Tissue Oxygen Consumption Research Articles

Acclimation to Hypoxia Does Not Improve Hypoxic Survival of the Immature Pig in Confined Atmosphere

Humans may be accidentally trapped in a confined atmosphere in which oxygen availability is limited. If acclimation would extend survival, hypoxic acclimation in confined spaces would be recommended. After hypoxic acclimation, an immature pig was transferred into an experimental sealed chamber. The O2, CO2, chamber temperature, and pressure changes due to the animal's breathing were recorded. Six days acclimation (n = 3) and 3 weeks of acclimation (n = 3) were compared to control pigs (n = 3). No signs of acute mountain sickness were noted in the pigs acclimated for 6 days, but some acute symptoms (which were resolved on the following day) were observed during the 3-week acclimation. The terminal partial pressure of inspired oxygen (PIO2; 3.5-3.6 kPa) was not affected by hypoxic acclimation. Oxygen consumption and CO2 production were similar in the three experimental groups. Our hypothesis that hypoxia acclimation would produce improved survival in a confined space was not supported by the results. It is possible that at very low inspired oxygen of 3.6 kPa, the oxygen consumption of critical life-supporting tissue reached the limit of viable cells in mammals. If this is right, no further improvement could be expected after hypoxic acclimation.

Metals - impact and implications

Impact of metal in vitro administration on rat tissue oxygen consumption is referred in the first part. Toxicological implications of in vivo metal administration to rats and the study of potential penetration of metal into the rat brain, which may eventually result in oxygen radical production are presented in second part.

Near-infrared spectroscopy during stagnant ischemia: a marker of ScvO2–SvO2 mismatch in septic patients with low cardiac output

Monitoring of oxygen saturation in the superior vena cava (ScvO2) was suggested as a simpler and cheaper assessment of the global DO2:VO2 ratio and was used successfully as a goal in treatment of patients with septic shock and severe sepsis [1]. In patients with low cardiac output (CO) the difference between SvO2 and ScvO2 is more expressed and problematically large confidence limits and poor correlation were found between the two values [2]. The thenar muscle oxygen saturation (StO2) measured with near-infrared spectroscopy (NIRS) during stagnant ischemia (cuff inflation-induced vascular occlusion) decreases slower in septic shock patients [3]. This may be due to slower muscle tissue oxygen consumption in sepsis. This phenomenon possibly contributes to the ScvO2–SvO2 mismatch in patients with low CO by adding more oxygenated venous blood to flow through the superior vena cava. The aim of present study was to determine the relationship between the StO2 deceleration rate and the ScvO2–SvO2 difference in septic patients with low CO.

High selenium diet protects against TNBS-induced acute inflammation, mitochondrial dysfunction, and secondary necrosis in rat colon

Objective We studied the protective effects of selenium in a rat model of 2,4,6-trinitrobenzene sulfonic acid (TNBS)–induced colitis to elucidate a possible mechanism of action. Method Rats were supplemented with sodium selenite for 21 d with a normal selenium diet (0.02 μg/g body weight), an intermediate selenium diet (ISD; 0.3 μg/g body weight), or a high selenium diet (HSD; 2 μg/g body weight). On day 22, colitis was induced with TNBS. Rats were sacrificed after 24 h and colonic tissue was removed for evaluation. Results Selenium supplementation (HSD) resulted in a significant increase in selenium in colonic tissue. Morphologically, the HSD resulted in the preservation of tissue architecture and attenuated neutrophil infiltration; no vasculitis or necrosis was detected. Biochemically, the HSD decreased tissue myeloperoxidase activity and protected the mitochondria in the colon of TNBS-treated animals as evaluated by preserving tissue oxygen consumption, mitochondrial DNA, and expression of cytochrome c. The HSD increased levels of nuclear respiratory factor-1 and mitochondrial transcription factor-A in normal colon tissue and under inflammatory conditions. The ISD resulted in only a minor protective effect. Conclusion The results indicate that tissue damage in TNBS-induced colitis is accompanied by the arrest of mitochondrial respiration, loss of mitochondrial DNA, and the expression of nuclear-encoded mitochondrial proteins. Selenium effectively protects colon mitochondria by upregulation of the expression of mitochondrial transcription factors nuclear respiratory factor-1 and mitochondrial transcription factor-A. Selenium prevented inflammatory and necrotic changes after induction of colitis. Selenium in a high dose is therefore a potential therapeutic agent in inflammatory bowel disease.

