Comparison of oxygen consumption rates in minimally transformed BALB/3T3 and virus-transformed 3T3B-SV40 cells

In the recent years, bioenergetics of tumor cells and particularly cell respiration have been attracting great attention because of the involvement of mitochondria in apoptosis and growing evidence of the possibility to diagnose and treat cancer by affecting the system of oxidative phosphorylation in mitochondria. In the present work, a comparative study of oxygen consumption in 3T3B-SV40 cells transformed with oncovirus SV40 and parental BALB/3T3 cells was conducted. Such fractions of oxygen consumption as “phosphorylating” respiration coupled to ATP synthesis, “free” respiration not coupled to ATP synthesis, and “reserve” or hidden respiration observed in the presence of protonophore were determined. Maximal respiration was shown to be only slightly decreased in 3T3B-SV40 cells as compared to BALB/3T3. However, in the case of certain fractions of cellular respiration, the changes were significant. “Phosphorylating” respiration was found to be reduced to 54% and “reserve” respiration, on the contrary, increased up to 160% in virus-transformed 3T3B-SV40 cells. The low rate of “phosphorylating” respiration and high “reserve” respiration indicate that under normal incubation conditions the larger part of mitochondrial respiratory chains of the virus-transformed cells is in the resting state (i.e. there is no electron transfer to oxygen). The high “reserve” respiration is suggested to play an important role in preventing apoptosis of 3T3B-SV40 cells.     

Oxygen consumption of human blood platelets. II. Effect of inhibitors on thrombin-induced oxygen burst

The effect of selected inhibitors on the thrombin-stimulated burst and the basal oxygen consumption of washed human platelets were investigated and compared with inhibition of the release reaction. Cyanide (0.2 mM) caused complete inhibition of the basal respiration, but only 15% inhibition of the thrombin-stimulated burst of oxygen consumption. Similar differential inhibitory effects were observed with oligomycin, antimycin, rotenone and N-ethylmaleimide. Prostaglandin E₁ (0.03 mM) and acetylsalicylic acid (0.8 mM) had little effect on basal respiration, but inhibited the thrombin-stimulated burst of oxygen consumption. N-Ethylmaleimide (0.4 mM) inhibited the release of calcium from platelets by 90%, while prostaglandin E₁, acetylsalicylic acid and the above mitochondrial inhibitors caused no more than 30% inhibition of the release reaction. Our results provide evidence that basal respiration and a portion of the thrombin-stimulated burst of oxygen consumption are involved in respiratory chain phosphorylation, and that this component of the thrombin-stimulated burst may be coupled to the maintenance of the release reaction.     

Oxygen consumption by Nephrops norvegicus (L.), (Crustacea: Decapoda) in relationship with its moulting stage

The study of the rate of oxygen consumption by Nephrops norvegicius (L.) at each moulting stage has been carried out in closed respiration chambers, using an oxygen electrode to measure the oxygen concentration.Although the mean respiration rate seems to be low when compared with other decapod crustaceans of similar weight, the general trends of the variation of the oxygen consumption during the moulting cycle are similar to those described for other species of decapods. It should be noted that, during the process of the ecdysis, there is no oxygen consumption.     

Partitioning of oxygen consumption between “maintenance” and “growth” in developing herring Clupea harengus (L.) embryos

Growth and oxygen consumption was measured in developing herring Clupea harengus (L.) embryos. By considering the variations in oxygen consumption with embryonic size and growth rate, an attempt was made to partition oxygen consumption between growth related and growth unrelated (i.e., “maintenance”) processes. The metabolic cost of growth was estimated as ≈ 150 ng O₂ · μg dry wt tissue formed⁻¹. This estimate compares favourably with the biochemical estimate of the costs of transport and net biosynthesis. The “maintenance” component was proportional to embryonic mass (77 ng O₂ · μg⁻¹· d⁻¹). Over the entire embryonic period, growth processes were responsible for ≈ 25% of the cumulated oxygen consumption.     

