Lung oxygen consumption and mitochondria of alveolar epithelial and endothelial cells.

We examined oxygen consumption by lung slices and measured the volume density of mitochondria of granular pneumocytes, alveolar type I cells, and alveolar capillary endothelial cells in several species. We found that lung oxygen consumption (mu-1 02 times h-1 times mg DNA-1) varies inversely with the log of animal body weight and with the species alveolar diameter and directly with the species respiratory rate. The volume density of granular pneumocyte mitochondria show a direct linear correlation with the lung's oxygen consumption and the species respiratory rate, and an inverse linear correlation with the species alveolar diameter. The volume density of mitochondria in type I alveolar epithelial cells and capillary endothelial cells, considered together, did not differ in the two species studied (mouse and rat). We conclude that there are interspecies differences in oxygen consumption by lung cells and that granular pneumocytes contribute to these differences. We suggest that, at least part of these differences, are related to interspecies differences in surfactant secretory activity.     

Effect in vivo of atrazine on haematology and O2 consumption in fish, Tilapia mossambica.

Studies were conducted on haematological constituents such as Red blood cells (RBC), White blood cells (WBC), Haemoglobin (Hb), Packed cell volume (PCV), Mean cell volume (MCV), Mean corpuscular haemoglobin (MCH), Mean corpuscular haemoglobin concentration (MCHC), Blood volume (BV), Blood water content (BWC) and Whole animal oxygen consumption (WAOC) in the fish exposed to sublethal concentration of atrazine. Significant changes were seen in the constituents of the blood and O2 consumption of fish suggesting the existence of respiratory distress in the fish as a consequence of atrazine toxicity.     

Proliferation-associated oxygen consumption and morphology of tumor cells in monolayer and spheroid culture.

The oxygen consumption rate, proliferative activity, and morphology of EMT6/Ro mouse mammary sarcoma cells in monolayer and multicellular spheroid culture have been investigated in a comparative study. During the transition of monolayer cells from the exponential into the plateau growth phase, there is a distinct decrease in the cellular volume that is associated with a corresponding decrease in the proliferative and respiratory activity of the cells. The decline in cell volume is mainly due to a decrease in the content of cytoplasm, whereas the size of the nucleus is only slightly reduced. A concomitant decrease in the number of mitochondria per cell obviously accounts for the reduction in cellular oxygen uptake. Despite a continuous decrease of cell proliferation from the surface to interior regions of EMT6 spheroids reflected by a gradient in tritiated thymidine labeling, volume-related oxygen consumption is rather uniform in viable regions of these aggregates. The finding can be explained by the results of the morphometric evaluation showing a uniform volume density of mitochondria, i.e., of oxygen-consuming sites within these spheroids.     

Bioenergetic function in cardiovascular cells: the importance of the reserve capacity and its biological regulation

The ability of the cell to generate sufficient energy through oxidative phosphorylation and to maintain healthy pools of mitochondria are critical for survival and maintenance of normal biological function, especially during periods of increased oxidative stress. Mitochondria in most cardiovascular cells function at a basal level that only draws upon a small fraction of the total bioenergetic capability of the organelle; the apparent respiratory state of mitochondria in these cells is often close to state 4. The difference between the basal and maximal activity, equivalent to state 3, of the respiratory chain is called the reserve capacity. We hypothesize that the reserve capacity serves the increased energy demands for maintenance of organ function and cellular repair. However, the factors that determine the volume of the reserve capacity and its relevance to biology are not well understood. In this study, we first examined whether responses to 4-hydroxynonenal (HNE), a lipid peroxidation product found in atherosclerotic lesions and the diseased heart, differ between vascular smooth muscle cells, adult mouse cardiomyocytes, and rat neonatal cardiomyocytes. In both types of cardiomyocytes, oxygen consumption increased after HNE treatment, while oxygen consumption in smooth muscle cells decreased. The increase in oxygen consumption in cardiomyocytes decreased the reserve capacity and shifted the apparent respiratory state closer to state 3. Neonatal rat cardiomyocytes respiring on pyruvate alone had a fourfold higher reserve capacity than cells with glucose as the sole substrate, and these cells were more resistant to mitochondrial dysfunction induced by 4-HNE. The integration of the concepts of reserve capacity and state-apparent are discussed along with the proposal of two potential models by which mitochondria respond to stress.     

