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PubMed1972

Phenotype of a temperature-sensitive, respiration-deficient (cyt) mutant of yeast.

A temperature-sensitive respiration-deficient mutant of yeast lacks hemoproteins and accumulates coproporphyrin III when cultivated at elevated temperatures. Cells grown at 20 C respired normally and contained cytochromes a, b, and c. Cells grown at 35 C showed respiration-deficient mutant characters; they did not respire, lacked cytochromes, and accumulated coproporphyrin III. Addition of protoporphyrin IX or protohemin IX to the culture medium restored the respiratory activity of this mutant during growth at 35 C. The activities of various enzymes, including succinate-2,6-dichlorophenol indophenol (DCPIP), reduced nicotinamide adenine dinucleotide (NADH(2))-DCPIP, succinate-cytochrome c, and NADH(2)-cytochrome c oxidoreductase, and cytochrome oxidase, and the cytochrome c content of cells cultured in various conditions were determined. Changes in the number and structure of mitochondria were associated with changes in respiratory activity.

Cell-Free System↗
PubMed1982

Modulation by copper of the products of nitrite respiration in Pseudomonas perfectomarinus.

A synthetic growth medium was purified with the chelator 1,5-diphenylthiocarbazone to study the effects of copper on partial reactions and product formation of nitrite respiration in Pseudomonas perfectomarinus. This organism grew anaerobically in a copper-deficient medium with nitrate or nitrite as the terminal electron acceptor. Copper-deficient cells had high activity for reduction of nitrate, nitrite, and nitric oxide, but little activity for nitrous oxide reduction. High rates of nitrous oxide reduction were observed only in cells grown on a copper-sufficient (1 micro M) medium. Copper-deficient cells converted nitrate or nitrite initially to nitrous oxide instead of dinitrogen, the normal end product of nitrite respiration in this organism. In agreement with this was the finding that anaerobic growth of P. perfectomarinus with nitrous oxide as the terminal electron acceptor required copper. This requirement was not satisfied by substitution of molybdenum, zinc, nickel, cobalt, or manganese for copper. Reconstitution of nitrous oxide reduction in copper-deficient cells was rapid on addition of a small amount of copper, even though protein synthesis was inhibited. The results indicate an involvement of copper protein(s) in the last step of nitrite respiration in P. perfectomarinus. In addition we found that nitric oxide, a presumed intermediate of nitrite respiration, inhibited nitrous oxide reduction.

Copper↗
PubMed2002

Respiration-dependent utilization of sugars in yeasts: a determinant role for sugar transporters.

In many yeast species, including Kluyveromyces lactis, growth on certain sugars (such as galactose, raffinose, and maltose) occurs only under respiratory conditions. If respiration is blocked by inhibitors, mutation, or anaerobiosis, growth does not take place. This apparent dependence on respiration for the utilization of certain sugars has often been suspected to be associated with the mechanism of the sugar uptake step. We hypothesized that in many yeast species, the permease activities for these sugars are not sufficient to ensure the high substrate flow that is necessary for fermentative growth. By introducing additional sugar permease genes, we have obtained K. lactis strains that were capable of growing on galactose and raffinose in the absence of respiration. High dosages of both the permease and maltase genes were indeed necessary for K. lactis cells to grow on maltose in the absence of respiration. These results strongly suggest that the sugar uptake step is the major bottleneck in the fermentative assimilation of certain sugars in K. lactis and probably in many other yeasts.

Anaerobiosis↗
PubMed2005

Genomic analysis of anaerobic respiration in the archaeon Halobacterium sp. strain NRC-1: dimethyl sulfoxide and trimethylamine N-oxide as terminal electron acceptors.

