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[Study of metabolism in critically ill patients by indirect calorimetry].

Indirect calorimetry was performed in 48 postsurgical critically ill patients including those with multiple organ failure. The patients were divided into two groups, organ failure group (OF group) and postsurgical control group (C group), according to the presence of postsurgical organ failure and severe infection. The following results were obtained. The ratio of energy expenditure to basal energy expenditure, reflecting a degree of hypermetabolism, was 1.44 +/- 0.38 in OF group and 1.26 +/- 0.23 in C group respectively. The change in respiratory quotient by caloric intake was greater in OF group than in C group, indicating that it is more important to maintain an adequate intake for the prevention of increased respiratory work and excess lipogenesis in OF group. When nitrogen intake as amino acids was sufficient (0.1-0.2 g/kg/day), nitrogen balance could be maintained around 0 by a caloric intake being equal to measured energy expenditure. A positive correlation was observed between arterial ketone body ratio reflecting energy charge in hepatocytes and respiratory quotient, indicating that in patients with impaired mitochondrial function in hepatocytes as shown by a decrease in ketone body ratio, glucose cannot be utilized effectively. These results let us conclude that calorimetry is indispensable in the management of critically ill patients and that adequate energy intake should be cautiously determined according to the calorimetry.

Calorimetry↗

Indirect calorimetry in postoperative patients with acute renal failure.

Oxygen consumption was measured and caloric expenditure calculated (indirect calorimetry) in ten adult patients with acute renal failure (ARF) following surgery. Eight patients recovered from ARF and six were discharged from the hospital. Caloric expenditure averaged 47 Kcal/kg/24 hours during the period of AFR. Attempts were made to match caloric replacement with expenditure based on indirect calorimetry, but was achieved in only one patient. Indirect calorimetry appears to be a practical method for guiding caloric replacement in these patients. Postoperative patients in ARF must undergo healing of incisions and bowel anastomoses, and contain infection. Since these patients are severely catabolic, it seems reasonable to treat them with adequate protein and calories and to use dialysis to control azotemia and water metabolism.

Acute Kidney Injury↗

Indirect calorimetry in the nutritional management of eating disorders.

The caloric prescription, a key component of the nutritional therapy of anorexia nervosa (AN) and bulimia nervosa (BN), may be empirically prescribed, or based on predicted resting energy expenditure (REE), yet adaptive changes in the metabolic rate may render both methods unreliable. Indirect calorimetry measurement of fasting REE was obtained in 32 patients with AN (n = 21) or BN (n = 11). Predicted REE was calculated according to the Harris-Benedict equation, and empiric caloric prescriptions were made by experienced physicians. In the AN group, mean measured REE was significantly lower than predicted REE (p = .00). The empiric caloric prescription was, as intended, significantly higher than the measured REE, but the two methods correlated significantly (r = .53, p < .05). The predicted REE overestimated caloric needs but was also highly correlated with measured REE (r = .69, p < .001). By regression analysis, measured REE could be calculated from predicted REE as follows: measured REE (Kcal/day) = (1.84 x Harris-Benedict predicted REE) - 1,435. In the BN group, mean measured REE was not significantly different from the empiric caloric prescription (p = .09) but was significantly lower than the Harris-Benedict predicted REE (p = .022). Neither correlated with measured REE in BN. Therefore, in BN indirect calorimetry is the only reliable method for determining caloric needs. In AN indirect calorimetry remains the preferred method, but when not available, we recommend the above equation to determine resting energy requirements.

Adolescent↗

A potential role for isothermal calorimetry in studies of the effects of thermodynamic non-ideality in enzyme-catalyzed reactions.

Attention is drawn to the feasibility of using isothermal calorimetry for the characterization of enzyme reactions under conditions bearing greater relevance to the crowded biological environment, where kinetic parameters are likely to differ significantly from those obtained by classical enzyme kinetic studies in dilute solution. An outline of the application of isothermal calorimetry to the determination of enzyme kinetic parameters is followed by considerations of the nature and consequences of crowding effects in enzyme catalysis. Some of those effects of thermodynamic non-ideality are then illustrated by means of experimental results from calorimetric studies of the effect of molecular crowding on the kinetics of catalysis by rabbit muscle pyruvate kinase. This review concludes with a discussion of the potential of isothermal calorimetry for the experimental determination of kinetic parameters for enzymes either in biological environments or at least in media that should provide reasonable approximations of the crowded conditions encountered in vivo.

