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PubMed1988

Evaluation of the isotopic equilibration between lactate and pyruvate.

When an isotopic tracer is infused for the purpose of determining the rate of turnover or oxidation of a substrate, it is assumed that the resulting isotopic enrichment by the tracer will reflect the kinetics of only the pool of interest. However, this may not be the case when carbon-labeled lactate is infused, since rapid isotopic exchange with the intracellular pyruvate and alanine pools could potentially occur. Therefore we have determined the extent of isotopic exchange occurring during the infusion of [3-13C]lactate into six anesthetized dogs. In the steady state, pyruvate enrichment was 91 +/- 2.2% (means +/- SE) of the lactate enrichment, and alanine enrichment was 81 +/- 3.3% of the pyruvate enrichment and 72 +/- 2.6% of the lactate enrichment. In contrast, when [3-13C]alanine was infused (n = 2), pyruvate (and lactate) enrichment was 9.9% of the alanine enrichment. We therefore conclude that there is rapid isotopic equilibration between lactate and pyruvate but that interaction with alanine reflects the true metabolic flux rates, rather than isotopic exchange. Consequently, lactate kinetics, as traditionally determined, more accurately reflect whole body pyruvate kinetics.

Animals↗
PubMed1983

Investigation of tubular handling of bicarbonate in man. A new approach utilizing stable carbon isotope fractionation.

Two alternative mechanisms have been proposed for tubular reabsorption of bicarbonate: (a) H+ secretion and CO2 reabsorption and (b) direct reabsorption of HCO-3. In an attempt to differentiate between the two mechanisms, the present study utilized the natural abundance of stable carbon isotopes (13C, 12C) in the urinary total CO2. This novel methodology used mass spectrometric analysis of 13C/12C ratios in urinary total CO2 under normal conditions and during acetazolamide treatment. Blood and respiratory CO2 were analyzed to yield reference values. The results demonstrate that alkaline urine is preferentially enriched with 13C relative to the blood. It is suggested that this fractionation results from reaction out of isotopic equilibrium in which HCO-3 converts to CO2 during the reabsorption process in the distal nephron. The presence of carbonic anhydrase in the proximal nephron results in rapid isotopic exchange between CO2 and HCO-3 and keeps them in isotopic equilibrium. The ratio of urinary 13C/12C increases strikingly after acetazolamide administration and consequent inhibition of carbonic anhydrase in the proximal tubule. Although it is possible that in the latter case high HCO-3 generates the CO2 (ampholyte effect), the isotope fractionation indicates that CO2 rather than HCO-3 is reabsorbed. In contrast, at low urinary pH and total CO2 values, the carbon isotope composition approaches that of blood CO2. This indicates rapid CO2 exchange between urine and blood, through luminal membrane highly permeable to CO2. These results could be anticipated by a mathematical model constructed to plot 13C concentration of urinary total CO2. It is concluded that the mechanism of HCO-3 reclamation in man (and, by inference, in other mammals as well) works by conversion of HCO-3 to CO2 and reabsorption of CO2.

Absorption↗
PubMed2002

Nuclear and isotopic techniques for addressing nutritional problems, with special reference to current applications in developing countries.

Nuclear and isotopic techniques are valuable tools in human nutritional research studies. Isotopes, both radioactive and nonradioactive, enable detailed evaluations of nutrient intake, body composition, energy expenditure, status of micronutrients, and nutrient bioavailability. In recent times, isotopic methods have been widely used in a number of coordinated research projects and technical cooperation projects of the International Atomic Energy Agency's Nutrition Programme. The doubly labeled water technique combines the use of the stable isotopes oxygen-18 and hydrogen-2 (deuterium) to measure total energy expenditure in free-living human subjects, and to investigate the magnitude and causes of both undernutrition and the emergence of obesity in developing countries. The deuterium dilution technique is a reliable tool to measure breastmilk intake and thereby infant growth and development. In collaboration with the World Health Organization's Growth Monitoring Program, this technique is being used to generate new data on growth standards for children in developing countries. This technique is also used in the measurement of body composition by the estimation of lean body mass and fat mass in individuals. Stable isotopes of iron and zinc have been successfully used to assess the nutritional impact of several nationwide food supplementation-programs conducted on pregnant and lactating women and children in both industrialized and developing countries. Isotopic techniques are especially suitable for monitoring changes in body composition, energy metabolism, and mineral status (with particular reference to osteoporosis) in the elderly. Nuclear methods have also served to develop models for a physiological reference man in Asia in support of radiological health and safety issues, for establishing elemental composition of foods, and for measurement of pollutants in the environment.

