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Phase equilibria of cholesterol/dipalmitoylphosphatidylcholine mixtures: 2H nuclear magnetic resonance and differential scanning calorimetry.

Deuterium nuclear magnetic resonance spectroscopy and differential scanning calorimetry are used to map the phase boundaries of mixtures of cholesterol and chain-perdeuteriated 1,2-dipalmitoyl-sn-glycero-3-phosphocholine at concentrations from 0 to 25 mol % cholesterol. Three distinct phases can be identified: the L alpha or liquid-crystalline phase, the gel phase, and a high cholesterol concentration phase, which we call the beta phase. The liquid-crystalline phase is characterized by highly flexible phospholipid chains with rapid axially symmetric reorientation; the gel phase has much more rigid lipid chains, and the motions are no longer axially symmetric on the 2H NMR time scale; the beta phase is characterized by highly ordered (rigid) chains and rapid axially symmetric reorientation. In addition, we identify three regions of two-phase coexistence. The first of these is a narrow L alpha/gel-phase coexistence region lying between 0 and about 6 mol % cholesterol at temperatures just below the chain-melting transition of the pure phospholipid/water dispersions, at 37.75 degrees C. The dramatic changes in the 2H NMR line shape which occur on passing through the phase transition are used to map out the boundaries of this narrow two-phase region. The boundaries of the second two-phase region are determined by 2H NMR difference spectroscopy, one boundary lying near 7.5 mol % cholesterol and running from 37 down to at least 30 degrees C; the other boundary lies near 22 mol % cholesterol and covers the same temperature range. Within this region, the gel and beta phases coexist. As the temperature is lowered below about 30 degrees C, the phospholipid motions reach the intermediate time scale regime of 2H NMR so that spectral subtractions become difficult and unreliable. The third two-phase region lies above 37 degrees C, beginning at a eutectic point somewhere between 7.5 and 10 mol % cholesterol and ending at about 20 mol %. In this region, the L alpha and beta phases are in equilibrium. The boundaries for this region are inferred from differential scanning calorimetry traces, for the boundary between the L alpha- and the two-phase region, and from a dramatic sharpening of the NMR peaks on crossing the boundary between the two-phase region and the beta-phase region. In this region, the technique of difference spectroscopy fails, presumably because the diffusion rate in both the L alpha- and beta-phase domains is so rapid that phospholipid molecules exchange rapidly between domains on the experimental time scale.

1,2-Dipalmitoylphosphatidylcholine↗

AC calorimetry of dimyristoylphosphatidylcholine multilayers: hysteresis and annealing near the gel to liquid-crystal transition.

The gel to liquid-crystal phase transition in dimyristoylphosphatidylcholine liposomes was studied with 0.4-Hz ac calorimetry. The ac heat capacity on heating scans exhibited a peak in the vicinity of 23.9 degrees C with a full width at half-maximum of 0.15-0.20 degrees C. The enthalpy change was 4.8 kcal/mol, in good agreement with conventional differential scanning calorimetry. On cooling scans, the peak shifted to lower temperature by 0.1-0.5 degrees C, the width increased to 0.25-0.40 degrees C, and the apparent enthalpy change was only 40% of that observed on heating. Both the heating and cooling heat capacities were stable for at least 20 min in quasi-isothermal conditions. Following a 1 h anneal at 10 degrees C, the heating scans were quite reproducible. The results have been interpreted in terms of the nucleation and subsequent annealing of small ordered domains in the bilayer on freezing the acyl chains. No peak associated with the pretransition was observed, as expected since the relaxation time for the degrees of freedom the produce the pretransition is much longer than the period of the 0.4-Hz temperature wave.

Calorimetry↗

Scanning calorimetry and Fourier-transform infrared studies into the thermal stability of cleaved bacteriorhodopsin systems.

