CARBON DIOXIDE AS A FACILITATING AGENT IN THE INITIATION AND GROWTH OF BUBBLES IN ANIMALS DECOMPRESSED TO SIMULATED ALTITUDES

1. Rats killed in a variety of ways (broken neck, nembutal, anoxia, electrocution) may undergo extensive bubble formation when subsequently decompressed from atmospheric pressure to simulated altitudes of 50,000 feet. On autopsy at sea level, large numbers of bubbles are found throughout the vascular system in the majority of animals. These bubbles appear to originate in small vessels deep within muscular regions, later spreading widely in arterial and venous systems. Dead rabbits and frogs also bubble profusely on decompression. 2. Bubble formation in dead animals is attributed primarily to the accumulation of CO₂, derived from residual cellular respiration after death, and from anaerobic glycolysis with attendant decomposition of bicarbonates in blood and tissue fluids. If anaerobic glycolysis is inhibited by using sodium iodoacetate as a lethal agent, bubble formation is greatly reduced or lacking on subsequent decompression. 3. Experiments in vitro suggest that high concentrations of CO₂ favor bubble formation by reducing the degree of mechanical disturbance necessary. 4. Administration of CO₂ in high concentrations to living frogs lowers the minimum altitude (pressure equivalent) at which bubble formation occurs, with exercise, in untreated animals. Pre-treatment with CO₂ also reduces the degree of muscular activity necessary for bubbles to form in frogs at higher altitudes. 5. Analyses have been made of the gas content of bubbles taken directly from the large veins of decompressed frogs and rats. In living animals the figures obtained indicate rapid equilibration with gas tensions in the blood. Bubbles taken from decompressed dead rats may contain 60–80 per cent CO₂. 6. The bearing of these experiments on the mechanisms of bubble initiation and growth in normal living animals is discussed. Reasons are given for suggesting that CO₂, due largely to its high dissolved concentration in localized active regions, may be an outstanding factor in the initiation and early growth of bubbles which in later stages are expanded and maintained principally by nitrogen.     

CO2 angiography: studies of gas filling of vessels and assessing the factors of influence on the injection process with a cardiovascular model]

Following the advent of digital subtraction angiography (DSA), carbon dioxide gas has become a useful contrast agent for arterial angiography. Former manual injection methods had precluded accurate dosing and reproducibility. An original gas injector was therefore developed and tested in a circulatory system model. This permitted an accurate evaluation of vascular gas-filling. it also proved possible to measure the injection parameters taking different influencing factors into account. It was shown that vessels up to 10 mm in diameter are virtually completely filled with gas. In vessels larger than 10 mm in diameter, a residual fluid level remains, which in turn reduces the possibility for complete vessel imaging. The injection flow is dependent primarily on the vascular circulation rate, vessel diameter and the inclination of the vessel. With respect to other factors examined, neither catheter size, number of holes nor the direction of the injection have any relevant influence.     

Methane bubbles in surface peat cores: in situ measurements

The quantification of greenhouse gas sources and sinks is important to understanding the impact of climate change. Methane (CH₄) is a potent greenhouse gas, which, on a global scale, is released largely as a product of anaerobic microbial decomposition and predominantly from wetlands. A zone of intense CH₄ production just below the water table is thought to contribute significantly to the overall flux from peat bogs. We describe the use of membrane inlet quadrupole mass spectrometry (QMS) to confirm the existence of bubbles, their gaseous concentrations and their localization at a fine spatial resolution within intact peat cores. We use the distribution of the noble gas argon (Ar) and the distinct QMS responses to dissolved and gaseous (bubble) phases to identify trapped bubbles with a resolution of 0.6 mm. Bubbles with CH₄ concentrations of up to 20 kPa were widely distributed in the upper 300 mm of the cores with ～11% of all profiles comprising bubbles. The dissolved concentrations responsible for the bubbles were on average 83±80 μm , indicating lower concentrations relative to other QMS studies. We suggest that if the distinction between dissolved and gaseous phases is not made in studies of CH₄ within peat profiles then the prominence of bubbles is likely to result in overestimates of dissolved CH₄ concentrations. Fluxes of CH₄ from peat as a result of drawdown or other perturbation are likely to be large, rapid and short lived because of bubble burst, and also larger than from peat without bubbles. We suggest that the dynamics of fluxes need to be modelled taking into account both gaseous and dissolved phases. Estimates of potential fluxes that assume CH₄ is dissolved are likely to overestimate fluxes if the gaseous phase has not been taken into account.     

