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PubMed1998

Surveillance of respirable crystalline silica dust using OSHA compliance data (1979-1995).

BACKGROUND: The objective of this work was to estimate the percentage of workers by industry that are exposed to defined concentrations of respirable crystalline silica dust. METHODS: An algorithm was used to estimate the percentage of total workers exposed to crystalline silica in 1993 at concentrations of at least 1, 2, 5, and 10 times the National Institute for Occupational Safety and Health (NIOSH) Recommended Exposure Limit (REL) of 0.05 mg/m3. Respirable crystalline silica air sampling data from regulatory compliance inspections performed by the Occupational Safety and Health Administration (OSHA), for the years 1979-1995, and recorded in the Integrated Management Information System (IMIS) were used to estimate exposures. Therefore, this work does not include industries such as mining and agriculture that are not covered by OSHA. The estimates are stratified by Standard Industrial Classification (SIC) codes. RESULTS: This work found that some of the highest respirable crystalline silica dust concentrations occurred in construction (masonry, heavy construction, and painting), iron and steel foundries (casting), and in metal services (sandblasting, grinding, or buffing of metal parts). It was found that 1.8% (13,800 workers) of the workers in SIC 174--Masonry, Stonework, Tile Setting, and Plastering--were exposed to at least 10 times the NIOSH REL. For SIC 162--Heavy Construction, Except Highway and Street Construction--this number is 1.3% (6,300 workers). SIC 172--Painting and Paper Hanging--which includes construction workers involved in sandblasting was found to have 1.9% (3,000 workers) exposed to at least 10 times the NIOSH REL. The industry that was found to have the highest percentage of workers (6%) exposed to at least the NIOSH REL was the cut stone and stone products industry. CONCLUSION: Not enough is being done to control exposure to respirable crystalline silica. Engineering controls should be instituted in the industries indicated by this work.

Algorithms↗
PubMed1999

Medical clearance for respirator use: sensitivity and specificity of a questionnaire.

BACKGROUND: OSHA regulations require that workers receive medical clearance prior to respirator use, and recently, a detailed questionnaire has been provided to assist with this purpose. However, there are limited published data on the sensitivity and specificity of self-administered questionnaires for identifying individuals who may safely wear a respirator. METHODS: We tested 474 consecutive workers at a Department of Energy complex. After completing the self-administered questionnaire, all workers received a standardized physician evaluation including interview, physical examination, and spirometry. The outcomes of the questionnaire assessment were compared to the outcomes of physician evaluation. RESULTS: Data for analysis were available from 413 of workers (87%). All workers received medical clearance; only 10 workers (2.4%) received work restrictions. The questionnaire demonstrated 100% sensitivity in identifying workers who required work restrictions, but had specificity of only 19%. Compared to physician evaluation, the questionnaire had modest sensitivity to the detection of chronic medical conditions. CONCLUSIONS: These data suggest that the rates of medical clearance for respirator use are very high, and that a self-administered questionnaire may be appropriate for medical clearance in certain settings. It is recommended that the policy of routine physician evaluation and spirometry for respirator clearance be re-examined.

Adult↗
PubMed2004

Partial protection by respirators on airways responses following exposure in a swine house.

BACKGROUND: Exposure to swine dust leads to an intense airway inflammation and increased bronchial responsiveness. The purpose of the present study was to evaluate the effect of a respiratory protection device during exposure in a swine house. METHODS: Twenty-two subjects, 11 with a respirator, were exposed. Symptoms, body temperature, nasal lavage, and a bronchial metacholine challenge were performed before and 7 hr after exposure. For exposure assessment a nasal sampler was evaluated. RESULTS: The subjects with a respirator showed an attenuated inflammatory nasal response. An increase in bronchial responsiveness was observed in both groups, significantly greater in the unprotected group. The use of respirators reduced endotoxin exposure by more than 90% (assessed by nasal samplers). CONCLUSION: The use of a respirator attenuated the inflammatory response compared with an unprotected group. The minor effect on bronchial responsiveness suggests that gases and/or ultrafine particles may also be important factors.

Adult↗
PubMed1996

The control of mitochondrial respiration in yeast: a possible role of the outer mitochondrial membrane.