UCP1 mRNA induction by RU486 in brown adipocytes is followed by marked induction of UCP1 protein levels

RU486 (mifepristone), a potent antagonist at progesterone and glucocorticoid receptors, is well known for its use in the termination of unwanted pregnancies, the potential development of oral contraceptives, treatment of certain cancers and other activities. Potentially, it could also play a role in thermogenesis and body weight control. RU486 acts on white adipose tissue and reverses hyperlipidic diet induced obesity in rats, or activates oxygen consumption and GDP-binding activity in brown adipose tissue (BAT), and it was suggested that BAT activation is mediated by sympathetic stimulation due to the RU486 effects on the central nervous system [1]. On the other hand, we showed, in a previous work, where RU486 was used as a tool for inhibiting progesterone effects, that it could have a possible direct effect, in vitro, on the uncoupling proteins expression in brown adipocytes [2]. Nevertheless, few studies have addressed the relationship between RU486 and brown adipose thermogenesis and they were focused mainly on the central and anti-glucocorticoid effects of RU486. Brown adipose tissue is highly relevant in the regulation of energy balance in small mammals, as rodents, due to the fact that it is the principal mediator of adaptive thermogenesis, which is mainly mediated by the uncoupling protein 1 (UCP1), an inner mitochondrial membrane protein which is able to catalyze the reentry of protons into the mitochondrial matrix, thus bypassing ATP synthase, uncoupling oxidative phosphorylation and producing heat. Thus, BAT can efficiently respond to environmental stimulus such as cold or an excessive caloric intake.

Po2profiles near arterioles and tissue oxygen consumption in rat mesentery

A scanning phosphorescence quenching microscopy technique, designed to prevent accumulated O(2) consumption by the method, was applied to Po(2) measurements in mesenteric tissue. In an attempt to further increase the accuracy of the measurements, albumin-bound probe was topically applied to the tissue and an objective-mounted pressurized bag was used to reduce the oxygen transport bypass through the thin layer of fluid over the mesentery. Po(2) was measured at multiple sites perpendicular to the blood/wall interface in the vicinity of 84 mesenteric arterioles (7-39 microm in diameter) at distances of 5, 15, 30, 60, 120, and 180 microm in seven anesthetized Sprague-Dawley rats, thereby creating Po(2) profiles. Interstitial Po(2) above and immediately beside arterioles was found to agree with known intravascular values. No significant difference in Po(2) profiles was found between small and large arterioles, indicating a small longitudinal Po(2) gradient in the precapillary mesenteric microvasculature. In addition, the Po(2) profiles were used to calculate oxygen consumption in the mesenteric tissue (56-65 nl O(2) x cm(-3) x s(-1)). Correction of these values for contamination with ambient oxygen yielded an oxygen consumption rate of 60-68 nl O(2) x cm(-3) x s(-1), the maximal limit for consumption in the mesentery. The results were compared with measurements made by other workers in regard to the employed techniques.

Impairments in microvascular reactivity are related to organ failure in human sepsis

Severe sepsis is a systemic inflammatory response to infection resulting in acute organ dysfunction. Vascular perfusion abnormalities are implicated in the pathology of organ failure, but studies of microvascular function in human sepsis are limited. We hypothesized that impaired microvascular responses to reactive hyperemia lead to impaired oxygen delivery relative to the needs of tissue and that these impairments would be associated with organ failure in sepsis. We studied 24 severe sepsis subjects 24 h after recognition of organ dysfunction; 15 healthy subjects served as controls. Near-infrared spectroscopy (NIRS) was used to measure tissue 1) microvascular hemoglobin signal strength and 2) oxygen saturation of microvascular hemoglobin (StO2). Both values were measured in thenar skeletal muscle before and after 5 min of forearm stagnant ischemia. At baseline, skeletal muscle microvascular hemoglobin was lower in septic than control subjects. Microvascular hemoglobin increased during reactive hyperemia in both groups, but less so in sepsis. StO2 at baseline and throughout ischemia was similar between the two groups; however, the rate of tissue oxygen consumption was significantly slower in septic subjects than in controls. The rate of increase in StO2 during reactive hyperemia was significantly slower in septic subjects than in controls; this impairment was accentuated in those with more organ failure. We conclude that organ dysfunction in severe sepsis is associated with dysregulation of microvascular oxygen balance. NIRS measurements of skeletal muscle microvascular perfusion and reactivity may provide important information about sepsis and serve as endpoints in future therapeutic interventions aimed at improving the microcirculation.