Adolescent↗
PubMed2005

Isotopic fractionation indicates anaerobic monochlorobenzene biodegradation.

The concentration and isotopic composition of monochlorobenzene (MCB) was monitored in the plume of an anaerobic, contaminated aquifer in Bitterfeld, Germany. An enrichment in the carbon isotopic composition of more than 4 delta units was found at the fringes of the plume relative to the center (-26.5 %), suggesting the occurrence of in situ biodegradation of MCB. A similar enrichment was measured in a detailed cross-section of the plume and in depth-specific samples obtained in a multilevel sampling well. The latter samples gave a good correlation of MCB concentrations and respective isotopic composition according to the Rayleigh equation. On the other hand, batch experiments using the aerobic MCB-degrading strains Ralstonia sp. DSM 8910, Acidovorax facilis UFZ B517, Rhodococcus erythropolis UFZ B528, and Pseudomonas veronii UFZ B547 showed that the known aerobic pathway initiated by dioxygenases does not result in a significant isotopic fractionation. Thus, a novel anaerobic pathway resulting in an isotopic fractionation appears to be the predominant process of MCB degradation in this aquifer. The study also clearly demonstrates the usefulness of isotopic fractionation analysis to prove biodegradation directly in the field, even when microcosm studies are not available and a metabolic pathway has not yet been elucidated.

Anaerobiosis↗
PubMed1995

Fractionation of nitrogen isotopes by mixed ruminal bacteria.

Mixed ruminal bacteria were cultured with glucose, cellulose or no carbohydrate, and ammonium bicarbonate or casein hydrolysate. Changes in amounts of bacterial ammonia and non-ammonia N were measured. Ratios of N isotopes expressed as delta 15N (delta 15N) were measured by isotope ratio mass spectrometry. When bacteria were cultured with glucose and ammonium bicarbonate, bacterial delta 15N decreased from .9 to -5.8/1000 and residual ammonia delta 15N increased from -1.4 to 12.7/1000. Fractionation of N isotope occurred during ammonia incorporation because the difference between delta 15N of ammonia and delta 15N of bacteria (delta 15N) was 18.8/1000 (P < .01). However, when casein hydrolysate was the N source, delta 15N between non-ammonia and bacteria averaged only 1.3/1000 (P > .1), indicating no fractionation of N isotopes occurred during utilization of amino acids. The amount of bacterial N was highest at 24 h of incubation when cellulose was the carbohydrate source. At that time, delta 15N between ammonia and bacteria was 8.9/1000 when ammonia was the N source, but delta 15N between non-ammonia and bacteria was 1.7/1000 when casein hydrolysate was the N source. Bacterial N decreased after 24 h when cellulose was the source of carbohydrate. Results indicate that fractionation of N isotopes occurred during ammonia incorporation, but not during incorporation of N from amino acids, deamination, and release of ammonia. Fractionation of N isotopes during incorporation of ammonia N may be used as a marker to study N metabolism by ruminal bacteria.

Amino Acids↗
PubMed1982

Use of isotope effects to elucidate enzyme mechanisms.

