenging during respiration. We speculate that respiring mitochondria, via the protonmotive pump, present a polarized, proton-rich surface that enhances nonenzymatic dismutation of extramitochondrial O2-. and that this is a previously unrecognized function of mitochondrial respiration with potential physiological ramifications.

Animals↗
PubMedDate not supplied

Effect of chlorpromazine on bovine sperm respiration.

Chlorpromazine (CPZ), an antipsychosis drug, causes the in vitro inhibition of sperm motility in various species, but the effect of this drug on sperm respiration is not well studied. This study was undertaken to evaluate the effect of this drug on ejaculated bovine sperm endogenous respiration. CPZ acted as a stimulator and an inhibitor of respiration at 0.1 and 0.75 mM CPZ, in the respiration medium, respectively. CPZ caused the release of sperm protein in the supernatant in a dose-dependent manner and the lactate dehydrogenase activity has also been demonstrated in the supernatant. Succinate and glycerol 3-phosphate supported the respiration among the respiratory substrates used in the present study, even after inhibition of endogenous respiration by 0.75 mM CPZ. Chlorpromazine has biphasic actions on bovine sperm respiration. The mechanism of action of the drug is discussed.

Animals↗
PubMed1988

The role of personal beliefs and social influences as determinants of respirator use among construction painters.

The purpose of this study was to identify and rank potential determinants of respirator use among painters in the construction industry. As part of a health survey of union painters, 169 male spray painters were questioned about their own beliefs concerning the consequences of wearing cartridge respirators, as well as about the perceived attitudes of others in the workplace toward respirators. Intended respirator use was more strongly associated with beliefs than was past use. The most important beliefs concerned discomfort or inconvenience. Other determinants that were associated with respirator use were respirator availability, cigarette smoking, and social influences. It was concluded that the most effective intervention strategy to increase respirator use would be multidimensional and that efforts to change only a worker's beliefs may have limited success.

Adult↗