Calorimetry↗

Dynamic monitoring of restricted eating disorders by indirect calorimetry: a useful cognitive approach.

OBJECTIVE: Outpatient treatment in restricted eating disorder: indirect calorimetry during dynamic monitoring. DESIGN: A retrospective observational study. SUBJECTS: Twenty seven women affected by restricted eating disorder (essentially anorexia nervosa) with a body mass index [weight (kg)/height (m2)] of 17.29+/-2.47 were studied. The sample was compared as itself control during rehabilitative way. INTERVENTIONS: Fat mass (FM) and fat free mass (FFM) were determined by anthropometry technique. REE/day and respiratory quotient (RQ,VCO2/VO2) were measured by indirect calorimetry using a Calorimeter Vmax 29n-Sensor Medics-California. Skinfold thickness and circumferences were also measured. Arm muscle area (AMA) and fat area were calculated by formulas reported in Frisancho. RESULTS: The data indicated a positive correlation between AMA, VO2/ml/min and resting energy expenditure (REE)/day values examined during follow-up of patients. The increase of these parameters indicated a good monitoring index correlated to a FFM recovery during psychonutritional rehabilitation. CONCLUSION: Indirect calorimetry represents a useful approach for determining REE and prescribing diets in these patients. Moreover, the combined use of anthropometric techniques allows to accurately assess and adjust therapy according to the patient's progress. This study shows that restricted eating disorders are characterized by a recovery of FFM related to improvement of body weight and REE/day. On the contrary, the increase of AFA revealed a recovery of fat-metabolism (corresponding to RQ decrease) and lipid/carbohydrates oxidation improvement, only in the presence, at the same time, of O2 consumption increase.

Adiposity↗

Validity of an abbreviated indirect calorimetry protocol for measurement of resting energy expenditure in mechanically ventilated and spontaneously breathing critically ill patients.

OBJECTIVE: To test a short indirect calorimetry protocol with five stable 1-min readings (5-min steady state) against the commonly used protocol of 30 1-min readings (30-min steady state) in critically ill patients with various modes of ventilation. DESIGN: A prospective clinical study. SETTING: A medical ICU of a university hospital. SUBJECTS: Forty-six mechanically ventilated patients (group A and B), and 16 spontaneously breathing patients (group C). INTERVENTION: Indirect calorimetry with the Deltatrac II MBM-200 Metabolic Monitor. RESULTS: Mechanically ventilated patients were classified into group A (controlled) and B (assisted) depending on the ventilation mode. All patients in group A, but only 48.8% of those in group B, received some form of analgosedation, and the doses were significantly higher in group A. The 30-min steady state test was 100.0%, 83.7%, and 75.0% successful in group A, B, and C, respectively. The corresponding rate for the 5-min steady state test was 100.0%, 81.4%, and 100.0%, respectively. The coefficient of determination (r2) for resting energy expenditure between the two protocols ranged between 0.972 and 0.994. The time required to collect the 5-min steady state data was 5.5+/-1.1, 9.9+/-5.7, and 6.5+/-3.3 min for group A, B, and C, respectively. CONCLUSION: Indirect calorimetry with 5-min steady state test correlated very well with the 30-min steady state test in both mechanically ventilated and spontaneously breathing patients. The time required varies depending on the mode of ventilation, and it is influenced by the level of sedation in mechanically ventilated patients. The abbreviated protocol may be more acceptable to spontaneously breathing patients.

Calorimetry, Indirect↗

Practical aspects of indirect calorimetry in laboratory animals.