A simple carbon dioxide injection system for photosynthetic studies

A simple carbon dioxide injection system has been developed for the maintenance of CO₂ concentrations in semiclosed cuvette systems suitable for photosynthesis and gaseous pollutant studies. The device injects small volumes of pure carbon dioxide into the cuvette in response to a signal from an infrared gas analyzer.     

Evaluating the efficiency of enzyme accelerated CO2 capture: chemical kinetics modelling for interpreting measurement results

In this paper, the efficiency of the carbonic anhydrase (CA) enzyme in accelerating the hydration of CO₂ is evaluated using a measurement system which consists of a vessel in which a gaseous flow of mixtures of nitrogen and CO₂ is bubbled into water or water solutions containing a known quantity of CA enzyme. The pH value of the solution and the CO₂ concentration at the measurement system gas exhaust are continuously monitored. The measured CO₂ level allows for assessing the quantity of CO₂, which, subtracted from the gaseous phase, is dissolved into the liquid phase and/or hydrated to bicarbonate. The measurement procedure consists of inducing a transient and observing and modelling the different kinetics involved in the steady-state recovery with and without CA. The main contribution of this work is exploiting dynamical system theory and chemical kinetics modelling for interpreting measurement results for characterising the activity of CA enzymes. The data for model fitting are obtained from a standard bioreactor, in principle equal to standard two-phase bioreactors described in the literature, in which two different techniques can be used to move the process itself away from the steady-state, inducing transients.     

Novel technique for component monitoring of CO2 exchange in plants

A novel technique designed for component monitoring of CO₂ exchange in plants is described. The system is based on application of self-clamping leaf chambers connected to an open gas-exchange measuring system and on automatic recording of CO₂ concentration. This technique was implemented in a commercially available instrument, PTM-48A Photosynthesis Monitor, which provides for long-term measurements of gas exchange and for discrimination of its separate components. Furthermore, many other plant functions can be monitored during plant growth and development under laboratory, greenhouse, and field conditions.     

Measurement of CO(2) Dissolved in Aqueous Solutions Using a Modified Infrared Gas Analyzer System

Total dissolved inorganic carbon (ΣCO₂) and aqueous carbon dioxide (H₂CO₃^*) in nutrient solutions may be measured by the injection of small gas or liquid samples (1 microliter to 8 milliliters) into a gas stripping column connected in-line with an infrared gas analyzer. The measurement of ΣCO₂ in solution requires sample acidification, while H₂CO₃^* and gaseous CO₂ are measured without the addition of lactic acid. The standard curve for ΣCO₂ was linear up to 300 nanomoles CO₂. Maximum sensitivity was approximately 300 picomoles. Measurements of H₂CO₃^* were independent of pH. Consequently, ΣCO₂ and H₂CO₃^* could be used to calculate the pH, HCO₃⁻, and CO₃²⁻ values of nutrient solutions. Injection and complete analyses required from 0.8 to 2 minutes.     

The contribution of entrapped gas bubbles to the soil methane pool and their role in methane emission from rice paddy soil in free-air [CO2] enrichment and soil warming experiments

Purpose

We attempted to determine the contribution of entrapped gas bubbles to the soil methane (CH₄) pool and their role in CH₄ emissions in rice paddies open to the atmosphere.

Methods

We buried pots with soil and rice in four treatments comprising two atmospheric CO₂ concentrations (ambient and ambient +200 μmol mol^?1) and two soil temperatures (ambient and ambient +2 °C). Pots were retrieved for destructive measurements of rice growth and the gaseous CH₄ pool in the soil at three stages of crop development: panicle formation, heading, and grain filling. Methane flux was measured before pot retrieval.

