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Energy balance in man measured by direct and indirect calorimetry.

In six 24-hr measurements of energy balance, direct and indirect calorimetry agreed within +/-3%, which is probably the range of experimental error. But in seven other 24-hr periods there was disagreement in the range of 8 to 23%, and these were usually days when the subjects ate much less than they spent metabolically. Our direct calorimeter is an insulated, water cooled suit. Continous measurements of O2 consumption and CO2 production provided data on metabolic expenditure (M) by indirect calorimetry. The 24-hr values for M matched the energy losses within +/-60 kcal (+/-3% of M) in four men who rested all day and lay down to sleep at night. Similar agreement was seen in one of the four who worked on a treadmill for 4 hr and stayed busy all day. but in another energy losses were 342 kcal greater than M (10% of M). When the experiments gave values for M minus the losses greater than +/-60 kcal, this is called "unmeasured energy". In further experiments, two subjects stayed awake for 24 hr, and their unmeasured energies were 279 and 393 kcal. The same two men, eating sparingly, also worked for 24 hr so as to double their resting metabolic expenditures; the unmeasured energies were even larger, 380 and 958 kcal. When they repeated the 24 hr of mild work, but ate nearly as much as they spent metabolically, one man was near energy balance, while the other showed an unmeasured energy of -363 kcal. Little heat storage was evident in these experiments; therefore, heat balance was present and energy balance should have been present. In the group of 13 experiments, it appeared that the greater the food deficit, the larger was the unmeasured energy (excess of metabolic expenditure over loss of energy).

Adult↗

Comparison of doubly labeled water, intake-balance, and direct- and indirect-calorimetry methods for measuring energy expenditure in adult men.

Energy expenditure (EE) of four adult men on a weight-maintenance diet was estimated by use of doubly labeled water, intake balance, and direct and indirect calorimetry. The doubly labeled water (2H218O) method was used to estimate free-living EE for 13 d. Metabolizable energy (ME) intake was used to estimate free-living EE for 1 wk. The subjects' 24-h EE was measured in a dual direct-indirect room calorimeter on 3 alternate days. Estimates of free-living EE as measured by ME intake and doubly labeled water indicate agreement between the two methods (mean difference +/- SEM, -1.04 +/- 0.63%). Measurements of EE with indirect and direct calorimetry are equivalent (mean difference 0.63 +/- 0.44%). The daily EE measured by doubly labeled water in these free-living adults over a 13-d period was 15.01% greater than the 24-h EE measured within the calorimeter.

Activities of Daily Living↗

Effects of gas leak around endotracheal tubes on indirect calorimetry measurement.

The use of uncuffed endotracheal tubes (ETT) in pediatric patients raises concern over the accuracy of indirect calorimetry measurement in the presence of a gas leak around an ETT. We examined the effects of ETT gas leak on respiratory gas measurements in a dog model. Mongrel dogs (n = 12) were sedated, paralyzed, intubated, and placed on mechanical ventilation. Leak was achieved by adjusting cuff volume. Oxygen consumption (VO2), CO2 production (VCO2), respiratory exchange ratio (RER), and resting energy expenditure (REE) were measured at each leak pressure (Pleak). Peak inspiratory pressure (PIP), Pleak, inspiratory and expiratory tidal volume (VTinsp, VTexp), VE, end tidal CO2 (ETCO2), and blood gases were recorded at each leak pressure. VO2, VCO, and REE decreased significantly with increasing gas leak. There was a linear relationship between VO2, VCO2, and REE with both TVratio (VTexp/VTinsp) and Pdiff (PIP-Pleak). Multiple regression equations based on TVratio and Pdiff were obtained which allowed correction of the measurement error in VO2, VCO2, and REE, with correlation coefficients (R2) of 0.71, 0.75, and 0.73, respectively. ETT gas leak affects measurements of VO2, VCO2, and REE, but not RER. Measurements made with TVratio greater than 0.80 required no correction. Measurements made with TVratio greater than 0.45 could be corrected to actual values in our model with regression equations based on TVratio and Pdiff. We conclude that indirect calorimetry measurements can be useful in the presence of tracheal gas leak around an ETT.