The chemical bond breaking steps are normally not rate limiting for enzymatic reactions. However, comparison of deuterium and tritium isotope effects on the same reaction, especially when coupled with 13C isotope effects for the same step measured with deuterated as well as unlabeled substrates, allows calculation of the intrinsic isotope effects on the bond breaking steps and thus a determination of the commitments to catalysis for the reactants. The variation in observed isotope effects as a function of reactant concentration can be used to determine kinetic mechanisms, while the pH variation of isotope effects can determine the stickiness of the reactants and which portions of the reactant mechanism are pH dependent. Finally the size of primary and secondary intrinsic isotope effects can be used to determine transition state structure.

Animals↗
PubMed1995

Stable isotope identification of lead sources in preschool children--the Omaha Study.

The objective was to determine, from analysis of the naturally occurring stable isotopes of lead, the relative contribution of food, handdust, housedust, soil and air lead to the absorbed (urinary) lead and the blood lead of children living in a former smelter city. A longitudinal 12 month study was conducted of 21 children, 2 - 3 years of age, living in central Omaha, balanced for race, gender and socioeconomic status. Field clean samples were collected monthly of 24 hour duplicate diet, handwipe and urine, with quarterly blood lead, annual environmental lead, weekly air for total lead and 206Pb, 207Pb and 208Pb by thermal ionization/mass spectrometry with a 205Pb spike in a Class II laboratory. Despite residence in a smelter city each child had a unique isotopic ratio of handwipe, blood and urine lead, the latter being identical. There was no correlation of handwipe isotopic ratio with proximity to a lead emission source or to the decade of the housing stock. The isotopic ratio of the annual mean handwipe lead predicted 43% of the variance of the annual mean blood and urine lead ratio (r2 = .43; p = .001). Handwipe lead ratios correlated (p < or = .05) with those of the windowsills and air ducts. The mean isotopic ratios of blood and urine lead were lower than those of handwipe and food, consistent with a contribution by endogenous bone lead. Clean catch urine provides a noninvasive index of blood lead isotopic ratio in children, as in adults.

Child, Preschool↗
PubMed1999

Nondispersive infrared spectrometry for 13CO2/12CO2-measurements: a clinically feasible analyzer for stable isotope breath tests in gastroenterology.

BACKGROUND: 13C-urea breath tests have become clinical routine for the diagnosis of Helicobacter pylori infection and other isotope breath tests have been invented e.g. for gastric emptying or quantitative liver function testing. Recently, isotope-selective nondispersive infrared spectrometers (NDIRS) have been developed for the analysis of the 13CO2/12CO2-enrichment in breath. In this study, we prospectively tested the validity of a newly developed NDIRS in comparison to isotope ratio mass spectrometry (IRMS). METHODS: 142 patients with dyspeptic symptoms were tested for Helicobacter pylori infection using the 13C-urea breath test. The isotope ratio analysis of the breath samples was performed in duplicate both using IRMS and NDIRS. RESULTS: The results of the baseline-corrected 13CO2-exhalation values between IRMS and NDIRS were in excellent agreement. The mean difference between both methods was 0.28 +/- 1.93 delta/1000. Evaluating the qualitative urea breath test results in reference to IRMS as the reference the NDIRS had a sensitivity of 97.8% and a specificity of 98.9%. CONCLUSION: The isotope-selective nondispersive infrared spectroscopy is going to become a reliable, but low-cost and easy-to-operate alternative to expensive isotope ratio mass spectrometry in the analysis of 13C-breath tests.

Adult↗
PubMed2002

Evaluation of exhibits from a murder case using the lead isotope method and scanning electron microscopy.

