Recovery Of 14CO2 Research Articles

Cholesteryl octanoate breath test. Preliminary studies on a new noninvasive test of human pancreatic exocrine function.

A new breath test for noninvasive assessment of pancreatic exocrine function in humans was developed. The test is based on the hydrolysis of cholesteryl-[1-14C]octanoate by pancreatic carboxyl ester lipase (cholesterol esterase) with subsequent absorption and hepatic metabolism of the liberated octanoate to 14CO2. The rate at which 14CO2 appears in breath appeared to be proportional to the rate of hydrolysis. The substrate is administered as a gum acacia stabilized emulsion of vegetable oil (18 g) containing cholesteryl octanoate (2 g; 4.4 microCi) dispersed in a 500-ml isotonic meal. Tests were performed in 6 healthy volunteers and 11 patients with pancreatic disease with varying degrees of steatorrhea. In healthy subjects, 14CO2 output was rapid with peak output occurring at 60-90 min in all subjects; cumulative output in 4 h averaged 30%. Duplicate studies indicated that the time-course of 14CO2 recovery was reproducible. The pattern of 14CO2 output in patients with pancreatic disease varied widely. Patients without steatorrhea (fecal fat less than or equal to 7 g/day) or with mild steatorrhea (fecal fat 7-11 g/day) had normal or near normal patterns of 14CO2 output, whereas patients with moderate or severe steatorrhea (fecal fat greater than 11 g/day) expired 14CO2 at a rate one-third to one-tenth that of the healthy volunteers. Addition of pancreatic enzyme supplementation to the test meal increased 14CO2 output in 6 of 6 patients with moderate or severe steatorrhea, suggesting that the activity of pancreatic carboxyl ester lipase was rate limiting in these patients. In an additional study in a healthy volunteer, 14CO2 and 13CO2 were measured simultaneously in breath after ingestion of a test meal containing cholesteryl-[1-13C]octanoate and 14C-octanoate. 14CO2 was expired more rapidly than 13CO2, suggesting that hydrolysis of the substrate may also be rate limiting in healthy volunteers. These studies indicate that severe pancreatic exocrine dysfunction can be detected with a simple breath test in 60-90 min.

Maternal Hypoxia Decreases Capillary Supply and Increases Metabolic Inefficiency Leading to Divergence in Myocardial Oxygen Supply and Demand

Maternal hypoxia is associated with a decrease in left ventricular capillary density while cardiac performance is preserved, implying a mismatch between metabolism and diffusive exchange. We hypothesised this requires a switch in substrate metabolism to maximise efficiency of ATP production from limited oxygen availability. Rat pups from pregnant females exposed to hypoxia (FIO2=0.12) at days 10-20 of pregnancy were grown to adulthood and working hearts perfused ex vivo. 14C-labelled glucose and 3H-palmitate were provided as substrates and metabolism quantified from recovery of 14CO2 and 3H2O, respectively. Hearts of male offspring subjected to Maternal Hypoxia showed a 20% decrease in cardiac output (P<0.05), despite recording a 2-fold increase in glucose oxidation (P<0.01) and 2.5-fold increase (P<0.01) in palmitate oxidation. Addition of insulin to Maternal Hypoxic hearts, further increased glucose oxidation (P<0.01) and suppressed palmitate oxidation (P<0.05), suggesting preservation in insulin signalling in the heart. In vitro enzyme activity measurements showed that Maternal Hypoxia increased both total and the active component of cardiac pyruvate dehydrogenase (both P<0.01), although pyruvate dehydrogenase sensitivity to insulin was lost (NS), while citrate synthase activity declined by 30% (P<0.001) and acetyl-CoA carboxylase activity was unchanged by Maternal Hypoxia, indicating realignment of the metabolic machinery to optimise oxygen utilisation. Capillary density was quantified and oxygen diffusion characteristics examined, with calculated capillary domain area increased by 30% (P<0.001). Calculated metabolic efficiency decreased 4-fold (P<0.01) for Maternal Hypoxia hearts. Paradoxically, the decline in citrate synthase activity and increased metabolism suggest that the scope of individual mitochondria had declined, rendering the myocardium potentially more sensitive to metabolic stress. However, decreasing citrate synthase may be essential to preserve local PO2, minimising regions of hypoxia and hence maximising the area of myocardium able to preserve cardiac output following maternal hypoxia.