Results

Bubbles that contained CH₄ accounted for a substantial fraction of the total CH₄ pool in the soil: 26–45 % at panicle formation and 60–68 % at the heading and grain filling stages. At panicle formation, a higher CH₄ mixing ratio in the bubbles was accompanied by a greater volume of bubbles, but at heading and grain filling, the volume of bubbles plateaued and contained ~35 % CH₄. The bubble-borne CH₄ pool was closely related to the putative rice-mediated CH₄ emissions measured at each stage across the CO₂ concentration and temperature treatments. However, much unexplained variation remained between the different growth stages, presumably because the CH₄ transport capacity of rice plants also affected the emission rate.

Conclusions

The gas phase needs to be considered for accurate quantification of the soil CH₄ pool. Not only ebullition but also plant-mediated emission depends on the gaseous-CH₄ pool and the transport capacity of the rice plants.     

Stability and reactivity of liposome‐encapsulated formate dehydrogenase and cofactor system in carbon dioxide gas‐liquid flow

Formate dehydrogenase from Candida boidinii (CbFDH) is potentially applicable in reduction of CO₂ through oxidation of cofactor NADH into NAD⁺. For this, the CbFDH activity needs to be maintained under practical reaction conditions, such as CO₂ gas‐liquid flow. In this work, CbFDH and cofactor were encapsulated in liposomes and the liposomal enzymes were characterized in an external loop airlift bubble column. The airlift was operated at 45°C with N₂ or CO₂ as gas phase at the superficial gas velocity U_G of 2.0 or 3.0 cm/s. The activities of liposomal CbFDH/cofactor systems were highly stable in the airlift regardless of the type of gas phase because liposome membranes prevented interactions of the encapsulated enzyme and cofactor molecules with the gas‐liquid interface of bubbles. On the other hand, free CbFDH was deactivated in the airlift especially at high U_G with CO₂ bubbles. The liposomal CbFDH/NADH could catalyze reduction of CO₂ in the airlift giving the fractional oxidation of the liposomal NADH of 23% at the reaction time of 360 min. The cofactor was kept inside liposomes during the reaction operation with less than 10% of leakage. All of the results obtained demonstrate that the liposomal CbFDH/NADH functions as a stable catalyst for reduction of CO₂ in the airlift. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

Experimental validation of a nonequilibrium model of CO<Subscript>2</Subscript> fluxes between gas,liquid medium,and algae in a flat-panel photobioreactor

Carbon dioxide (CO₂) availability strongly affects the productivity of algal photobioreactors, where it is dynamically exchanged between different compartments, phases, and chemical forms. To understand the underlying processes, we constructed a nonequilibrium mathematical model of CO₂ dynamics in a flat-panel algal photobioreactor. The model includes mass transfer to the algal suspension from a stream of bubbles of CO₂-enriched air and from the photobioreactor headspace. Also included are the hydration of dissolved CO₂ to bicarbonate ion (HCO₃⁻) as well as uptake and/or cycling of these two chemical forms by the cells. The model was validated in experiments using a laboratory-scale flat-panel photobioreactor that controls light, temperature, and pH and where the concentration of dissolved CO₂, and partial pressure of CO₂ in the photobioreactor exhaust are measured. First, the model prediction was compared with measured CO₂ dynamics that occurred in response to a stepwise change in the CO₂ partial pressure in the gas sparger. Furthermore, the model was used to predict CO₂ dynamics in photobioreactors with unicellular, nitrogen-fixing cyanobacterium Cyanothece sp. The metabolism changes dramatically during a day, and the distribution of CO₂ is expected to exhibit a pronounced diurnal modulation that significantly deviates from chemical equilibrium.     

