OLT1177, a ß-sulfonyl nitrile compound, safe in humans, inhibits the NLRP3 inflammasome and reverses the metabolic cost of inflammation.

Activation of the NLRP3 inflammasome induces maturation of IL-1ß and IL-18, both validated targets for treating acute and chronic inflammatory diseases. Here, we demonstrate that OLT1177, an orally active ß-sulfonyl nitrile molecule, inhibits activation of the NLRP3 inflammasome. In vitro, nanomolar concentrations of OLT1177 reduced IL-1ß and IL-18 release following canonical and noncanonical NLRP3 inflammasome activation. The molecule showed no effect on the NLRC4 and AIM2 inflammasomes, suggesting specificity for NLRP3. In LPS-stimulated human blood-derived macrophages, OLT1177 decreased IL-1ß levels by 60% and IL-18 by 70% at concentrations 100-fold lower in vitro than plasma concentrations safely reached in humans. OLT1177 also reduced IL-1ß release and caspase-1 activity in freshly obtained human blood neutrophils. In monocytes isolated from patients with cryopyrin-associated periodic syndrome (CAPS), OLT1177 inhibited LPS-induced IL-1ß release by 84% and 36%. Immunoprecipitation and FRET analysis demonstrated that OLT1177 prevented NLRP3-ASC, as well as NLRP3-caspase-1 interaction, thus inhibiting NLRP3 inflammasome oligomerization. In a cell-free assay, OLT1177 reduced ATPase activity of recombinant NLRP3, suggesting direct targeting of NLRP3. Mechanistically, OLT1177 did not affect potassium efflux, gene expression, or synthesis of the IL-1ß precursor. Steady-state levels of phosphorylated NF-κB and IkB kinase were significantly lowered in spleen cells from OLT1177-treated mice. We observed reduced IL-1ß content in tissue homogenates, limited oxidative stress, and increased muscle oxidative metabolism in OLT1177-treated mice challenged with LPS. Healthy humans receiving 1,000 mg of OLT1177 daily for 8 d exhibited neither adverse effects nor biochemical or hematological changes.

Evaluating passive physiological data collection during Spravato treatment.

Spravato and other drugs with consciousness-altering effects show significant promise for treating various mental health disorders. However, the effects of these treatments necessitate a substantial degree of patient monitoring which can be burdensome to healthcare providers and may make these treatments less accessible for prospective patients. Continuous passive monitoring via digital devices may be useful in reducing this burden. This proof-of-concept study tested the MindMed Session Monitoring System™ (MSMS™), a continuous passive monitoring system intended for use during treatment sessions involving pharmaceutical products with consciousness-altering effects. Participants completed 129 Spravato sessions with MSMS at an outpatient psychiatry clinic specializing in Spravato treatment. Results indicated high rates of data quality and self-reported usability among participants and health care providers (HCPs). These findings demonstrate the potential for systems such as MSMS to be used in consciousness-altering treatment sessions to assist with patient monitoring.

Long-term propranolol use in severely burned pediatric patients: a randomized controlled study.

OBJECTIVE: To determine the safety and efficacy of propranolol given for 1 year on cardiac function, resting energy expenditure, and body composition in a prospective, randomized, single-center, controlled study in pediatric patients with large burns. BACKGROUND: Severe burns trigger a hypermetabolic response that persists for up to 2 years postburn. Propranolol given for 1 month postburn blunts this response. Whether propranolol administration for 1 year after injury provides a continued benefit is currently unclear. METHODS: One-hundred seventy-nine pediatric patients with more than 30% total body surface area burns were randomized to control (n = 89) or 4 mg/kg/d propranolol (n = 90) for 12 months postburn. Changes in resting energy expenditure, cardiac function, and body composition were measured acutely at 3, 6, 9, and 12 months postburn. Statistical analyses included techniques that adjusted for non-normality, repeated-measures, and regression analyses. P < 0.05 was considered significant. RESULTS: Long-term propranolol treatment significantly reduced the percentage of the predicted heart rate and percentage of the predicted resting energy expenditure, decreased accumulation of central mass and central fat, prevented bone loss, and improved lean body mass accretion. There were very few adverse effects from the dose of propranolol used. CONCLUSIONS: Propranolol treatment for 12 months after thermal injury, ameliorates the hyperdynamic, hypermetabolic, hypercatabolic, and osteopenic responses in pediatric patients. This study is registered at clinicaltrials.gov: NCT00675714.