Activity and respiration in the anemone, Metridium senile (L.) exposed to salinity fluctuations

Activity and respiration in the anemone, Metridium senile (L.), were monitored under both constant and fluctuating salinity conditions. During constant exposure to 50% sea water it was found that the animals retracted the tentacles and that the rate of oxygen consumption decreased by ≈50%. The same response was elicited from animals in 100% sea water in a contracted state. Animals exposed to continually fluctuating salinities were found to retract the tentacles, contract the body wall, and produce amounts of mucus during periods of decreasing salinities. These reactions were reversed during exposure to increasing salinity. Oxygen consumption never ceased entirely in animals exposed to dilute sea water and it was found that during declining oxygen tension M. senile regulated its oxygen consumption until the environmental oxygen tension fell to ≈30% saturation.     

The combined effects of temperature,salinity, and declining oxygen tension on oxygen consumption in the marine pulmonate Amphibola crenata (Gmelin, 1791)

Oxygen consumption of Amphibola crenata (Gmelin) was measured in various salinity-temperature combinations (< 0.1‰ to 41‰ salinity and 5 to 30°C) in air, and following exposure to declining oxygen tensions. In all experimental conditions, respiration varied with the 0.44 power of the body weight (sd = 0.14). The aquatic rate was consistently higher than the aerial rate of oxygen consumption, although at 30 °C the two rates were similar. Oxygen consumption increased with temperature up to 25 °C in all salinities; the lowest values were recorded at temperatures below 10 °C and at 30 °C in the most dilute medium. At all exposure temperatures, the oxygen consumption of Amphibola decreased regularly with salinity down to 0.1 ‰, and following exposure to concentrated sea water (41‰). Salinity had the least effect at 15 °C which was the acclimation temperature. In general, all of the temperature coefficients (Q₁₀ values) were low, < 1.65. However, Q₁₀ values above 2.8 were recorded at a salinity of 17.8‰ between 10 and 15 °C. Oxygen consumption of all size classes of Amphibola was more temperature dependent in air than in water and small individuals show a greater difference between their aerial and aquatic rates than larger snails. The rates of oxygen consumption in declining oxygen tensions were expressed as fractions of the rates in air saturated sea water at each experimental salinity-temperature combination. The quadratic coefficient B₂ becomes increasingly more negative with both decreasing salinity and temperatures up to 20 °C. At higher temperatures (25 and 30 °C) the response is reversed such that O₂ uptake in snails becomes increasingly independent of declining oxygen tensions at higher salinities. On exposure to a salinity of 4‰, Amphibola showed no systematic response to declining oxygen tension with respect to temperature. The ability of Amphibola to maintain its rate of oxygen consumption in a wide range of environmental conditions is discussed in relation to its potential for invading terrestrial habitats and its widespread distribution on New Zealand's intertidal mudflats.     

Increased oxygen consumption during the uptake of water by the eversible vesicles of Petrobius brevistylis

The oxygen consumption of individuals of Petrobius brevistylis has been measured before, during, and after the uptake of fluid by the insects eversible abdominal vesicles. There was no difference in consumption between fresh animals and those that had been dehydrated or those that had been allowed to rehydrate after an exposure to a NaCl solution. However, during the process of taking up fluid there was over a 200% increase in the respiration rate. While the increase in the rate of oxygen consumption was related to the concentration of the fluid from which the animals were taking up water, the increase in the amount of oxygen consumed was related to the concentration of the substrate solution and the amount of uptake. Part of the oxygen increase is ascribed to the posture adopted by the animals during the uptake of fluid and the remainder to the active transport processes that may be occurring.     