We have investigated anaerobic respiration of the archaeal model organism Halobacterium sp. strain NRC-1 by using phenotypic and genetic analysis, bioinformatics, and transcriptome analysis. NRC-1 was found to grow on either dimethyl sulfoxide (DMSO) or trimethylamine N-oxide (TMAO) as the sole terminal electron acceptor, with a doubling time of 1 day. An operon, dmsREABCD, encoding a putative regulatory protein, DmsR, a molybdopterin oxidoreductase of the DMSO reductase family (DmsEABC), and a molecular chaperone (DmsD) was identified by bioinformatics and confirmed as a transcriptional unit by reverse transcriptase PCR analysis. dmsR, dmsA, and dmsD in-frame deletion mutants were individually constructed. Phenotypic analysis demonstrated that dmsR, dmsA, and dmsD are required for anaerobic respiration on DMSO and TMAO. The requirement for dmsR, whose predicted product contains a DNA-binding domain similar to that of the Bat family of activators (COG3413), indicated that it functions as an activator. A cysteine-rich domain was found in the dmsR gene, which may be involved in oxygen sensing. Microarray analysis using a whole-genome 60-mer oligonucleotide array showed that the dms operon is induced during anaerobic respiration. Comparison of dmsR+ and DeltadmsR strains by use of microarrays showed that the induction of the dmsEABCD operon is dependent on a functional dmsR gene, consistent with its action as a transcriptional activator. Our results clearly establish the genes required for anaerobic respiration using DMSO and TMAO in an archaeon for the first time.

Anaerobiosis↗
PubMed2005

Functional domains of NosR, a novel transmembrane iron-sulfur flavoprotein necessary for nitrous oxide respiration.

Bacterial nitrous oxide (N2O) respiration depends on the polytopic membrane protein NosR for the expression of N2O reductase from the nosZ gene. We constructed His-tagged NosR and purified it from detergent-solubilized membranes of Pseudomonas stutzeri ATCC 14405. NosR is an iron-sulfur flavoprotein with redox centers positioned at opposite sides of the cytoplasmic membrane. The flavin cofactor is presumably bound covalently to an invariant threonine residue of the periplasmic domain. NosR also features conserved CX3CP motifs, located C-terminally of the transmembrane helices TM4 and TM6. We genetically manipulated nosR with respect to these different domains and putative functional centers and expressed recombinant derivatives in a nosR null mutant, MK418nosR::Tn5. NosR's function was studied by its effects on N2O respiration, NosZ synthesis, and the properties of purified NosZ proteins. Although all recombinant NosR proteins allowed the synthesis of NosZ, a loss of N2O respiration was observed upon deletion of most of the periplasmic domain or of the C-terminal parts beyond TM2 or upon modification of the cysteine residues in a highly conserved motif, CGWLCP, following TM4. Nonetheless, NosZ purified from the recombinant NosR background exhibited in vitro catalytic activity. Certain NosR derivatives caused an increase in NosZ of the spectral contribution from a modified catalytic Cu site. In addition to its role in nosZ expression, NosR supports in vivo N2O respiration. We also discuss its putative functions in electron donation and redox activation.

Amino Acid Sequence↗
PubMed1969

Respiration-enhancing effect of tuberculin on splenic cells of hypersensitive guinea pigs.

Oxygen utilization in splenic cells of normal and BCG-sensitized guinea pigs was measured in a Warburg respirometer in the absence and in the presence of Old Tuberculin (OT). In the absence of OT, normal and sensitive cells respired at the same rate; in the presence of OT, normal cell respiration remained unchanged, whereas the respiration of sensitive cells increased significantly. This enhancement of oxygen utilization in sensitive cells was induced with OT but not with Purified Protein Derivative and it was most pronounced during the first hour of the interaction between the cells and the antigen. When normal splenic cells were treated with an extract of hypersensitive cells, their respiration was enhanced in the presence of OT. Oxygen utilization in cells of desensitized animals did not differ from that in cells of normal animals. The absence of reactivity to OT in splenic cells and in the skin of desensitized guinea pigs suggested that both phenomena are expressions of antigen-induced biological changes in cells of animals with delayed hypersensitivity.

Animals↗
PubMed1969

Influence of oxygen on development of nitrate respiration in Bacillus stearothermophilus.