Randomized controlled trial to determine the efficacy of long-term growth hormone treatment in severely burned children.

BACKGROUND: Recovery from a massive burn is characterized by catabolic and hypermetabolic responses that persist up to 2 years and impair rehabilitation and reintegration. The objective of this study was to determine the effects of long-term treatment with recombinant human growth hormone (rhGH) on growth, hypermetabolism, body composition, bone metabolism, cardiac work, and scarring in a large prospective randomized single-center controlled clinical trial in pediatric patients with massive burns. PATIENTS AND METHODS: A total of 205 pediatric patients with massive burns over 40% total body surface area were prospectively enrolled between 1998 and 2007 (clinicaltrials.gov ID NCT00675714). Patients were randomized to receive either placebo (n = 94) or long-term rhGH at 0.05, 0.1, or 0.2 mg/kg/d (n = 101). Changes in weight, body composition, bone metabolism, cardiac output, resting energy expenditure, hormones, and scar development were measured at patient discharge and at 6, 9, 12, 18, and 24 months postburn. Statistical analysis used Tukey t test or ANOVA followed by Bonferroni correction. Significance was accepted at P < 0.05. RESULTS: RhGH administration markedly improved growth and lean body mass, whereas hypermetabolism was significantly attenuated. Serum growth hormone, insulin-like growth factor-I, and IGFBP-3 was significantly increased, whereas percent body fat content significantly decreased when compared with placebo, P < 0.05. A subset analysis revealed most lean body mass gain in the 0.2 mg/kg group, P < 0.05. Bone mineral content showed an unexpected decrease in the 0.2 mg/kg group, along with a decrease in PTH and increase in osteocalcin levels, P < 0.05. Resting energy expenditure improved with rhGH administration, most markedly in the 0.1 mg/kg/d rhGH group, P < 0.05. Cardiac output was decreased at 12 and 18 months postburn in the rhGH group. Long-term administration of 0.1 and 0.2 mg/kg/d rhGH significantly improved scarring at 12 months postburn, P < 0.05. CONCLUSION: This large prospective clinical trial showed that long-term treatment with rhGH effectively enhances recovery of severely burned pediatric patients.

Mortality related to gender, age, sepsis, and ethnicity in severely burned children.

Several studies have noted gender differences in adult mortality related to thermal injury, however, little is published on gender-related outcomes of burn patients 17 years of age or less. The aim of this study was to evaluate the relationships between mortality, gender, prepubertal and during puberty, ethnic origin, and age, with or without identified sepsis in severely burned children. Seven hundred forty-seven children admitted to our burn hospital from March 1985 to January 2005 with burns greater than 40% total body surface area were studied. Mortality associated with identified sepsis, gender, age, and ethnic origin were outcomes of interest. Two hundred sixty (35%) of the patients studied were girls and 487 (65%) were boys. No significant difference could be shown between girls and boys for the number of operations, time from burn to hospital admission, or the presence of identifiable inhalation injury. Nearly 60% of the male nonsurvivors and 48% of the female nonsurvivors in this study had identifiable sepsis at postmortem. The mortality rate was higher in infants and toddlers, age 0 to 2.9 years, compared with children and adolescents, age 3 to 17 years; however, there was no significant difference in rate of mortality between genders, prepuberty versus puberty, those with septic episodes, or ethnic origin. Burn mortality among infants and toddlers, children, and adolescents with greater than 40% total body surface area burns with or without identified sepsis could not be shown to be gender or ethnic origin dependent.

Identification of factors contributing to hepatomegaly in severely burned children.