Mitochondrial respiration in yeast (S. cerevisiae) is regulated by the level of glucose in the medium. Glucose is known to inhibit respiration by repressing key enzymes in the respiratory chain. We present evidence that the early events in this inhibition include the closure of VDAC channels, the primary pathway for metabolite flow across the outer membrane. Aluminum hydroxide is known to inhibit the closure of VDAC. Addition of aluminum acetylacetonate to yeast cells, which should elevate the aluminum hydroxide concentrations in the cytoplasm, caused the inhibition of cell respiration by glucose to be delayed for up to 100 min. No significant effect of aluminum was observed in cells grown on glycerol. Yeast cells lacking the VDAC gene were also unresponsive to the addition of aluminum salt in the presence of glucose. Therefore, the closure of VDAC channels may be an early step in the inhibition of the respiration of yeast by glucose.

Aluminum Hydroxide↗
PubMed2003

Rapid detection of respiring Escherichia coli O157:H7 in apple juice, milk, and ground beef by flow cytometry.

BACKGROUND: Rapid and simple methods to detect viable pathogenic microbes in foods and drinks are required. Flow cytometry was used for the rapid detection of respiring Escherichia coli O157:H7 cells in apple juice, milk, and ground beef. METHODS: CTC (5-cyano-2,3-ditolyl tetrazolium chloride) was used to estimate the respiratory activity of bacteria. Fluorescein isothiocyanate (FITC)-labeled anti-E. coli O157:H7 direct antibody (FA) was used for the specific detection of target cells. Food samples were inoculated with starved E. coli O157:H7 and E. coli K-12 cells, and analyzed by both fluorescent microscopy and flow cytometry after double staining with FA and CTC. RESULTS: Respiring E. coli O157:H7 cells in food samples showed strong fluorescence of both FA (green) and CTC (red); thus, they could be clearly and specifically distinguished from respiring E. coli K-12 or inactive cells. A good correlation was achieved in flow cytometric analysis between the numbers of inoculated viable E. coli O157:H7 and those detected in milk and apple juice. The detection threshold for this flow cytometry for E. coli O157:H7 in milk, apple juice, and ground beef was 10(3) cells/ml (milk and apple juice) or 10(3) cells/g (ground beef) of sample when the total bacterial number in the sample was 10(6) cells/ml. CONCLUSIONS: Respiring E. coli O157:H7 in food samples can be detected specifically within a few hours. Flow cytometry with FA-CTC double staining can be used to examine food contamination with various pathogenic microbes demonstrating physiologic activity through the use of a suitable fluorescent antibody.

Animals↗
PubMed1993

Inhibition of glycolysis and mitochondrial respiration of Ehrlich ascites carcinoma cells by methylglyoxal.

The effect of methylglyoxal (MG) on the aerobic glycolysis of Ehrlich ascites carcinoma (EAC) cells has been tested. Methylglyoxal inhibited glucose utilization and glucose 6-phosphate (G6P) and L-lactate formation in whole EAC cells. Methylglyoxal strongly inactivated glyceraldehyde 3-phosphate dehydrogenase (GA3PD) of the malignant cells, whereas MG has little inactivating effect on this enzyme from several normal sources. Methylglyoxal also inactivated only the particulate hexominase of the EAC cells, but this inactivation was less pronounced than the effect on GA3PD. Methylglyoxal has little inactivating effect on glucose 6-phosphate dehydrogenase (G6PD), and no effect on L-lactate dehydrogenase (LDH) of the malignant cells. Glucose-dependent L-lactic acid formation of EAC-cell-free homogenate was strongly inhibited by MG, but when GA3PD of normal cells was added to this homogenate, significant lactate formation was observed even in the presence of MG. Methylglyoxal also inhibited the respiration of EAC-cell mitochondria. Respiration of mitochondria isolated from liver and kidney of normal mice, however, remained unaffected. As a consequence of the inhibition of glycolysis and mitochondrial respiration, the ATP level of the EAC cells was drastically reduced. Studies reported herein strongly suggest that the tumoricidal effect of MG is mediated at least in part through the inhibition of mitochondrial respiration and inactivation of GA3PD, and this enzyme may play an important role in the high glycolytic capacity of the malignant cells.

Adenosine Triphosphate↗
PubMed1980

Respiration in rat cerebral astrocytes from primary culture.