Animals↗

[Practical aspects of indirect calorimetry in post-anesthesia recovery].

The application of indirect calorimetry among intensive care patients is usually considered as a complicate technique, owing to the difficulties to interpret the bulk of obtained data. As a matter of fact, the variability of energy expenditure (EE) among these patients leads to prefer continuous measurements. This variability is related to interindividual variations as well as variations in the same patient (variations due to body temperature, activity state, diet-induced thermogenesis...). A computer program of data storage and calculations was developed. It allows: on-line storage as well as numerical and graphic display of VO2, VCO2, RQ and EE; menu-driven storage of all occurring events; automatic suppression of the recordings made after ventilator alarms; graphic printing and metabolic calculations. The two main indications for indirect calorimetry are the determination of actual EE and VO2 monitoring in acute states.

Adult↗

[Fatty liver in indirect calorimetry controlled total parenteral nutrition].

The authors have investigated the relationship between total parenteral nutrition (TPN) under indirect calorimetry and the development of fatty infiltrations of the liver in 21 surgical and traumatized patients. TPN was monitored by indirect calorimetry in order to balance lipid and amino acid intake. Glucose intake was adapted only when daily glucose utilization was higher than 150 g. Non protein respiratory quotient (npRQ) and respiratory quotient (RQ) remained below 0.9 during the whole study. The need for lipids was always significantly higher than the lipid intake up to the 5th day after operation or trauma. The need for carbohydrates was significantly lower than the intake of carbohydrates up to the 9th day after operation or trauma. In addition, the utilization of lipids was higher than the utilization of carbohydrates. After 2 weeks of TPN, no fatty infiltration of the liver could be detected by computer tomography and chemical analysis. The calorimetric monitoring of the TPN regimen and the individual adaptation of substrate intake might be responsible for the protection of the liver observed in this study.

Blood Glucose↗

[Heat production in the early stages of axolotl growth according to direct and indirect calorimetry findings].

The level of energetic metabolism of the growing axolotls was studied by the methods of direct and indirect calorimetry. Just after the hatching the heat production of the larvae calculated by the gas exchange exceeds markedly the level of heat production measured by the calorimeter. During the subsequent growth the level of heat production changes as a transitional process, so that therafter indirect calorimetry gives constantly higher values of energetic metabolism than direct one.

Ambystoma↗

An MRI calorimetry technique to measure tissue ultrasound absorption.

High-intensity focused ultrasound (US) surgery guided by magnetic resonance imaging (MRI) is a very promising form of minimally invasive thermal therapy. To apply this technique optimally, the interaction mechanisms of high-intensity US with tissue need to be better understood, in particular, the variation of ultrasound absorption with frequency and temperature. However, agreement on the value of measured tissue US absorption is poor, largely because of intrinsic experimental complications of prior investigations. A new approach toward measuring tissue US absorption, based on a form of MRI calorimetry, is proposed here, which allows non-invasive energy measurement through spatial temperature mapping with MRI. A modified two-dimensional spoiled gradient-echo sequence has been implemented to map temperature based on proton resonance frequency (PRF) shift. Validation experiments show excellent agreement of MRI measured energy with that delivered by a calibrated source. MRI calorimetry of US heating of tissue-mimicking polyethylene glycerol material has been performed. Using a hydrophone measurement of the incident US field, its US absorption coefficient was measured as 0.032 cm-1. As this approach can be applied over a range of frequencies, tissues, and temperatures, it should provide a much improved means of measuring absolute tissue US absorption coefficients to improve US therapy planning, future transducer design, and US dosimetry models.

Absorption↗

A study of the growth for the microalga Chlorella vulgaris by photo-bio-calorimetry and other on-line and off-line techniques.