Respiratory activity of bacteria Acinetobacter calcoaceticus TM-31 during assimilation of alkane hydrocarbons

The respiratory activity of Acinetobacter calcoaceticus TM-31 with resect to alkane hydrocarbons was studied. The dynamics of oxygen consumption by the cells while assimilating n-hexadecane was assayed by a modified technique using an oxygen electrode. The dependence of cell respiratory activity on the amount of n-hexadecane within the concentration range of 0.03-0.66% was determined. It was demonstrated that the cells also displayed respiratory activity towards other medium-chain n-alkanes: hexane, octane, decane, tridecane, and heptadecane. Thus, we demonstrated the possibility of determining alkanes by measuring the respiratory activities of microorganisms.     

Growth-related changes of oxygen consumption rates of tumor cells grown in vitro and in vivo

Growth-related changes of oxygen consumption rates of tumor cells, grown in vitro or in vivo, were investigated. For in vitro investigations, L929 and DS-carcinosarcoma cells were cultured in artificial media. For in vivo studies, DS-carcinosarcoma cells were implanted into the abdominal cavity of Sprague-Dawley rats (ascites tumor, containing malignant cells, leukocytes, lymphocytes, and macrophages). Oxygen uptake was measured photometrically. Parameters of the extracellular medium judged to possibly influence the respiratory activity of tumor cells were monitored at different growth stages (glucose, lactate, and amino acid levels, oxygen and carbon dioxide partial pressures, and pH values). The results obtained clearly show that the oxygen uptake of tumor cells grown in vitro decreased as quiescence developed. In contrast, the respiratory activity of in vivo DS-carcinosarcoma ascites cells increased as tumor growth reached plateau phase. The differences observed cannot be attributed solely to changes of the environmental conditions monitored. It is likely that an increased respiration rate of activated host cells might profoundly contribute to the elevation of the respiratory capacity of DS-carcinosarcoma ascites tumors grown in vivo. These data provide evidence that solid tumors in vivo can increase their O2 uptake at an enhanced O2 availability not only due to an enlarged tumor volume with adequate O2 supply but also due to an elevation of the respiratory activity of different cell populations within a tumor.     

The red blood cells in growing guinea pigs. (Cavia cobaya). II. Communication: erythropoiesis and red blood cells in the postnatal development period

Bone-marrow smears of 175 guinea pigs aged 1-27 days and venous blood samples of 351 animals aged 1-25 days were prepared for cell counting. A significant increase of erythroblasts were found between life day 1 and 2; normoblasts increased in number synchronously with a decrease of erythroblasts after the 5th day. The percentage of the erythroid bone marrow increased from 10 to 14 during the developmental period. Beyond the perinatal period the red blood picture is characterized by the following changes: a decrease of erythrocyte count, hematocrit, hemoglobin, mean corpuscular volume, and mean corpuscular hemoglobin; a constant mean corpuscular hemoglobin concentration; an increase of the reticulocyte count. The decrease of the red cell count is compensated by a decreasing oxygen affinity attained by an important increase of 2,3-DPG. Nevertheless, the stimulus for a raising erythropoiesis remains constant which can be shown by the growing percentage of erythroid cells and reticulocytes. The difference between the human postnatal development and that of the guinea pig becomes obvious. Cell counts in dependence of body masses in postnatally growing guinea pigs, veil the perinatal finding of the increase in erythrocytes up to the 5th day and the decrease of the mean corpuscular volume after the 3rd day.     