Intercellular Diffusion Limits to CO(2) Uptake in Leaves : Studies in Air and Helox

We studied plants of five species with hypostomatous leaves, and six with amphistomatous leaves, to determine the extent to which gaseous diffusion of CO₂ among the mesophyll cells limits photosynthetic carbon assimilation. In helox (air with nitrogen replaced by helium), the diffusivities of CO₂ and water vapor are 2.3 times higher than in air. For fixed estimated CO₂ pressure at the evaporating surfaces of the leaf (pⁱ), assimilation rates in helox ranged up to 27% higher than in air for the hypostomatous leaves, and up to 7% higher in the amphistomatous ones. Thus, intercellular diffusion must be considered as one of the processes limiting photosynthesis, especially for hypostomatous leaves. A corollary is that CO₂ pressure should not be treated as uniform through the mesophyll in many leaves. To analyze our helox data, we had to reformulate the usual gas-exchange equation used to estimate CO₂ pressure at the evaporating surfaces of the leaf; the new equation is applicable to any gas mixture for which the diffusivities of CO₂ and H₂O are known. Finally, we describe a diffusion-biochemistry model for CO₂ assimilation that demonstrates the plausibility of our experimental results.     

Continuous monitoring of rhizosphere respiration after labelling of plant shoots with 14CO2

The present work describes an original method to follow rate of ¹⁴CO₂ and total CO₂ production from rhizosphere respiration after plant shoots had been pulse-labelled with ¹⁴CO₂. We used a radioactivity detector equipped with a plastic cell for flow detection of beta radiation by solid scintillation counting. The radioactivity detector was coupled with an infrared gas analyser. The flow detection of ¹⁴CO₂ was compared to trapping of ¹⁴CO₂ in NaOH and counting by liquid scintillation. First, we demonstrated that NaOH (1 M) trapped 95% of the CO₂ of a gaseous sample. Then, we determined that the counting efficiency of the radioactivity flow cell was 41% of the activity of gaseous samples as determined by trapping in NaOH (1 M) and by counting by static liquid scintillation. The sensitivity of the ¹⁴CO₂- flow detection was 0.08 Bq mL⁻¹ air and the precision was 2.9% of the activity measured compared to 0.9% for NaOH trapping method. We presented two applications which illustrate the relevance of ¹⁴CO₂-flow detection to investigations using 14C to trace photoassimilates within the plant-soil system. First, we examined the kinetics of 14CO₂ production when concentrated acid is added to NaH¹⁴CO₃. This method is the most commonly used to label photoassimilates with ¹⁴C. Then, we monitored ¹⁴CO₂ activity in rhizosphere respiration of 5-week old maize cultivated in soil and whose shoots had been pulse-labelled with ¹⁴CO₂. We conclude that alkali traps should be used for a cumulative determination of ¹⁴CO₂ because they are cheap and accurate. On the other hand, we demonstrated that the flow detection of ¹⁴CO₂ had a finer temporal resolution and was consequently a relevant tool to study C dynamics in the rhizosphere at a short time scale. This revised version was published online in June 2006 with corrections to the Cover Date.     

Membrane oxygenators: current developments in design and application

Cardiopulmonary bypass (CPB) procedures require a blood-gas exchanger (oxygenator) to temporarily replace the respiratory function of the lungs. In the past the majority of CPB procedures have been carried out with bubble oxygenators which effect gas exchange by dispersion of bubbles into the blood. Membrane oxygenators, on the other hand, utilize a hydrophobic gas permeable membrane between the blood and gas phases.Bubble oxygenators are being superseded by membrane types for CPB due to improvements in membrane technology and mass transfer efficiency. These advances are reviewed in this paper and are illustrated by reference to the gas exchange and operating characteristics of a number of clinical oxygenators designed for adult CPB.Membrane oxygenatorsare also being used for long-term support in the treatment of acute respiratory failure. Operated in a partial bypass circuit, the oxygenator may have to function for several days or weeks. In one particular treatment method, the rate of spontaneous breathing is controlled by the partial or total removal of the metabolic CO₂ production by the membrane oxygenator. For this method, known as extracorporeal CO₂ removal (ECCO₂R), the oxygenator must be optimized for CO₂ transfer at low blood flow rates. The suitability of clinical oxygenators for ECCO₂R is discussed in terms of gas exchange and functionality over a prolonged operation.     