Hepatomegaly is a common postmortem observation in severely burned children, with the liver often tripling in size when compared with normal livers for age, weight, and sex. Lesions identified at autopsy include deposition of large and small fat droplets in the hepatocyte, congestion, centrilobular necrosis, and cholestasis. The present study was designed to identify the primary causes of hepatomegaly in severely burned children postmortem. For this purpose, 41 autopsies were reviewed and, when available, blood and tissue samples were studied. Histopathologic findings showed that large intrahepatocytic fat droplets within hepatocytes and cholestasis were important contributors to hepatomegaly. Liver density and wet/dry weight ratios significantly decreased with increasing liver size. Hepatocyte volume increased with increasing liver size (P < 0.001) as did total fat content (P < 0.001). The liver enzymes, alanine aminotransferase and aspartate aminotransferase, remained normal except within 5 to 10 days of injury and 5 to 10 days of death. Triglycerides made up 4% to 70% of the total fat, with the percentage of triglycerides increasing with the severity of hepatomegaly. Saturated fatty acids represented about 85% of the total fatty acids in normal-sized livers, whereas in the largest livers (400% of predicted), only 25% of the fatty acids were saturated. This study provides evidence that 85% to 90% of the hepatomegaly observed in severely burned children postmortem is associated with hepatocyte enlargement, which includes up to 19% intracellular fat. Increases in extracellular protein, intracellular glycogen, and fluid accumulation may make a minor contribution to postburn hepatomegaly.

Gene expression profiles of livers from thermally injured rats.

The liver plays an important role in a severe thermal injury by modulating immune function, inflammatory processes and the acute phase response, which are an orchestrated attempt to restore homeostasis. Using high-density oligonucleotide arrays, we examined the gene expression profile in the livers of rats between 2 and 240 h after a 40% total body surface area (TBSA) burn. Alterations in gene expression unique to a thermal injury were identified. Approximately 39 genes out of 8700 genes on each array across all the time points showed a significant change in expression patterns. Real time reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analyses verified significant changes in early growth response-1 (Egr-1) messenger RNA (mRNA) and protein levels corresponding to the array data. Significant increases in serum levels of alpha-2-macroglobulin that correspond to changes in its mRNA levels were observed at 6 and 24 h after burn, p<0.05. The genomic pattern for liver in the hypermetabolic phase after the burn injury involves transcription factors, stress and inflammatory responses, cytoskeletal and extracellular matrix modifications, and regulation of cell proliferation and differentiation. During the initial phase of thermal injury gene expression profiles in the liver may provide some insight into how cellular protection mechanisms and systemic hypermetabolism are initiated and controlled. The genome wide changes observed may provide a rational therapeutic strategy to improve burn care.

Recombinant human growth hormone increases thyroid hormone-binding sites in recovering severely burned children.

Thyroxine (T4), Tri-iodothyronine (T3), and total serum protein levels are reduced in severely burned children. T4 and T3 are carried on serum transport proteins via thyroid hormone-binding sites (THBS). Treatment of bums with recombinant human growth hormone (rhGH) increases albumin (Alb) and prealbumin (PreAlb), which bind nearly 30% of circulating T4 and T3. This study investigated the effect of rhGH on THBS sites in burned children. Records of 11 acutely burned children with a total body surface area burned >40% who were randomized to either a daily subcutaneous injection of rhGH at 0.05 mg/kg/day or placebo for 6 months after discharge from hospital were reviewed. Thyroxine uptake percentage (TU%), Total T4 levels (TT4), free thyroxine index, Alb and PreAlb, and height and weight measurements taken at discharge and 6 months later were compared in both groups. In the six children who received rhGH, mean TU% had decreased from 41 +/- 1 to 33 +/- 1% by 6 months postdischarge, (P < 0.001), mean TT4 increased from 5.8 +/- 0.3 to 8.1 +/- 0.8 microg/dL, (P < 0.02), mean Alb increased from 2.0 +/- (0.6) to 3.5 +/- (0.1) g/dL (P < 0.0001), and mean PreAlb increased from 8.7 +/- 0.7 to 16.5 +/- 2.1 mg/dL, (P < 0.006). There were no significant changes in the five children in the placebo (control group), and height and weight did not significantly change in either group. rhGH significantly increases THBS in severely burned children, possibly through increases in serum Alb and PreAlb. The increases in circulating thyroxine observed in this group may be involved in the attenuation of growth arrest.

Extended hypermetabolic response of the liver in severely burned pediatric patients.