Widespread Distribution of Soluble Di-Iron Monooxygenase (SDIMO) Genes in Arctic Groundwater Impacted by 1,4-Dioxane

Soluble di-iron monooxygenases (SDIMOs), especially group-5 SDIMOs (i.e., tetrahydrofuran and propane monooxygenases), are of significant interest due to their potential role in the initiation of 1,4-dioxane (dioxane) degradation. Functional gene array (i.e., GeoChip) analysis of Arctic groundwater exposed to dioxane since 1980s revealed that various dioxane-degrading SDIMO genes were widespread, and PCR-DGGE analysis showed that group-5 SDIMOs were present in every tested sample, including background groundwater with no known dioxane exposure history. A group-5 thmA-like gene was enriched (2.4-fold over background, p < 0.05) in source-zone samples with higher dioxane concentrations, suggesting selective pressure by dioxane. Microcosm assays with (14)C-labeled dioxane showed that the highest mineralization capacity (6.4 ± 0.1% (14)CO2 recovery during 15 days, representing over 60% of the amount degraded) corresponded to the source area, which was presumably more acclimated and contained a higher abundance of SDIMO genes. Dioxane mineralization ceased after 7 days and was resumed by adding acetate (0.24 mM) as an auxiliary substrate to replenish NADH, a key coenzyme for the functioning of monoxygenases. Acetylene inactivation tests further corroborated the vital role of monooxygenases in dioxane degradation. This is the first report of the prevalence of oxygenase genes that are likely involved in dioxane degradation and suggests their usefulness as biomarkers of dioxane natural attenuation.

Octanoate and Nonaoate Oxidation Increases 50–80% Over the First Two Days of Life in Piglet Triceps Brachii and Gracilis Muscle Strips ,

An in vitro muscle strip incubation system was developed to measure the rate of catabolism of 1 mmol/L [1-14C]octanoate, 1 mmol/L [1-14C]nonanoate, 1 mmol/L [9-14C]nonanoate, and 10 mmol/L [U-14C]glucose by measuring the recovery of14CO2. Muscle strips (13 mm × 1.5 mm, ~50 mg) were isolated from triceps brachii and gracilis muscles of newborn and 2-d-old, small (<950 g) and large (>1450 g) piglets. The position of the14C label in the substrate affected the rate and amount of recovery in14CO2. Therefore, comparisons were made between age groups (0 vs. 2 d old) within substrates but limited across substrates to comparisons of [1-14C]–labeled fatty acids. The medium-chain fatty acid (MCFA) oxidation rates [pmol/(h · mg)] in muscle strips isolated from piglets from the 2 weight groups (<950 and >1450 g) did not differ (P> 0.99), there was a trend towards a difference between triceps brachii and gracilis muscle (P= 0.09; data not shown), and there were no significant interactions involving pig weight or muscle type; therefore, results were pooled across these factors. During the first 2 d of life, MCFA oxidation [pmol/(h · · mg muscle strip)] increased (P< 0.05) 50–80%, but the glucose oxidation rate did not change (P> 0.82). By d 2, the oxidation rate of nonanoate as represented by the one carbon was 25% greater than for octanoate (P< 0.05). The conversion of [9-14C]nonanoate to14CO2indicated that muscle had the capacity to oxidize the propionyl-CoA produced by β-oxidation of nonanoate and that odd-chain C-9 MCFA provided anabolic carbon to the citric acid cycle.