Oxidation of the energetic substrates by the body is associated with oxygen consumption, carbon dioxide production, and heat release specific to the nature of the energetic substrates being oxidized. Therefore, measurement of respiratory exchanges (indirect calorimetry) is a powerful method to investigate heat production of a living organism. In this article, we review the elementary principles of indirect calorimetry and describe the operating principle of the two most typical devices used to perform indirect measurements of energy expenditure in the laboratory animal: the closed-circuit and the open-circuit. We then discuss some practical aspects of the day-to-day use of these devices: respective advantages and limitations of each technique, data processing, calibration, correction for body-size, and computation of the energy expended for activity. In the second part, we review some of the standard formulas of indirect calorimetry that offer the possibility to obtain more precise information such as the rate of oxidation of carbohydrates (CHO), lipids and proteins if some hypotheses are made on the intensity of lipogenic, ketogenic, and gluconeogenic processes. Finally, a practical example of the measurement of energetic cost of activity and thermic effect of food in the rat is given.

Animals↗

Isothermal acid-titration calorimetry for evaluating the pH dependence of protein stability.

A new method, which can be called as isothermal acid-titration calorimetry (IATC), was proposed for evaluating the enthalpy of protein molecules as a function of pH using isothermal titration calorimetry (ITC). This measurement was used to analyze the acid-denaturation of bovine ribonuclease A. The enthalpy change by acid-denaturation of this protein was estimated as 310 kJ/mol at pH 2.8 and 40 degrees C. This value agreed well with the enthalpy change obtained by differential scanning calorimetry. The midpoint pH and proton binding-number difference observed by IATC agreed well with those of the acid transition of the three-dimensional structure monitored by circular dichroism spectrometry. The van't Hoff enthalpy of the transition was derived from the temperature dependence of the midpoint pH and the proton binding-number difference. It agreed well with the calorimetric enthalpy change directly observed by IATC, strongly indicating that there was no stable intermediate state during the acid transition of this protein.

Algorithms↗

Calorimetry of enzyme-catalyzed reactions.

This mini-review shows the valuable contributions of Professor Julian Sturtevant to the current applications of calorimetry to the study of enzyme-catalyzed reactions. The more recent applications of calorimetric techniques such as isothermal titration calorimetry and flow calorimetry to the study of enzyme kinetics, as well as the advantages on using calorimetric techniques in the determination of kinetic parameters of enzymes, is also discussed here.

Calorimetry↗

Resting energy expenditure in severely burned children: analysis of agreement between indirect calorimetry and prediction equations using the Bland-Altman method.

INTRODUCTION: Knowledge of a child's resting energy expenditure (REE) is essential in optimizing nutritional support for severely burned children. The provision of adequate nutritional support is vital in order to avoid the consequences of malnutrition or overfeeding. Nutritional requirements for severely burned children are often based on equations for estimates of REE. The accuracy of the predictive equations of REE has been questioned and many authors have advocated the measurement of REE. This study tests the hypothesis that estimates of REE vary significantly from measured REE (MREE) in a population of severely burned children, and are not accurate for determining nutritional requirements. METHODS: In 91 severely burned children aged between 3 and 18 years, REE was measured by indirect calorimetry (MREE) at the height of the hypermetabolic response and compared with predicted equations (PREE) from the Food and Agriculture/World Health Organization/United Nations University (FAO/WHO/UNU), Schofield-HW and Harris-Benedict. Agreement between indirect calorimetry and predicted equations was assessed following the Bland-Altman method. RESULTS: In the entire cohort group, predicted REE from all three equations were significantly lower compared to MREE (p<0.05). There was poor agreement between the MREE and predicted using all three equations. The Schofield-HW equation showed the lowest mean MREE-PREE difference: 635+/-526 kcal/day (limits of agreement -608 and 1878 kcal/day; 95% confidence interval for the bias 525-745 kcal/day). Additionally, all three equations under predicted REE and were not significantly different from one another (p=0.98). CONCLUSIONS: Until more accurate predicted equations are developed, we recommend indirect calorimetry measurements for determining resting energy expenditure in severely burned children.

Adolescent↗

Multifrequency calorimetry of the folding/unfolding transition of cytochrome c.