The study of the bioenergetic characteristics of the red blood cells of Black Sea fish: the common stingray (<Emphasis Type="Italic">Dasyatis pastinaca</Emphasis> L.) and black scorpionfish (<Emphasis Type="Italic">Scorpaena porcus</Emphasis> L.)

The oxygen consumption rate in red blood cell suspensions of two Black Sea fish species, a cartilaginous fish, the common stingray (Dasyatis pastinaca L.) and the teleost black scorpionfish (Scorpaena porcus L.) has been studied. The proposed stimulants of activators and inhibitors of the mitochondria electron transport chain had very predictable responses, indicating that mitochondria in fish erythrocytes have a classical set of respiratory enzymes. Despite the fact that the basic respiratory activity of common stingray erythrocytes was greater than those of the scorpionfish, the responses of common stingray red blood cells to the exposure during investigation of the respiratory activity of the mitochondria have an inverse relationship. The oxygen consumption rates in suspensions of scorpionfish erythrocytes in response to the stimulant were higher according to both the amplitude and the duration of the response. Investigations have shown the high energy potential of the red blood cell mitochondria of the scorpionfish and stingray. This may be the energy basis for maintaining the high intracellular concentrations of ATP required not only to keep an adequate level of intracellular metabolism, but also to provide a special mode of blood flow through the capillary beds.     

Physiological responses of mules on prolonged exposure to high altitude (3 650 m)

Eight healthy male animals were inducted and kept for 2 1/2 years at 3 650 m altitude and subjected to normal work schedules. Physiological measurements viz. heart rate, blood pressure, minute ventilation, oxygen consumption, respiration rate, hemoglobin, packed cell haematocrit volume and eosinophil count were made on these animals at periodic intervals. On acute induction to an altitude of 3 650 m these animals demonstrated a sudden increase in tidal volume, a decrease in Rf and no change in V_E, suggesting a decreased dead space/tidal volume ratio at altitude.However, all these changes stabilised within 3 weeks but on prolongation of stay, the physical state of these animals was adversely affected. The respiratory adjustments occurring on return to sea level appear to be a response to thermal stress. The initial increase in heart rate and blood pressure stabilised by the 2nd week.     

Ventilation and circulation during exercise inOctopus vulgaris

Summary Ventilation frequency, volume, oxygen uptake, and oxygen transport by the blood have been studied in unrestrained octopus,Octopus vulgaris before, during and after recovery from 20 min of enforced activity. Exercise increased oxygen consumption 2.8 fold. The percentage utilisation of oxygen from the branchial water is maintained or increased at around 35% during activity and the calculated ventilation volume increases by 3 times. Prior to exercise the hemocyanin in arterial blood is 98% saturated and there is 83% utilisation of the oxygen in the blood. During activity there is remarkably little change in blood parameters so that the hemocyanin in the arterial blood remains at 96% saturation and oxygen utilisation is 90%. Cardiac output was calculated to have risen 2.5 fold during activity. As theP _{O 2} gradients across the gill do not change significantly during exercise the major adaptation which can account for an increase in oxygen consumption must be a 3 fold increase in the transfer factor. At rest 22% of the total CO₂ present in the blood is excreted during its passage through the gills and this rises to 32% during activity. There is no accumulation of CO₂ and only a slight acidification of the blood during activity. A significant respiratory and metabolic acidosis is avoided and the hemocyanin continues to function normally.     

Respiratory activity of the bacteriumAcinetobacter calcoaceticus TM-31 during assimilation of alkane hydrocarbons

The respiratory activity ofAcinetobacter calcoaceticus TM-31 with resect to alkane hydrocarbons was studied. The dynamics of oxygen consumption by the cells while assimilatingn-hexadecane was assayed by a modified technique using an oxygen electrode. The dependence of cell respiratory activity on the amount ofn-hexadecane within the concentration range of 0.03–0.66% was determined. It was demonstrated that the cells also displayed respiratory activity towards other medium-chainn-alkanes: hexane, octane, decane, tridecane, and heptadecane. Thus, we demonstrated the possibility of determining alkanes by measuring the respiratory activities of microorganisms.     