Tissue distribution of lysine metabolism in commercial turkeys

The essential amino acid lysine is typically the first or second limiting amino acid in diets fed to growing turkeys. However, there is little information about how and where lysine is degraded. Therefore, an investigation of the tissue distribution of lysine catabolism was performed using liver, kidney, enterocytes, breast, thigh, heart and lung of four and eight week old turkeys. The tissues were assayed for the initial enzyme activity of the primary pathway of lysine oxidation, lysine ketoglutarate reductase (LKR). Additionally, lysine oxidation (LOX) was determined based on the recovery of 14CO2 from radioactive lysine. All tissues assayed had measureable LKR activity except lung. Liver had the highest activity (5.2 mmol/min*g liver). Intestine, kidney and liver oxidized lysine (P&lt;0.05) and liver had the highest lysine oxidation (15.9 μmol/min*g tissue). A significant week * tissue interaction and week effect were detected. A positive correlation (P&lt;0.05) was observed between LKR activity and LOX. In conclusion, it is clear that extra hepatic tissues contribute to whole body lysine metabolism. (Support: HATCH WVA 470, Virginia Poultry Grower Cooperative)

Effect of artichoke extract (Cynara scolymus L.) on palmitic‐1‐14C acid oxidation in rats

Studies on the effect of the artichoke extract (AE) on oxidation of palmitic-1-14C acid administered intravenously to rats at a dose 25 and 50 mg/kg bw demonstrated marked enhancement of both 14CO2 expiration rate and 14CO2 recovery in the expired air. The extract suppressed accumulation of palmitic-1-14C acid in serum lipids and epididymal fat pad tissue as well. The effects of the extract on 14CO2 expiration rate, 14CO2 recovery, as well as accumulation of palmitic-1-14C acid were dose dependent. Total14CO2 recovery in expired air during 60 min was elevated by 17.3% (p < 0.05) and 52.1% (p < 0.001) in rats administered the extract at a dose of 25 and 50 mg/kg, respectively. The rats supplemented with the AE at a dose of 25 and 50 mg/kg bw were characterized by 10.0% (not significant) and 19% (p < 0.05) decrease in( 14)C radioactivity of serum lipids as well as reduction of epididymal fat tissue 14C radioactivity by 8.7 and 17.5% (p < 0.05), respectively, in comparison with the control rats. Thus, the results demonstrate that the AE possess stimulatory properties with respect to oxidation of palmitic acid administered to rats, and provide new information on the mechanism of antilipemic activity of the extract associated with activation of lipid oxidation in the organism.

Degradation of 14C-Zinc Ammonium Acetate in Soils as Influenced by Soil Types, Soil Sterilization, and Carriers

ABSTRACT Zinc ammonium acetate (ZAA), typically applied to soils in anhydrous ammonia as a carrier, has been used to improve corn (Zea maysL.) productivity. This study aimed to determine the fate of ZAA in soils as influenced by soil type (sandy, silt, and clay loam), sterilization (sterile and non-sterile), and two carriers (H2O and NH4OH). A 16 d laboratory incubation experiment with 14C-ZAA showed that total recovery of carbon-14 (14C) from 14CO2 trap and soil extraction by CaCl2 ranged from 72% to 94% in the first 8 d for sterilized soils. However, < 17% 14C was found in non-sterilized soils. Most 14C recovered in sterilized soil was associated with soil extraction, and relatively little was found in the CO2 traps. All sterilized soils provided similar 14C recoveries except the sandy loam. Slightly more 14C was extracted from the soil when NH4OH was the ZAA carrier rather than water. Conversely, recovery of 14CO2 continued to increase during the 16 d incubation, but started faster when water was the ZAA carrier. Microbial activity appeared to be instrumental in the assimilation and disappearance of ZAA.

A Model for the Effect of Rhizodeposition on the Fate of Phenanthrene in Aged Contaminated Soil