Differential scanning calorimetry and Fourier-transform infrared spectroscopy have been used to characterize the thermal stability of bacteriorhodopsin (BR) cleaved within different loops connecting the helical rods. The results are compared to those of the native protein. We show that the denaturation temperature and enthalpy of BR cleaved at peptide bond 71-72 or 155-156 are lower than those of the intact protein, and that these values become even lower for the BR cleaved at both peptide bonds. The effect of cleavage on the denaturation temperature and enthalpy values seems to be additive as has been previously suggested [Khan, T. W., Sturtevant, J. M., & Engelman, D. M. (1992) Biochemistry 31, 8829]. The thermal denaturation of all the samples was irreversible and scan-rate dependent. When cleaved at the 71-72 bond BR follows quantitatively the predictions of the two-state kinetic model at pH 9.5, with an activation energy of 374 kJ/mol, similar to that of native BR. Calorimetry experiments with different populations of intact and cleaved BR provide direct evidence for some intermolecular cooperativity upon denaturation. The denatured samples maintain a large proportion of alpha helices and beta structure, a fact which seems to be related to their low denaturation enthalpy as compared to that of water-soluble, globular proteins.

Bacteriorhodopsins↗

Thermodynamics of reversible and irreversible unfolding and domain interactions of glucoamylase from Aspergillus niger studied by differential scanning and isothermal titration calorimetry.

The stability of three forms of glucoamylase from Aspergillus niger has been investigated by differential scanning and isothermal titration calorimetry: Glucoamylase 1 (GA1), which consists of a catalytic domain and a starch-binding domain (SBD) connected by a heavily O-glycosylated linker region; glucoamylase 2 (GA2), which lacks SBD; and a proteolytically cleaved glucoamylase (GACD), which contains the catalytic domain and part of the linker region. The structures of the catalytic domain with part of the linker region and of SBD are known from crystallography and NMR, respectively, but the precise spatial arrangement of the two domains in GA1 is unknown. To investigate the stability of the three glucoamylase forms, we unfolded the enzymes thermally by differential scanning calorimetry (DSC). Aggregation occurs upon heating GA1 and GA2 at pH values between 2.5 and 5.0, whereas no aggregation is observed at higher pH (5.5-7.5). At all pH values, the catalytic domain of GA1 and GA2 unfolds irreversibly, while SBD unfolds reversibly in the pH range 5. 5-7.5 where aggregation does not occur. The unfolding of the catalytic domain of all glucoamylase forms seems to follow an irreversible one-step mechanism with no observable reversible intermediates on the experimental time scale. SBD of GA1 unfolds reversibly, and the ratio between the van't Hoff and calorimetric enthalpies is 1.4 +/- 0.1. Assignment of peaks of the DSC profile to the domains at pH 7.5 is achieved by using two different ligands: Acarbose, a very strong inhibitor that binds exclusively to the catalytic domain, and beta-cyclodextrin, a small starch analogue of which 2 molecules bind solely to the two binding sites present in SBD. Differences are seen in the unfolding processes of GA1 and GA2 since the former unfolds with one peak at all pH values, while the calorimetric trace of the latter can be resolved into more peaks depending on pH and the chemical composition of the buffers. In general, peaks corresponding to unfolding of GA2 are more complex than the peaks of GA1 and GACD. Some part of GA2 unfolds before the rest of the molecule which may correspond to the linker region or a particular early unfolding part of the catalytic domain. This leads to the conclusion that the structure of the GA2 molecule has a larger cooperative unfolding unit and is less stable than the structures of GA1 and GACD and that the C-terminal part of the linker region has a destabilizing effect on the catalytic domain.

Acarbose↗

On the value of c: can low affinity systems be studied by isothermal titration calorimetry?

