Mathematical Modelling of the Role of Intra- and Extracellular Activity of Carbonic Anhydrase and Membrane Permeabilities of HCO3, H2O and CO2 in 18O Exchange

Abstract

Numerical simulation of ¹⁸O exchange between CO₂, H₂O and HCO₃ ^? explains the ratio mass 46/mass 44 (=C¹⁸O¹⁶O/C¹⁶O₂) measured by mass spectrometry to approximately 1% relative mean difference. In the presence of intact red blood cells the reaction takes place extra- and intracellularly at different reaction rates. Because CO₂ hydration/dehydration is accelerated intracellularly by carbonic anhydrase, a difference in intra- and extracellular concentration of labelled reactants occurs. The ensuing transfer of reactants across the cell membrane depends on their membrane permeabilities which are approximately P_{CO 2} ? 2 cm/s, P_{H 2 O} ? 0.002 cm/s and P_{HCO 3} ? 0.00015 cm/s (values from the literature). We found that the numerical simulation is affected most by P_{HCO 3} . The influence of the other permeabilities is at least ten times less. Therefore within the range of normal literature values, P_{HCO 3} is the only permeability that has a rate limiting effect on ¹⁸O exchange. This is why, in turn, P_{HCO 3} can be derived from an experimentally determined time course of mass 46/mass 44 by a fitting procedure. Another crucial parameter that can be estimated from the experimental results is the intraerythrocytic carbonic anhydrase activity A_i .     

A new Technique for Measurement of Myocardial O2-Utilization

The coronary reserve of mammalian hearts is significantly heterogeneous. From this phenomenon we conclude on a heterogeneous energy-demand, i.e., ATP-demand and therefore on a nonuniform ischemic tolerance of cardiomyocytes.

Our aim was to establish a method for direct quantification of cardiomyocyte energy production. According to Langendorff, 10 isolated rabbit hearts were perfused with oxygen enriched Krebs-Henseleit solution. Using the stable oxygen isotope ¹⁸O₂, respiratory water of 7 hearts was labelled, and samples were taken from the coronary venous effluent. Defined cardiac tissue areas were lyophylized. Using the guanidine-hydrochloride technique, μl-water samples from the tissue and the coronary venous effluent were quantitatively converted to CO₂. With the help of mass-spectrometry the δ¹⁸O/¹⁶O-isotope ratios were determined and related to the SMOW-scale.

Compared to control hearts, a significant shift to higher SMOW-values within respiratory water from ¹⁸O-labelled hearts was detected (mean = 14.61%). Control hearts (maximal scatter = 1.42%) as well as ¹⁸O-labelled hearts (4.06%) showed high reproducibilit, of the method. With the employed experimental setup, significant regional differences were not detected. The kinetics of the coronary venous effluent showed a tendency toward saturation with H₂ ¹⁸O within 2 min after starting the ¹⁸O₂-perfusion. After 50 s perfusion increase in SMOW-values reached statistical significance.     

Half-Width Integral Method for Gas Concentration Measuring in Tunable Diode Laser Absorption Spectroscopy

ABSTRACT

We analyze the influence of random errors and absorption shape fitting errors on gas concentration measurement in tunable diode laser absorption spectroscopy. We then propose a new data processing method according to the characteristic of random errors and fitting absorption shape. This method only uses the integral value within the half width at half maximum of the fitting absorption shape to determine the gas concentration in actual measurements. Meanwhile, the O₂ transition at 13,150.197 cm^?1 is selected to measure the O₂ concentration using this method. Compared with the traditional method, the method established in this paper can accurately measure O₂ concentration.     

The Utilization of 18O Labelled Potassium Phosphate in Studies Concerning Fertilizer Phosphorus Assimilation by Plants

The utilization of ¹⁸O labelled potassium phosphate KH₂P¹⁸O₄, instead of radiophosphorus, ³²P, labelled phosphate in biochemical and ecological studies concerning fertilizer phosphorus assimilation by plants is presented.