Respiration was measured polarographically in astrocytes from dissociated neonatal rat cerebra grown in primary culture. Cells grown in a modified Eagle's minimal essential medium containing 20% fetal calf serum for 14 to 21 days respired at a rate of 34.23 n at. equiv 0 mg protein-1 min-1. Cells which were grown in this medium for 14 days and the grown in serum-free medium containing 0.25 mM dibutyrylcyclic-AMP (dbcAMP) for three to six days, had respiratory rates 20% higher than in cells from untreated control cultures (P = 0.005). Maximal inhibition by oligomycin required 200 pmole per mg protein for dbcAMP-treated cells and 80 pmole per mg protein for untreated cells. Oligomycin inhibited the respiration of both dbcAMP-treated and untreated cells by about 70%. At a concentration of 175 nmole per mg protein, 2,4-dinitrophenol (DNP) produced a maximal stimulation of respiration, which was 191% of the resting rate in dbcAMP-treated cells and 218% of the resting rate in untreated cells. The maximal DNP-stimulated respiratory rates were the same in dbcAMP-treated and untreated cells.

Animals↗
PubMed2003

Quantification of priming and CO2 respiration sources following slurry-C incorporation into two grassland soils with different C content.

The fate of incorporated slurry-C was examined in a laboratory experiment using two UK grassland soils, i.e. a Pelostagnogley (5.1 %C) and a Brown Earth (2.3 %C). C3 and C4 slurries were incorporated into these two wet-sieved (C3) soils (from 4-10 cm depth). Gas samples were collected 0.2, 1, 2, 3, 4, 6, 9, 20, 30 and 40 days after slurry application and analyzed for CO2 concentration and delta13C content. Slurry incorporation into the soil strongly increased soil CO2 respiration compared with the unamended soil. Total (40 day) cumulative CO2 flux was higher for the Pelostagnogley than the Brown Earth. The 13C natural abundance tracer technique enabled quantification of the sources of respired CO2 and priming effects (days 0-9). Proportionally more slurry-derived C was respired from the Pelostagnogley (46%) than the Brown Earth (36%). The incorporated slurry-C was lost twice as fast as the native soil C in both soils. Slurry incorporation induced a priming effect, i.e. additional release of soil-derived C, most pronounced in the Pelostagnogley (highest C content). The majority of respired soil-derived C (>70%) was primed C. The study indicated that potential reductions in ammonia volatilisation following slurry injection to grasslands might be negated by enhanced loss of primed soil C (i.e. pollution swapping).

Agriculture↗
PubMed2006

Glycolytic sequence and respiration of Debaryomyces hansenii as compared to Saccharomyces cerevisiae.

The fermentation and respiration activities of Debaryomyces hansenii were compared with those of Saccharomyces cerevisiae grown to stationary phase with high respiratory activity. It was found that: (a) glucose consumption, fermentation and respiration were lower than for S. cerevisiae; (b) fasting produced a much smaller decrease of respiration; (c) glucose consumed and not transformed to ethanol was higher; (d) in S. cerevisiae, full oxygenation prevented ethanol production but this effect was reversed by CCCP, whereas D. hansenii still showed some ethanol production under aerobiosis, which was moderately increased by CCCP. ATP levels were similar in the two yeasts. Levels of glycolytic intermediaries after glucose addition, and enzyme activities, indicated that the main difference and limiting step to explain the lower fermentation of D. hansenii is phosphofructokinase activity. Respiration and fermentation, which are lower in D. hansenii, compete for the re-oxidation of reduced nicotinamide adenine nucleotides; this competition, in turn, seems to play a role in defining the fermentation rates of the two yeasts. The effect of CCCP on glucose consumption and ethanol production also indicates a role of ADP in both the Pasteur and Crabtree effects in S. cerevisiae but not in D. hansenii. D. hansenii shows an alternative oxidase, which in our experiments did not appear to be coupled to the production of ATP.

Alcohol Dehydrogenase↗
PubMed1992

Effect of benzoic acid on metabolic fluxes in yeasts: a continuous-culture study on the regulation of respiration and alcoholic fermentation.