We have used a combination of lead isotopes and scanning electron microscopy to determine the relationships between different exhibits in a murder case. Samples involved lead projectiles removed from the deceased's head and a pillow, lead-rich scrapings and particles (gunshot residues) from spent cartridges and a silencer, and particles from a pillowcase. The lead projectiles had the same isotopic composition. with the lead being derived from the same dominantly geologically old source(s). The lead smear from the silencer had the same isotopic composition as the projectiles, and the lead was probably from the same source. The particles from the spent cartridges had varying elemental compositions ranging from PbO to PbCuZn +/- Ba with or without Si and are consistent with derivation from the primer. The lead isotopic compositions of the particles from the spent cartridges show some variations, but these are markedly different from those of the projectiles, indicating lead from a mixture of geologically old and geologically young lead. The particles from the pillowcase were extremely small (usually <50 microm size) and showed varying isotopic compositions, some consistent with the gunshot residue from the cartridges. As the exhibits had been handled extensively prior to the present investigation, including some being sent to North America, there is a high likelihood that handling was not done in clean room environments and may have been contaminated. In this instance, as we were concerned about contamination, especially of the pillowcase, we felt contamination negated use of the results for assistance in proving the innocence or guilt of the accused. A combination of high-precision lead isotope measurements with scanning electron microscopy provides a powerful tool for forensic investigations if precautions are taken in handling the exhibits.

Firearms↗
PubMedDate not supplied

[Variability of some isotopic characteristics of carbon in man].

Variability of some carbon isotope characteristics in man (i.e. carbon isotope composition of hair, blood, nails, expiratory carbon dioxide) was studied. Slow (with the period about 20-30 days) oscillations near some average level with a variable amplitude were observed in the isotope composition of the hair carbon. The maximum deviation from the mean level in one person tested was 6%. In the isotope composition of carbon dioxide respired under physical load and under conditions of aerobic metabolism during short periods (up to 15 minutes) there were also found significant variations from the mean level, the maximal ones being 6/1000. An attempt was made to explain the changes observed of isotope characteristics in terms of the principal carbon isotope fractionation cell model proposed earlier.

Carbon Dioxide↗
PubMed2006

[Terrestrial plant stable carbon isotope composition and global change].

Stable carbon isotope analysis is a rapid and reliable technique developed in recent years, and has been widely applied to reconstruct the sequences of atmospheric CO2 concentration changes, clarify the hysteresis effect and junior effect of temperature and precipitation on tree growth, and distinguish the distributions of plants with different photosynthetic pathways. The water use efficiency (WUE) of different plant functional groups and the variations of plant WUE with tempo-spatial and climatic changes can be also indicated by determining plant carbon isotope composition. In this paper, the effects of environmental factors, e.g., atmospheric CO2 concentration, air temperature, precipitation, and altitude on terrestrial plant carbon isotope composition were discussed, and the advances and applications of carbon isotope technique in global change research were summarized. Furthermore, the existing and disputed problems in carbon isotope analysis were discussed, and the future trends of carbon isotope technique in global change research were prospected, aimed to widen people's knowledge and promote the development of this technique.

Air↗
PubMed1990

Isotopic diversity as an unexplored mind-matter dimension.

The existence of several stable isotopes for most chemical elements is a well known and widely used fact of chemistry. Nevertheless, despite all the attention given to the stable and radioactive isotopes of individual elements, there have been very few, if any, attempts to look at isotopicity as a single phenomenon in its own right. Some possible links between isotopicity and various structural and functional levels of organization of the material and informational realm are put forward in this article, which presents isotopicity as an independent and informationally active level of spontaneous pattern formation and self-organization, giving a flexible link between morphogenetic potentialia and the microscopic operational level of physico-chemical processing. A dynamical parallel is drawn between Anderson localization induced by an isotopic disorder and holographic mental processing interpretable as self-focusing of mental patterns and spontaneous creativity.

Biology↗
PubMed1994

13CO2 isotopic laser used through the operating channel of laser laparoscopes: a comparative study of power and energy density losses.