The folding-unfolding transition of Fe(III) cytochrome c has been studied with the new technique of multifrequency calorimetry. Multifrequency calorimetry is aimed at measuring directly the dynamics of the energetic events that take place during a thermally induced transition by measuring the frequency dispersion of the heat capacity. This is done by modulating the folding/unfolding equilibrium using a variable frequency, small oscillatory temperature perturbation (approximately 0.05-0.1 degrees C) centered at the equilibrium temperature of the system. Fe(III) cytochrome c at pH 4 undergoes a fully reversible folding/unfolding transition centered at 67.7 degrees C and characterized by an enthalpy change of 81 kcal/mol and heat capacity difference between unfolded and folded states of 0.9 kcal/K*mol. By measuring the temperature dependence of the frequency dispersion of the heat capacity in the frequency range of 0.1-1 Hz it has been possible to examine the time regime of the enthalpic events associated with the transition. The multifrequency calorimetry results indicate that approximately 85% of the excess heat capacity associated with the folding/unfolding transition relaxes with a single relaxation time of 326 +/- 68 ms at the midpoint of the transition region. This is the first time that the time regime in which heat is absorbed and released during protein folding/unfolding has been measured.

Animals↗

Determination of the onset of crystallization of N1-2-(thiazolyl)sulfanilamide (sulfathiazole) by UV-Vis and calorimetry using an automated reaction platform; subsequent characterization of polymorphic forms using dispersive Raman spectroscopy.

This work describes the use of UV/visible spectroscopy and calorimetry to follow the onset of crystallization of a commercially available compound, N(1)-2-(thiazolyl)sulfanilamide (sulfathiazole), during crystallization reactions performed using an automated reaction platform. Sulfathiazole has been the subject of numerous publications through which considerable confusion about the morphic form is apparent. This work does not attempt to investigate exhaustively the polymorph issue, but rather to exploit the use of the HEL auto-MATE for monitoring the onset of crystal formation. Real-time calorimetry and UV-Vis spectroscopy are compared as tools for determining the onset of crystallization. Subsequently, differential scanning calorimetry, dispersive Raman, and infrared spectroscopy analysis serve to identify the crystal forms generated by the HEL auto-MATE. A solvent-anti-solvent matrix and several bench-top crystallization experiments were performed to supplement the investigation in terms of generating the desired polymorphs.

Ammonia↗

Thermodynamics of ion binding to phosphatidic acid bilayers. Titration calorimetry of the heat of dissociation of DMPA.

The heat of dissociation of the second proton of 1,2-dimyristoylphosphatidic acid (DMPA) was studied as a function of temperature using titration calorimetry. The dissociation of the second proton of DMPA was induced by addition of NaOH. From the calorimetric titration experiment, the intrinsic pK0 for the dissociation reaction could be determined by applying the Gouy-Chapman theory. pK0 decreases with temperature from ca. 6.2 at 11 degrees C to 5.4 at 54 degrees C. From the total heat of reaction, the dissociation enthalpy, delta Hdiss, was determined by subtracting the heat of neutralization of water and the heat of dilution of NaOH. In the temperature range between 2 and 23 degrees C, delta Hdiss is endothermic with an average value of ca. 2.5 kcal.mol-1 and shows no clear-cut temperature dependence. In the temperature range between 23 and 52 degrees C, delta Hdiss calculated after subtraction of the heat of neutralization and dilution is not the true dissociation enthalpy but includes contributions from the phase transition enthalpy, delta Htrans, as the pH jump induces a transition from the gel to the liquid-crystalline phase. The delta Cp for the reaction enthalpy observed in this temperature range is positive. Above 53 degrees C, the pH jump induces again only the dissociation of the second proton, and the bilayers stay in the liquid-crystalline phase. In this temperature range, delta Hdiss seems to decrease with temperature. The thermodynamic data from titration calorimetry and differential scanning calorimetry as a function of pH can be combined to construct a complete enthalpy-temperature diagram of DMPA in its two ionization states.

Calorimetry, Differential Scanning↗

Van't Hoff and calorimetric enthalpies from isothermal titration calorimetry: are there significant discrepancies?