Calorimetry and other on-line techniques are used for the first time as complement to the traditional off-line methods in order to follow the growth of the green Chlorella vulgaris microalgae. A 2-L photo-bio-reactor was adapted from a commercial calorimeter used previously to study heterotrophic microbial growth. An external source of light was added to favor the photosynthesis of the autotrophic cells. Heterotrophic growth was also tested with external glucose in the broth. A third mode, mixotrophic, allowed faster autotrophic plus heterotrophic growth. Calorimetric measurements were performed considering the corresponding calibrations in order to consider only the energy involved during the microalgal growth. The three different modes of Chlorella cultures were energetically characterized. Besides calorimetry, the weight of diluted nitric acid added to maintain the pH of the culture was correlated with the cellular growth and the nitrogen composition of the algae. Additionally, the on-line infrared spectroscopy proved to be an efficient technique to follow the composition of the broth in glucose, nitrates, and phosphates. These results were compared and complemented with some classic off-line techniques used to track this kind of cultures.

Biomass↗

Characterization of the thermal properties of microcrystalline cellulose by modulated temperature differential scanning calorimetry.

The purpose of this study was to characterize the thermal properties of microcrystalline cellulose (MCC) and to investigate the influence of water on these properties. Differential scanning calorimetry (DSC), modulated temperature differential scanning calorimetry (MTDSC), thermomechanical analysis (TMA), and scanning electron microscopy (SEM) were used to characterize MCC. Three reproducible step transitions were detected in the dry material at 132, 159, and 184 degrees C; for these transitions the magnitude of the heat capacity change varied by a factor of two. Exposure of MCC to water lowers the transition temperature in a manner comparable to a glass transition. The effect of water was different for samples equilibrated to different atmospheric humidities versus water added by granulation. A change in the physical properties of MCC after granulation with high amounts of water was observed. In conclusion, it appears that MCC has glass transitions, which come in reproducible triplets, and these transitions are affected by the presence of water. Also, for the materials studied, the transition temperatures are not affected by particle size and pulp source.

Calorimetry, Differential Scanning↗

Investigation of molecular interactions between paclitaxel and DPPC by Langmuir film balance and differential scanning calorimetry.

Molecular interactions between paclitaxel and dipalmitoylphosphatidyl choline (DPPC) were investigated by Langmuir film balance and differential scanning calorimetry (DSC). Both the lipid monolayer at the air-water interface and that in the lipid bilayer vesicles (liposomes) were employed as model cell membranes. Thermodynamic and kinetic analyses of the DPPC/pacltaxel monolayer system and the paclitaxel penetration into the DPPC monolayer showed that DPPC and paclitaxel can form a nonideal miscible system in the lipid monolayer over a wide range of the DPPC/paclitaxel molar ratios. Paclitaxel exerts an area-condensing effect on the DPPC monolayer at small molecular areas and an area-expanding effect at large molecular areas on the pi-A behavior of the DPPC monolayer, which can be explained by the intermolecular forces and geometric accommodation between paclitaxel and DPPC. Based on a calculation of the excess free energy of the mixed monolayer system, the most stable state of the system occurs at the monolayer composition of 5% paclitaxel. Penetration kinetics showed that the paclitaxel penetration into the DPPC monolayer increases with increasing the drug concentration in the subphase, but there is a limit of approximately 500 ng/mL. Any further increase in paclitaxel concentration had no additional significant effects on the drug penetration. Differential scanning calorimetry showed that paclitaxel caused broadening of the main phase transition. There was no significant change in the peak melting temperature of the DPPC bilayers, which demonstrated that paclitaxel was localized in the outer hydrophobic cooperative zone of the bilayer.

1,2-Dipalmitoylphosphatidylcholine↗

Evaluation of glassy-state dynamics from the width of the glass transition: results from theoretical simulation of differential scanning calorimetry and comparisons with experiment.