Isothermal titration calorimetry (ITC) allows the determination of DeltaG degrees, DeltaH degrees, and DeltaS degrees from a single experiment and is thus widely used for studying binding thermodynamics in both biological and synthetic supramolecular systems. However, it is widely believed that it is not possible to derive accurate thermodynamic information from ITC experiments in which the Wiseman "c" parameter (which is the product of the receptor concentration and the binding constant, K(a)) is less than ca. 10, constraining its use to high affinity systems. Herein, experimental titrations and simulated data are used to demonstrate that this dogma is false, especially for low affinity systems, assuming that (1) a sufficient portion of the binding isotherm is used for analysis, (2) the binding stoichiometry is known, (3) the concentrations of both ligand and receptor are known with accuracy, and (4) there is an adequate level of signal-to-noise in the data. This study supports the validity of ITC for determining the value of K(a) and, hence, DeltaG degrees from experiments conducted under low c conditions but advocates greater caution in the interpretation of values for DeltaH degrees. Therefore, isothermal titration calorimetry is a valid and useful technique for studying biologically and synthetically important low affinity systems.

Calorimetry↗

Maltodextrin-anionic surfactant interactions: isothermal titration calorimetry and surface tension study.

Interactions between maltodextrin (DE = 10) and an anionic surfactant (sodium dodecyl sulfate, SDS) were studied in a buffer solution (pH 7.0, 10 mM NaCl, 20 mM Trizma, 30.0 degrees C) using isothermal titration calorimetry (ITC), surface tension, differential scanning calorimetry (DSC), and turbidity techniques. ITC measurements indicated that the binding of SDS to maltodextrin was exothermic and that, on average, one SDS monomer bound per 24 glucose units of maltodextrin at saturation. Surface tension measurements indicated that there was a critical surfactant concentration ( approximately 0.05 mM SDS) below which surfactant and maltodextrin did not interact and that the amount of surfactant bound to the maltodextrin above this concentration increased with increasing maltodextrin concentration. Turbidity measurements indicated that the solutions remained transparent at all maltodextrin (0-1 wt %) and SDS (0-20 mM) concentrations studied, which suggested that phase separation did not occur. DSC measurements indicated that no phase transitions occurred between 10 and 110 degrees C for maltodextrin solutions (0.5 wt %) in the presence or absence of surfactant. A phase diagram was developed to describe the interactions between SDS and maltodextrin.

Anions↗

Solid-state stability testing of drugs by isothermal calorimetry.

A new technique has been developed to calculate rapidly the solid-state room-temperature degradation rate of drugs and drug candidates. The technique utilizes measurements of the initial rate of heat output at several elevated temperatures by isothermal calorimetry and the degradation rate of the compound determined at a single elevated temperature by chromatography. The activation energies and degradation rates at 25 degrees C calculated by conventional methods and by isothermal calorimetry are compared and discussed. The compounds studied were phenytoin, triamterene, digoxin, tetracycline, theophylline, diltiazem, and several proprietary ICI compounds.

Calorimetry↗

Determination of the glass transition temperatures of some new methyl methacrylate copolymers using modulated temperature differential scanning calorimetry (MTDSC).

PURPOSE: The purpose of this study was to determine the glass transition temperatures of new graft copolymers using Modulated Temperature Differential Scanning Calorimetry (MTDSC), and to assess the differences between starch and cellulosic derivatives of methyl methacrylate and between two different drying methods used in their preparation. METHODS: Graft copolymers of methyl methacrylate were synthesized and dried by oven or freeze-drying. Surface area measurements and different thermal analysis techniques (Differential Scanning Calorimetry (DSC), Thermogravimetric analysis (TGA) and MTDSC) were used to characterize these copolymers. Results. DSC was not sensitive enough to identify the T(g)s of the copolymers, however they were clearly identifiable by MTDSC. T(g) values obtained may depend on the method of preparation that also altered their physical characteristics e.g. specific surface area. Cellulose derivatives showed lower T(g)s than starch derivatives. The results also depended on the drying method used, thus, freeze dried products had slightly lower T(g)s than oven dried products. CONCLUSIONS: MTDSC represents a useful thermal technique that allows the identification of glass transitions in these new copolymers with higher sensitivity and resolution than conventional DSC, separating the transition from overlapping phenomena such as decomposition or dehydration. The Tg of this new class of copolymers appeared to be dependent on polymer composition and drying method used.