Effect of Oxygen on the Rate of Photorespiration in Detached Tobacco Leaves

The rate of carbon dioxide exchange in both light and darkness by detached tobacco leaves placed at various oxygen concentrations was measured by an Infra-Red CO₂ Analyzer and a Clark oxygen electrode. It was observed that during illumination oxygen had two different effects. One was to stimulate carbon dioxide evolution and the other to inhibit carbon dioxide absorption. Concentration of carbon dioxide at compensation point was found to be a linear function of oxygen concentration and this has been explained as due mainly to an increased evolution of carbon dioxide. Such an evolution during illumination has been called photorespiration. Increased concentrations of oxygen also had a stimulating effect on the magnitude of the initial post-illumination burst of carbon dioxide in darkness, but no effect on the subsequent steady rates. These data have been explained as due to the suspension of regular respiration in darkness and its replacement by a different process, tentatively called photorespiration. A second effect of oxygen was to reduce the efficiency (called “carboxylation efficiency”) with which a leaf was able to remove carbon dioxide from the atmosphere.     

Electrode measurement of oxygen tension with 1-ms time resolution

A continuous flow device utilizing a Clark oxygen electrode was constructed; this device had a dead time and resolution of 1 ms. Mixing was tested by observing the neutralization of acid with base, and at the maximal flow rate, the mixing was 94% complete within 1 ms and better than 98% complete within 2 ms after initial mixing. Observation of the oxygenation of hemoglobin gave data which agreed with previous data obtained by a stopped-flow optical experiment. The respiration of phosphorylating submitochondrial particles was measured utilizing this device. The burst of respiration in submitochondrial particles was triphasic, with a very rapid burst lasting some 60 ms, followed by a longer burst of respiration lasting more than 4 s.     

A coral polyp model of photosynthesis, respiration and calcification incorporating a transcellular ion transport mechanism

A numerical simulation model of coral polyp photosynthesis, respiration and calcification was developed. The model is constructed with three components (ambient seawater, coelenteron and calcifying fluid), and incorporates photosynthesis, respiration and calcification processes with transcellular ion transport by Ca-ATPase activity and passive transmembrane CO₂ transport and diffusion. The model calculates dissolved inorganic carbon and total alkalinity in the ambient seawater, coelenteron and calcifying fluid, dissolved oxygen (DO) in the seawater and coelenteron and stored organic carbon (CH₂O). To reconstruct the drastic variation between light and dark respiration, respiration rate dependency on DO in the coelenteron is incorporated. The calcification rate depends on the aragonite saturation state in the calcifying fluid (Ωa _cal). Our simulation result was a good approximation of “light-enhanced calcification.” In our model, the mechanism is expressed as follows: (1) DO in the coelenteron is increased by photosynthesis, (2) respiration is stimulated by increased DO in the light (or respiration is limited by DO depletion in the dark), then (3) calcification increases due to Ca-ATPase, which is driven by the energy generated by respiration. The model simulation results were effective in reproducing the basic responses of the internal CO₂ system and DO. The daily calcification rate, the gross photosynthetic rate and the respiration rate under a high-flow condition increased compared to those under the zero-flow condition, but the net photosynthetic rate decreased. The calculated calcification rate responses to variations in the ambient aragonite saturation state (Ωa _amb) were nonlinear, and the responses agreed with experimental results of previous studies. Our model predicted that in response to ocean acidification (1) coral calcification will decrease, but will remain at a higher value until Ωa _amb decreases to 1, by maintaining a higher Ωa _cal due to the transcellular ion transport mechanism and (2) the net photosynthetic rate will increase.     

Electrode measurement of oxygen tension with 1-ms time resolution.

A continuous flow device utilizing a Clark oxygen electrode was constructed; this device had a dead time and resolution of 1 ms. Mixing was tested by observing the neurtralization of acid with base, and at the maximal flow rate, the mixing was 94% complete within 1 ms and better than 98% complete within 2 ms after initial mixing. Observation o of the oxygenation of hemoglobin gave data which agreed with previous data obtained by a stopped-flow optical experiment. The respiration of phosphorylating submitochondrial particles was measured utilizing this device. The burst of respiration in submitochondrial particles was triphasic, with a very rapid burst lasting some 60 ms, followed by a longer burst of respiration lasting more than 4 s.     

