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PubMed2001

Estimation of available energy of dietary fibres by indirect calorimetry in rats.

BACKGROUND: Knowledge of energetic availability of dietary fibres is important for human nutrition. But up to now results are often different and depend on the methods used. Estimation of metabolisable energy of dietary fibres (mainly by balance technique) is a time-consuming procedure and needs special technical effort. AIM OF THE STUDY: Validation of the experimental design for short-term studies by using indirect calorimetry with feeding below maintenance requirement to evaluate the energetic availability of dietary fibres and their influence on absorption velocity of carbohydrates (CHO). METHODS: Energy expenditure and CHO oxidation (including short-chain fatty acids as fermentation products) were estimated in Wistar rats over 23 h after being fed a basal diet for the first day (300 KJ/kg0.75, 20% protein, 3% fat, 77% CHO) followed by supplementation with either microcrystalline cellulose, the soluble rye fibre arabinoxylan, apple pectin, amylomaize starch (with 48% of resistant starch) or gelatinised wheat starch (200 KJ/kg0.75 each) as control for the following days. Energetic availability was determined by comparing the increase of CHO oxidation after addition of gelatinised wheat starch with that of the dietary fibres tested. RESULTS: In comparison to wheat starch (100%), the following energetic availability of the dietary fibres was found: microcrystalline cellulose 14%, arabinoxylan 33%, pectin 39%, amylomaize starch 62%. The time-course of CHO oxidation indicated that microcrystalline cellulose enhances, whereas the soluble rye fibre slows down the velocity of CHO absorption due to the different consistency of the intestinal contents modified by the kind and properties of the used dietary fibres. After intake of arabinoxylan or pectin, CHO oxidation remained at a higher level during the experimental period elucidating an increased activity of fermentation to short-chain fatty acids. CONCLUSIONS: Short-term experiments in rats using indirect calorimetry are a suitable method for comparative estimation of the energetic availability of dietary fibres. Results are partly in agreement with values estimated by long-term in vivo methods.

Animals↗
PubMed1995

[Intraoperative calorimetry in aortic bifurcation reconstruction].

Oxygen uptake (VO2) and carbon dioxide elimination (VCO2) can be measured with an indirect calorimeter, this method is well established in routine monitoring of ICU patients to evaluate metabolic state as a reflection of stress. In various experimental studies it was demonstrated that anaesthetics can influence whole-body metabolism. The purpose of this study was to examine whether indirect calorimetry can be used intraoperatively during routine anaesthesia and whether presumable changes in metabolism can be detected immediately. Abdominal aortic cross-clamping changes circulation, nutritional supply of the lower extremities and thus VO2 and VCO2. We therefore used this operation for our study. METHOD. Eleven patients, mean age 64 years, undergoing reconstruction of the aortic bifurcation, were studied. After premedication with piritramid and atropine, total intravenous anaesthesia (TIVA) was performed with fentanyl and midazolam after an induction with thiopental. Patients were ventilated with a Servo-Ventilator 900 D and a constant FiO2 of 0.5, without N2O. Routine monitoring consisted of ECG, pulsoximetry, CVP and continuous AP. VO2 and VCO2 were measured with a Deltatrac (Datex), and data were registered every minute. For statistical evaluation we used a Wilcoxon-Ranksum test for matched pairs, p < 0.05 was considered significant. Data from specific time (5 min after intubation, 5 min before clamping; 5, 10 and 15 min after clamping, before declamping and 5 and 10 min after declamping and at the end of surgery) were calculated. In addition to absolute values, we compared the measured VO2 and VCO2 to baseline (5 min before clamping = MP2). RESULTS. Mean operating time was 139 min +/- 37; aortic cross-clamping time for the first extremity was 38 min and 55 min for the second. As expected, there was a significant decrease in VO2 (90% of baseline) and VCO2 (75% of baseline) during aortic cross-clamping. After declamping VO2 again rose to 110% of baseline, or to 103% for the second limb. VCO2 increased to only 90% and 82%, respectively. At the end of surgery VO2 reached baseline, whereas VCO2 remains at 83%. The respiratory quotient VCO2: VO2 was markedly reduced from 0.95 +/- 0.156 to 0.73 +/- 0.06 during surgery. The Deltatrac showed every change in VO2 without delay; changes in VCO2 seem to occur somewhat retarded. DISCUSSION. Aortic cross-clamping leads to a marked decrease in VO2 and VCO2 reflecting the temporary reduction in whole-body metabolism. Declamping results in a compensatory rise, especially in VO2. VCO2 seems to increase less after declamping, perhaps due to the CO2 pool of the organism or to a change in metabolism from carbohydrate to mainly fat oxidation. The results of this study demonstrate that indirect calorimetry can easily be performed during anaesthesia and surgery. Preconditions are a non-rebreathing system without airleak, constant FiO2 < 0.6 and no use of nitrous oxide.