HYPOTHESIS: The acute phase response is a cascade of events contributing to hypermetabolism and substrate catabolism. It was believed to persist for only a short time after injury. There is now evidence that systemic catabolism and hypermetabolism associated with thermal injury persevere for a long time. We hypothesize that the proinflammatory hepatic acute phase response perseveres for an extended time and enhances hypermetabolism longer than previously believed. DESIGN: Prospective study. SETTING: Intensive Care Burn Unit, Shriners Hospital for Children. PATIENTS: Twenty-three children (aged 1-16 years) sustaining a severe thermal injury (>/=40% total-body surface area) who remained in the intensive care unit longer than 30 days. MAIN OUTCOME MEASURES: Patient demographics, nutritional support, incidence of sepsis, inhalation injury, mortality, and levels of serum constitutive proteins, type I and type II acute phase proteins, free fatty acids, proinflammatory cytokines, insulin-like growth factor (IGF) I, IGF binding protein-1, IGF binding protein-3, and hepatocyte growth factor. RESULTS: After thermal injury, constitutive hepatic protein levels decreased 2- to 3-fold 80 days after burn, whereas acute phase protein levels increased. Free fatty acid levels were increased 5 days after burn. Proinflammatory cytokine levels (interleukin [IL] 1, IL-6, IL-8, IL-10, and tumor necrosis factor) and IGF binding protein-1 levels were elevated for 40 days after burn, whereas serum IGF-I and IGF binding protein-3 levels were decreased. Hepatocyte growth factor levels were increased immediately after burn but rapidly returned to the normal range. CONCLUSIONS: Despite adequate nutritional support, a severe thermal injury induces the proinflammatory acute phase response for a prolonged period. Thus, the liver with the hepatic acute phase response plays a more important role during catabolism after burn than previously believed. Pharmacologic agents that improve hepatic function may be an effective approach to attenuate hypermetabolism after trauma.

Dietary fat composition alters pulmonary function in pigs.

OBJECTIVES: We investigated the effect of various dietary fats on pulmonary surfactant composition and lung function changes that occur before and after endotoxin infusion in pigs. METHODS: Eighteen pigs were assigned to three groups (n = 6 per group) to receive a diet of protein (20% of calories), carbohydrate (20% of calories), and fat (40% of calories). In one group the fat content consisted entirely of palmitic acid. In the second group, fat came from Intralipid, which provided predominantly linoleic acid. The third group was fed fish oil. Pigs were maintained on these diets for 21 d before the experiment. Cardiovascular and pulmonary functions were determined on day 22. Pigs then were infused with endotoxin (80 mg. kg(-1). min(-1)) until the pulmonary arterial pressure reached a pressure similar to that found in trauma victims (45 to 50 mmHg). Cardiovascular and pulmonary function tests were then repeated, the animals killed, and the lungs removed for study. RESULTS: Compliance was reduced in the linoleate and fish-oil groups compared with the palmitate group before and after endotoxin. Compliance changes in pigs fed the linoleate and fish-oil diets were consistent with significant increases in lung wet:dry weight ratios, increased CO(2) retention, histologic evidence of vascular congestion, intra-alveolar edema, and alveolar septa thickening. Changes in surfactant phosphatidylcholine composition between groups were consistent with the notion that increased unsaturated fatty acids could affect surfactant function. CONCLUSIONS: We concluded that the common practice of providing calories in the form of polyunsaturated fatty acids to critically ill patients carries the risk of being detrimental to lung function.

Influence of demographics and inhalation injury on burn mortality in children.

BACKGROUND: Pulmonary failure has emerged as one of the leading causes of mortality in burned children due, in part, to the success in reducing the incidence of sepsis, early surgery and fluid resuscitation, and new advances in nutritional support. To evaluate the effect of pulmonary injury, age, gender, race, and burn size on mortality, the records of 3179 burned children admitted to our burn center from 1985 to 2001 were reviewed. In this population, 1246 were admitted within 14 days of injury with burns greater than 20% of their total body surface area (TBSA). METHODS: Lethal burn areas (LAs) for a thermal injury only or burn plus inhalation injury were estimated from best fit probit curve within 95% confidence limits. Data analysis was by chi(2)-test, t-test, or Fisher's exact test where appropriate. RESULTS: The lethal burn area for a 10% mortality rate with and without concomitant inhalation injury was a 50 and 73% TBSA burn, respectively. Children up to the age of 3 with >/=20% TBSA burns had a higher rate of mortality (9.9%) compared to those 3-12 years of age (4.9%) and 13-18 years of age (4.2%). Children with 21-80% TBSA burns showed a significant difference in mortality (P<0.05) between those with burn plus inhalation injury (13.9%) and burn only (2.9%), while those with 81-100% TBSA burns showed no significant difference between burn only and burn plus inhalation injury. CONCLUSION: Inhalation injury remains one of the primary contributors to burn mortality. Children under the age of 3 years, however, are at a higher risk both with and without inhalation injury.