A denitrifying mutant of Bacillus stearothermophilus NCA 2184, strain 2184-D, was used to explore the development of nitrate respiration in relation to oxygen respiration. Aerobically grown wild-type cultures could acquire the ability to use nitrate as a result of selection of nitrate-respiring mutants by the presence of nitrate and a reduced oxygen tension. Fluctuation analysis has revealed that the frequency of occurrence of the nitrate-respiring mutant is about 7.5 x 10(-8) per bacterium per generation. Nitrate reductase and nitrite reductase appeared to be induced sequentially in strain 2184-D by the addition of nitrate. The formation of both of these enzymes was repressed by oxygen so that cells grown aerobically with nitrate possessed a low basal level of nitrate reducatase and exhibited no denitrification. The rate of synthesis of nitrate reductase increased quickly after addition of nitrate and removal of oxygen. It then declined to a lower steady-state level. Cells grown anaerobically with nitrate retained approximately 30 to 40% of the respiratory activity of aerobically grown cells. Aeration of anaerobically grown cells in the presence of amino acids increased the respiratory activity to normal aerobic levels. This aeration promoted rapid degradation of the existing nitrate reductase with or without the added amino acids.

Bacillus↗
PubMed2005

Age related reference ranges for respiration rate and heart rate from 4 to 16 years.

BACKGROUND: Clinical vital signs in children (temperature, heart rate, respiration rate, and blood pressure) are an integral part of clinical assessment of degree of illness or normality. Despite this, only blood pressure and temperature have a reliable evidence base. The accepted ranges of heart and respiration rate vary widely. METHODS: This study examined 1109 children aged 4-16 years in their own schools. Age, sex, height, weight, and resting respiration rate and heart rate were recorded. The data were used to produce age related reference ranges for everyday clinical use. RESULTS: Reference intervals are presented for the range of heart rate and respiration rate of healthy resting children aged 4-16 years. The recorded values are at variance with standard quoted ranges in currently available texts.

Adolescent↗
PubMed1991

Ventilatory function after exposure to various respirable hazards in a population of former coal miners.

The ventilatory function of 406 male former coal miners who had presented at the Cook County Hospital occupational medicine clinic between January 1976 and April 1987 was studied to determine whether subsequent exposure to respiratory hazards after leaving the coal mines adversely affected lung function. The miners were divided into five exposure groups based on their exposure to respirable hazards. These were coal dust only, coal dust plus asbestos dust, coal dust plus silica dust, coal dust plus another respirable hazard and coal dust plus two other respirable dust exposures. Duration of employment in coal mines, race, smoking history, and mean age were not significantly different between the various exposure groups. No significant difference was found in the per cent of predicted forced expiratory volume in one second (FEV1), per cent of predicted forced vital capacity (FVC), and FEV1/FVC when the coal dust only group was compared with each of the other four exposure groups using ANOVA. Among former coal miners who present for a respiratory disability determination, therefore, exposure to respirable hazards subsequent to employment in coal mines is not associated with a statistically significant deterioration in ventilatory function.

Coal Mining↗
PubMed1996

Biological indicators of exposure to total and respirable aluminium dust fractions in a primary aluminium smelter.

OBJECTIVES: The study attempts to define biological indicators of aluminium uptake and excretion in workers exposed to airborne aluminium compounds in a primary aluminium smelter. Also, this study defines the total and respirable aluminium dust fractions in two different potrooms, and correlates their concentrations with biological indicators in this group of workers. METHODS: Air was sampled at defined work sites. Non-destructive and conventional techniques were used to find total and respirable aluminium content of the dust. Blood and urine was collected from 84 volunteers employed at various work stations throughout the smelter and from two different cohorts of controls matched for sex, age, and socioeconomic status. Aluminium in serum samples and urine specimens was measured by flameless atomic absorption with a PE 4100 ZL spectrometer. RESULTS: The correlation of aluminium concentrations in serum and urine samples with the degree of exposure was assessed for three arbitrary exposure categories; low (0.036 mg Al/m3), medium (0.35 mg Al/m3) and high (1.47 mg Al/m3) as found in different areas of the smelter. At medium and high exposure, the ratio of respirable to total aluminium in the dust samples varied significantly. At high exposure, serum aluminium, although significantly raised, was still within the normal range of an unexposed population. The workers with low exposure excreted aluminium in urine at levels significantly higher than the controls, but still within the normal range of the population. However, potroom workers with medium and high exposure had significantly higher urinary aluminium than the normal range. CONCLUSIONS: It is concluded that only urinary aluminium constitutes a practical index of occupational exposure at or above 0.35 mg Al/m3, and that the respirable fraction of the dust may play a major role in the biological response to exposure to aluminium in a smelter environment.