The enthalpy of a reaction is most often determined through one of two means; it can be determined directly using calorimetry or indirectly by measuring the temperature dependence of the equilibrium constant (i.e., the van't Hoff method). Recently, discrepancies have been noted between the enthalpy measured by calorimetry, and the enthalpy determined by the van't Hoff method,. This has been suggested to indicate that the binding reaction is more complex than the simple one-to-one binding model used to describe the data. To better understand possible discrepancies between and, we have undertaken both experimental studies using isothermal titration calorimetry to measure the binding energetics of Ba(2+) binding 18-crown-6 ether and 2'-CMP binding RNase A, along with a simulation of a system involving a molecule in conformational equilibrium coupled with binding. We find that when experimental setup and analysis are correctly performed, no statistically significant discrepancies between and exist even for the linked system.

Barium↗

Interaction of the local anesthetics dibucaine and tetracaine with sarcoplasmic reticulum membranes. Differential scanning calorimetry and fluorescence studies.

The local anesthetics dibucaine and tetracaine inhibit the (Ca2+ + Mg2+)-ATPase from skeletal muscle sarcoplasmic reticulum [DeBoland, A. R., Jilka, R. L., & Martonosi, A. N. (1975) J. Biol. Chem. 250, 7501-7510; Suko, J., Winkler, F., Scharinger, B., & Hellmann, G. (1976) Biochim. Biophys. Acta 443, 571-586]. We have carried out differential scanning calorimetry and fluorescence measurements to study the interaction of these drugs with sarcoplasmic reticulum membranes and with purified (Ca2+ + Mg2+)-ATPase. The temperature range of denaturation of the (Ca2+ + Mg2+)-ATPase in the sarcoplasmic reticulum membrane, determined from our scanning calorimetry experiments, is ca. 45-55 degrees C and for the purified enzyme ca. 40-50 degrees C. Millimolar concentrations of dibucaine and tetracaine, and ethanol at concentrations higher than 1% v/v, lower a few degrees (degrees C) the denaturation temperature of the (Ca2+ + Mg2+)-ATPase. Other local anesthetics reported to have no effect on the ATPase activity, such as lidocaine and procaine, did not significantly alter the differential scanning calorimetry pattern of these membranes up to a concentration of 10 mM. The order parameter of the sarcoplasmic reticulum membranes, calculated from measurements of the polarization of the fluorescence of diphenylhexatriene, is not significantly altered at the local anesthetic concentrations that shift the denaturation temperature of the (Ca2+ + Mg2+)-ATPase.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Isothermal calorimetry study of calcium caseinate and whey protein isolate edible films cross-linked by heating and gamma-irradiation.

The contribution of thermal and radiative treatments as well as the presence of some excipients, namely glycerol, carboxymethylcellulose (CMC), pectin, and agar, on the formation of protein-protein interactions as well as the formation and loss of protein-water interactions was investigated by means of differential scanning calorimetry in an isothermal mode. Protein-water interactions were assessed through measurement of the heat of the wetting parameter. Isothermal calorimetry measurements pointed out that gamma-irradiation does not favor protein-water interactions, as reflected by its endothermic contribution (P < or = 0.05) to the heat of wetting values. Although significant (P < or = 0.05), the effect of the thermal treatment on endothermic responses using isothermal calorimetry was found to be somewhat lower. Among excipients added to biofilm formulations, glycerol generated the most important losses of protein-water interactions, as inferred by its significant (P < or = 0.05) endothermic impact on the heat of wetting values.

Agar↗

Thermogenesis associated with fermentable carbohydrate in humans, validity of indirect calorimetry, and implications of dietary thermogenesis for energy requirements, food energy and body weight.