The effects of chamber pressurization on soil-surface CO2 flux and the implications for NEE measurements under elevated CO2

Soil and ecosystem trace gas fluxes are commonly measured using the dynamic chamber technique. Although the chamber pressure anomalies associated with this method are known to be a source of error, their effects have not been fully characterized. In this study, we use results from soil gas-exchange experiments and a soil CO₂ transport model to characterize the effects of chamber pressure on soil CO₂ efflux in an annual California grassland. For greater than ambient chamber pressures, experimental data show that soil-surface CO₂ flux decreases as a nonlinear function of increasing chamber pressure; this decrease is larger for drier soils. In dry soil, a gauge pressure of 0.5 Pa reduced the measured soil CO₂ efflux by roughly 70% relative to the control measurement at ambient pressure. Results from the soil CO₂ transport model show that pressurizing the flux chamber above ambient pressure effectively flushes CO₂ from the soil by generating a downward flow of air through the soil air-filled pore space. This advective flow of air reduces the CO₂ concentration gradient across the soil–atmosphere interface, resulting in a smaller diffusive flux into the chamber head space. Simulations also show that the reduction in diffusive flux is a function of chamber pressure, soil moisture, soil texture, the depth distribution of soil CO₂ generation, and chamber diameter. These results highlight the need for caution in the interpretation of dynamic chamber trace gas flux measurements. A portion of the frequently observed increase in net ecosystem carbon uptake under elevated CO₂ may be an artifact resulting from the impact of chamber pressurization on soil CO₂ efflux.     

RuBPCO kinetics and the mechanism of CO2 entry in C3 plants

Abstract. The CO₂ partial pressure in the chloroplasts of intact photosynthetic C₃ leaves is thought to be less than the intercellular CO₂ partial pressure. The intercellular CO₂ partial pressure can be calculated from CO₂ and H₂O gas exchange measurements, whereas the CO₂ partial pressure in the chloroplasts is unknown. The conductance of CO₂ from the intercellular space to the chloroplast stroma and the CO₂ partial pressure in the chloroplast stroma can be calculated if the properties of photosynthetic gas exchange are compared with the kinetics of the enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBPCO). A discrepancy between gas exchange and RuBPCO kinetics can be attributed to a deviation of CO₂ partial pressure in the chloroplast stroma from that calculated in the intercellular space. This paper is concerned with the following: estimation of the kinetic constants of RuBPCO and their comparison with the CO₂ compensation concentration; their comparison with differential uptake of ¹⁴CO₂ and ¹²CO₂; and their comparison with O₂ dependence of net CO₂ uptake of photosynthetic leaves. Discrepancy between RuBPCO kinetics and gas exchange was found at a temperature of 12.5 °C, a photosynthetic photon flux density (PPFD) of 550 μmol quanta m^?2 s^?1, and an ambient CO₂ partial pressure of 40 Pa. Consistency between RuBPCO kinetics and gas exchange was found if CO₂ partial pressure was decreased, temperature incresed and PPFD decreased. The results suggest that a discrepancy between RuBPCO kinetics and gas exchange is due to a diffusion resistance for CO₂ across the chloroplast envelope which decreases with increasing temperature. At low CO₂ partial pressure, the diffusion resistance appears to be counterbalanced by active CO₂ (or HCO₃) transport with high affinity and low maximum velocity. At low PPFD, CO₂ partial pressure in the chloroplast stroma appears to be in equilibrium with that in the intercellular space due to low CO₂ flux.     

Continuous Measurements of the Free Dissolved CO(2) Concentration during Photosynthesis of Marine Plants: Evidence for HCO(3) Use in Chondrus crispus

An experimental system consisting of a gas exchange column linked to an assimilation chamber has been developed to record continuously the free dissolved CO₂ concentration in seawater containing marine plants. From experiments performed on the red macroalga Chondrus crispus (Rhodophyta, Gigartinales), this measurement is in agreement with the free CO₂ concentration calculated from the resistance to CO₂ exchanges in a biphasic system (gas and liquid) as earlier reported. The response time of this apparatus is short enough to detect, in conditions of constant pH, a photosynthesis-caused gradient between free CO₂ and HCO₃⁻ pools which half-equilibrates in 25 seconds. Abolished by carbonic anhydrase, the magnitude of this gradient increases with decreasing time of seawater transit from the chamber to the column apparatus. But its maximum magnitude (0.35 micromolar CO₂) is negligible compared to the difference between air and free CO₂ (11.4 micromolar CO₂). This illustrates the extent of the physical limiting-step occurring at the air-water interface when inorganic carbon consumption in seawater is balanced by dissolving gaseous CO₂. The direction of this small free CO₂/HCO₃⁻ gradient indicates that HCO₃⁻ is consumed during photosynthesis.     