Calorimetry, Differential Scanning↗

Some mathematical and technical issues in the measurement and interpretation of open-circuit indirect calorimetry in small animals.

Indirect calorimetry is increasingly used to investigate why compounds or genetic manipulations affect body weight or composition in small animals. This review introduces the principles of indirect (primarily open-circuit) calorimetry and explains some common misunderstandings. It is not widely understood that in open-circuit systems in which carbon dioxide (CO2) is not removed from the air leaving the respiratory chamber, measurement of airflow out of the chamber and its oxygen (O2) content paradoxically allows a more reliable estimate of energy expenditure (EE) than of O2 consumption. If the CO2 content of the exiting air is also measured, both O2 consumption and CO2 production, and hence respiratory quotient (RQ), can be calculated. Respiratory quotient coupled with nitrogen excretion allows the calculation of the relative combustion of the macronutrients only if measurements are over a period where interconversions of macronutrients that alter their pool sizes can be ignored. Changes in rates of O2 consumption and CO2 production are not instantly reflected in changes in the concentrations of O2 and CO2 in the air leaving the respiratory chamber. Consequently, unless air-flow is high and chamber size is small, or rates of change of O2 and CO2 concentrations are included in the calculations, maxima and minima are underestimated and will appear later than their real times. It is widely appreciated that bigger animals with more body tissue will expend more energy than smaller animals. A major issue is how to compare animals correcting for such differences in body size. Comparison of the EE or O2 consumption per gram body weight of lean and obese animals is misleading because tissues vary in their energy requirements or in how they influence EE in other ways. Moreover, the contribution of fat to EE is lower than that of lean tissue. Use of metabolic mass for normalisation, based on interspecific scaling exponents (0.75 or 0.66), is similarly flawed. It is best to use analysis of covariance to determine the relationship of EE to body mass or fat-free mass within each group, and then test whether this relationship differs between groups.

Animals↗

Indirect calorimetry in patients with active respiratory infection--prevention of cross-infection.

BACKGROUND: Measurement of resting energy expenditure in patients with active respiratory infection is hampered by the risk of contamination of indirect calorimeters and connecting tubes that cannot be sterilized. This validation study tested whether the use of disposable standard-bore air tubes and an antibacterial filter, by reducing air flow, introduces an error due to recirculation of expired air in the canopy. METHODS: Eleven healthy volunteers underwent indirect calorimetry twice in random order, using a Deltatrac calorimeter either with standard wide-bore tubing or with a disposable standard-bore air tube and filter. Methods were compared by Bland-Altman plots and by calculating trends over time. RESULTS: The new tube and filter reduced air flow by 40%. Measured resting energy expenditure did not differ significantly between methods, with limits of agreement -135 to +188 kcal/d. Carbon dioxide flow (VCO(2)) and respiratory quotient (RQ), but not oxygen flow decreased slowly over time with both methods. CONCLUSIONS: The use of an air filter and standard-bore tubes do not introduce a systematic error into indirect calorimetry. Although trends in VCO(2) and RQ are consistent with minor recirculation of exhaled air, this modified Deltatrac system can be safely and reliably used to measure resting energy expenditures in patients with active tuberculosis and other airborne infection.

Adult↗

Differential scanning calorimetry of nuclei reveals the loss of major structural features in chromatin by brief nuclease treatment.