The purpose of this study is to investigate the quantitative relationship between the width of the glass transition, DeltaTg, and glass fragility or activation energy for structural relaxation. The ultimate objective is the estimation of structural relaxation time as a function of temperature from the width of the glass transition region, allowing characterization of glass dynamics by a single simple measurement. The Moynihan correlation indicates that activation energy for structural relaxation should be inversely proportional to the width of the glass transition, but recent experimental studies suggest this relationship is a poor approximation for glasses of pharmaceutical interest. The present study is an effort to better understand the validity of the Moynihan correlation by selected experimental studies and a theoretical analysis of those factors that impact the glass transition width. Experimental data for glass transition widths for (poly)vinylpyrrolidone, sucrose, and trehalose are obtained using a variety of procedures, and relaxation time data are obtained using the thermal activity monitor. The theoretical analysis begins by simulating the temperature dependence of the heat capacity by breaking the cooling and heating scans into a large number of temperature steps followed by isothermal holds, during which relaxation of the material is calculated. Here, the modified VTF equation is used for relaxation time and the generalized Kohlraush-Williams-Watts stretched exponential function is used to describe the relaxation kinetics. Simulations are performed for materials of varying fragility and varying "stretched exponential" constants, beta, and the width of the glass transition region, DeltaTg, is evaluated from the simulated heat capacity versus temperature curves as one would do with experimental data. Agreement between the theoretical simulations and experimental DeltaTg data is excellent. The simulations demonstrate that although the Moynihan correlation is not valid for variable beta, a modification of the Moynihan correlation which includes variation in beta is a good approximation. Thus, an estimate of fragility may be obtained from glass transition width data provided an estimate of beta is available. Furthermore, it is shown that a first approximation for beta may be obtained from the magnitude (i.e., height) of the differential scanning calorimetry thermal overshoot. We also find that using the modified VTF equation to evaluate the temperature dependence of the structural relaxation time at the glass transition, and integrating this expression to lower temperatures, it is possible to obtain an evaluation of the relaxation time constant, tau(beta), in the glass at any temperature, using only the DeltaTg and beta values obtained from a single differential scanning calorimetry scan. These estimated time constants correlate very well with the values directly measured with the thermal activity monitor.

Algorithms↗

Bilayer to micelle transition of DMPC and alcohol ethoxylate surfactants as studied by isoperibol calorimetry.

The interaction of dimyristoylphosphatidylcholine (DMPC) with non-ionic surfactants has been studied using isoperibol calorimetry. Phospholipid-surfactant systems were formed in the isoperibol calorimeter with varying amounts of surfactant and the change in enthalpy on formation was measured. Solubilization of the phospholipid lamellae was assessed as a decrease in the enthalpy of reaction of co-films containing DMPC and increasing amounts of three linear alcohol ethoxylate surfactants: C(10)H(21)(OCH(2)CH(2))(3)OH, C(10)H(21)(OCH(2)CH(2))(5)OH, or C(12)H(25)(OCH(2)CH(2))(7)OH. The isoperibol calorimetry data for DMPC/surfactant/water systems were consistent with a theoretical three-stage model for the solubilization of phospholipids by surfactants, whereby phospholipid bilayers are transformed into mixed micelles with increasing amounts of surfactant. The results indicate that: (i) the interaction between phospholipid and surfactants results in a non-linear correlation between the enthalpy of reaction and the surfactant concentration; (ii) the structural stage of the lamellar to micelle transition (mixed bilayers, mixed micelles, or both) can be determined from calorimetric data; (iii) phase boundaries in the solubilization process (bilayer saturation, micelle saturation) can be identified as break points in the enthalpy-concentration curve; and (iv) increasing the hydrophilicity of the surfactant results in a decrease of the surfactant concentration producing the onset of solubilization.

Alcohols↗

Sertaconazole/hydroxypropyl-beta-cyclodextrin complexation: isothermal titration calorimetry and solubility approaches.

Complexation of sertaconazole (SN) with hydroxypropyl-beta-cyclodextrin (HP-beta-CD) was characterized by phase-solubility diagram measurements and isothermal calorimetry (ITC) in aqueous medium, and by differential scanning calorimetry (DSC), Raman spectroscopy and X-ray diffractometry in solid state. The strongest interaction was observed at pH 1.2, at which two different 1:1 complexes can be formed depending on the hydrophobic ring of the drug involved in the process. At pH 5.8 and 7.4 the likelihood of 1:2 stoichiometry increases as a consequence of the simultaneous complexation of the nonprotonized imidazolyl and the dichlorophenyl groups. In the presence of 20% HP-beta-CD, SN solubility is enhanced by a factor of 116, 107, and 5 at pH 1.2, 5.8, and 7.4, respectively. Complexation enthalpy recorded by ITC showed the same tendency which confirms the practical interest of this technique for fast screening of the potential of CDs as drug solubilizers. Solubility and dissolution rate of the drug from compacts prepared with freeze-dried complexes were significantly greater than those obtained with SN powder or compacts made with physical blends.