Aged↗
PubMed2006

Direct calorimetry of free-moving eels with manipulated thyroid status.

In birds and mammals, the thyroid gland secretes the iodothyronine hormones of which tetraiodothyronine (T4) is less active than triiodothyronine (T3). The action of T3 and T4 is calorigenic and is involved in the control of metabolic rate. Across all vertebrates, thyroid hormones also play a major role in differentiation, development and growth. Although the fish thyroidal system has been researched extensively, its role in thermogenesis is unclear. In this study, we measured overall heat production to an accuracy of 0.1 mW by direct calorimetry in a free-moving European eel (Anguilla anguilla L.) with different thyroid status. Hyperthyroidism was induced by injection of T3 and T4, and hypothyroidism was induced with phenylthiourea. The results show for the first time at the organismal level, using direct calorimetry, that neither overall heat production nor overall oxygen consumption in eels is affected by hyperthyroidism. Therefore, we conclude that the thermogenic metabolism-stimulating effect of thyroid hormones (TH) is not present with a cold-blooded fish species like the European eel. This supports the concept that TH does not stimulate thermogenesis in poikilothermic species.

Anguilla↗
PubMed2003

Elucidating thermodynamic parameters for electron transfer proteins using isothermal titration calorimetry: application to the nitrogenase Fe protein.

Establishing thermodynamic parameters for electron transfer reactions involving redox proteins is essential for a complete description of these important reactions. While various methods have been developed for measuring the Gibbs free energy change (Delta G(HR) or E(m)) for the protein half-reactions, deconvolution of the respective contributions of enthalpy (Delta H(HR)) and entropy (Delta S(HR)) changes is much more challenging. In the present work, an approach is developed using isothermal titration calorimetry (ITC) that allows accurate determination of all of these thermodynamic parameters for protein electron transfer half-reactions. The approach was validated for essentially irreversible and reversible electron transfer reactions between well-characterized mediators and between mediators and the protein cytochrome c. In all cases, the measured thermodynamic parameters were in excellent agreement with parameters determined by electrochemical methods. Finally, the calorimetry approach was used to determine thermodynamic parameters for electron transfer reactions of the nitrogenase Fe protein [4Fe-4S](2+/+) couple in the absence or presence of MgADP or MgATP. The E(m) value was found to change from -290 mV in the absence of nucleotides to -381 mV with MgATP and -423 mV with MgADP, consistent with earlier values. For the first time, the enthalpy (Delta H(HR)) and entropy (Delta S(HR)) contributions for each case were established, revealing shifts in the contribution of each thermodynamic parameter induced by nucleotide binding. The results are discussed in the context of current models for electron transfer in nitrogenase.

Adenosine Diphosphate↗
PubMed1992

Fluorescence probes in biochemistry: an examination of the non-fluorescent behavior of dansylamide by photoacoustic calorimetry.

Photoacoustic calorimetry is shown to be a simple, precise, and accurate method for the quantification of the photophysics of a fluorescence probe, e.g., dansylamide, in a variety of solvents. This technique, which is described in detail, provides a direct measurement of the energy that is released nonradiatively following photostimulation, and can therefore be used to indirectly determine the amount of energy released via luminescent pathways. Photoacoustic calorimetry combined with established absorption and fluorescence methodologies provides a complete arsenal for characterizing the photophysical properties of many systems. Comparison of the photoacoustic signal for dansylamide versus standard compounds (ferrocene, tetraphenylethylene, 8-anilinonaphthalene-1-sulfonate, and/or 5,5'-dithiobis(2-nitrobenzoic acid) in 12 different solvents gave fh values (fraction of each absorbed 337.1-nm photon returned as heat) from a low of 0.530 in 1,4-dioxane to a high of 0.973 in water. The trend noted with solvent polarity is different and more revealing than that determined by the more classical approach of examining either the wavelength of the emission maximum or the fluorescence quantum yield.

Calorimetry↗
PubMed1979

Thermal stability of fatty acid-serum albumin complexes studied by differential scanning calorimetry.