Matrix metalloproteinase expression in cytokine stimulated human dermal fibroblasts.

In this study, we investigated the effect of inflammatory cytokines on matrix metalloproteinase (MMP-1) and TIMP-1 production in human dermal fibroblasts, which play a pivotal role in wound healing, ranging from the synthesis and remodeling of extracellular matrix (ECM) to the synthesis of growth factors. The balance of MMPs and TIMPs is crucial in directing successful wound repair. Human adult dermal fibroblasts were seeded in six well plates (7.5 x 10(4) cells/ml) in complete media. Eighty to ninety percent confluent cells were treated with interleukin-1beta (IL-1beta), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) (10 ng/ml) for 6h in serum free media with suitable controls run in triplicate. Supernatants were assayed for pro-MMP-1 & TIMP-1. Extracted total RNA was used for reverse transcription polymerase chain reaction (RT-PCR) with sequence specific primers for MMP-1, TIMP-1 and beta-actin. Signal intensity was normalized to the internal control (beta-actin). Statistical analysis used ANOVA. MMP-1 and TIMP-1 mRNA expression were markedly increased with IL-6 and TNF-alpha treatment and remains unchanged with IL-1beta. Pro-MMP-1 protein levels are unchanged with TNF-alpha and significantly increased with IL-1beta and IL-6 treatment. However, TNF-alpha significantly increases TIMP-1 protein levels. Data suggests differential regulation of MMP-1 and TIMP-1 protein levels by the cytokines found in stimulated dermal fibroblasts. Further characterization of this response will provide an understanding of the mechanisms of pathogenesis of extracellular matrix (ECM) and the potential role of metalloproteinases in tissue remodeling after injury.

The influence of age and gender on resting energy expenditure in severely burned children.

INTRODUCTION: Recent evidence suggests that female severely burned children have higher endogenous anabolic hormone levels and a shorter ICU stay compared with males. The purpose of this study was to analyze the influence of age and gender on resting energy expenditure (REE) in severely burned children from acute hospitalization through 12 months postburn. METHODS: A total of 100 pediatric patients with > 40% total body surface area (TBSA) burn were enrolled in a prospective study and followed by indirect calorimetry measurements. The REE was expressed as actual REE kcal/d, percent of predicted REE, and REE/ body mass index (BMI). Statistical analysis was performed by Student t test and one-way ANOVA for repeated measures. Significance was accepted at P < 0.05. RESULTS: The measured REE was significantly higher in males versus females at all time points (P < 0.05). The percent of predicted REE was significantly higher in males versus females during the acute hospitalization, at discharge, 6 and 9 months postburn (P < 0.05). The REE/BMI showed a significant difference between males and females at the acute and discharge time period (P < 0.05). In children 3 to 9.9 years of age, the measured REE and the percent of predicted REE were significantly higher in males versus females during the acute study, at discharge and 6 months postburn (P < 0.05). The measured REE at discharge, 9 and 12 months postburn for children >10 years of age was significantly higher in males compared with females (P < 0.05). CONCLUSION: Data show that female children exert a decreased hypermetabolic response compared with male children, which may improve burn outcomes in females.

The use of beta-adrenergic blockade in preventing trauma-induced hepatomegaly.