2-Hydroxypropyl-beta-cyclodextrin↗

Quantitative crystallinity determinations for beta-lactam antibiotics by solution calorimetry: correlations with stability.

The solution calorimetry method is based on the observation that amorphous forms are normally significantly higher in energy than are crystalline forms. The utility and validity of the calorimetric method were investigated for cephalothin sodium, cefazolin sodium, cefamandole nafate, and cefamandole sodium. Amorphous, partially crystalline, and crystalline forms were prepared and characterized by X-ray diffraction (powder), by solution calorimetry, and, for cephalothin sodium, by the thermal decomposition rate at 50 degrees. Qualitatively, there was a good correlation between calorimetric crystallinity and the (less precise) crystallinity derived from X-ray data. The energy and structure of the amorphous state depend on the history of the sample; even samples of the same crystalline polymorph, containing no amorphous phase, may differ in energy. Thus, the absolute value of the crystallinity (X-ray or calorimetric) depends on the choice of amorphous and crystalline standards. The heat of solution is a precise (+/- 1%) and unambiguous measure of the relative crystallinity; and provided amorphous and crystalline standards are appropriately chosen, the calorimetric crystallinity correlates well with chemical stability.

Absorption↗

BisANS binding to tubulin: isothermal titration calorimetry and the site-specific proteolysis reveal the GTP-induced structural stability of tubulin.

Interactions of bisANS and ANS to tubulin in the presence and absence of GTP were investigated, and the binding and thermodynamic parameters were determined using isothermal titration calorimetry. Like bisANS binding to tubulin, we observed a large number of lower affinity ANS binding sites (N1 = 1.3, K1 = 3.7 x 10(5) M(-1), N2 = 10.5, K2 = 7 x 10(4)/M(-1)) in addition to 1-2 higher affinity sites. Although the presence of GTP lowers the bisANS binding to both higher and lower affinity sites (N1 = 4.3, N2 = 11.7 in absence and N1 = 1.8, N2 = 3.6 in presence of GTP), the stoichiometries of both higher and lower affinity sites of ANS remain unaffected in the presence of GTP. BisANS-induced structural changes on tubulin were studied using site-specific proteolysis with trypsin and chymotrypsin. Digestion of both alpha and beta tubulin with trypsin and chymotrypsin, respectively, has been found to be very specific in presence of GTP. GTP has dramatic effects on lowering the extent of nonspecific digestion of beta tubulin with trypsin and stabilizing the intermediate bands produced from both alpha and beta. BisANS-treated tubulin is more susceptible to both trypsin and chymotrypsin digestion. At higher bisANS concentration (>20 microM) both alpha and beta tubulins are almost totally digested with enzymes, indicating bisANS-induced unfolding or destabilization of tubulin structure. Again, the addition of GTP has remarkable effect on lowering the bisANS-induced enhanced digestion of tubulin as well as stabilizing effect on intermediate bands. These results of isothermal titration calorimetry, proteolysis and the DTNB-kinetics data clearly established that the addition of GTP makes tubulin compact and rigid and hence the GTP-induced stabilization of tubulin structure. No such destabilization of tubulin structure has been noticed with ANS, although, like bisANS, ANS possesses a large number of lower affinity binding sites. On the basis of these results, we propose that the unique structure of bisANS, which in absence of GTP can bind tubulin as a bifunctional ligand (through its two ANS moieties), is responsible for the structural changes of tubulin.

Anilino Naphthalenesulfonates↗

Low-affinity binding determined by titration calorimetry using a high-affinity coupling ligand: a thermodynamic study of ligand binding to protein tyrosine phosphatase 1B.