Microcosm data were used to develop a deterministic model to describe how rhizodeposition affects the fate of phenanthrene in aged contaminated soil. Microbial mineralization and soil sequestration of 14C-phenanthrene were compared in microcosms amended weekly with phenolic-rich mulberry root extracts versus unamended controls. Mineralization was higher in the amended soils simulating the rhizosphere (57.7 +/- 0.9%) than in controls simulating bulk (unplanted) soils (53.2 +/- 0.7%) after 201 days (p < 0.05). Humin was the main soil sink for the residual 14C-label. Whereas the total 14C-label associated with humin remained constant in biologically active soils (at about 30%), it increased up to 80% after 201 days in sterile controls. The initial phenanthrene extraction with n-butanol (commonly used to assess bioavailability) slightly underestimated the fraction thatwas mineralized (assessed by 14CO2 recovery). Changes in the unextractable fraction (determined by combustion in a biological oxidizer) suggested the presence of two soil sequestration domains: (1) irreversibly bound residue, and (2) an intermediate transition phase that is unextractable by solvents at a given point in time but could become bioavailable due to physicochemical or biological transformations of the binding matrix. The fate of phenanthrene was accurately modeled by considering the transfer of the 14C label between different soil compartments as first-order kinetic processes. Model simulations suggested that the system was approaching a stable end-point after 201 days of simulated rhizoremediation, and corroborated that microorganisms have a significant impact on the fate of phenanthrene in soil.

Oxygen‐Enhanced Biodegradation of Phenoxy Acids in Ground Water at Contaminated Sites

The effects of adding oxygen to anaerobic aquifer materials on biodegradation of phenoxy acid herbicides were studied by laboratory experiments with aquifer material from two contaminated sites (a former agricultural machinery service and an old landfill). At both sites, the primary pollutants were phenoxy acids and related chlorophenols. It was found that addition of oxygen enhanced degradation of the six original phenoxy acids and six original chlorophenols. Inverse modeling on 14C 4-chloro-2-methylphenoxypropanoic acid (MCPP) degradation curves revealed that increasing the oxygen concentrations from <0.3 mg/L up to 7 to 8 mg/L shortened the lag phases (from approximately 150 d to 5 to 25 d) and increased first-order degradation rate constants by 1 order of magnitude (from approximately 5 x 10(-2) d(-1) to up to 30 x 10(-2) d(-1)). Additionally, the degree of MCPP mineralization was increased (30% to 50% mineralized at low oxygen concentrations and 50% to 70% mineralized at high oxygen concentrations, based on 14CO2 recovery). These positive effects on degradation were observed even at relatively low oxygen concentrations (2 mg/L). Furthermore, effects related to the addition of oxygen on the general geochemistry were studied. An oxygen consumption of 2.2 to 2.6 mg O2/g dw was observed due to oxidation of solid organic matter and, to some extent (0.5% to 11% of the total oxygen consumption), water-soluble compounds such as Fe2+, dissolved Mn, nonvolatile organic carbon, and NH4+. Overall, the results suggest that stimulated biodegradation by addition of oxygen might be a feasible remediation technology at herbicide-contaminated sites, although oxygen consumption by the sediment could limit the applicability.

Controlled Release of Nitrate and Sulfate to Enhance Anaerobic Bioremediation of Phenanthrene in Marine Sediments

Experimental measurements demonstrated that rates of in situ microbial anaerobic biodegradation of phenanthrene in undisturbed marine sediments were enhanced when controlled-release electron acceptors (i.e., nitrate and sulfate (for comparison)) were employed. The experimental method used whole and interval cores injected with radiolabeled 14C-phenanthrene, which were incubated, sacrificed, and processed for 14CO2 recovery to determine degradation rates. Nitrocellulose and CaSO4 were formulated to release the electron acceptors into the sediments at rates consistent with bacterial utilization (i.e., that avoided inhibition observed previously at high soluble nitrate (although not sulfate) concentrations). The controlled-release of both compounds, measured in collateral experiments, enhanced the natural anaerobic phenanthrene biodegradation rates by factors up to 2-3. Biodegradation via sulfate reduction was most rapid in the early stages (24 days) of experiments, consistent with reports that many marine bacteria in submerged sediments are sulfate reducers. In comparison, in longer experiments (after 42 days), the anaerobic biodegradation rates were observed at least as high with the addition of nitrocellulose (over 40% of added phenanthrene recovered as 14CO2). Both nitrate and sulfate reduction was observed during anaerobic incubation, although the presence of nitrate seemed to reduce the sulfate reduction. The studied forms of the controlled-release nitrate and sulfate may provide capping amendments to decontaminate marine harbor sediments.

Mineralization of soil-aged isoproturon and isoproturon metabolites by Sphingomonas sp. strain SRS2.

