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Objective: The purposes of this experiment were to determine the repeatability of acute mountain sickness (AMS), AMS symptoms, and physiological responses across 2 identical hypoxic exposures., Methods: Subjects (n = 25) spent 3 nights at simulated altitude in a normobaric hypoxia chamber: twice at a partial pressure of inspired oxygen (PIO2) of 90mmHg (4000 m equivalent; "hypoxia") and once at a PIO2 of 132 mmHg (1000 m equivalent; "sham") with 14 or more days between exposures. The following variables were measured at hours 0 and 12 of each exposure: AMS severity (ie, Lake Louise score [LLS]), AMS incidence (LLS ≥3), heart rate, oxygen saturation, blood pressure, and the fraction of exhaled nitric oxide. Oxygen saturation and heart rate were also measured while subjects slept., Results: The incidence of AMS was not statistically different between the 2 exposures (84% vs 56%, P > .05), but the severity of AMS (ie, LLS) was significantly lower on the second hypoxic exposure (mean [SD], 3.1 [1.8]) relative to the first hypoxic exposure (4.8 [2.3]; P < .001). Headache was the only AMS symptom to have a significantly greater severity on both hypoxic exposures (relative to the sham exposure, P < .05). Physiological variables were moderately to strongly repeatable (intraclass correlation range 0.39 to 0.86) but were not associated with AMS susceptibility (P > .05)., Conclusions: The LLS was not repeatable across 2 identical hypoxic exposures. Increased familiarity with the environment (not acclimation) could explain the reduced AMS severity on the second hypoxic exposure. Headache was the most reliable AMS symptom., (Copyright © 2014 Wilderness Medical Society. Published by Elsevier Inc. All rights reserved.)

Human enamel has been found to be a coating with excellent mechanical performance, and has undergone extensive investigation and discussion. However, most of the reported studies consider the enamel as a homogeneous anisotropic biocomposite. The current study illustrated the graded properties of the biocomposite from its functional load-bearing direction. Within the thickness of the enamel, from the outer surface towards the enamel-dentin junction (EDJ), the elastic modulus (E(x)) and hardness (H(x)) of enamel exist in an exponential relationship with normalized thickness (x) as E(x)=111.64x(0.18) (R(2)=0.94) and H(x)=4.41x(0.16) (R(2)=0.87) GPa, respectively. Moreover, the creep ability of enamel increases towards the EDJ. The graded properties of the biocomposite can be explained by both microstructural and compositional changes along the thickness of the material towards the EDJ. Finite element analysis indicates that the graded properties of enamel have important roles in reducing the enamel-dentin interface stresses and maintaining the integrity of the multilayer tooth structure. The results provide a new angle to understand the excellent mechanical behaviour of the multilayer tooth structure and may inspire the development of new functionally graded materials and coating structures., (Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Aims: Cold stress has been shown to produce dramatic increases in 2-fluoro-2-deoxy-D-Glucose ((18)FDG) accumulation by brown adipose tissue (BAT) in rodents. However, neither the effects of other types of stress on (18)FDG accumulation nor the effects of stressors on the accumulation of tracers of other aspects of energy metabolism have been evaluated. In this report we studied the effects of cold stress, burn injury and cutaneous wounds on murine BAT at the macroscopic, microscopic and metabolic level., Main Methods: Glucose metabolism was studied with (18)FDG, fatty acid accumulation was evaluated with trans-9(RS)-(18)F-fluoro-3,4(RS,RS)-methyleneheptadecanoic acid (FCPHA) and tricarboxcylic acid cycle (TCA) activity was evaluated with (3)H acetate., Key Findings: All three stressors produced dramatic changes in BAT at the macroscopic and microscopic level. Macroscopically, BAT from the stressed animals appeared to be a much darker brown in color. Microscopically BAT of stressed animals demonstrated significantly fewer lipid droplets and an overall decrease in lipid content. Accumulation of (18)FDG by BAT was significantly (p<0.01) increased by all 3 treatments (Cold: ~16 fold, burn ~7 Fold and cutaneous wound ~14 fold) whereas uptake of FDG by white fat was unchanged. This effect was also demonstrated non invasively by μPET imaging. Although less prominent than with (18)FDG, BAT uptake of FCPHA and acetate were also significantly increased by all three treatments. These findings suggest that in addition to cold stress, burn injury and cutaneous wounds produce BAT activation in mice., Significance: This study demonstrates brown fat activated by several stressors leads to increased uptake of various substrates., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Infection is the most common and most serious complication of a major burn related to burn size. Recent studies have demonstrated that statin treatment can decrease mortality in murine or human sepsis. In the current study mice were anesthetized and subjected to a dorsal 30% TBSA scald burn. Simvastatin or placebo were administered by intraperitoneal injection once daily or every 12h. On post burn day 7 cecal ligation and puncture with a 21-gauge needle (CLP) was performed under ketamine/xylazine anesthesia, the two different dosing schedules were continued and survival was monitored. In other groups of mice, interleukin-6 (IL-6) levels in blood were measured in mice at 7 days after injury. A simvastatin dependent improvement in survival was observed in the burn sepsis model. This protection was found to be dose and time dependent. In addition, statin treatment reduced the elevation in IL-6 levels of mice burned 7 days previously. However, IL-6 levels in burned mice with or without statin treatment were elevated by CLP to the same degree. The results of these studies suggest that statin treatment reduces mortality in mice with burns and CLP and that this effect may not be mediated via IL-6 levels., (Copyright © 2010 Elsevier Ltd and ISBI. All rights reserved.)