BACKGROUND: Animal studies and theory show fermentable carbohydrate (FC) intake causes appreciably thermogenesis, but a similar occurrence in humans is controversial. HYPOTHESES: (a) That indirect calorimetry (IDC) is a valid method to assess thermogenesis during fermentation. (b) That a consistent and rigorous approach to the analysis of published IDC data from human studies will establish a representative thermogenic response to FC. (c) That conventional estimates of food energy and energy requirements can mismatch appreciably, more especially when thermogenesis is ignored. PURPOSE: To derive information and understanding of IDC, thermogenesis and energy balance in relation to food energy and energy requirement estimates. METHODS: (a) The validities of IDC equations that estimate the heat of reaction and carbohydrate utilization were assessed for various types of FCs under various circumstances. (b) Pooled analysis of eight published randomized cross-over studies in humans with elevation of FC intake. Studies were analysed for the first time or reanalysed according to a consistent approach with appropriate corrections for confounders. (c) Some 1500 regular and 'special' diet compositions were examined to assess the extent to which Atwater general food energy factors and updated estimates of energy requirements mismatch due to variation in substrate-associated thermogenesis and substrate-associated faecal+urinary energy losses. Impact of such mismatches on BMI was assessed under conditions of all else being equal. RESULTS: (a) Indirect calorimetry was valid, providing robust estimates of heat production during various types of fermentation; only small correction factors were necessary. By contrast, IDC equations for carbohydrate utilization sometimes applied poorly to FC. (b) A best estimate of thermogenesis in humans due to fermentation, above that due to oral glucose as a reference standard, was 0.39 (s.e.m. 0.14) kJ per kJ net metabolizable energy (NME; P<0.05, n=8 studies, total 72 humans) compared with 0.34 kJ/kJ from theory. Six sources of bias were identified; all had potential to underestimate FC thermogenesis. (c) Mismatches in energy availability and requirement estimates were often marked and translated into long-term differences in body mass index from approximately 20 to 33 kg/m(2) in average-height middle-aged initially obese women, and from approximately 22 to a non-survivable 13 kg/m(2) in initially slim women. CONCLUSIONS: (a) Indirect calorimetry is valid for the present purpose. (b) Thermogenesis in response to FC is real in humans and is comparable to that in animals and in theory. (c) Mismatches between estimates of energy requirements and dietary energy as metabolizable energy means the two expressions are not directly comparable, which has implications for the expression of food energy, energy requirements and the conduct and interpretation of research related to body weight.

Body Weight↗

Simplification of the method of assessing daily and nightly energy expenditure in children, using heart rate monitoring calibrated against open circuit indirect calorimetry.

Total daily energy expenditure (TDEE) can be assessed in children using several methods: the double-labelled water technique (DLW); indirect whole-body calorimetry; activity diary; accelerometry and heart rate (HR) monitoring. This last, low-cost and convenient technique has been validated against the DLW technique that is normally considered the reference method. The main advantages of HR monitoring are its social acceptability, and the examination of the time spent in specific activities or intensive exercise sessions. The aim of this study was to assess a new method for computing energy expenditure (EE) with the HR monitoring technique in children, using an open-circuit ventilated hood indirect calorimetry system. Eleven healthy children participated in this study, with a mean age of age 8.9+/-3 years. Seven of them were studied again 6 months later, so that 17 measurements were available for analysis. The calibration period was used to collect simultaneous data from HR and EE measured by indirect calorimetry (IC). Measurements were made when the children were resting (REE=Resting Energy Expenditure), during 30 min of a post-prandial period, and during two different activity levels on a cyclergometer (two periods of 15 min at 20 and 60 W output). Results of EE during a longer calibration period (2.5 h) were compared with those of a shorter period (1.5 h), for a group of six subjects. Results of nightly EE measurements of five subjects were recorded using IC and compared with the HR predicted method. Results of EE measurements estimated with different regression equations were compared to EE measured by IC. The short calibration period (1.5 h) gave similar results to a longer period (2.5 h), with a mean difference of less than 3%. The polynomial third-order relationship between HR and EE gave the highest correlation coefficient, the smallest mean square error and the best agreement (Bland-Altman method). EE predicted with this model gave the best results during the total calibration period, the post-prandial period and the medium and high activity periods. Comparison between measured and predicted nightly EE showed that REE - REE/10 was the best formula to assess nightly EE, and the mean difference was less than 4%. This study demonstrates the validity of a shortening of the calibration period and the reliability of the REE - REE/10 formula to compute nightly EE. The best model for computing daily EE is a polynomial third-order regression equation.

Adolescent↗