Differential scanning calorimetry has been used to study the thermal stability of bovine serum albumin as affected by binding of fatty acids (lauric acid and stearic acid) and sodium dodecyl sulfate (SDS). All the ligands stabilized the protein molecules in a similar manner, but to different levels. A maximum increase in denaturation temperature of 30 degrees C was obtained with lauric acid. The thermograms indicate the presence of several ligand-albumin complexes having different heat stabilities. Variations in pH in 0.9% NaCl affected the heat stability of both ligand-poor and ligand-rich albumin, the former being more sensitive to variations in pH within the physiological range. Variations in NaCl concentration affected the thermal stabilities at neutral pH, expecially at low salt concentrations. While ligand-rich albumin was somewhat destabilized by increasing NaCl concentrations, ligand-poor albumin was strongly stabilized. The potential use of differential scanning calorimetry in ligand-albumin research is discussed.

Animals↗
PubMed1971

Flow calorimetry of the sorption of butanols to elastin preparations and comparison with surface areas determined by krypton-85 adsorption.

1. The apparent surface areas of elastin samples as well as of several other fibrous protein preparations (collagen, keratin, polymeric stroma of aorta) were determined using two different approaches: (a) the Brunauer-Emmett-Teller method with 85Kr and (b) microflow calorimetry with n- and tert.-butanol as adsorbents in a heptane stream. 2. Areas of heat signals obtained by flow calorimetry for the adsorption and desorption of n- and tert.-butanol were substantially equivalent; desorption was more protracted than adsorption, the difference between the speed of desorption and adsorption increased with decreasing chain length of the alcohols (methanol, ethanol, n-propanol and n-butanol). 3. An inverse linear relationship was found between the energy change recorded during the adsorption process and the chain length of the alcohols (methanol, ethanol, n-propanol, n-butanol). 4. Heats of adsorption of tert.-butanol were systematically found to be significantly lower than those of n-butanol with all the protein samples investigated. 5. The apparent surface areas of the protein samples determined with tert.-butanol were on the average of the same order or only slightly higher than those obtained with 85Kr. Results obtained with n-butanol were significantly higher. The difference between surface areas obtained with n- and tert.-butanol depended on the nature of the protein sample, on its method of preparation and to some extent on the residual humidity of the sample. 6. The results could be explained on the basis of the hydrophobic theory of elastin structure (see ref. 4) and confirmed our former conclusions (see ref. 3) concerning the significantly higher surface areas of elastin samples purified by different procedures as compared to collagen or to keratin. They also confirmed the accessibility of the surface of elastic fibers to the molecular probes used in the polymeric stroma of aorta.

Adsorption↗
PubMed1994

Interaction of F-actin with phosphate analogues studied by differential scanning calorimetry.

The thermal unfolding of F-actin and the changes induced in it by the binding of phosphate analogues were studied by differential scanning calorimetry. It is shown that the conformation of actin is drastically altered by interaction with beryllium fluoride or aluminium fluoride, while the effects of vanadate and phosphate are negligible. The effect of beryllium fluoride on the F-actin structure, as reflected in a significant increase of the actin thermal stability, is much more pronounced in the presence of Mg2+ than in the case of F-actin polymerized by KCl or LiCl in the absence of Mg2+. It is concluded that differential scanning calorimetry is a very convenient method for probing the conformational changes in F-actin caused by the interaction with phosphate analogues.

Actins↗
PubMed1995

Tight binding affinities determined from thermodynamic linkage to protons by titration calorimetry.

A general titration calorimetry method is described that can be used to determine the affinity of tight binding interactions with proteins. The method is based on the thermodynamic linkage between ligand binding and coupled protonation reactions. The protons linked to a given ligand-binding reaction are measured by titration calorimetry, and integration of the resulting data set yields the pH dependence of the binding affinity based on thermodynamic relationships developed elsewhere. When the pH dependence of the binding affinity is combined with the absolute affinity determined independently at a pH at which the affinity can be conveniently measured, the absolute binding affinity over the entire pH range is determined. The method is well suited for determining high-affinity binding interactions of protein antigens with antibodies, but is applicable to any macromolecular ligand-binding reaction that is coupled to protonation.

Antigen-Antibody Reactions↗
PubMed1995

Validation of the shrinkage temperature of animal tissue for bioprosthetic heart valve application by differential scanning calorimetry.

Shrinkage temperature is most often used to report the degree of cross-linking in glutaraldehyde-fixed animal tissue for use in bioprosthetic heart valve fabrication. Present practice utilizes the measurement of hydrothermal shrinkage observed when a sample is subjected to a temperature programme. This measurement at best gives a general indication of the efficiency of the treatment, i.e. the extent of cross-linking in the tissue. When differential scanning calorimetry has been used, the ambiguity arising from the scant reporting of the protocols used does not permit easy comparison of experimental results. This report addresses the considerations necessary to obtain optimum results in the differential scanning calorimetry experiment for the determination of shrinkage temperature in biological tissue. The shrinkage temperature of two previously unreported tissue types, porcine pericardium and equine pericardium, are provided and compared with those of bovine pericardium and porcine aortic valve leaflets.