Some Factors Governing Respiration,Glucose Metabolism and Iodoacetate Sensitivity in Tetrahymena pyriformis*

SYNOPSIS. The physiologic response of Tetrahymena pyriformis W to glucose, as measured by the respiratory rate in a buffered suspension, was found to be altered by exposure of the culture to this carbohydrate in the peptone-based growth medium. Glucose-grown cells had an elevated respiratory rate in the presence of glucose in the Tris-buffered suspension, while cells not grown with the glucose supplement required sodium and potassium for the response. Orthophosphate elevated the rate of oxygen consumption independently of the glucose effect. The endogenous respiratory of glucose and non-glucose cultured cells were significantly diffe The 2 types of cells responded differently to the cations, sod and potassium, and were not identical in sensitivity to iodoacet Iodoacetate poisoning was found to be due to inhibition of “uncompetitive” type, potentiated by glucose, orthophosphate, the absence of potassium.     

Data consistency,yield, maintenance,and hysteresis in batch cultures of Candida lipolytica cultured on n-hexadecane

Candida lipolytica was cultured batchwise using n-hexadecane as the main carbon source. Biomass production, n-hexadecane consumption, oxygen consumption, and carbon dioxide evolution were measured to follow the fermentation. The consistency of the measured data was examined using integrated and instantaneous available electron and carbon balances. Values of the “true” growth yield, η_max, and maintenance coefficient, m_e were estimated using three different sets of data (biomass and n-hexadecane, oxygen and biomass, and CO₂ and biomass), and the results were compared with estimates obtained from literature data. Hysteresis patterns were observed in plots of specific rates of oxygen consumption and carbon dioxide evolution versus specific growth rate.     

Regulation of soybean nitrogen fixation in response to rhizosphere oxygen: I. Role of nodule respiration

Nitrogen fixation (acetylene reduction) rates of nodules on intact field-grown soybean (Glycine max) subjected to altered oxygen concentration (0.06-0.4 cubic millimeter per cubic millimeter) returned to initial rates during an 8-hour transitory period. Hydroponically grown soybean plants also displayed a transitory (1-4 hours) response to changes in the rhizosphere oxygen concentration after which the fixation rates returned to those observed under ambient oxygen concentrations. It was hypothesized that soybean nodules contain a regulatory mechanism which maintains a stable oxygen concentration inside nodules at a sufficiently low concentration to allow nitrogenase to function. A possible physiological mechanism which could account for this regulation is adjustment in nodule respiration activity such that nodule oxygen concentration and nitrogen fixation are maintained at stable levels. Experiments designed to characterize the non-steady-state oxygen response and to test for the presence of nodule respiratory control are presented. Non-steady-state acetylene reduction and nodule respiration (oxygen uptake) rates measured after alterations in the external oxygen concentration indicated that the regulatory mechanism required 1 to 4 hours to completely adjust to changes in the external oxygen concentration. Steady-state nodule respiration, however, did not respond to alterations in the rhizosphere oxygen concentration. It was concluded that soybean nodules can adjust to a wide range of rhizosphere oxygen concentrations, but the mechanism which controls nitrogen fixation rates does not involve changes in the nodule respiration rate.     

Respiration of intact and damaged plants ofSpirodela oligorhiza SCHL

The wound effect (“Wundeffekt”) was investigated inSpirodela oligorhiza by comparing respiration in intact plants and plants cut into halves and quarters. The effect of damage was not evident immediately after injury but after 18 hours the respiratory rate of cut plants was twice to three times that of intact plants. The increased respiration rate is, therefore, not the direct result of mechanical injury.     