OBJECTIVE: The objective of this study was to test the hypothesis that hepatomegaly in burned children can be attenuated or reversed by blocking lipolysis and reducing free fatty acids delivered to the liver. SUMMARY BACKGROUND DATA: Accelerated lipolysis in severely burned children has been shown to play an important role in the accumulation of hepatic TGs. Severely burned children who survive 10 days or more after injury commonly have enlarged livers often twice or more normal size for their sex, age, and weight. METHODS: Ninety-eight children, 2 to 18 years of age, with burns covering more than 40% of their body surface and who received either propranolol (beta-adrenergic blockade) or placebo were studied. Liver weights were measured by ultrasonic scanning. Body composition changes were identified by dual-image x-ray absorptiometry and validated by whole-body potassium-40 scintillation counting. Discarded abdominal cutaneous adipose tissue was collected before and after propranolol or placebo for microarray analysis. RESULTS: In 80% of severely burned children studied not receiving propranolol, liver sizes increased by 100% or more while 86% of burned children receiving propranolol showed a decrease or no change in liver size over the same period of time after injury. Gene expression patterns of adipose tissue after propranolol treatment showed that all of the identified genes related to lipid metabolism were down-regulated. CONCLUSIONS: Data reported here support the hypothesis that beta-adrenergic blockade can reduce delivery of fatty acids to the liver and hepatic congestion commonly found in severely burned children by inhibiting lipolysis and reducing hepatic blood flow.

Body composition changes with time in pediatric burn patients.

BACKGROUND: Major trauma and burns are associated with whole body catabolism which can persist for 1 or more years after injury. This study investigates body composition in massively burned children for up to 2 years. METHODS: Twenty-five pediatric patients with greater than 40% total body surface area burns were studied. At discharge, 6, 12, 18, and 24 months after burn height, weight, body composition, resting energy expenditure (REE), serum growth hormone, insulin-like growth factor-I (IGF-I), IGF binding protein-3 (IGFBP-3), insulin, cortisol, parathyroid hormone, and thyroid hormones were measured. Tukey's test was used for analysis. Significance was accepted at p < 0.05. RESULTS: Lean body mass, fat mass, bone mineral content, height, and weight increased significantly during the second year after burn. Percent predicted REE decreased significantly, whereas IGFBP-3 and parathyroid hormone levels increased significantly over time. Insulin and T3 uptake were significantly higher at discharge. CONCLUSIONS: Body composition of severely burned children significantly improved in the second year compared with the first year after injury. This demonstrates a need for long-term rehabilitation in these burn patients.

Beneficial effects of extended growth hormone treatment after hospital discharge in pediatric burn patients.

OBJECTIVE: To study the efficacy of growth hormone given to severely burned children from discharge to 12 months after burn and for 12 months after the drug was discontinued. SUMMARY BACKGROUND DATA: We have previously shown that low-dose recombinant human growth hormone (rhGH), given to children after a severe thermal injury, successfully improved lean muscle mass, bone mineral content, and growth. The aim of the present study was to investigate long-term functional improvements after treatment. METHODS: Forty-four pediatric patients with over 40% total body surface area burns were studied for 24 months after burn. Patients were randomized to receive either rhGH (0.05 mg/kg body weight) or placebo. Height, weight, body composition, serum hormones, resting energy expenditure, cardiac function, muscle strength, and number of reconstructive procedures performed were measured during rhGH treatment and for 12 months after treatment was discontinued. Statistical analysis used Tukey's multiple comparison test. Significance was accepted at P < 0.05. RESULTS: Height, weight, lean body mass, bone mineral content, cardiac function, and muscle strength significantly improved during rhGH treatment compared with placebo (P < 0.05). This treatment significantly increased GH, IGF-I, and IGFBP-3, whereas serum cortisol decreased (P < 0.05). The number of operative reconstructive procedures was significantly lower with rhGH (P < 0.05). Improvements in height, bone mineral content, and IGF-1 concentrations persisted after rhGH treatment (P < 0.05). No side effects with rhGH were observed. CONCLUSIONS: Administration of rhGH for 1 year after burn was safe and improved recovery. These salutary effects continued after rhGH treatment was discontinued.

Oxidative and heat stress gene changes in hypertrophic scar fibroblasts stimulated with interleukin-1beta.