Animals↗
PubMed1996

[Water calorimetry measurements in a 85 MeV clinical proton beam].

The importance of water calorimetry in clinical proton beam dosimetry has increased for various reasons: there has been an increasing interest in proton therapy from the radiotherapy world; existing dosimetry protocols for clinical proton beams recommend calorimetry as the primary dosimetry method (ie, AAPM and ECHED); water has recently become the reference material for dose specification. For the water calorimetric dose evaluation, the water calorimeter developed in Ghent was used. Ionometry was performed following the ECHED protocol. The study resulted in a calorimetric to ionometric dose ratio of 0.974 +/- 0.009. The discrepancy should in our opinion be attributed to the (Wle)p,air value of 35.2 J/C implemented in the protocol, possibly next to ion chamber dependent effects for which indications are found in the ionometry measurements.

Calorimetry↗
PubMed2005

Ligand binding to one-dimensional lattice-like macromolecules: analysis of the McGhee-von Hippel theory implemented in isothermal titration calorimetry.

The theory developed by McGhee and von Hippel for ligand binding to a one-dimensional lattice-like macromolecule provides a closed analytical form in the Scatchard representation. The application of such theory has been complicated by two facts: (1) it has been practically reduced to binding techniques, such as equilibrium dialysis, in which the partition between bound and free concentrations of all reactant species are directly accessible and experimentally determined, but infrequently applied to other binding techniques, such as calorimetry or spectroscopy, in which the direct observable is a magnitude proportional to the advance of the binding reaction monitored along the titration experiment, and (2) Scatchard analysis, developed as a quantitative graphical method, is currently outdated and used only qualitatively because of its weaknesses, limitations, and deficiencies. However, a general exact method for applying such theory to titration techniques in a correct and precise manner, without any limitation, can be delineated. In this article, the theory of cooperative ligand binding to linear lattice-like macromolecules has been implemented in isothermal titration calorimetry for the first time. This technique provides a complete thermodynamic characterization of ligand binding, but it has been barely used properly for this type of system. The description, the analysis of the formalism, and practical guidelines are presented, with considerations for experimental design and data analysis.

Calorimetry↗
PubMed2006

Differential scanning calorimetry as a tool to estimate binding parameters in multiligand binding proteins.

The stability of proteins and their interactions with other molecules is a topic of special interest in biochemistry because many cellular processes depend on that. New methods and approaches are constantly developed to elucidate the energetics of biomolecular recognition. In this sense, the application of the theory of macromolecular unfolding linked to ligand binding to differential scanning calorimetry (DSC) has proved to be a useful tool to simultaneously characterize the energetics of unfolding and binding. Although the general theory is well known, the applicability of DSC to study the interaction of biomolecules is not common. In the current work, we estimated the binding parameters of 8-anilinonaphthalene-1-sulfonic acid to human serum albumin using DSC. This model system was chosen due to both the complex stoichiometry and the moderate binding constants. From DSC curves acquired at different ligand concentrations, we obtained the number of bound ligands, the binding constants, and the binding enthalpy for each independent binding site. Compared with those parameters determined by titration calorimetry, the results highlight the potentiality of DSC to estimate binding parameters in multiligand binding proteins.

Anilino Naphthalenesulfonates↗
PubMed2005

Calorimetry and mass spectrometry study of oxidized calmodulin interaction with target and differential repair by methionine sulfoxide reductases.

Calmodulin is known to be a target for oxidation, which leads to conversion of methionine residues to methionine sulfoxides. Previously, we reported that both methionine sulfoxide reductases MsrA and MsrB were able to reduce methionine sulfoxide residues in oxidized calmodulin. In the present study, we have made use of the interaction between calmodulin and RS20, a peptide model for calmodulin targets, to probe the structural consequences of oxidation and mode of repair both by MsrA and MsrB. Isothermal titration calorimetry and differential scanning calorimetry showed that oxidized calmodulin interacts with RS20 via its C-terminal domain only, resulting in a non-productive complex. As shown by spectrofluorometry, oxidized calmodulin treated with MsrA exhibited native binding affinity for RS20. In contrast, MsrB-treatment of oxidized calmodulin resulted in 10-fold reduced affinity. Mass spectrometry revealed that the sulfoxide derivative of methionine residue 124 was differentially repaired by MsrA and MsrB. This provided a basis for rationalizing the difference in binding affinities of oxidized calmodulin reported above, since Met124 residue had been shown to be critical for interaction with some targets. This study provides the first evidence that in an oxidized polypeptide chain MetSO residues might be differentially repaired by the two Msr enzymes.