The aim of the study was to determine the effect of aging of the herbicide isoproturon and its metabolites monodesmethyl-isoproturon and 4-isopropyl-aniline in agricultural soil on their availability to the degrading bacterium Sphingomonas sp. strain SRS2. The 14C-ring-labeled isoproturon, monodesmethyl-isoproturon, and 4-isopropyl-aniline were added to sterilized soil and stored for 1, 49, 71, or 131 d before inoculation with strain SRS2. The availability of the compounds was estimated from the initial mineralization and the amount of 14CO2 recovered after 120 d of incubation. Aging in soil for 131 d reduced the initial mineralization of isoproturon and monodesmethyl-isoproturon and, in the case of isoproturon, also reduced the recovery of 14CO2. Initial mineralization and recovery of 14CO2 from aged 4-isopropyl-aniline were slightly reduced, but less 14CO2 was generally produced than with isoproturon or monodesmethyl-isoproturon. Thus, recovery of 14CO2 from 14C-isoproturon and 14C-monodesmethyl-isoproturon was 50.7 to 64.4% of the initially added 14C, while recovery from 14C-4-isopropyl-aniline was only 11.7 to 17.0%. Sorption measurements revealed similar Freundlich constants (K(f)) for isoproturon and monodesmethyl-isoproturon, whereas K(f) for 4-isopropyl-aniline was more than fivefold greater. The findings imply that in soil, partial degradation of isoproturon to 4-isopropyl-aniline may lead to reduced mineralization of the herbicide due to sorption of the aniline moiety.

Mineralization of aged atrazine, terbuthylazine, 2,4‐D, and mecoprop in soil and aquifer sediment

The effect of aging of the herbicides atrazine, terbuthylazine, 2,4-D, and mecoprop on their bioavailability to degrading microorganisms was studied in soil and aquifer sediment. 14C-ring-labeled herbicide (2.5 mg/kg) was added to sterilized soil or aquifer sediment and stored at 10 degrees C for up to 103 d before inoculation with either the atrazine and terbuthylazine-degrading Pseudomonas sp. strain ADP (atrazine-degrading Pseudomonas) or an enriched culture able to mineralize 2,4-D and mecoprop. The initial mineralization rate and recovery of 14CO2 after 62 to 113 d of incubation were used as measures of the availability of the compounds to the microorganisms. Aging in soil reduced the initial mineralization of atrazine. Thus, only 17% of the added 14C-atrazine had been mineralized after 21 h of incubation when aged for 88 d as compared with 33% when the atrazine had been aged for 1 d. 14CO2 recovery was only 58% after 88 d of aging as compared with 81% when aged for 1 d. A similar effect of aging was seen with terbuthylazine. With 2,4-D, the effect of aging in soil on mineralization by the enriched culture was much smaller. Aging had no effect on mineralization of mecoprop in soil or on mineralization of any of the herbicides in aquifer sediment.

Hexahydro-1,3,5-trinitro-1,3,5-triazine mineralization by zerovalent iron and mixed anaerobic cultures.

Soil microcosms were used to evaluate the potential benefits of an integrated microbial-Fe0 system to treat groundwater contamination by RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine). Microcosms amended with both Fe0 filings and municipal anaerobic sludge mineralized RDX faster and to a greater extent than separate treatments, with up to 51% 14CO2 recovery after 77 d. The nitroso byproducts 1,3-dinitro-5-nitroso-1,3,5-triazacyclohexane (MNX), 1,3-dinitroso-5-nitro-1,3,5-triazacyclohexane (DNX), and 1,3,5-trinitroso-1,3,5-triazacyclohexane (TNX) were detected in all microcosms, although these compounds never accumulated above 5% of the added RDX on a molar basis. A soluble intermediate that was tentatively identified as methylenedinitramine [(O2NNH)2CH2] was relatively persistent, although it accumulated to a much lower extent in combined-treatment reactors than in sets with Fe0 or sludge alone. Some of the radiolabel was bound to soil and Fe0 and could not be extracted with CH3CN. This fraction, which was recovered by combustion with a biological oxidizer, was also found at lower concentrations in combined-treatment reactors. This work suggests that permeable reactive Fe0 barriers might be an effective approach to intercept and degrade RDX plumes and that treatment efficiency might be enhanced by biogeochemical interactions through bioaugmentation.