A quantitative framework for the design of acellular hemoglobins as blood substitutes: Implications of dynamic flow conditions

The delivery of oxygen to tissue by cell-free carriers eliminates intraluminal barriers associated with red blood cells. This is important in arterioles, since arteriolar tone controls capillary perfusion. We describe a mathematical model for O 2 transport by hemoglobin solutions and red blood cells flowing through arteriolar-sized tubes to optimize values of p50, Hill number, hemoglobin molecular diffusivity and concentration. Oxygen release is evaluated by including an extra-luminal resistance term to reflect tissue oxygen consumption. For low consumption (i.e., high resistance to O 2 release) a hemoglobin solution with p50 = 15 mmHg, n = 1, D HBO2 = 3 × 10 − 7 cm 2/s delivers O 2 at a rate similar to that of red blood cells. For high consumption, the p50 must be decreased to 5 mmHg. The model predicts that regardless of size, hemoglobin solutions with higher p50 will present excess O 2 to arteriolar walls. Oversupply of O 2 to arteriolar walls may cause constriction and paradoxically reduced capillary perfusion.

Hyperthyroidism in the developing rat testis is associated with oxidative stress and hyperphosphorylated vimentin accumulation

Hyperthyroidism was induced in rats and somatic indices and metabolic parameters were analyzed in testis. In addition, the morphological analysis evidenced testes maturation and intense protein synthesis and processing, supporting the enhancement in vimentin synthesis in hyperthyroid testis. Furthermore, vimentin phosphorylation was increased, indicating an accumulation of phosphorylated vimentin associated to the cytoskeleton, which could be a consequence of the extracellular-regulated kinase (ERK) activation regulating the cytoskeleton. Biomarkers of oxidative stress demonstrated an increased basal metabolic rate measured by tissue oxygen consumption, as well as, increased TBARS levels. In addition, the enzymatic and non-enzymatic antioxidant defences appeared to respond according to the augmented oxygen consumption. We observed decreased total glutathione levels, with enhancement of reduced glutathione, whereas most of the antioxidant enzyme activities were induced. Otherwise, superoxide dismutase activity was inhibited. These results support the idea that an increase in mitochondrial ROS generation, underlying cellular oxidative damage, is a side effect of hyperthyroid-induced biochemical changes by which rat testis increase their metabolic capacity.

Experimental small bowel preservation using Polysol: A new alternative to University of Wisconsin solution, Celsior and histidine-tryptophan-ketoglutarate solution?

To evaluate the potential of Polysol, a newly developed preservation solution, in cold storage of small bowel grafts, compared with the current standards, University of Wisconsin solution (UW), Celsior and histidine-tryptophan-ketoglutarate solution (HTK). Male Wistar rats were used as donors. Small bowels were retrieved, flushed and then stored in the respective 4 solutions for 18 h at 4 centigrade. Functional integrity of the grafts was evaluated by isolated reperfusion with oxygenated Krebs-Henseleit buffer at 37 centigrade for 30 min in all 4 groups. Polysol preservation exhibited the highest tissue ATP concentration and the lowest release of LDH. Malondialdehyde, an index for tissue lipid peroxidation, was also the lowest in Polysol. Tissue oxygen consumption was significantly higher in Polysol than in the others. Of interest, UW-storage promoted 10-fold higher apoptosis than in the others. Moreover, electron microscopy revealed that the mucosal villi/micro-villi formation and the cell organelles, including mitochondria, were both significantly better preserved in Polysol, while deleterious alterations were apparent in the others, most notably in UW. Although Celsior and HTK exhibited the better trend of results than UW in some parameters, but could not reach the over-all superiority to UW. Cold storage using Polysol resulted in significantly better integrity and function of small bowel grafts than UW. Hence, Polysol may be a novel alternative for the small bowel preservation.

Forearm and Calf Tissue Oxygenation in Term Neonates Measured with Near-Infrared Spectroscopy

Peripheral tissue oxygenation has been studied with near-infrared spectroscopy (NIRS) on either the forearm or calf with questionable comparability. The aim was to compare forearm and calf tissue oxygenation in healthy term neonates measured with NIRS. Fractional oxygen extraction, tissue oxygenation index, and mixed venous oxygenation were similar in both extremities, whereas oxygen delivery and oxygen consumption of calf tissue were higher.