Adult↗
PubMed2000

Two patients with occupational asthma who returned to work with dust respirators.

OBJECTIVES: To assess the efficacy of dust respirators in preventing asthma attacks in patients with occupational asthma (asthma induced by buckwheat flour or wheat flour). METHODS: The effect of the work environment was examined in two patients with occupational asthma with and without the use of a commercially available mask or a dust respirator. Pulmonary function tests were performed immediately before and after work and at 1 hourly intervals for 14 hours after returning to the hospital. RESULTS: In patient 1, environmental exposure resulted in no symptoms during and immediately after work, but coughing, wheezing, and dyspnoea developed after 6 hours. Peak expiratory flow rate (PEFR) decreased by 44% 7 hours after leaving the work environment, showing only a positive late asthmatic reaction (LAR). In patient 2, environmental exposure resulted in coughing and wheezing 10 minutes after initiation during bread making, and PEFR decreased by 39%. After 7 hours, PEFR decreased by 34%. The environmental provocation tests in both patients were repeated after wearing a commercial respiratory. This resulted in a complete suppression of LAR in patient 1 and of immediate asthmatic reaction (IAR) and LAR in patient 2. CONCLUSIONS: Two patients with asthma induced by buckwheat flour or wheat flour in whom asthmatic attacks could be prevented with a dust respirator are reported. Dust respirators are effective in preventing asthma attacks induced by buckwheat flour and wheat flour.

Adult↗
PubMed1987

Inhibition of yeast respiration and fermentation by benomyl, carbendazim, isocyanates, and other fungicidal chemicals.

The inhibition of yeast (Saccharomyces cerevesiae) metabolism by fungicidal chemicals was investigated. Glucose- or ethanol-dependent yeast respiration was measured with an oxygen electrode, and manometric determination of carbon dioxide release was used to measure fermentation. Both respiration and fermentation were inhibited more by benomyl than by identical molar concentrations of its breakdown product, carbendazim. Butyl isocyanate, another benomyl breakdown product, inhibited respiration more but inhibited fermentation less than the parent compound. Of the isocyanates tested, hexyl isocyanate was the most inhibitory towards both activities. Captan was more active and iprodione less active than benomyl. Because benomyl rapidly broke down to carbendazim when it was prepared in 80% ethanol, only 59% of the dissolved benomyl was intact when it was added to yeast to determine its effect on respiration or fermentation.

Benomyl↗
PubMed2006

Mitochondrial Ca2+-induced K+ influx increases respiration and enhances ROS production while maintaining membrane potential.

We recently demonstrated a role for altered mitochondrial bioenergetics and reactive oxygen species (ROS) production in mitochondrial Ca(2+)-sensitive K(+) (mtK(Ca)) channel opening-induced preconditioning in isolated hearts. However, the underlying mitochondrial mechanism by which mtK(Ca) channel opening causes ROS production to trigger preconditioning is unknown. We hypothesized that submaximal mitochondrial K(+) influx causes ROS production as a result of enhanced electron flow at a fully charged membrane potential (DeltaPsi(m)). To test this hypothesis, we measured effects of NS-1619, a putative mtK(Ca) channel opener, and valinomycin, a K(+) ionophore, on mitochondrial respiration, DeltaPsi(m), and ROS generation in guinea pig heart mitochondria. NS-1619 (30 microM) increased state 2 and 4 respiration by 5.2 +/- 0.9 and 7.3 +/- 0.9 nmol O(2).min(-1).mg protein(-1), respectively, with the NADH-linked substrate pyruvate and by 7.5 +/- 1.4 and 11.6 +/- 2.9 nmol O(2).min(-1).mg protein(-1), respectively, with the FADH(2)-linked substrate succinate (+ rotenone); these effects were abolished by the mtK(Ca) channel blocker paxilline. DeltaPsi(m) was not decreased by 10-30 microM NS-1619 with either substrate, but H(2)O(2) release was increased by 44.8% (65.9 +/- 2.7% by 30 muM NS-1619 vs. 21.1 +/- 3.8% for time controls) with succinate + rotenone. In contrast, NS-1619 did not increase H(2)O(2) release with pyruvate. Similar results were found for lower concentrations of valinomycin. The increase in ROS production in succinate + rotenone-supported mitochondria resulted from a fully maintained DeltaPsi(m), despite increased respiration, a condition that is capable of allowing increased electron leak. We propose that mild matrix K(+) influx during states 2 and 4 increases mitochondrial respiration while maintaining DeltaPsi(m); this allows singlet electron uptake by O(2) and ROS generation.