Differential scanning calorimetry revealed that chromatin melts in four distinct transitions in intact HeLa nuclei at 60 degrees C, 76 degrees C, 88 degrees C, and 105 degrees C. Calorimetry of whole cells was characterized by the same four transitions along with another at 65 degrees C, which was probably RNA. Isolated chromatin, however, melted in only two transitions at 72 degrees C and 85 degrees C. Very brief digestion of HeLa nuclei with either micrococcal nuclease or DNase I resulted in the conversion of a structure that melted at 105 degrees C to one that melted at 88 degrees C. Further digestion with micrococcal nuclease to the level of the mononucleosome did not result in any further substantial changes in either enthalpy or melting temperatures. In contrast, further DNase I digestion that resulted in cleavage within the nucleosome produced a pronounced shift in melting temperatures to broad transitions at 62 degrees C and 78 degrees C.

Calorimetry, Differential Scanning↗

Isothermal titration calorimetry reveals a zinc ion as an atomic switch in the diadenosine polyphosphates.

Diadenosine polyphosphates (diadenosine 5',5'''-P(1),P(n)-polyphosphate (Ap(n)A)) are 5'-5'''-phosphate-bridged dinucleosides that have been proposed to act as signaling molecules in a variety of biological systems. Isothermal titration calorimetry was used to measure the affinities of a variety of metal cations for ATP, diadenosine 5',5'''-P(1),P(3)-triphosphate (Ap(3)A), diadenosine 5',5'''-P(1),P(4)-tetraphosphate (Ap(4)A), and diadenosine 5',5'''-P(1),P(5)-pentaphosphate (Ap(5)A). The binding of Mg(2+), Ca(2+), and Mn(2+) to ATP is shown to take place with the beta,gamma-phosphates (primary site) and be endothermic in character. The binding of Ni(2+), Cd(2+), and Zn(2+) to ATP is found to take place at both the primary site and at a secondary site identified as N-7 of the adenine ring. Binding to this second site is exothermic in character. Generally, the binding of metal cations to diadenosine polyphosphates involves a similar primary site to ATP. No exothermic binding events are identified. Critically, the binding of Zn(2+) to diadenosine polyphosphates proves to be exceptional. This appears to involve a very high affinity association involving the N-7 atoms of both adenine rings in each Ap(n)A, as well as the more usual endothermic association with the phosphate chain. The high affinity association is also endothermic in character. A combination of NMR and CD evidence is provided in support of the calorimetry data demonstrating chemical shift changes and base stacking disruptions entirely consistent with N-7 bridging interactions. N-7 bridging interactions are entirely reversible, as demonstrated by EDTA titration. Considering the effects of Zn(2+) on a wide variety of dinucleoside polyphosphate-metabolizing enzymes, we examine the possibility of Zn(2+) acting as an atomic switch to control the biological function of the diadenosine polyphosphates.

Adenosine Triphosphate↗

The multidrug efflux regulator TtgV recognizes a wide range of structurally different effectors in solution and complexed with target DNA: evidence from isothermal titration calorimetry.

TtgV modulates the expression of the ttgGHI operon, which encodes an efflux pump that extrudes a wide variety of chemicals including mono- and binuclear aromatic hydrocarbons, aliphatic alcohols, and antibiotics of dissimilar chemical structure. Using a 'lacZ fusion to the ttgG promoter, we show that the most efficient in vivo inducers were 1-naphthol, 2,3-dihydroxynaphthalene, 4-nitrotoluene, benzonitrile, and indole. The thermodynamic parameters for the binding of different effector molecules to purified TtgV were determined by isothermal titration calorimetry. For the majority of effectors, the interaction was enthalpy-driven and counterbalance by unfavorable entropy changes. The TtgV-effector dissociation constants were found to vary between 2 and 890 mum. There was a relationship between TtgV affinity for the different effectors and their potential to induce gene expression in vivo, indicating that the effector binding constant is a major determinant for efficient efflux pump gene expression. Equilibrium dialysis and isothermal titration calorimetry studies indicated that a TtgV dimer binds one effector molecule. No evidence for the simultaneous binding of multiple effectors to TtgV was obtained. The binding of TtgV to a 63-bp DNA fragment containing its cognate operator was tight and entropy-driven (K(D) = 2.4 +/- 0.35 nm, DeltaH = 5.5 +/- 0.04 kcal/mol). The TtgV-DNA complex was shown to bind 1-napthol with an affinity comparable with the free soluble TtgV protein, K(D) = 4.8 +/- 0.19 and 3.0 +/- 0.15 mum, respectively. The biological relevance of this finding is discussed.