Cerebral blood flow during cardiopulmonary bypass in pediatric cardiac surgery: the role of transcranial Doppler – a systematic review of the literature

BackgroundTranscranial Doppler Ultrasound (TCD) is a sensitive, real time tool for monitoring cerebral blood flow velocity (CBFV). This technique is fast, accurate, reproducible and noninvasive. In the setting of congenital heart surgery, TCD finds application in the evaluation of cerebral blood flow variations during cardiopulmonary bypass (CPB).MethodologyWe performed a search on human studies published on the MEDLINE using the keyword "trans cranial Doppler" crossed with "pediatric cardiac surgery" AND "cardio pulmonary by pass", OR deep hypothermic cardiac arrest", OR "neurological monitoring".DiscussionCurrent scientific evidence suggests a good correlation between changes in cbral blood flow and mean cerebral artery (MCA) blood flow velocity. The introduction of Doppler technology has allowed an accurate monitorization of cerebral blood flow (CBF) during circulatory arrest and low-flow CPB. TCD has also been utilized in detecting cerebral emboli, improper cannulation or cross clamping of aortic arch vessels. Limitations of TCD routine utilization are represented by the need of a learning curve and some experience by the operators, as well as the need of implementing CBF informations with, for example, data on brain tissue oxygen delivery and consumption.ConclusionIn this light, TCD plays an essential role in multimodal neurological monitorization during CPB (Near Infrared Spectroscopy, TCD, processed electro encephalography) that, according to recent studies, can help to significantly improve neurological outcome after cardiac surgery in neonates and pediatric patients.

Neuropeptide Y Deficiency Attenuates Responses to Fasting and High-Fat Diet in Obesity-Prone Mice

Neuropeptide Y (NPY) stimulates feeding and weight gain, but deletion of the NPY gene does not affect food intake and body weight in mice bred on a mixed genetic background. We reasoned that the orexigenic action of NPY would be evident in C57Bl/6J mice susceptible to obesity. NPY deficiency has no significant effect in mice fed a normal rodent diet. However, energy expenditure is elevated during fasting, and hyperphagia and weight gain are blunted during refeeding. Expression of agouti-related peptide (AGRP) in the hypothalamus is increased in NPY knockout (NPYko) than wild-type mice, but unlike wild type there is no further increase in AGRP when NPYko mice are fasted. Moreover, NPYko mice have higher oxygen consumption and uncoupling protein-1 expression in brown adipose tissue during fasting. The failure of an increase in orexigenic peptides and higher thermogenesis may contribute to attenuation of weight gain when NPYko mice are refed. C57Bl/6J mice lacking NPY are also less susceptible to diet-induced obesity (DIO) as a result of reduced feeding and increased energy expenditure. The resistance to DIO in NPYko mice is associated with a reduction in nocturnal feeding and increased expression of anorexigenic hypothalamic peptides. Insulin, leptin, and triglyceride levels increase with adiposity in both wild-type and NPYko mice.

Real‐time In Situ Monitoring of Human Prostate Photodynamic Therapy with Diffuse Light

Photodynamic therapy (PDT) requires oxygen to cause cellular and vascular tumor damage. Tissue oxygen concentration, in turn, is influenced by blood flow and blood oxygenation. Real-time clinical measurement of these hemodynamic quantities, however, is rare. This paper reports the development and application of a probe, combining diffuse reflectance spectroscopy (DRS) for measurement of tumor blood oxygenation and diffuse correlation spectroscopy (DCS) for measurement of tumor blood flow. The instrument was adapted for clinical use during interstitial prostate PDT. Three patients with locally recurrent prostate cancer received 2 mg/ kg motexafin lutetium (MLu) 3 h before illumination and a total light dose of 100 J/cm(2) at 150 mW/cm. Prostrate blood oxygen saturation (StO2) decreased only slightly (approximately 3%) after treatment. On the other hand, prostate blood flow and total hemoglobin concentration over the course of PDT decreased by 50% and 15%, respectively, suggesting MLu-mediated PDT has an anti-vascular effect. While it is certainly impossible to draw definite conclusions from measurements of only three patients, the observed differences in tumor blood flow and blood oxygenation responses during PDT can, in principle, be used to choose among tissue oxygen consumption models and therefore emphasize the potential clinical value for simultaneous monitoring of both parameters.