Animals↗
PubMed2005

Multithermal titration calorimetry: a rapid method to determine binding heat capacities.

Herein a new method that allows binding DeltaCp to be determined with a single experiment is presented. Multithermal titration calorimetry (MTC) is a simple extension of isothermal titration calorimetry (ITC) that explicitly takes into account the thermal dependences of DeltaH and the binding constant. Experimentally, this is accomplished by performing a single stepwise titration with ITC equipment, allowing temperature re-adjustments of the system at intermediate states of the titration process. Thus, from the resulting multitherm, DeltaCp can also be determined. The experimental feasibility of MTC was tested by using the well-characterized lysozyme-chitotriose complex as a model system.

Algorithms↗
PubMed2006

The reactivity of sodium borohydride with various species as characterized by adiabatic calorimetry.

The reactivity of sodium borohydride in the presence of other species has been examined by adiabatic calorimetry. In combination with water, sodium borohydride exhibits an exotherm at room temperature accompanied by generation of gas (presumed to be hydrogen). Addition of potassium hydroxide to a sodium borohydride-water mixture is found to stabilize the solution and require a higher temperature for reaction to occur. However, if iron oxide is also included, reaction takes place near room temperature. Very rapid reaction was found when a metal chloride was brought in contact with a solution containing sodium borohydride, water, and potassium hydroxide. When sodium borohydride was added to an oxygenated hydrocarbon, reaction at room temperature also took place, but to a more limited extent. Peak temperatures above 200 degrees C and maximum pressures in excess of 2000 psia were observed in most cases. Kinetics extracted from the calorimetry data are presented for some of the sodium borohydride combinations.

Borohydrides↗
PubMed2006

Enthalpy and conformational volume changes of mammalian oxy-hemoglobins investigated by pulsed photoacoustic calorimetry.

Enthalpy and conformational volume changes induced by laser photo-dissociation reactions of mammalian oxy-hemoglobin, such as human, bovine, pig, horse and rabbit oxy-hemoglobins, are investigated by pulsed photoacoustic calorimetry. Generally, the response time of the photoacoustic calorimetry is restricted by the width of the laser pulse and the bandwidth of the acoustic detector. Considering the time window of the experimental system, the enthalpy and conformational volume changes detected should be caused by the tertiary relaxation of the heme proteins. In order to calculate the enthalpy and conformational volume changes, the quantum yields of the photo-dissociation products of oxy-hemoglobins must be measured and taken into account. Finally, the enthalpy and conformational volume changes of the oxy-hemoglobins connected with the tertiary relaxation are obtained, which show that for all measured mammalian oxy-hemoglobins the values of enthalpy changes are in the range of 30.0-46.8 kcal/mol and volume changes are of 2.3-7.8 ml/mol although the quantum yields for the different species may have much bigger differences. A possible explanation of the results is presented.

Algorithms↗
PubMed1997

Resting energy expenditures in Asian women measured by indirect calorimetry are lower than expenditures calculated from prediction equations.

OBJECTIVE: Measured resting energy expenditure (REE) and REEs calculated using the Harris-Benedict equation, Food and Agriculture Organization/World Health Organization/United Nations University (FAO/WHO/UNU) equations (FAO equations), and the Liu equation were compared in Asian women. DESIGN: REEs were measured using indirect calorimetry in the morning after an overnight fast and compared with REEs calculated using the Harris-Benedict equation, the FAO equations, and the Liu equation. Height, weight, and 3-day diet records were also obtained. SUBJECTS: Thirty-six healthy, free-living Asian women aged 19 to 52 years and living in the United States were recruited from Washington State University, Pullman, and completed the study. STATISTICAL ANALYSES: Paired t tests, stepwise regression, one-way analysis of variance, and Pearson correlation coefficients were used for the statistical analyses. Significance was set at P < or = .05. RESULTS: A significant correlation was found between measured REE and REE derived from the Harris-Benedict equation (R = 0.67, P < .0001), the FAO equations (R = 0.70, P < .0001), and the Liu equation (R = 0.70, P < .0001). However, measured REE was significantly lower than REE calculated using the Harris-Benedict and FAO equations by 8.5% (P < .001) and 5.4% (P < .01), respectively. No significant difference was noted between measured REE and REE derived from the Liu equation. APPLICATIONS: Caution must be taken when predicting REE of Asian women using the Harris-Benedict equation or the FAO equation. Indirect calorimetry or an equation specific to Asians, such as the Liu equation, is recommended when an accurate estimate is necessary.

Adult↗