Oxygen-related processes in red blood cells exposed to tert-butyl hydroperoxide

The correlation between the oxidative processes in tert-butyl hydroperoxide (tBHP)-exposed red blood cells and the reactions of oxygen consumption and release were investigated. Red blood cell exposure to tBHP resulted in transient oxygen release followed by oxygen consumption. The oxygen release in red blood cells was associated with intracellular oxyhaemoglobin oxidation. The oxygen consumption proceeded in parallel with free radical generation, as registered by chemiluminescence, but not to membrane lipid peroxidation. The oxygen consumption was also observed in membrane-free haemolyzates. The order of the organic hydroperoxide-induced reaction of oxygen release with respect to the oxidant (tBHP) was estimated to be 0.9 +/- 0.1 and that of the oxygen consumption reaction was determined to be 2.4 +/- 0.2. The apparent activation energy values of the oxygen release and oxygen consumption were found to be 107.5 +/- 18.5 kJ/mol and 71.0 +/- 12.5 kJ/mol, respectively. The apparent pKa value for the functional group(s) regulating the cellular oxyHb interaction with the oxidant in tBHP-treated red blood cells was estimated to be 6.7 +/- 0.2 and corresponded to that of distal histidine protonation in haemoprotein. A strong dependence of tBHP-induced lipid peroxidation on the oxygen concentration in a red blood cell suspension was observed (P50 = 32 +/- 3 mmHg). This dependence correlated with the oxygen dissociation curve of cellular haemoglobin. The order of the membrane lipid peroxidation reaction with respect to oxygen was found to be 0.5 +/- 0.1. We can conclude that the intensity of the biochemical process of membrane lipid peroxidation in tBHP-exposed erythrocytes is controlled by small changes in such physiological parameters as the oxygen pressure and oxygen affinity of cellular haemoglobin. Neither GSH nor oxyhaemoglobin oxidation depended on oxygen pressure.     

Monitoring the rate of oxygen consumption in plant cell suspensions

A method is described that allows the rate of oxygen consumption to be monitored in plant cell suspensions. The method utilized oxygen electrodes placed in beakers of plant cells subjected to various treatments. The voltage readings from calibrated electrodes were converted to % oxygen (100% equals air equilibration) and the rate of oxygen consumption was estimated by calibration graphs made with no cells present. This system simultaneously monitors one to sixteen or more samples, allowing comparison of treatments on identically treated cells. We have used this method to study the respiratory burst of plant cells produced in response to viable or heat-killed bacteria. Because the system was computer-monitored and open to the atmosphere, data could be collected over several hours. Various factors that affected the measurement of dissolved oxygen concentration with this technique were explored and considered. This revised version was published online in June 2006 with corrections to the Cover Date.     

The respiratory behavior of Lewis carcinoma cells--existence of a cyanide-resistant respiration

The respiratory O2 consumption of vegetative, non-necrotic, Lewis lung carcinoma cells was found to be very low compared with that of non-tumor tissues and was highly resistant to cyanide. However, the resistant rate was inhibited by salicylhydroxamic acid (SHAM) in either isolated cells or tissue fragments. In addition, this compound did not affect their cytochrome-c oxidase activity. The results support the existence of an alternative oxidase system that significantly contributes to oxygen uptake in Lewis carcinoma cells. To our knowledge, this is the first report showing significant SHAM-sensitivity of tumor respiration and perhaps of higher animal cell respiration.     

The reduction of Alamar Blue by peripheral blood lymphocytes and isolated mitochondria

Alamar Blue is a widely used nontoxic indicator of cell proliferative activity, which penetrates quickly through the biological membranes and can be easily reduced by intracellular enzymes. Accumulation of reduced fluorescent form of Alamar Blue during short-term culture of human peripheral blood lymphocytes may be used as a cell viability test since it was prevented by disruption of plasma membrane by digitonin. The inhibition of Alamar Blue reduction by NaN3 indicates that its metabolism is associated with mitochondrial activity. A compaative study of Alamar Blue reduction and oxygen consumption on isolated rat liver mitochondria shows, that the Alamar Blue reduction is not associated with the activity of specific complex of respiratory chain and it seems to be an integral indicator of oxidation-reduction activity of respiratory chain components.     