Bacterial Proteins↗

Metabolic effects of altering the 24 h energy intake in man, using direct and indirect calorimetry.

1. The metabolic effects of increasing or decreasing the usual energy intake for only 1 d were assessed in eight adult volunteers. Each subject lived for 28 h in a whole-body calorimeter at 26 degrees on three separate occasions of high, medium or low energy intake. Intakes (mean +/- SEM) of 13830 +/- 475 (high), 8400 +/- 510 (medium) and 3700 +/- 359 (low) kj/24 h were eaten in three meals of identical nutrient composition. 2. Energy expenditure was measured continuously by two methods: direct calorimetry, as total heat loss partitioned into its evaporative and sensible components: and indirect calorimetry, as heat production calculated from oxygen consumption and carbon dioxide production. For the twenty-four sessions there was a mean difference of only 1.2 +/- 0.14 (SEM) % between the two estimates of 24 h energy expenditure, with heat loss being less than heat production. Since experimental error was involved in both estimates it would be wrong to ascribe greater accuracy to either one of the measures of energy expenditure. 3. Despite the wide variation in the metabolic responses of the subjects to over-eating and under-eating, in comparison with the medium intake the 24 h heat production increased significantly by 10% on the high intake and decreased by 6% on the low intake. Mean (+/- SEM) values for 24 h heat production were 8770 +/- 288, 7896 +/- 297 and 7495 +/- 253 kJ on the high, medium and low intakes respectively. The effects of over-eating were greatest at night and the resting metabolic rate remained elevated by 12% 14 h after the last meal. By contrast, during under-eating the metabolic rate at night decreased by only 1%. 4. Evaporative heat loss accounted for an average of 25% of the total heat loss at each level of intake. Changes in evaporative heat loss were +14% on the high intake and -10% on the low intake. Sensible heat loss altered by +9 and -5% on the high and low intakes respectively. 5. It is concluded that (a) the effects on 24 h energy expenditure of over-feeding for only 1 d do not differ markedly from those estimated by some other workers after several weeks of increasing the energy intake: (b) the resting metabolic rate, measured at least 14 h after the last meal, can be affected by the previous day's energy intake; (c) the zone of ambient temperature within which metabolism is minimal is probably altered by the level of energy intake.

Adult↗

Whole-body calorimetry studies in adult men. 1. The effect of fat over-feeding on 24 h energy expenditure.

Eight young men of normal weight were maintained for 1 week on a weight-maintenance diet followed by a 1-week period of over-feeding with extra fat designed to increase energy intake by 50%. Two 36 h calorimetry sessions with low and high physical activities were included in each feeding period. Faecal and urine collections permitted checks on energy malabsorption and nitrogen excretion. Over-feeding led to increases in body-weight, faecal energy and N excretion and in estimated N retention. Faecal energy outputs on the maintenance and over-feeding diets were 5 and 4.4% of the respective gross energy intakes. Energy expenditure on fat over-feeding increased by 5.6% on the low-activity regimen and 6.4% on the high-activity regimen. This amounted, in terms of the extra energy intake, to 9 and 11% on the inactive and active schedules respectively. The increase affected day- and night-time rates of energy expenditure plus the basal metabolic rate. Individuals with a low percentage body fat showed the greatest response to over-feeding. Nutrient-balance studies derived from calorimetry suggested that fat over-feeding led to substantial fat deposition with no evidence of sparing of carbohydrate oxidation. The theoretical cost of depositing dietary fat was exceeded, suggesting that regulatory thermogenic mechanisms may have been stimulated to a small extent.