OBJECTIVE: To evaluate the power transmission, spot size, power density, and energy density of a new isotopic carbon dioxide (13CO2) laser compared with a conventional CO2 laser. METHODS: Experiments were performed in a laboratory using a conventional CO2 laser and an isotopic 13CO2 laser. Two laparoscopes with 5-mm and 7.5-mm operating channels were connected to a standard coupler and to each of the lasers. Standardized measurements were made of power transmission and spot size using both nitrogen purge gas and CO2 purge gas at rates of 1-20 L/minute. Power density and energy density were calculated for continuous mode and ultrapulse mode transmission, respectively. RESULTS: The isotopic 13CO2 laser power transmission was higher and proportional to input power, while spot size was smaller compared with the conventional laser and was insensitive to power level or purge rate. Power density and energy density were markedly higher with the isotopic 13CO2 laser, reaching the threshold for complete ablation, and were much more predictable. The 7.5-mm operating channel generally had superior operating results compared with the 5-mm channel because of the smaller spot size and higher power transmission. CONCLUSIONS: The isotopic 13CO2 laser is associated with much higher power density and energy density capabilities than are conventional CO2 lasers. At surgery, we have noted less thermal injury, faster ablation, more precise and predictable tissue effects, and greater control of tissue effect with the isotopic 13CO2 laser than with other CO2 lasers. These results are attributed to the improved beam propagation through CO2 insufflation gas measured in the laboratory. Thermal injury can be varied according to the required surgical situation and can be kept to an absolute minimum at high-pulse energy.

Carbon Dioxide↗
PubMed1993

Isotope effects and alternative substrate reactivities for tryptophan 2,3-dioxygenase.

Tryptophan 2,3-dioxygenase (EC 1.13.1.12) is a hemoprotein which catalyzes the first step in the oxidative degradation of tryptophan. The reaction is believed to proceed by addition of O2 across the 2,3-bond of the indole ring, followed by decomposition of the resultant dioxetane to give N-formylkynurenine. A primary D2O isotope effect of 4.4 on Vmax/Km was observed at the pH optimum, pH 7.0. This implies that abstraction of the indole proton is at least partially rate-determining. An inverse secondary isotope effect of 0.96 was observed for L-[2-3H]tryptophan at this pH. The secondary isotope effect signals the formation of the C-O bond at C-2. As the rate of proton abstraction increased with increasing pH, the D2O isotope effect decreased to 1.2 at pH 8.5 and the secondary isotope effect increased to 0.92. The rate-determining steps therefore change with increasing pH, and bond formation at C-2 becomes more rate-limiting. The secondary isotope effect did not change significantly with varying O2 concentration so that substrate binding is primarily ordered with O2 binding first. The specificity of the enzyme towards substituted tryptophans shows that substitution of the phenyl ring of the indole is sterically unfavorable. Steric hindrance is highest at the 4- and 7-positions, while the 5- and 6-positions are less sensitive. 6-Fluoro-L-tryptophan was more reactive than tryptophan, and the increased reactivity can be explained by an electronic effect that enhances of the rate of C-O bond formation at C-2.

Animals↗
PubMed2007

Using hooves for high-resolution isotopic reconstruction of bovine dietary history.

The objective of this study was to ascertain whether sequential sampling and isotopic analysis of bovine hooves could be used to reconstruct the dietary history of cattle. A controlled, on-farm experiment was conducted in which cattle were switched from a barley-based diet to an isotopically different diet incorporating maize, urea and seaweed (the isotopic spacing between diets was 13.6 per thousand for delta(13)C and 8.0 per thousand for delta(15)N) and maintained on that diet for 168 days. Postmortem sampling of the cleaned anterior wall of the lateral, left front claw was carried out on five individuals using a micro-drilling technique. From the first 60 mm of each claw, up to 41 samples with a spacing between them of less than 1 mm were collected. Bands were less than 1 mm deep and had a mean width of 1.2 mm. The hoof keratin showed a rapid increase followed by a slower increase in its delta(13)C and delta(15)N values following the diet switch, suggesting that C and N in hoof keratin originate from more than one pool. However, the response of the N isotope composition of the hoof was somewhat delayed compared with that of C. Estimated mean hoof growth rates for these cattle were 10.5 +/- 2.3 mm per month and 6.7 +/- 1.0 mm per month (+/-SD, n = 5) when receiving the barley-based transition diet and the maize-based experimental diet, respectively. These values are considerably higher than previous estimates obtained by visual methods and they suggest that diet may have a greater influence on hoof growth rates than seasonality. These results demonstrate that hooves are a suitable incremental tissue for high-resolution isotopic reconstruction of the dietary history of bovine animals.