A competition-based method is used for the determination of the thermodynamic parameters for a low-affinity ligand binding reaction by isothermal titration calorimetry. This method is based on the coupling of a high-affinity ligand to the binding of the low-affinity ligand. Results are presented for the binding of a nonhydrolyzable phosphotyrosine analog phosphonodifluoromethyl phenylalanine (F2Pmp)-containing peptide (Ac-Asp-Ala-Asp-Glu-F2Pmp-Leu-NH2), arsenate, and inorganic phosphate to the intracellular human protein tyrosine phosphatase 1B(PTP1B). The binding constants are 3.3 x 10(6), 4.3 x 10(3), and 48 M-1 for the F2Pmp-containing peptide, arsenate, and inorganic phosphate, respectively. The binding of arsenate and inorganic phosphate to PTP1B is enthalpy driven. This is in contrast to the binding of the F2Pmp-containing peptide which is mainly driven by entropy. The calorimetrically determined binding constants are in agreement with the Ki values determined by enzyme inhibition studies. This demonstrates that isothermal titration calorimetry can be used to quantitatively determine the thermodynamic parameters for the interactions between proteins and low-affinity ligands if a proper coupling ligand can be identified.

Amino Acid Sequence↗

A simplified procedure of direct calorimetry for bedside monitoring of the resting metabolic rate.

A simplified procedure of direct calorimetry (SPDC) for determination of resting metabolic rate of respiratory uncompromised subjects in a supine position is presented. This procedure was based on computer-assisted measurements of heat losses due to evaporation, radiation, conduction, and convection. The subject's total loss of mass was recorded hydraulically with a beam scale and afterwards transformed into a digital electric signal. Differences between dry bulb temperature and mean skin temperature were measured by semiconductor thermistors and also transformed into digital signals. With special software an interfaced personal computer assisted in performing SPDC and in calculating heat losses due to evaporation, radiation, and conduction. In a thermoneutral environment, six healthy volunteers were investigated to determine the mean convective heat transfer coefficient (hc) from the difference in an individual between the metabolic energy transformation (M) measured by indirect calorimetry (IC) and the sum of heat losses by radiation, conduction, and evaporation. The room-specific value of hc of 2.12 (SD 0.22) W.m-2.degrees C-1 was in good agreement with data in the literature. Compared to the results of M from a second series of IC, the total heat loss (THL) measured by SPDC in a thermoneutral environment was calculated as 100.5 (SD 6.0)%. The THL by SPDC performed three times at 3-h intervals on ten other volunteers revealed a mean difference of 0.22 (SD 1.74) W.m-2. Thus, SPDC would seem to be a valid and reproducible method under conditions of thermal neutrality.

Adult↗

Titration calorimetry as a binding assay for lipid-binding proteins.

Titration calorimetry has been evaluated as a method for obtaining binding constants and thermodynamic parameters for the cytosolic fatty acid- and lipid-binding proteins. An important feature of this method was its ability to accurately determine binding constants in a non-perturbing manner. The equilibrium was not perturbed, since there was no requirement to separate bound and free ligand in order to obtain binding parameters. Also, the structure of the lipid-protein complex was not perturbed, since native ligands were used rather than non-native analogues. As illustrated for liver fatty acid-binding protein, the method distinguished affinity classes whose dissociation constants differed by an order of magnitude or less. It also distinguished endothermic from exothermic binding reactions, as illustrated for the binding of two closely related bile salts to ileal lipid-binding protein. The main limitations of the method were its relatively low sensitivity and the difficulty working with highly insoluble ligands, such as cholesterol or saturated long-chain fatty acids. However, the signal-to-noise ratio was improved by manipulating the buffer conditions, as illustrated for oleate binding to rat intestinal fatty acid binding protein. Binding parameters are reported for oleate interactions with several wild-type and mutant lipid-binding proteins from intestine. Where possible, the binding parameters obtained from calorimetry were compared with results obtained from fluorescence and Lipidex binding assays of comparable systems.

Animals↗