Effect of platelet-activating factor on cold-preserved liver grafts.

Platelet-activating factor (PAF) may play an important role in graft injury in liver transplantation. Livers excised from male Wistar rats were preserved in University of Wisconsin solution for 6 h and then perfused with Krebs-Henseleit solution containing vehicle (bovine serum albumin) or PAF. Impairment of parenchymal cells was assessed by reference to tissue adenosine triphosphate levels, oxygen consumption and alanine aminotransferase activity in the effluent. The effect on non-parenchymal cells was evaluated by measurement of purine nucleoside phosphorylase and alanine aminotransferase levels in the effluent. Administration of as little as 1.0 ng kg-1 PAF caused a significant decrease in adenosine 5'-triphosphate concentration and oxygen consumption (P < 0.05), although non-parenchymal cell injury was not affected. PAF can therefore cause liver graft dysfunction with hepatocytes as the main target, even in the absence of microcirculatory disturbance secondary to interaction between blood cells and endothelial cells.

Hyperhomocysteinemia Alters The Cardiac Metabolism by Impairing Nitric Oxide Bioavailability Through Oxidative Stress

Hyperhomocysteinemia (HHcy) has been considered a vascular disease which is associated with increased levels of reactive oxygen species resulting in scavenging of nitric oxide (NO). However, little is known of the impact on cardiac function and myocardial metabolism. L-Homocysteine was intravenously infused into conscious adult male dogs (titrated to give plasma homocysteine up to 10μM over 120 minutes). There was no change in any hemodynamic parameters between baseline and HHcy. Veratrine induced NO dependent coronary vasodilation (Bezold-Jarisch reflex) was reduced by 29% which was restored by simultaneous infusion of ascorbic acid. Myocardial substrate use was calculated as coronary sinus-arterial difference times coronary blood flow. Acute HHcy significantly increased glucose uptake (1.7±1.2 to 6.8±1.2 μmol/min) and decreased FFA uptake (6.0±1.6 to 3.1±1.0 μmol/min). Lactate uptake was also increased (3.9±3.0 to 11.2±6.1 μmol/min). Myocardial oxygen consumption (MVO2) of LV tissue from chronic HHcy dogs (fed with methionine for 2 weeks) was also measured in vitro in response to bradykinin (BK). BK-induced reduction in MVO2 was significantly decreased by HHcy and this effect was completely inhibited with coincubation with ascorbic acid, tempol, or apocynin. In conclusion, HHcy modulates the substrate use of heart by inhibiting NO bioavailability through the generation of superoxide. The progression of the disease associated with HHcy should also be evaluated based on alterations in the regulation of cardiac metabolism and substrate use. Supported by HL-PO1-43023 grant.

Compression-induced changes in the physiological state of the breast as observed through frequency domain photon migration measurements

We use optical spectroscopy to characterize the influence of mammographic-like compression on the physiology of the breast. We note a reduction in total hemoglobin content, tissue oxygen saturation, and optical scattering under compression. We also note a hyperemic effect during repeated compression cycles. By modeling the time course of the tissue oxygen saturation, we are able to obtain estimates for the volumetric blood flow (1.64+/-0.6 mL/100 mL/min) and the oxygen consumption (1.97+/-0.6 micromol/100 mL/min) of compressed breast tissue. These values are comparable to estimates obtained from previously published positron emission tomography (PET) measurements. We conclude that compression-induced changes in breast physiological properties are significant and should be accounted for when performing optical breast imaging. Additionally, the dynamic characteristics of the changes in breast physiological parameters, together with the ability to probe the tissue metabolic state, may prove useful for breast cancer detection.

Norepinephrine but not hypoxia stimulates HIF-1α gene expression in brown adipocytes

The cellular response to hypoxic stress is mainly mediated via activation of the transcription factor hypoxia-inducible factor-1α (HIF-1α). In the present study, the sympathetically controlled brown adipose tissue was used to investigate the effect of norepinephrine on HIF-1α gene expression. Norepinephrine increased HIF-1α mRNA levels in cultured brown adipocytes, whereas the hypoxia-mimic cobalt was without effect. Cold exposure of mice increased HIF-1α gene expression in brown adipose tissue. In UCP1-ablated mice, which are incapable of inducing thermogenic oxygen consumption in brown adipose tissue, cold exposure generated a significantly higher elevation of HIF-1α mRNA levels than in wild-type. These results demonstrate that cold-induced HIF-1α gene expression is independent of thermogenic oxygen consumption leading to hypoxia, but is consistent with a norepinephrine regulation of HIF-1α gene expression. Thus, by elevating HIF-1α gene expression, norepinephrine may mediate an increased potential to respond to hypoxia in brown adipose tissue and possibly in other tissues.