Addition of benzoate to the medium reservoir of glucose-limited chemostat cultures of Saccharomyces cerevisiae CBS 8066 growing at a dilution rate (D) of 0.10 h-1 resulted in a decrease in the biomass yield, and an increase in the specific oxygen uptake rate (qO2) from 2.5 to as high as 19.5 mmol g-1 h-1. Above a critical concentration, the presence of benzoate led to alcoholic fermentation and a reduction in qO2 to 13 mmol g-1 h-1. The stimulatory effect of benzoate on respiration was dependent on the dilution rate: at high dilution rates respiration was not enhanced by benzoate. Cells could only gradually adapt to growth in the presence of benzoate: a pulse of benzoate given directly to the culture resulted in wash-out. As the presence of benzoate in cultures growing at low dilution rates resulted in large changes in the catabolic glucose flux, it was of interest to study the effect of benzoate on the residual glucose concentration in the fermenter as well as on the level of some selected enzymes. At D = 0.10 h-1, the residual glucose concentration increased proportionally with increasing benzoate concentration. This suggests that modulation of the glucose flux mainly occurs via a change in the extracellular glucose concentration rather than by synthesis of an additional amount of carriers. Also various intracellular enzyme levels were not positively correlated with the rate of respiration. A notable exception was citrate synthase: its level increased with increasing respiration rate. Growth of S. cerevisiae in ethanol-limited cultures in the presence of benzoate also led to very high qO2 levels of 19-21 mmol g-1 h-1. During growth on glucose as well as on ethanol, the presence of benzoate coincided with an increase in the mitochondrial volume up to one quarter of the total cellular volume. Also with the Crabtree-negative yeasts Candida utilis, Kluyveromyces marxianus and Hansenula polymorpha, growth in the presence of benzoate resulted in an increase in qO2 and, at high concentrations of benzoate, in aerobic fermentation. In contrast to S. cerevisiae, the highest qO2 of these yeasts when growing at D = 0.10 h-1 in the presence of benzoate was equal to, or lower than the qO2 attainable at mu(max) without benzoate. Enzyme activities that were repressed by glucose in S. cerevisiae also declined in K. marxianus when the glucose flux was increased by the presence of benzoate.(ABSTRACT TRUNCATED AT 400 WORDS)

Benzoates↗
PubMed1995

Involvement of cyanide-resistant respiration in cell-type proportioning during Dictyostelium development.

Involvement of cyanide (CN)-resistant respiration in cell-type proportioning was analyzed using the developmental system of Dictyostelium discoideum. When migrating slugs were vitally stained with rhodamine 123, which is known to stain actively respirating mitochondria coupled with an elevated electronic potential of the inner membrane, the posterior prespore region was stained more strongly than the anterior prestalk region. Application of benzohydroxamic acid (BHAM) and propyl gallate, specific inhibitors of CN-resistant respiration, to starved Dictyostelium cells induced formation of unique cell masses, in which almost all of the cells differentiated into stalk-like cells with a large vacuole and thick cell wall. BHAM was also found to enhance the expressions of prestalk-specific genes such as ecmA and ecmB in the unique cell mass in a position-dependent manner. In contrast, the expression of a prespore-specific gene, Dp87, was almost completely inhibited by BHAM. Taken together these results strongly suggest the involvement of CN-resistant respiration in the proportion regulation of cell types differentiating during the Dictyostelium development.

Animals↗
PubMed1994

Activated THP-1 cells depress mitochondrial respiration in Hep G2 cells infected with influenza B virus.