The 30–60 mg dry plant samples were heated under 10^?2 mm Hg vacuum in Pyrex ampoules to form carbon dioxide (C¹⁶O¹⁸O) for ¹⁸O mass spectromietric analyses. Potassium phosphate labelled with 1.36 ¹⁸O atom per cent is sufficient to obtain a significant increase of the ¹⁸O plant content, the added ¹⁸O quantity in 0.5 kg soil being 1.4 × 10^?2 g ¹⁸O in this case.     

Experimental and ab initio structure of BrN02

Abstract

The ν₂ fundamental bands of different isotopomers of BrN0₂ (⁷⁹Br¹⁵N¹⁶O₂, ⁸¹Br¹⁵N¹⁶O₂, ⁷⁹Br¹⁴N¹⁸O₂, and ⁷⁹Br¹⁴N¹⁶O¹⁸O) located around 13 µm were recorded using highresolution Fourier transform infrared spectrometry. More than 8000 lines of all these isotopomers were reproduced using a Watson-type A-reduced Hamiltonian with a rootmean-square deviation of better than 7×10^?4 cm^?1 for the four isotopomers. Rotational and centrifugal distortion constants for the ν₂=1 states as well as for the vibrational ground states of these isotopomers were determined. For the first time, an analysis of the ground-state rotational constants obtained in this study combined with the constants obtained in our previous work on the ν₂ bands of ⁷⁹Br¹⁴N¹⁶O₂, and ⁸¹Br¹⁴N¹⁶O₂, has allowed us to calculate the r_m, structure of nitryl bromide. The structural parameters obtained were r_m(Br–N)=2.0118(l6) Å, r_m(N–O)=l.l956(12) Å and α(O–N–O)=131.02(12)Å. A new ab initio structure of nitryl bromide calculated at the CCSD(T)/SDBaug-cc-pVQZ level of theory is presented and was found to be in fair agreement with the experimental structure.     

Regime Shift Signatures from Stable Oxygen Isotopic Records of Otoliths of Atlantic Cod (Gadus morhua)

Abstract

The sudden collapse of Atlantic cod (Gadus morhua) may relate to ocean climate, or regime shifts as demonstrated in production of Pacific salmon. This paper reports the results of stable oxygen isotope ratio analyses (¹⁸O/¹⁶O or δ¹⁸O_A) from 91 otoliths of cod over a period of about 20 years. Seasonal δ¹⁸O_A variations of individual otoliths started at an initial value of about ?0.5 to 0‰ VPDB, and then reached a stable level in the range of +2.5 to +3.5‰ VPDB after 4–5 years. The initial low values correspond to the natal sources of mature cod, while the higher δ¹⁸O_A values represent the water conditions before the cod was caught. This pattern of δ¹⁸O_A variation was observed over the life history of all cod examined. Furthermore, the calculated isotopic temperatures agreed with those obtained from summer bottom trawl survey, indicating that δ¹⁸O_A of otoliths could be used as a thermometer in determining the ambient seawater temperature where the cod lived. Comparison of long-term δ¹⁸O_A records and biological and meteorological observations suggested that decadal-scale ecosystem changes did occur in the late 1970s and early 1990s in Atlantic Canada, comparable to regime shifts occurred in the North Pacific.     

Isotope effect for transport and magnetic properties of La 0.35 Pr 0.35 Ca 0.3 MnO 3 thin films

The effect of ¹⁶ O → ¹⁸ O isotope substitution on electrical resistivity, magnetoresistance, and ac magnetic susceptibility was studied for La_0.35Pr_0.35Ca_0.3MnO₃ epitaxial thin films deposited onto LaAlO₃ and SrTiO₃ substrates. For the films on LaAlO₃, the isotope substitution resulted in the reversible transition from a metal-like to insulating state. The applied magnetic field ( H ≥ 2 T) transformed the sample with ¹⁸O back to the metallic state. The films on SrTiO₃ remained metallic at low temperatures for both ¹⁶O and ¹⁸O, but the shift of the resistivity peak corresponding to onset of metallic state exceeded 63 K after ¹⁶ O → ¹⁸ O substitution. The temperature dependence of both resistivity and magnetic susceptibility was characterized by hysteresis, especially pronounced in the case of the films on LaAlO₃. Such a behavior gives certain indications of the phase separation characteristic of interplay between ferromagnetism and charge ordering. Received 11 February 2000 and Received in final form 13 September 2000     