Effects of hyperbaric oxygen treatment on liver functions, oxidative status and histology in septic rats

The liver is thought to be responsible for multiple organ failure during sepsis. Increase in tissue oxygen consumption is a major component of the septic response. Hyperbaric oxygen (HBO) therapy provides more oxygenation in the whole body. This study examined the effect of HBO alone or in combination with cefepime (CEF) on the liver in septic rats. We divided 90 male rats into six groups; control, HBO, sepsis (SEP), SEP+HBO, SEP+CEF, and SEP+CEF+HBO. Sepsis was induced with an intraperitoneal injection of Escherichia coli (2.1 x 10(9) cfu). A total of six HBO sessions were performed at 2 atm absolute for 90 min at 6-h intervals. CEF was administered intraperitoneally at a dose of 50 mg/kg twice daily. Animals were killed 48 h after sepsis induction. Their liver and blood were removed for biochemical and histopathological analysis. Liver thiobarbituric acid reactive substances as well as serum alanine transaminase, aspartate transaminase and alkaline phosphatase levels increased while the activity of the antioxidant enzymes superoxide dismutase and catalase decreased significantly in septic rats. These parameters returned to nearly control levels in the SEP+CEF+HBO group. Histological observations supported these findings: Hepatocellular degeneration was observed and intensive polymorphonuclear cell infiltration appeared in all fields of septic animal livers. HBO alone could not sufficiently reverse these histopathological changes, but most liver sections presented normal histology when it was combined with CEF. HBO may be a useful adjuvant therapy modality to improve the efficacy of sepsis treatment.

Clinical study of a formula for delayed rapid fluid resuscitation for patients with burn shock

Objective: To explore a suitable formula of delayed rapid fluid resuscitation for patients with burn shock. Methods: Twenty patients with burns over 40% of total body surface area (TBSA) admitted 4–8 h after injury were studied. Plasma was used as colloid in 9 cases in the infused patients (the plasma group, PG) and gelofusine was used as colloid in 11 cases in the infused patients (the gelofusine group, GG). Rapid fluid resuscitation was administered under strict hemodynamic monitoring immediately after admission. Hemodynamic indexes including pulmonary arterial pressure (PAP), pulmonary artery wedge pressure (PAWP), cardiac output (CO), pulmonary vascular resistance (PVR), and systemic vascular resistance (SVR), hemorheological parameters such as blood viscosity and plasma viscosity, and tissue oxygenation indices oxygen delivery (DO 2), oxygen consumption (VO 2), oxygen extraction (O 2ext), lactic acid (LA) and base deficiency of arterial blood (AD) were determined in order to monitor function in or damage to important viscera. Results: The amount of fluid rapidly infused in the first 2 h after hospitalization accounted for 38.8 ± 6.0% of the required fluid amount for the first 24 h as calculated from the formula. When the amount of fluid infused into the patient before admission to our hospital was added, the total amount accounted for 48.3 ± 5.0% of the fluid amount for the first 24 h. The actual amount of fluid infused in the first 24 h was 31.4 ± 8.9% more than that of the amount calculated with the Evans’ formula. The amount of fluid infused in the second 24 h was nearly equal to the amount calculated with the Evans’ formula. After fast fluid replacement therapy, all the parameters determined were markedly improved. Conclusion: It is suggested on the basis of our study that the fluid amount for delayed rapid fluid resuscitation in patients with burn shock should be calculated as follows: (1) in the first 24 h, the amount of fluid (ml) is equal to TBSA (%) × body weight (kg) × 2.6. The ratio between colloid and electrolytes is 1:1, each of the two fluid types is 1.3 ml per (%) TBSA per kg body weight. The volume of water required is 2000 ml. Half of the total amount of fluid is proposed should be infused in the first 2 h after hospitalization under strict monitoring of hemodynamic indices. (2) In the second 24 h, the amount of fluid (ml) is equal to TBSA (%) × body weight (kg) × 1. The ratio between colloid and electrolytes 1:1, each of the two is 0.5 ml. Water volume is 2000 ml.