Animals↗
PubMed1982

Effects of butyrate on ouabain-sensitive respiration of hamster brown adipocytes.

Brown adipose tissue is an important site of cold-induced nonshivering thermogenesis in many mammals. The plasma membrane-bound Na+-K+-ATPase has been shown to be significantly involved in this thermogenesis although its exact role is unknown at present. Evidence that coupling of oxidative phosphorylation to electron transport may become loosened during thermogenesis has prompted an investigation of potential roles of the Na+-K+ pump that would be compatible with altered respiratory coupling. One such role is that of modulating norepinephrine (NE)-induced lipolysis and hence provision of free fatty acids to the mitochondria. Under such conditions, inhibition of the pump would reduce NE-induced respiration by limiting substrate availability. If, in fact, the primary role of the pump in NE-induced thermogenesis is to facilitate substrate availability, provision of exogenous substrate should bypass this involvement and ameliorate the ouabain inhibition of respiration. In the present study, this possibility was examined by determining the effect of an exogenous substrate, butyrate, on the contribution of the Na+-K+ pump to NE-stimulated respiration of isolated hamster brown adipocytes. Although exogenous butyrate was able to serve as a substrate for brown adipocyte respiration, its presence had no significant effect on the ouabain sensitivity of NE-induced rates of oxygen consumption. That is, ouabain (1 mM) inhibited the NE-evoked thermogenesis of the adipocytes by 77.7 +/- 6.5% in the absence of butyrate (2 mM) and by 73.4 +/- 9.9% in its presence. It appears, therefore, that the contribution of the Na+-K+ membrane pump to brown fat thermogenesis does not simply reflect modulation of NE-evoked lipolysis.

Adipose Tissue, Brown↗
PubMed1989

Enhanced glucose transport in response to inhibition of respiration in Clone 9 cells.

An acceleration of ATP synthesis by anaerobic glycolysis provides important compensation for interference with respiration in a variety of cells. Effective compensation for an inhibition of respiration, however, can occur in cells in which glucose entry is rate limiting only if sufficient glucose becomes available through an enhancement of transport. We present here a detailed study of the effects of inhibition of respiration in Clone 9 cells, a continuous cell line characterized by low internal glucose concentrations (less than 10% that of the external medium) and minimal stores of glycogen. Exposure of these cells to 5 mM cyanide results in a 90% fall in cell ATP and a twofold rise in cell Na+ within 20 min. By the end of 1 h, however, there is a 4.5- to 7-fold increase in cytochalasin B-inhibitable glucose transport that is accompanied by a parallel increase in the rate of lactate production, a partial recovery of cell ATP, and no further rise in cell Na+. The acute fall in ATP resulting from a submaximally effective concentration of cyanide (0.5 mM) is moreover followed by a time-dependent recovery of cell ATP to near-normal levels and subsequent resistance to challenge with even 5 mM cyanide. The stimulation of facilitative glucose transport resulting from exposure to cyanide is attributable to an increase in maximal velocity rather than to a change in Km and persists for more than 2 h after removal of the inhibitor. These results demonstrate that, in these cells characterized by low internal glucose concentrations, regulation of glucose entry is of central importance in ATP homeostasis and that a major component of the adaptive response to an inhibition of respiration is a time-dependent increase in glucose transport.

3-O-Methylglucose↗
PubMed1980

Stereospecific stimulation of brown adipocyte respiration by catecholamines via beta 1-adrenoreceptors.