Physical,optical and radiative properties of CaSO4–B2O3–P2O5 glasses doped with Sm3+ ions

Trivalent rare earth ions doped borosulfophosphate glasses are in high demand owing to their several unique attributes that are advantageous for applications in diverse photonic devices. Thus, Sm³⁺ ion doped calcium sulfoborophosphate glasses with composition of 25CaSO₄–30B₂O₃–(45?x)P₂O₅–xSm₂O₃ (where x?=?0.1, 0.3, 0.5, 0.7 and 1.0 mol%) were synthesized using melt-quenching technique. X-ray diffraction confirmed the amorphous nature of the prepared glass samples. Differential thermal analyses show transition peaks for melting temperature, glass transition and crystallization temperature. The glass stability is found in the range 91?°C to 116?°C which shows increased stability with addition of Sm₂O₃ concentration. The Fourier transform infrared spectral measurements carried out showed the presence of vibration bands due to PO linkage, BO_3, BO₄, PO₄, POP, OPO, SOB, and BOB unit. Glass density showed increase in value from 2.179 to 2.251?g cm^?3 with increase in Sm₂O₃ concentration. The direct, indirect band gap and Urbach energy calculated were found to be within 4.368–4.184?eV, 3.641–3.488?eV and 0.323–0.282?eV energy ranges, respectively. The absorption spectra revealed ten prominent peaks centered at 365, 400, 471, 941, 1075, 1228, 1375, 1477, 1528 and 1597?nm corresponding to ⁴D_3/2,⁶H_5/2→⁴I_11/2,⁶P_3/2, ⁶F_11/2, ⁶F_9/2, ⁶F_7/2, ⁶F_5/2, ⁶F_3/2, ⁶H_15/2 and ⁶F_1/2 transitions respectively. Photoluminescence spectra monitored at the excitation of 398?nm exhibits four emission bands positioned at 559, 596,643 and 709?nm corresponding to ⁴G_5/2→⁶H_5/2, ⁶H_7/2, ⁶H_9/2 and ⁶H_11/2 transitions respectively. The nephelauxetic parameters calculated showed good influence on the local environment within the samarium ions site and the state of the SmO bond. The Judd–Ofelt intensity parameters calculated for all glass samples revealed that Ω_6?>?Ω_4?>?Ω₂. The emission cross-section and the branching ratios values obtained for ⁴G_5/2→⁶H_7/2 transition indicate its suitability for LEDs and solid-state laser application.     

Breath water-based doubly labelled water method for the noninvasive determination of CO2 production and energy expenditure in mice

ABSTRACT

We explored a novel doubly labelled water (DLW) method based on breath water (BW-DLW) in mice to determine whole body CO₂ production and energy expenditure noninvasively. The BW-DLW method was compared to the DLW based on blood plasma. Mice (n?=?11, 43.5?±?4.6?g body mass (BM)) were administered orally a single bolus of doubly labelled water (1.2?g H₂¹⁸O kg BM^?1 and 0.4?g ²H₂O kg BM^?1, 99 atom% (AP) ¹⁸O or ²H). To sample breath water, the mice were placed into a respiration vessel. The exhaled water vapour was condensed in a cold-trap. The isotope enrichments of breath water were compared with plasma samples. The ²H/¹H and ¹⁸O/¹⁶O isotope ratios were measured by means of isotope ratio mass spectrometry. The CO₂ production (RCO₂) was calculated from the ²H and ¹⁸O enrichments in breath water and plasma over 5 days. The isotope enrichments of breath water vs. plasma were correlated (R²?=?0.89 for ²H and 0.95 for ¹⁸O) linearly. The RCO₂ determined based on breath water and plasma was not different (113.2?±?12.7 vs. 111.4?±?11.0?mmol?d^–1), respectively. In conclusion, the novel BW-DLW method is appropriate to obtain reliable estimates of RCO₂ avoiding blood sampling.     