INTRODUCTION: Cutaneous wounds that involve loss of tissue heal through a complex process of generating granulation tissue to initially cover the wound, followed by epithelialization, and contraction. Normal healing requires a delicate balance between cellular, matrix, and vascularity build up and breakdown. Defects in the regulation of this balance can alter normal scar formation through fibroblastic hyperproliferation, which is characteristic of hypertrophic scar formation. METHODS: Primary fibroblasts cultures from hypertrophic scar or adjacent normal skin were seeded in growth medium. Half of each was stimulated with interleukin (IL)-1beta for 6 h and half served as control nonstimulated fibroblasts. Supernatants were tested for expressed proteins and the fibroblasts for changes in gene expression. RESULTS: Comparison between normal skin and hypertrophic scar fibroblasts stimulated with IL-1beta indicate that 15 genes increased and 8 genes decreased expression. When normal skin was stimulated with IL-1beta, there were increases in the expression of heat shock transcription factor-1, hsp70, and IL-6 When hypertrophic scar was stimulated with IL-1beta, there was a decrease in nuclear factor-kappaB, GADD45-alpha, p53, p53 binding protein, and Cox-2 genes. These genes may play specific but different roles in controlling the cellular response to cell stress and apoptosis. CONCLUSION: Data presented suggest that oxidative and heat stress proteins, stimulated through IL-1beta, may be important mediators of abnormal scarring.

Endogenous anabolic hormones and hypermetabolism: effect of trauma and gender differences.

OBJECTIVE: Protein degradation, negative nitrogen balance and compromised structure of essential organs have been associated with resistance and decreased production of anabolic hormones. In turn, increased levels of anabolic hormones are associated with improved survival. The aims of the present study were to determine the pattern of anabolic hormones, resting energy expenditure and cytokines in severely thermally injured pediatric patients and to compare these parameters in female and male patients. METHODS: Sixty-five children (1 to 16 years of age) sustaining a severe thermal injury (> or =40% TBSA) were included into the study. Patients were further divided into females (n = 22) and males (n = 43). Patient demographics, nutritional support, incidence of sepsis, inhalation injury, and mortality were noted. Resting energy expenditure was measured during hospital course by indirect calorimetry. Blood was drawn 0, 10, 20, and 40 days postburn and serum insulin-like growth factor-I (IGF-I), insulin-like growth factor binding protein-1 and -3 (IGFBP-1, and -3), growth hormone, insulin, and cytokines were measured. RESULTS: There were no significant differences between females and males for demographics, nutritional intake, or concomitant injuries. In both groups, endogenous anabolic agents were drastically decreased by 3- to 5-fold up to 40 days posttrauma. Females had significantly higher levels of IGF-I, IGFBP-3, growth hormone, and insulin when compared with males, P < 0.05. Increased levels of anabolic hormones were associated with decreased stay on the ICU (females 36 +/- 22 days versus males 53+/- 39 days), decreased serum IL-1beta and TNF-alpha as well as resting energy expenditure, P < 0.05. CONCLUSION: Data indicate that despite adequate nutritional support, severe thermal injury leads to decreased anabolic hormones over a prolonged period of time. Female patients had significantly increased levels of anabolic hormones, which are associated with decreased proinflammatory mediators and hypermetabolism, leading to a significant shorter ICU stay compared with male patients.

Gene profiling in muscle of severely burned children: age- and sex-dependent changes.

BACKGROUND: Thermal injury is associated with a pronounced catabolic response in skeletal muscle. This study identifies gene expression changes in skeletal muscle of thermally injured girls and boys using high-density oligonucleotide arrays. METHODS: Six burned children with a mean age of 8.3 +/- 1.3 years and TBSA burn size covering 51 +/- 6% admitted to our hospital with in 48 h of injury and six cleft lip and cleft palate patients were studied. Total RNA was isolated, in vitro transcribed, and hybridized to HG-U95 Av.2 Affymetrix arrays. Messenger RNA expression patterns of controls and burn patients were compared using Affymetrix GeneChip Analysis Suite 5.2 and dChip. RESULTS: Statistical analysis of the 12,625 genes on each array showed a significant increase in the expression of 77 genes in burn children and a decrease in 21 genes when compared to controls (P < 0.05). We found three genes in burned males and two genes in burned females with decreased expression in muscle compared to controls. Chromosomes 1, 2, 7, 12, and 16 showed genes with increased expression in muscle from burned children, while chromosomes 3, 7, 8, 19, and 22 had genes with decreased expression. Categories of genes affected were related to metabolism, proliferation, transcription/translation, immune response, stress response, angiogenesis, and signal transduction. CONCLUSIONS: Genes that are differentially expressed in skeletal muscle of burned children, but whose function in muscle is unknown, include those related to various transcription factors and those known to encode proteins involved in signaling pathways. Further analysis is required to achieve the ultimate goal of making functionally relevant conclusions about the molecular pathology of burn injury.