Regulation of respiration by catecholamines was studied in adipocytes isolated from interscapular brown adipose tissue of warm-acclimated rats by rapid digestion of collagenase. (-)-Norepinephrine stimulated adipocyte respiration 10-12 times above basal values in less than 3 min. (Vmax = 410 +/- 29.5 nmol O2 . min-1 . 10(-6) cells-1). Stimulated respiration remained stable for at least 20 min, provided that cells were incubated in balanced salt media containing bicarbonate. The maximal capacity of total brown adipose tissue for norepinephrine-stimulated respitarion was estimated at 1.5 ml O2/min per rat. beta-Adrenergic agonists increased calorigenesis stereospecifically with an order of potency expected for respiratory stimulation via adrenoceptors of the beta 1-subtype: (-)-isoproterenol (1/2 Vmax = 2 nM) greater than (-)-norepinephrine (1/2 Vmax = 20 nM) approximately equal to (-)-epinephrine (1/2 Vmax = 40 nM) greater than corresponding (+)-stereoisomers. The alpha-adrenergic agonist phenylephrine (1/2 Vmax = 5 microM) stimulated adipocyte respiration as rapidly and as effectively as beta-agonists. Although alpha-adrenoreceptors are present in brown adipose tissue, studies with alpha- and beta-adrenergic antagonists revealed that norepinephrine elicits thermogenesis at physiological concentrations (less than or equal to 1 microM) predominantely via beta 1-adrenergic pathways.

Acclimatization↗
PubMed1983

Effect of changes in thyroid status on tissue respiration in fetal and newborn sheep.

The effect of both exogenous and endogenous changes in thyroid status on in vitro tissue respiration and thermogenic enzymes (sodium-potassium-adenosine triphosphatase and alpha-glycerophosphate dehydrogenase) was studied in fetal and newborn sheep. Oxygen consumption of liver and brain increased from 25 +/- 4.1 and 58.5 +/- 2.8 microliters O2 X 100 mg-1 X h-1, respectively, in tissues from unthyroidectomized fetal animals at 136-140 days gestation to 60 +/- 4.2 and 72 +/- 1.5 microliters O2 X 100 mg-1 X h-1 in tissues from unthyroidectomized newborn lambs between birth and 7 days of age. The physiological changes in thyroid function that normally occur at birth resulted in a mean (+/- SE) plasma triiodothyronine (T3) concentration of 563 +/- 39 ng/dl in the newborn lambs compared with 39 +/- 8 ng/dl in the fetal animals. Kidney respiration and thermogenic enzyme activities in the several tissues studied did not change. Liver, kidney, and brain respiration and thermogenic enzymes from T3-treated thyroidectomized fetal and newborn lambs were not increased (compared with untreated thyroidectomized animals) despite a marked increase in plasma T3 concentrations. Conclusions are that 1) liver, kidney cortex, and frontal brain cortex in the fetal and newborn lamb are relatively insensitive to the calorigenic effect of thyroid hormones, and 2) a perinatal increase in hepatic and cerebral respiration occurs in newborn animals (compared with fetal animals) but is probably not due solely to perinatal increases in thyroid hormones.

Animals↗
PubMed1987

Effects of thyroid state on respiration of perfused rat and guinea pig hearts.

The effects of thyroid state on the respiration of the isolated heart were investigated using retrograde perfused rat and guinea pig hearts. In both species, hypothyroidism caused a marked depression in circulating thyroid hormone concentrations and in the respiration of the isolated, retrograde perfused heart. The effects on myocardial respiration could be attributed to changes in the contraction frequency and in the oxygen consumption per beat, with little contribution from basal respiration. Treatment of animals with thyroxine elevated plasma thyroid hormones to a similar extent in rats and guinea pigs. In the latter, thyroxine treatment was associated with substantial increases in the contraction frequency and the oxygen consumption per beat of the isolated heart. In contrast, only small changes were apparent in the retrograde perfused rat heart, observations that were confirmed in rat hearts perfused at near physiological work loads. It was concluded that rat hearts isolated from normal animals function at near maximal thyroid state, in contrast to the guinea pig heart, which requires higher circulating concentrations of thyroid hormones to attain maximal responses.

Animals↗