Adult↗

Absorbed dose to water determination with ionization chamber dosimetry and calorimetry in restricted neutron, photon, proton and heavy-ion radiation fields.

Absolute dose measurements with a transportable water calorimeter and ionization chambers were performed at a water depth of 20 mm in four different types of radiation fields, for a collimated (60)Co photon beam, for a collimated neutron beam with a fluence-averaged mean energy of 5.25 MeV, for collimated proton beams with mean energies of 36 MeV and 182 MeV at the measuring position, and for a (12)C ion beam in a scanned mode with an energy per atomic mass of 430 MeV u(-1). The ionization chambers actually used were calibrated in units of air kerma in the photon reference field of the PTB and in units of absorbed dose to water for a Farmer-type chamber at GSI. The absorbed dose to water inferred from calorimetry was compared with the dose derived from ionometry by applying the radiation-field-dependent parameters. For neutrons, the quantities of the ICRU Report 45, for protons the quantities of the ICRU Report 59 and for the (12)C ion beam, the recommended values of the International Atomic Energy Agency (IAEA) protocol (TRS 398) were applied. The mean values of the absolute absorbed dose to water obtained with these two independent methods agreed within the standard uncertainty (k = 1) of 1.8% for calorimetry and of 3.0% for ionometry for all types and energies of the radiation beams used in this comparison.

Calorimetry↗

Theory and validity of indirect calorimetry during net lipid synthesis.

A critical examination is made of the validity of indirect calorimetry when the nonprotein respiratory quotient is greater than 1. The different published stoichiometries for lipogenesis from glucose are excluded as a source of uncertainty in the interpretation of gaseous exchange measurements. The validity of indirect calorimetry is proved independently by an algebraic approach which, in contrast to previous attempts, makes minimal assumptions about stoichiometries. Although equations relating the respiratory quotient to the heat equivalent of oxygen are found valid, there is uncertainty in using these equations to predict accurately carbohydrate utilization and fat oxidized or synthesized. Reference tables interrelating respiratory data, the heat equivalent of oxygen, and net fuel utilization or synthesis for specified fuels are provided. A suggested framework for calculating energy expenditure in terms of ATP gain is given as an appendix.

Adipose Tissue↗

Resting energy expenditure in children in a pediatric intensive care unit: comparison of Harris-Benedict and Talbot predictions with indirect calorimetry values.

The use of prediction equations has been recommended for calculating energy expenditure. We evaluated two equations that predict energy expenditure, each of which were corrected for two different stress factors, and compared the values obtained with those calculated by indirect calorimetry. The subjects were 55 critically ill children on mechanical ventilation. Basal metabolic rates were calculated with the Harris-Benedict and Talbot methods. Measured resting energy expenditure was 4.72 +/- 2.53 MJ/d. The average difference between measured resting energy expenditure and the Harris-Benedict prediction with a stress factor of 1.5 was -0.98 MJ/d, with an SD delta of 1.56 MJ/d and limits of agreement from -4.12 to 2.15; for a stress factor of 1.3 the average difference was -0.22 MJ/d, with an SD delta of 1.57 MJ/d and limits of agreement from -3.37 to 2.93. The average difference between measured resting energy expenditure and the Talbot prediction with a stress factor of 1.5 was -0.23 MJ/d, with an SD delta of 1.36 MJ/d and limits of agreement from -2.95 to 2.48; for a stress factor of 1.3, it was 0.42 MJ/d, with an SD delta of 1.24 MJ/d and limits of agreement from -2.04 to 2.92. These limits of agreement indicate large differences in energy expenditure between the measured value and the prediction estimated for some patients. Therefore, neither the Harris-Benedict nor the Talbot method will predict resting energy expenditure with acceptable precision for clinical use. Indirect calorimetry appears to be the only useful way of determining resting energy expenditure in these patients.

Adult↗