Influence of air pollution and site conditions on trends of carbon and oxygen isotope ratios in tree ring cellulose

Oxygen and carbon isotopic compositions of tree ring cellulose (δ¹³C_cell and δ¹⁸O_cell) were measured for pines growing at four sites in east Germany. Three sites differed markedly in soil water availability within a short distance and the fourth site served as a reference. The choice of the sites was guided by the desire to detect effects of air pollution on the long-term trend of isotopic compositions and to examine the influence of soil water availability on the relationship between the carbon and oxygen isotope ratios. Locations in east Germany are particularly well suited for the study of pollution effects because there was a steady increase in environmental contamination until the German Reunification in 1990, followed by a sharp decline due to the implementation of stricter environmental standards.

The long-term trend of δ¹³C_cell showed an extraordinary increase in the period 1945–1990 and a rapid decrease after 1990, whereas δ¹⁸O_cell remained nearly constant. The increase of δ¹³C_cell is explained by secondary fractionation caused by phytotoxicity of SO₂. Two effects are mainly responsible for the secondary fractionation under SO₂ exposure: increase of dark respiration, and changes in photosynthate allocation and partitioning. Both effects do not influence δ¹⁸O_cell. Furthermore, a significant positive correlation between the year-to-year variations of carbon and oxygen isotope ratios (δ¹³C_resid and δ¹⁸O_resid) has been found for all sites. The slopes of the relationship between δ¹³C_resid and δ¹⁸O_resid differ insignificantly. It is concluded that this relationship is not influenced by soil water availability but by climatic variables.     

Nuclear Magnetic Resonance Spectroscopy Study of a Disaccharidic Crown Ether.

Abstract

The disaccharidic anhydro derivative 6-O-(5,6-anhydro-3-deoxy-1,2-O-isopropylidene α-D-glucofuranos-3-yl)-1,2-O-isopropylidene-3-O-n-dodecyl-α-D-glucofuranose (1) led to the disaccharidic crown ether 2 in 40% yield when treated at low concentration with 2.5 eq. of KOH in toluene-Me₂SO. Compound 2 structure was proved through a detailed NMR analysis (¹H, ¹³C, ¹H-¹H and ¹³C-¹H 2D correlations). This structural elucidation indicated that compound 2 resulted from the intramolecular attack of the C-5-O^?alkoxide group, generated in the basic medium, on the C-6′ carbon of the 5′,6′-anhydro group.     

Infrared Spectra of the Anion-Radicals of Benzophenone and Some of Its Isotopic Isomers

Abstract

Band assignment of the infrared spectra of the anion-radicals of benzophenone-d_O, -d₁₀ and ¹⁸O has been given on the basis of isotopic shifts. The frequency of the ν_C=O band decreases by ca. 270 cm^?1 with the transformation of neutral keton into anion-radical, due to the strong electron-withdrawing character of the carbonyl group; frequencies of the other modes decrease slightly, down to 30 cm^?1.     

Oscillator strengths for the Schumann-Runge bands of 16O18O

Oscillator strenghts were measured for the (2-0)–(15-0) Schumann-Runge bands of ¹⁶O¹⁸O. Individual rotational lines were studied at a resolution of ≈0.05Å. Band oscillator strenghts decrease with increasing rotation at a rate intermediate between those for ¹⁶O₂ and ¹⁸O₂. Our measurements are in good agreement with oscillator strenghts calculated using potential curves and dipole moments derived from ¹⁶O₂ measurements, and support conclusions that ¹⁶O¹⁸O plays only a minor role in the photodissociation of atmospheric O₂. Dipole moments deduced from our oscillator strenghts agree well with those obtained for ¹⁶O₂, ¹⁸O₂, as well as with recent ab initio and semi-empirical determinations.     

A diode laser spectrometer for symmetry selective detection of ozone isotopomers

A tunable diode laser absorption spectrometer operating in the 10 μm range is described, which for the first time permits simultaneous detection and quantification of the five naturally most abundant ozone isotopomers: ¹⁶O₃, ¹⁶O¹⁶O¹⁸O, ¹⁶O¹⁸O¹⁶O, ¹⁶O¹⁶O¹⁷O, and ¹⁶O¹⁷O¹⁶O. Ozone samples of 25 μmole size are analysed with an estimated accuracy of 6‰ (2σ). This level of accuracy is demonstrated by comparing spectroscopically determined isotopologue enrichments of ¹⁶O₂ ¹⁷O and ¹⁶O₂ ¹⁸O with mass spectrometer measurements. Samples for the comparison were generated from natural oxygen in an electric discharge under two different pressure conditions. The precision obtained is sufficient to study both the isotopic and the symmetry dependence of the unique oxygen isotope anomaly in ozone formation, which shows isotopomer specific fractionation values well in the 10% range. PACS 42.60.By; 42.55.Px; 33.20.Ea     