Influenza B virus has been aetiologically linked to Reye Syndrome (RS), but the mechanism(s) by which this pathogen could disrupt liver metabolism and produce the hepatic mitochondrial injury characteristic of the syndrome are unknown. In this study, two mechanisms by which infection of hepatocytes with influenza B virus could disrupt cellular metabolism were investigated. (1) virus-induced increase in pro-oxidant iron with subsequent iron-induced lipid peroxidation (LP) and (2) increased membrane permeability. Hep G2 cells, a well-differentiated continuous human liver cell line derived from a hepatoblastoma, were infected with allantoic-fluid derived influenza B Lee/40 virus (AFDV) at a multiplicity of infection of 10 for 24 h; productive infection was confirmed by both haemagglutination of chick erythrocytes and by plaque assay. Infection of Hep G2 cells preloaded with 59Fe-transferrin resulted in increased release of 59Fe (153 +/- 17% of controls, P < 0.03). However, the iron released did not result in increased LP (assessed by thiobarituric acid reactive substances; TBARS). To confirm that this lack of of increase in TBARS was not due to insensitivity of the cell line to pro-oxidant iron, cells were exposed to 15 microM iron ascorbate for 60 min. Production of TBARS was increased (122 +/- 4% of controls, P < 0.0003). Release of 51Cr from infected cells was also increased (128 +/- 12% of controls, P < 0.05); thus the infected cells exhibited a generalized increase in membrane permeability. However, infection did not depress mitochondrial respiration (as assessed by the formation of MTT-f3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide-formazan. To determine if the combination of viral infection and soluble products of activated macrophages would affect mitochondrial respiration, infected hepatocytes were exposed to the supernatant fluid from THP-1 cells which had previously been incubated with lipopolysaccharide at 100 ng ml-1 for 18 h. This supernate did depress the formation of MTT-f (81 +/- 5% of controls, P < 0.03). We conclude that influenza B virus does productively infect Hep G2 cells, and does increase hepatocyte membrane permeability. This effect does not impair mitochondrial respiration directly. However, infection does act in concert with soluble products of activated macrophages to depress hepatic mitochondrial respiration. Whether this interaction can be explained by virus-induced permeability changes and/or other effects of infection deserves further investigation.

Animals↗
PubMed2001

Mechanisms of the deleterious effects of tamoxifen on mitochondrial respiration rate and phosphorylation efficiency.

Tamoxifen (TAM), the widely prescribed drug in the prevention and therapy of breast cancer, is a well-known modulator of estrogen receptor (ER) that also inhibits the proliferation of different cell types that lack the ER. However, the ER-independent action mechanisms of TAM and its side effects have not been yet clarified. Mitochondria are essential in supporting the energy-dependent regulation of cell functions. Changes in mitochondria result in bioenergetic deficits leading to the loss of vital functions to cell survival. Therefore, this study describes the effects of TAM on mitochondrial bioenergetics, contributing to a better understanding of the biochemical mechanisms underlying the multiple antiproliferative and toxic effects of this drug. TAM at concentrations above 20 nmol/mg protein, preincubated with isolated rat liver mitochondria at 25 degrees C for 3 min, significantly depresses, in a dose-dependent manner, the phosphorylation efficiency of mitochondria as inferred from the decrease in the respiratory control and ADP/O ratios, the perturbations in mitochondrial transmembrane potential (DeltaPsi), the fluctuations associated with mitochondrial energization, and the phosphorylative cycle induced by ADP. Furthermore, TAM at up to 40 nmol/mg protein stimulates the rate of state 4 respiration and at higher concentrations it strongly inhibits state 3 and uncouples the mitochondrial respiration. The stimulation of state 4 respiration parallels the decrease of DeltaPsi as a consequence of proton permeability. The TAM-stimulatory action of ATPase is also observed in intact mitochondria, suggesting that TAM promotes extensive permeability to protons due to destructive effects in the structural integrity of the mitochondrial inner membrane. These multiple effects of TAM on mitochondrial bioenergetic functions, causing changes in the respiration, phosphorylation efficiency, and membrane structure, may explain the cell death induced by this drug in different cell types, its anticancer activity in ER-negative cells, and its side effects.

Adenosine Diphosphate↗
PubMed2003

Metabolic interactions between methanogenic consortia and anaerobic respiring bacteria.

Most types of anaerobic respiration are able to outcompete methanogenic consortia for common substrates if the respective electron acceptors are present in sufficient amounts. Furthermore, several products or intermediate compounds formed by anaerobic respiring bacteria are toxic to methanogenic consortia. Despite the potentially adverse effects, only few inorganic electron acceptors potentially utilizable for anaerobic respiration have been investigated with respect to negative interactions in anaerobic digesters. In this chapter we review competitive and inhibitory interactions between anaerobic respiring populations and methanogenic consortia in bioreactors. Due to the few studies in anaerobic digesters, many of our discussions are based upon studies of defined cultures or natural ecosystems.

Acetates↗
PubMed1992

Amyotrophic lateral sclerosis with ophthalmoplegia and multisystem degeneration in patients on long-term use of respirators.