Widespread potential for microbial MTBE degradation in surface-water sediments.

Microorganisms indigenous to stream and lake bed sediments, collected from 11 sites throughout the United States, demonstrated significant mineralization of the fuel oxygenate, methyl-tert-butyl ether (MTBE). Mineralization of [U-14C]MTBE to 14CO2 ranged from 15 to 66% over 50 days and did not differ significantly between sediments collected from MTBE contaminated sites and from sites with no history of MTBE exposure. This result suggests that even the microbial communities indigenous to newly contaminated surface water systems will exhibit some innate ability to attenuate MTBE under aerobic conditions. The magnitude of MTBE mineralization was related to the sediment grain size distribution. A pronounced, inverse correlation (p < 0.001; r2 = 0.73) was observed between the final recovery of 14CO2 and the percentage content of silt and clay sized grains (grain diameter < 0.125 mm). The results of this study indicate that the microorganisms that inhabit the bed sediments of streams and lakes can degrade MTBE efficiently and that this capability is widespread in the environment. Thus aerobic bed sediment microbial processes may provide a significant environmental sink for MTBE in surface water systems throughout the United States and may contribute to the reported transience of MTBE in some surface waters.

Acetogenic Microbial Degradation of Vinyl Chloride

Under methanogenic conditions, microbial degradation of [1,2-14C]vinyl chloride (VC) resulted in significant (14 ± 3% maximum recovery) but transient recovery of radioactivity as 14C-acetate. Subsequently, 14C-acetate was degraded to 14CH4 and 14CO2 (18 ± 2% and 54 ± 3% final recoveries, respectively). In contrast, under 2-bromoethanesulfonic acid (BES) amended conditions, 14C-acetate recovery remained high (27 ± 1% maximum recovery) throughout the study, no 14CH4 was produced, and the final recovery of 14CO2 was only 35 ± 4%. These results demonstrate that oxidative acetogenesis may be an important mechanism for anaerobic VC biodegradation. Moreover, these results (1) demonstrate that microbial degradation of VC to CH4 and CO2 may involve oxidative acetogenesis followed by acetotrophic methanogenesis and (2) suggest that oxidative acetogenesis may be the initial step in the net oxidation of VC to CO2 reported previously under Fe(III)-reducing, SO4-reducing, and humic acids-reducing conditions.

Role for Acetotrophic Methanogens in Methanogenic Biodegradation of Vinyl Chloride

Under methanogenic conditions, stream-bed sediment microorganisms rapidly degraded [1,2-14C]vinyl chloride to 14CH4 and 14CO2. Amendment with 2-bromoethanesulfonic acid eliminated 14CH4 production and decreased 14CO2 recovery by an equal molar amount. Results obtained with [14C]ethene, [14C]acetate, or 14CO2 as substrates indicated that acetotrophic methanogens were responsible for the production of 14CH4 during biodegradation of [1,2-14C]VC.

A Plant-Soil-Atmosphere Microcosm for Tracing Radiocarbon from Photosynthesis through Methanogenesis

We designed a CO2-controlled cuvette and stripping system to trace a 14CO2 pulse-label from photosynthetic assimilation by wetland plants (in this study Orontium aquaticum L.) to its release as 14CH4 by microbial respiration. The system maintained cuvette CO2 concentrations to within ±5 Pa of the set-point, and it allowed continuous recovery of 14CO2 and 14CH4 for 17 d without damage to the enclosed plant. The first emissions of 14CH4 were detected <12 h after photosynthetic assimilation of the label. The 14CH4 flux increased linearly from 0.12 Bq min-1 at 12 h to 3.0 Bq min-1 at 5 d, then plateaued at =2 Bq min-1. We could not distinquish between 14CH4 produced by aceticlastic methanogenesis vs. that produced by CO2 reduction. Radiocarbon activity in the soil dissolved inorganic C pool peaked on the first day then declined slowly. We did not detect radiocarbon activity in soil solution pools of several low molecular weight organic acids (acetate, formate, lactate, and propionate), but the label was detected in the bulk dissolved organic C pool. We argue that radiocarbon will be useful for investigating the contribution of root exudates to methanogenic metabolism, but data interpretation will require separation of the relative contributions of CO2 reduction and aceticlastic methanogenesis to overall 14CH4 emissions. Processes such as CH4 oxidation and acetogenesis must also be considered in quantitative estimates of photosynthetic support of methanogenesis.