Isotopic Assessment of Sources and Processes Affecting Sulfate and Nitrate in Surface Water and Groundwater of Luxembourg

Abstract

Surface water and deep and shallow groundwater samples were taken from selected parts of the Grand-Duchy of Luxembourg to determine the isotopic composition of nitrate and sulfate, in order to identify sources and/or processes affecting these solutes. Deep groundwater had sulfate concentrations between 20 and 40mg/L, δ³⁴S_sulfate values between ?3.0 and ?20.0‰, and δ¹⁸O_sulfate values between +1.5 and +5.0‰ nitrate was characterized by concentrations varying between <0.5 and 10mg/L, δ¹⁵N_nitrate values of ～?0.5‰, and δ¹⁸O_nitrate values ～+3.0‰. In the shallow groundwater, sulfate concentrations ranged from 25 to 30mg/L, δ³⁴S_sulfate values from ?20.0 to +4.5‰, and δ¹⁸O_sulfate values from ～+0.5 to +4.5‰ nitrate concentrations varied between ～10 and 75mg/L, δ¹⁵N_nitrate values between +2.5 and +10.0‰, and δ¹⁸O_nitrate values between +1.0 and +3.0‰. In surface water, sulfate concentrations ranged from 10 to 210mg/L, δ³⁴S_sulfate values varied between ?9.3 and +10.9‰, and δ¹⁸O_sulfate values between +3.0 and +10.7‰ were observed. Nitrate concentrations ranged from 10 to 40mg/L, δ¹⁵N_nitrate values from +6.5 to +12.0‰, and δ¹⁸O_nitrate values from ?0.4 to +4.0‰. Based on these data, three sulfate sources were identified controlling the riverine sulfate load. These are soil sulfate, dissolution of evaporites, and oxidation of reduced S minerals in the bedrock. Both groundwater types were predominantly influenced by sulfate from the two latter lithogenic S sources. The deep groundwater and a couple shallow groundwater samples had nitrate derived mainly from soil nitrification. All other sampling sites were influenced by nitrate originating from sewage and/or manure. A decrease in nitrate concentration observed along one of the rivers was attributed to denitrification. It appears that sulfate within Luxembourg's aquatic ecosystem is mainly of lithogenic origin, whereas nitrate is often derived from anthropogenic activities.     

18O/16O and 13C/12C Fractionation During the Reaction of Carbonates with Phosphoric Acid: Effects of Cationic Substitution and Reaction Temperature

Abstract

The factors for ¹⁸O/¹⁶O fractionation between carbonates and CO₂ gas produced by the dissolution of the carbonates in phosphoric acid (sealed vessel method) have been investigated as a function of reaction temperature (20–90°C) and cationic substitution in the solid. Synthetic CaCO₃, Ca_0.75 Mn_0.25 CO₃, MnCO₃, BaCO₃ and SrCO₃ powders, and a natural kutnahorite sample were used as solids. The δ¹⁸O values of the gaseous CO₂ liberated by the reaction with phosphoric acid decrease with increasing temperature and seem to be a linear function of T(°K)^?2. The slopes are specific for different carbonates. No temperature-depended ¹³C/¹²C fractionation seems to exist.     

A Modified Technique for the Preparation of SO2 from Sulphates and Sulphides for Sulphur Isotope Analyses

Abstract

A modified technique for the conversion of sulphates and sulphides to SO₂ with the mixture of V₂O₅—SiO₂ for sulphur isotopic analyses is described. This technique is more suitable for routine analysis of large number of samples. Modification of the reaction vessel and using manifold inlet system allows to analyse up to 24 samples every day. The modified technique assures the complete yield of SO₂, consistent oxygen isotope composition of the SO₂ gas and reproducibility of δ³⁴S measurements being within 0.10‰. It is observed, however, oxygen in SO₂ produced from sulphides differs in δ¹⁸O with respect to that produced from sulphates.     