A Direct Confirmation of the Standard Method of Estimating Intercellular Partial Pressure of CO(2)

The partial pressure of CO₂ inside leaves of several species was measured directly. Small gas exchange chambers were clamped above and below the same section of an amphistomatous leaf. A flowing gas stream through one chamber allowed normal CO₂ and water vapor exchange. The other chamber was in a closed circuit consisting of the chamber, an infrared gas analyzer, and a peristaltic pump. The CO₂ in the closed system rapidly reached a steady pressure which it is believed was identical to the CO₂ pressure inside the leaf, because there was no flux of CO₂ across the epidermis. This measured partial pressure was in close agreement with that estimated from a consideration of the fluxes of CO₂ and vapor at the other surface.     

Automatic gas tank switching system for CO2 incubators

Summary The use and construction of an automatic gas tank switching system are described. This device monitors the gas pressure in a CO₂ incubator gas system and automatically switches to a reserve tank when the main supply tank is depleted. The unit contains an alarm system that signals either a loss of power or of gas pressure in the supply system. This research was supported by National Cancer Institute Contract No 1-CP-33226, grants CA 13058 and CA 14680 and an institutional grant to the Michigan Cancer Foundation by the United Foundation of Greater Detroit.     

Spiracle activity in moth pupae—The role of oxygen and carbon dioxide revisited

After decades of intensive research, the actual mechanism behind discontinuous gas exchange in insects has not been fully understood. One open question concerns the actual way (closed, flutter, and open) of how spiracles respond to tracheal gas concentrations. As the results of a classic paper [Burkett, B.N., Schneiderman, H.A., 1974. Roles of oxygen and carbon dioxide in the control of spiracular function in cecropia pupae. Biological Bulletin 147, 274-293] allow ambiguous interpretation, we thus reexamined the behavior of the spiracles in response to fixed, controlled endotracheal gas concentrations.The tracheal system of diapausing pupae of Attacus atlas (Saturniidae, Lepidoptera) was flushed with gas mixtures varying in PO₂ and PCO₂ while the behavior of the spiracles was monitored using changes in the pressure signal. This novel pressure based technique proved to be superior to classic visual observation of single spiracles. A two-dimensional map of the spiracle behavior in response to endotracheal PO₂ and PCO₂ was established. Typically, it contained two distinct regions only, corresponding to “closed” and “open” spiracles. A separate “flutter” region was missing. Because fluttering is commonly observed in moth pupae, we suggest that the intermittent spiracle opening during a flutter phase is an effect of non-steady-state conditions within the tracheal system. For low PCO₂ the minimum PO₂ resulting in open spiracles was linearly dependent upon PCO₂. Above a threshold of 1-1.5 kPa CO₂ the spiracles were open irrespective of PO₂. We propose a hypothetical spiracular control model, which is simple and explains the time course of endotracheal partial pressures during all phases of discontinuous gas exchange.     

Technique for Measuring 14CO2 Uptake by Soil Microorganisms In Situ

Uptake of ¹⁴CO₂ in soils due to algae or sulfur-oxidizing bacteria was examined by incubation of soil samples with gaseous ¹⁴CO₂ and subsequent chemical oxidation of biologically fixed radioactive isotope to ¹⁴CO₂ for detection with a liquid scintillation counting system. The ¹⁴CO₂ was added to the soil in the gas phase so that no alteration of the moisture or ionic strength of the soil occurred. Wet oxidation of radioactive organic matter was carried out in sealed ampoules, and the ¹⁴CO₂ produced was transferred to a phenethylamine-liquid scintillation counting system with a simply constructed apparatus. The technique is inexpensive and efficient and does not require elaborate traps since several possible interfering factors were found to have no harmful effects. Experiments in coal mine regions and in geothermal habitats have demonstrated the ecological applicability of this technique for measurement of CO₂ fixation by sulfur-oxidizing bacteria and soil algae.