Energy coupling for membrane hyperpolarization in Lemna: respiration rate,ATP level and membrane potential at low oxygen concentrations

Respiration rate, ATP content and membrane potential of Lemna have been measured as a function of the concentration of dissolved oxygen. Kinetic analysis showed that within the range from 1 μM to 20 μM O₂, the respiration rate of isolated mitochondria and intact plants was a hyperbolic function of the oxygen concentration. The apparent Michaelis constant (K _m ) for the oxygen of respiration of intact plants (1.15±0.08 μM) is close to that for isolated mitochondria (1.07±0.06 μM), so that diffusion of oxygen within the tissue was obviously not rate-limiting under the applied experimental conditions. The ATP level decreased in parallel with the respiration rate when the oxygen concentration was reduced. In contrast, the hyperpolarization of the membrane potential above the diffusion potential had already decreased at oxygen concentrations where the respiration rate and ATP level remained practically unchanged and was completely abolished at oxygen concentrations above the K _m of respiration. This result is discussed according to the current models for electrogenic pumps. It is concluded that ATP cannot be the fuel for the electrogenic process under investigation.     

Compensatory and adaptive rearrangement in the human respiratory system under acute hypoxic conditions

Relationships between the parameters of external respiration (minute volume and respiration rate) and those of internal, tissue respiration (oxygen consumption, arteriovenous oxygen difference and efficiency of oxygen uptake) were studied during a period of acute hypoxia and upon its completion. The subjects were exposed to hypoxia for 25 min using oxygen-nitrogen hypoxic gas mixtures (HGMs) differing in oxygen content (8 and 12%, HGM-8 and HGM-12, respectively). From the third to the fifth minutes of exposure to HGM-8, the respiration minute volume (RMV) was found to increase by 51 ± 33% as compared to the background value; however, the body’s oxygen consumption (OC) was 35 ± 22% reduced. Afterwards, OC grew to reach, from the 20th to the 25th min of hypoxia, 108 ± 21% of the background value and 181% of the value determined from the third to the fifth minutes of hypoxia. OC growth was accompanied by an insignificant RMV increase (by 12%) as compared to the level determined from the third to the fifth minutes of hypoxia, whereas the efficiency of oxygen uptake from the arterial blood increased by 75% for the same period. RMV growth from the third to the fifth minutes of hypoxia occurred as expense result of a higher breathing depth; at the same time, the respiration rate decreased as compared to the background value. By the period from the 20th to the 25th min of exposure to HGM-8, the respiration rate increased by 21% as compared to the period from the third to the fifth minutes of hypoxia. The efficiency of oxygen uptake from the arterial blood remained higher than the background value for at least 5 min after completion of the exposure to HGM-8. During the same period, the ventilation equivalent, an indicator of the efficiency of external respiration, i.e., of oxygen supply to the body, was significantly lower than the background value. During the exposure to HGM-12, RMV increased to a lesser extent than on exposure to HGM-8, however, the efficiency of oxygen uptake was higher during exposure to HGM-12; therefore, OC was also higher in the latter case. Therefore, the assumption that, during hypoxia, intensified external respiration (ventilatory response) itself compensates oxygen deficiency in inhaled air is revised. Ventilatory response is only a portion of the entire functional system of respiration (both external and tissue respiration). The role of ventilatory response is important for conditioning the tissue respiration rearrangement to eliminate deficiency of oxygen consumption during hypoxia. The retained higher oxygen uptake from the arterial blood during the period after completion of hypoxic treatment testifies to the adaptive implication of changes in tissue respiration; the same is confirmed by a reduced ventilation equivalent after hypoxia, which is indicative of the growing efficiency of external respiration, i.e., of an improved oxygen supply to the body.     