Animal Feed↗
PubMed1987

Isotopic characterisation of kerogen-like material in the Murchison carbonaceous chondrite.

Isotopic data for C, H and N in acid-resistant residues from carbonaceous chondrites show substantial variability during stepwise pyrolysis and/or combustion. After subtraction of contributions due apparently to inorganic C grains, of probably circumstellar origin, considerable isotopic variability remains, attributable to the kerogen-like organic fraction. That variability may be interpreted in terms of three or four distinct components, based on C, H and N isotopes. The relative proportions of those components vary significantly from sample to sample. The different isotopic components are tentatively identified in terms of specific chemical/structural moieties within the kerogen-like material. This combination of chemical, structural and isotopic information suggests a complex for meteoritic organic matter. At least three components within the organic populations as a whole still carry a signature of apparently interstellar D-enrichment. Part, at least, of the interstellar carrier consisted of reactive entities, not solely polymers.

Carbon↗
PubMed1993

Cosmogenic and nucleogenic isotopic changes in Mars: their rates and implications to the evolutionary history of Martian surface.

We present calculations of rates of production of several nuclides in the Martian atmosphere and in the regolith due to nuclear interactions of cosmic ray and radiogenic particles and consider their implications to the evolutionary history of Mars. Nuclides selected are those which, considering their chemical properties, may be useful as tracers for delineating the past histories of the Martian atmosphere and regolith. Calculations are presented for different assumed atmospheric pressures. The regolith production rates for the present thin Martian atmosphere (approximately 20 g cm-2) are expected to be fairly robust because they are based primarily on observed cosmogenic effects in the Moon, for which semiempirical estimates of nuclide production rates have been provided earlier by Reedy (1981). Uncertainties which arise in the calculations of nuclide production rates for an earlier hypothetical Martian atmosphere of approximately 300-500 g cm-2 thickness are discussed. Compared to cosmic ray production rates, the nucleogenic production rates are smaller by several orders of magnitude. However, the nucleogenic production extends to much deeper levels, whereas the cosmogenic production is essentially confined to the top 750-1000 g cm-2 depth. Important examples of nucleogenic production are discussed. Isotopes of neon and argon appear to be very promising for delineating relative magnitudes of a number of planetary processes related to the temporal changes in the thickness of the atmosphere, as well as their release from the regolith. However, quantification of the processes would require higher-precision isotopic data for the atmosphere and also direct measurements of isotopic ratios in the Martian regolith, along with supplementary information on changes in the isotopic compositions of hydrogen, carbon, and nitrogen, which are affected by a variety of mechanisms of escape of gases from the atmosphere. Cosmogenic effects are minimal in these cases. We show that although we can at present draw but limited inferences, the planet Mars presents a unique opportunity to use cosmogenic nuclides as tools to delineate the evolutionary history of the planet as a whole, as well as its regolith and the atmosphere. This arises because of two factors: minimal degassing of the planet, and a fairly intense chemical weathering history of the upper surface. Consequently, an appreciable fraction of some of the isotopes of volatile elements is contributed by nuclear reactions.

Argon↗
PubMed1983

Use of enriched stable isotopes to determine bioavailability of trace elements in humans.

Use of stable isotopes with analysis by thermal ionization mass spectrometry can determine trace element availability from dietary sources safely and with a high degree of accuracy. Stable isotopes of zinc (70Zn), copper (65Cu) and iron (58Fe) were fed with semipurified diets to humans. Excretion of isotopes was determined by isotope dilution in fecal composites collected for 15 days following the feedings. A thermal ionization, magnetic sector mass spectrometer was used to measure isotopic ratios. Total mineral content of fecal composites was determined by atomic absorption spectrophotometry. Mean absorption of zinc, iron and copper in six elderly men was 17.3, 8.7 and 26.0% respectively. Results agreed closely with absorption determined simultaneously in the same subjects using radioisotopes.

Aged↗