Bacterial degradation of emulsified crude oil and the effect of various surfactants.

A Rhodococcus sp. 094 bacterium was tested for its ability to oxidize alkanes in crude oil emulsified by nonionic chemical and biological surfactants. Oxidation rates were measured in a 3-h period by Warburg respirometry. 14CO2 recovery was measured from the [1-14C]hexadecane spiked crude oil. Response to emulsified oil depended on the physiological state of the bacteria (i.e., cells harvested in the exponential and stationary growth phases) were tested. Oxidation rates by cells in the exponential growth phase were negatively affected by surfactant amendment. Oxidation rates by cells in the stationary growth phase were in some cases stimulated by surfactants. The stimulatory effect depended on both the chemical structure and the physicochemical properties (i.e., hydrophilic-lipophilic balance (HLB)) of the surfactants. Surfactants with intermediate HLB values (8-12) gave the best results. Neither the biosurfactants nor the commercial oil-spill dispersants tested had any significant stimulatory effect.

Recovery of 13CO2 and 14CO2 in human bicarbonate studies: a critical review with original data.

1. In order to establish biological and/or methodological explanations for the wide variability in recovery (50-100%) of labelled CO2 after administration of [13C]bicarbonate or [14C]bicarbonate, 34 human bicarbonate studies involving 480 subjects were analysed, and potential methodological issues were investigated in the laboratory. 2. Overall, continuous infusion studies reported a higher recovery than bolus studies (84 +/- 11% versus 69 +/- 12%; P < 0.001). No significant differences in recovery were found between 14C and 13C studies, children and adults, obese and lean subjects, or rest and exercise (steady state). Higher recoveries were found during feeding than during fasting (84 +/- 8% versus 74 +/- 7%; P < 0.001). Different methods used to analyse the results (0-10%) and different study protocols, which include differences in the duration of infusions and background drift in 13C enrichment (0-10%), contribute to the variability. 3. The laboratory studies suggest multiple sources of potential error, including loss of CO2 from the scintillation fluid (up to > 30%, but only in 14C studies in which the scintillation fluid is not alkalized), diffusion of CO2 through syringes and tubing (0 to > 10%), non-linearity of CO2 analysers (up to 8%), inaccuracies in the measurement of bicarbonate concentrations (13C studies) or the strength of CO2-trapping agents (14C studies; 0-8%). 4. It is concluded that much of the variability in the recovery of labelled bicarbonate is likely to be attributable to methodological differences, and that attention to these will ensure better interpretation of metabolic studies that involve oxidation of carbon-labelled substrates.

Nutritional influence on the recovery of 14CO2 in critically ill trauma patients.

Several factors, including age, nutritional and metabolic status, and underlying pathological conditions, should be considered in estimating the correction factor for labeled CO2 retention. We have evaluated the nutritional influence of this correction factor in a group of severely injured (injury severity score = 33 +/- 5), hypermetabolic, and highly catabolic adult trauma patients. Primed-constant infusion of NaH14CO3 was used, and the breath 14CO2 radioactivity was measured. The experiment was conducted once in the fasting condition during the early catabolic "flow" phase of injury and again during and after a week of intravenous feeding. The mean value of the 14CO2 recovery (85.2 +/- 2.7%) in the basal fasting condition was higher than the value of 81% commonly used in normal subjects. During intravenous nutritional support the recovery was 100.8 +/- 1.7%. Recovery of tracer increased linearly with O2 uptake and CO2 production. The linear expression obtained between recovery and CO2 production could estimate the recovery within 3% in trauma patients, and that obtained by multiple regression with CO2 production and O2 uptake would predict the recovery within 1.5%.