Respiration in air and water of three mangrove snails

The oxygen consumption of three species of Malaysian mangrove gastropods was measured in air and sea water at the temperatures commonly recorded in the mangrove. The experiments in air were carried out after the animals had regained fluid lost from the mantle due to handling. Fluid loss can have considerable effects on rate of oxygen consumption. Nerita arliculata (Gould) was found throughout the mangrove and experiences from 50 to 92% aerial exposure. It has a gill and a ratio of aerial to aquatic respiration rates of 2.7 at 28°C. 50% of the animals can survive underwater for 72 h at 28°C. The other two species, Cerithidea obtusa (Lamarck) and Cassidula aurisfelis (Brugière) experience over 95% aerial exposure, have their mantle cavities modified as lungs and have air : water respiration rate ratios of 5.5 and 6.0, respectively. 50% can survive from 48 to 36 h underwater at 28°C. Acclimated animals have Q₁₀'s of about 1.6 in air and 1.4 in water. The respiratory physiology of the snails is compared with that of rocky shore species.     

Bathymetric adaptations of reef-building corals at davies reef,great barrier reef,Australia. II. Light saturation curves for photosynthesis and respiration

Light saturation (P-I) curves for oxygen production and consumption were constructed in the laboratory for corals collected from depths of 1 to 45 m on the southeastern (seaward) side of Davies Reef, Great Barrier Reef, Australia. This depth range represents a gradient of from 82.6 to 2.9% of surface light, which was measured as photosynthetic photon flux density (PPFD) between 400 and 700 nm. Adaptative changes in photokinetic parameters were observed over the entire range of light intensities. The compensation intensity (I_c), I_k, the intensity at which photosynthesis was 95% light saturated (I_0.95), and the respiration rate (?R), all decreased with decreasing light intensity. The maximal rate of photosynthesis when normalized on the basis of coral chlorophyll-a content (P_ma^g) tended to decrease with decreasing irradiance but the change was not statistically significant. The $\text{P}\text{R}$ ratio ($\text{P}{}_{\mathrm{m}}{}^{\mathrm{g}}\text{?R}$), the initial slope of a light saturation curve (α), and the maximum rate of photosynthesis when normalized by coral protein content (P_mp^g), all increased with decreasing light intensity. The logarithms of the values for each variable parameter were proportional to the logarithms of the fraction of the surface PPFD (T) transmitted to the depth at which the corals grew. This enabled changes in these parameters to be accurately estimated for corals growing at any depth on the reef. Comparison of experimental data with published values for “saturating” irradiance and P_ma^g, suggests that the endosymbiotic algae within reef-building corals are photosynthetically intermediate between classical “sun” and “shade” plants.     

BIOCHEMICAL COMPOSITION AND METABOLIC ACTIVITY OF SCRIPPSIELLA TROCHOIDEA (DINOPHYCEAE) RESTING CYSTS1

The composition and metabolic activity of cysts of the marine dinoflagellate Scrippsiella trochoidea (Stein) Loeblich were examined during dormancy, quiescence, and germination. On a per cell basis, newly formed cysts contained an order of magnitude more carbohydrate but significantly less protein and chlorophyll a than did exponentially growing vegetative cells. Loss of lipid and carbohydrate from cysts during the initial dormancy period reflected a respiration rate estimated to be 10% of the respiratory activity in vegetative cells. Among older, quiescent cysts the calculated respiration rate decreased further to approximately 1.5% of the vegetative rate and appeared to proceed largely at the expense of carbohydrate reserves. These estimated rates of respiration were in good agreement with direct measurements of cyst oxygen consumption. The transfer of quiescent cysts to conditions permissive for germination resulted in a rapid increase in respiration rate, as evidenced by carbohydrate loss and O₂ consumption. The increased respiratory activity was followed by an increase in protein content and, later, by an increase in chlorophyll a content and photosynthetic capacity. Just prior to germination the P/R ratio became greater than 1, and the estimated chlorophyll-specific photosynthetic activity reached 75% of the rate in vegetative cells. Complete restoration of photosynthetic and respiratory capacity apparently was not achieved until after excystment. These data confirm the common assumption that dinoflagellate cysts represent true “resting” cells, containing extensive energy reserves and displaying greatly reduced metabolic activity.