Rotational spectra of 16O3 and of the five 18O isotopic species

The pure rotational spectrum of all of the isotopic species (with ¹⁸O in place of ¹⁶O) of the molecule of ozone has been studied in the range of 18 to 382 GHz. The Watson Hamiltonian developed up to terms in P⁶ is used. For each substituted form, the Watson molecular parameters, measured frequencies, calculated frequencies, and associated uncertainties are given. By using the distortion parameters, the Coriolis coupling constant and the force constants are calculated. The equilibrium structure is obtained from the particular isotopic substitution ¹⁶O₃ ? ¹⁸O₃.     

Measurements of O-Enriched Ozone Isotopomer Abundances Using High-Resolution Fourier Transform Far-IR Spectroscopy

The distribution of ozone isotopomers in ozone mixtures produced by electric discharge in mixtures of ¹⁶O₂ and ¹⁸O₂ at 77 K was measured by high-resolution FTIR spectroscopy. It was of key importance to assess not only the total amount of isotopomers of a certain mass but also the relative amounts of corresponding asymmetric and symmetric ozone species of the same mass given as the ratios [¹⁶O¹⁶O¹⁸O]/[¹⁶O¹⁸O¹⁶O] and [¹⁶O¹⁸O¹⁸O]/[¹⁸O¹⁶O¹⁸O]. For many purposes both ratios have been assumed to have the statistical value 2.00. Pure rotational spectra in the far-IR region (30–100 cm⁻¹) were recorded for three different ¹⁸O-enriched ozone mixtures, all at 0.00185 cm⁻¹ resolution. All the spectra were corrected for thermal emission. Linestrengths for individual lines in a particular spectrum were measured by means of a fitting technique taking into account contributions from all other lines in the spectrum. For this purpose theoretical linestrengths for all six ozone species containing ¹⁶O and ¹⁸O obtained from a quantum-number-dependent dipole operator were used. The ratios between observed and theoretical linestrengths were used to determine the abundances of individual isotopomers in a particular ozone mixture. For one of the ozone samples the abundances of all six ozone species were determined within 1% relative uncertainty. For the three ozone mixtures studied, the ratio between asymmetric and symmetric species of mono-¹⁸O ozone were determined to 1.99(2), 2.01(2), and 2.10(6). The ratio between asymmetric and symmetric species of di-¹⁸O ozone were determined to 2.51(4), 2.42(10), and 2.46(3).     

On the potential of Raman‐spectroscopy‐based carbonate mass spectrometry

The potential for using Raman spectroscopy to measure stable oxygen isotope ratios (¹⁸O/¹⁶O) in carbonates is evaluated by measuring the Raman spectra and isotope ratios of a suite of 60 synthesized, ¹⁸O‐enriched calcite crystals ranging in composition from natural abundance (0.2 mole‐% ¹⁸O) to 1.2 mole‐% ¹⁸O. We determined the Raman‐inferred isotopic ratios (R_Raman) by fitting curves to the ν₁ symmetric stretching peak at 1086 cm⁻¹ and the smaller satellite peak, associated with the ν₁ stretching mode of singly substituted carbonate groups (C¹⁶O₂¹⁸O) at 1065 cm⁻¹. The ratio of the two peak areas shows a 1:1 correspondence with the ¹⁸O/¹⁶O ratios derived from standard mass spectrometry methods, confirming that the relative intensities of the ν₁ symmetric stretching peaks is a direct measure of the isotopic ratio in the carbonates. The 1‐sigma uncertainties of the R_Raman values of the individual crystals were 0.00079 (384‰ PDB) and 0.00043 (210‰ PDB) for the four‐crystal sample means. This level of uncertainty is much too high to provide significant estimates of natural variability; however, there are multiple prospects for improving the accuracy and precision of the technique. Carbon isotope ratios in carbonates cannot be measured by our approach, but our results highlight the potential of Raman‐based isotope ratio measurement for C and other elements in minerals and organic compounds. Copyright © 2012 John Wiley & Sons, Ltd.