Changes in haemorheology in the racing greyhound as related to oxygen delivery

Arterial blood samples were obtained from six greyhounds during rest, immediately before, and after a 704-m (7/16th mile) race. Measurements were made of various haematological (red cell count, haemoglobin, packed cell volume, white cell count, plasma proteins) and haemorheological variables. Blood and plasma viscosity were determined at high wall shear stresses (67-200 dynes.cm-2, 670-2000 microN.cm-2) in a 20-microns glass capillary device which was designed to take the diameter dependence of blood viscosity (Fahraeus-Lindqvist effect) into account. Compared to values at rest, substantial haemoconcentration occurred before the race, mainly due to splenic discharge of red cells. Additional haemoconcentration was found after the race. The increase of effective blood viscosity caused by elevation of packed cell volume was greater than the increase in O2 binding capacity resulting from the elevated haemoglobin concentration, suggesting that the haemoconcentration observed in the exercising greyhound does not enhance O2 delivery to skeletal muscle. The main physiological effect of red cell discharge from the contracting spleen appeared to be a consequence of the volume rather than the composition of the circulating blood.     

A reduction in the in situ rates of oxygen and glucose consumption of cells in EMT6/Ro spheroids during growth

The rates of consumption of oxygen and glucose by EMT6/Ro cells in multicellular spheroids were measured at various times during normal growth. In situ spheroid cellular consumption rates were similar to those of exponentially growing single cells up to a spheroid diameter of 150 micron. Further growth resulted in decreases in the rates of both oxygen and glucose consumption which were correlated with the increase in spheroid diameter and cell number. At a diameter of 1300 micron, both rates of cellular consumption had decreased by a factor of 2.5. The rates of consumption per unit of nonnecrotic spheroid volume decreased in a similar manner. Measurements with single cells demonstrated that the rate of oxygen consumption was coupled with glucose concentration, and vice versa. The rates of consumption for cells dissociated from small spheroids indicated that there was some effect of the spheroid environment. As the spheroids grew, however, association in the spheroid structure accounted for a smaller proportion of the total observed reduction in the rates of nutrient consumption. The presence of central necrosis also appeared to have no effect on the rates of consumption of these nutrients. Spheroid-derived cells showed a decrease in cell volume with growth as the cells accumulated in a quiescent state. Measurements with single cells demonstrated that oxygen and glucose consumption were correlated with cell volume and with the development of nonproliferating cells. We conclude that the observed decrease in oxygen and glucose consumption with growth in spheroids is largely due to the progressive accumulation of cells in a quiescent state characterized by an inherently lower cellular rate of nutrient utilization.     

The activity of several components of the innate immune system in diploid and triploid turbot

The use of triploid fish may be of interest in research, e.g. study of how this condition affects the size and activity of cells. In addition, triploid fish are sterile and production of triploids in fish species that are marketed after reaching sexual maturity may be of economic interest. In the present study, the effects of triploidy on the activity of several components of the innate immune system of turbot (Psetta maxima L) were determined. Triploid turbot had bigger cells (erythrocytes and neutrophils) but the number of blood erythrocytes, leucocytes and thrombocytes was lower than in diploid fish. The differential cell count was similar in both types of fish. The respiratory burst and the phagocytic activities were higher in neutrophils of triploid turbot. However, because the number of neutrophils was higher in diploids, the total respiratory burst activity and the phagocytosis per microliter of blood was similar in both types of fish. No differences were found in serum complement, lysozyme or bactericidal activities. The results indicate that the activities of the humoral components of the innate immune system tested are similar in diploid and triploid fish and that the lower leucocyte number found in triploids is compensated for by higher cell activity.