We describe two patients with sporadic amyotrophic lateral sclerosis (ALS), who had developed progressive external ophthalmoplegia of a predominantly supranuclear type while they survived on respirators, and displayed histopathological abnormalities both typical and atypical of ALS. Patient 1 was a 43-year-old man with ALS of 5-year duration, who had initially exhibited fulminant ALS, and remained on a respirator for 4 years. Patient 2 was a 51-year-old man with ALS of 13-year duration, who remained on a respirator for 8 years. Both patients died in a "totally locked-in state". Autopsy of both patients revealed not only histopathological abnormalities consistent with ALS, but also multisystem degeneration which involved the pontine tegmentum, substantia nigra, Clarke's dorsal nuclei and spinocerebellar tracts. In addition, Patient 2 displayed intracytoplasmic neuronal basophilic inclusion bodies which exhibited marked immunoreactivity to anti-ubiquitin antibodies. Our case reports indicate that the longer survival which is possible through the use of respirators may make one subgroup of ALS patients prone to develop atypical clinical and neuropathological features which are not observed during the natural course of ALS.

Adult↗
PubMed1991

Defects in resting metabolic rates and mitochondrial respiration in Kwashiorkor and dietary obese rats.

Resting metabolic rates have been measured and compared with hepatic mitochondrial respiration in Kwashiorkor and diet-induced obese weaned rats. In Kwashiorkor, resting metabolic rate was 21% lower than the value of controls, while that of the obese rats was 14% higher than in control animals. The resting metabolic rate for Kwashiorkor animals was 50% of the predicted basal metabolic rate (BMR), whereas that of the obese rats was 23% higher than the predicted BMR. The mitochondrial oxygen consumption patterns, using malate plus glutamate or succinate as respiratory substrates, revealed that the resting respiration (state 4) was 23.9% higher in Kwashiorkor and 29.1% higher in obese animals, while the active (state 3) respiration was 34.8% lower in Kwashiorkor and 43.3% lower in obese rats compared to controls. The respiratory control ratios (RCR) were 51.1% and 43.8% in Kwashiorkor and obese rats, respectively, relative to the values in control rats. It is concluded from these studies that Kwashiorkor disease and diet-induced obesity appear to interfere with oxygen utilization at the level of state 3 mitochondrial respiration, which is markedly decreased when compared to the values for control animals.

Animals↗
PubMed1990

Menaquinone is an obligatory component of the chain catalyzing succinate respiration in Bacillus subtilis.

The question was investigated as to whether the bacterial menaquinone (MK) is a component of the electron transport chain catalyzing succinate respiration in Bacillus subtilis. Three different methods were applied, and the following consistent results were obtained. (i) Solvent extraction of MK from the bacterial membrane caused total inhibition of the respiratory activities with succinate and NADH, while the activity of succinate dehydrogenase remained unaffected. The respiratory activities were restored on incorporation of vitamin K1 into the membrane preparation. (ii) The membrane fraction of a B. subtilis mutant containing 15% of the wild-type amount of MK, respired succinate and NADH at reduced activities. Wild-type activities were restored on fusion of the preparation to liposomes containing vitamin K1. (iii) The membrane fraction of B. subtilis catalyzed succinate oxidation by various water-soluble naphtho- or benzoquinones at specific activities exceeding to that of succinate respiration. The results suggest that MK is involved in succinate respiration, although its redox potential is unfavorable.

Bacillus subtilis↗
PubMed1981

The muc+ gene of plasmid pKM101 prevents respiration shutoff in far ultraviolet-irradiated Salmonella typhimurium.

The plasmid PKM101 is known to protect Escherichia coli and Salmonella typhimurium against killing by far UV irradiation and to enhance UV-induced mutagenesis. The muc+ gene of the plasmid is responsible for both of these effects. This paper shows that respiration of S. typhimurium shuts off about an hour after UV irradiation and that pKM101 prevents the shutoff. Plasmids which contained Tn5 translocatable elements, either in (and having produced a muc mutation) or flanking the muc+ gene, have been introduced into S. typhimurium. The muc mutant plasmid, which does not protect its host against UV killing and does not enhance UV induced mutagenesis, also does not protect against UV induced respiration shutoff. Likewise, plasmids in which the Tn5 translocatable elements flank the muc+ gene protect against shutoff of respiration. Thus the muc+ gene of pKM101 is responsible for protection against UV induced shutoff of respiration in S. typhimurium.

DNA Repair↗