Endotoxemia plays an important role in the pathogenesis of sepsis and is accompanied by dysregulated apoptosis of immune and non-immune cells. Treatment with statins reduces mortality in rodent models of sepsis and endotoxemia. Inhibition of protein isoprenylation, including farnesylation, has been proposed as a mechanism to mediate the lipid-lowering-independent effects of statins. Nonetheless, the effects of the inhibition of isoprenylation have not yet been studied. To investigate the role of farnesylation, we evaluated the effects of farnesyltransferase inhibitor and statin on survival following lipopolysaccharide (LPS) challenge in mice. Both simvastatin (2mg/kg BW) and FTI-277 (20mg/kg BW) treatment improved survival by twofold after LPS injection, as compared with vehicle alone (p<0.01). LPS-induced cleavage (activation) of caspase-3, an indicator of apoptotic change, and increased protein expression of proapoptotic molecules, Bax and Bim, and activation of c-Jun NH(2)-terminal kinase (JNK/SAPK) in the liver and spleen were attenuated by both simvastatin and FTI-277. These results demonstrate that farnesyltransferase inhibitor as well as statin significantly reduced LPS-induced mortality in mice. Our findings also suggest that inhibition of protein farnesylation may contribute to the lipid-lowering-independent protective effects of statins in endotoxemia, and that protein farnesylation may play a role in LPS-induced stress response, including JNK/SAPK activation, and apoptotic change. Our data argue that farnesyltransferase may be a potential molecular target for treating patients with endotoxemia., (Copyright 2009 Elsevier Inc. All rights reserved.)

Tropoelastin is the monomer building block of the biopolymer elastin, which is responsible for elasticity in arteries, lung and skin. Previous studies have shown that, in contrast to predictions made based on primary sequence, tropoelastin has little regular secondary structure in aqueous solution and displays considerable flexibility. This investigation defines the level of residual structure present in tropoelastin and uses the naturally-occurring structure-inducing osmolyte trimethylamine N-oxide to examine the potential for regular structure in tropoelastin. Tropoelastin is defined as a thermodynamically unfolded premolten globule, which can account for its ability to elastically deform.

The Rio Tinto site is recognised as a terrestrial Mars analogue because of the presence of jarosite and related sulfates which have recently been identified by the NASA Mars Exploration Rover "Opportunity" in the El Capitan region of Meridiani Planum on Mars. It has long been known that acidophilic microbial action is responsible for the deep blood-red colour of the water in Rio Tinto, where the pH varies from about 1.5 to 3.0 and the water is rich in iron and sulfur. Following recent Raman spectroscopic characterisation of the mineral phases of the Rio Tinto system, we report here a study of the biological components found in several specimens of deposited minerals and near the waterside that were collected during a GeoRaman VI Conference organized field trip in 2006. Key biosignatures were found for carotenoids, scytonemin and mycosporine-like amino acids, which are indicative of the biological colonisation of exposed mineral substrates; information from this study will be useful for targeting Martian sites using a miniaturized Raman instrument where the biosignatures of relict or extant life could remain in the geological record.

A rapid method for measuring 3-methylhistidine (3MH) in rat and human urine with higher sensitivity and precision than any previously reported method is described using internal standard [1-(13)C]3MH (M+1) and negative chemical ionization (NCI) gas chromatography/mass spectrometry (GC/MS). Internal standard [1-(13)C]3MH (M+1) was added to rat and human urine samples, hydrolyzed, and absorbed onto cation exchange columns. The column eluent was dried and derivatized for GC/MS analysis. Quantification of 3MH levels was accomplished by monitoring the m/z 204 fragment. The m/z 204 fragment was chosen due to the fragment's abundance and stability as determined by analysis of [methyl-(2)H(3), (18)O(2)]3MH (M+7) and [methyl-(13)C]3MH (M+1) fragmentation patterns under NCI conditions. This method shows excellent linearity (0.9989) over the range studied (0-0.5 mol), high recovery (95.9%), and low coefficient of variation (4.7%). The described method is sensitive enough to detect 6.8 pmol amount of urinary 3MH with a precision of 9.1%. The in vivo utility of this method to quantify urinary 3MH was tested in a burn injury rat model and on urine specimens from pediatric burn patients. Data obtained from the urine of burn-injured rats and pediatric burn patients match previously reported trends and validate the in vivo utility of this method.

Hyperglycemia, glucose intolerance and elevated insulin levels frequently occur in burned patients; however, the mechanism(s) for this insulin resistance has not been fully elucidated. One possible mechanism could involve alterations in the phosphorylation of serine 307 of the insulin receptor substrate-1 (IRS-1) via activation of stress kinase enzymes, including SAPK/JNK. In the present study we examined the time course of the effect of burn injury to mice on: levels of IRS-1 protein, phosphorylation of serine 307 of IRS-1, SAPK/JNK kinase levels and activity and Akt kinase activity in hind limb skeletal muscle. Burn injury produced a reduction in hind limb muscle mass 24 h after injury, and, which persisted for 168 h. At 24 h after injury, there was a dramatic ( approximately 9-fold) increase in phosphorylation of IRS-1 serine 307 followed by a more moderate elevation thereafter. Total IRS-1 protein was slightly elevated at 24 h after injury and decreased to levels below sham treated animals at the later times. Burn injury did not appear to change total SAPK/JNK protein content, however, enzyme activity was increased for 7 days after injury. Akt kinase activity was decreased in skeletal muscle following burn injury; providing a biochemical basis for burn-induced insulin resistance. These findings are consistent with the hypothesis that burn-induced insulin resistance may be related, at least in part, to alterations in the phosphorylation of key proteins in the insulin signaling cascade, including IRS-1, and that changes in stress kinases, such as SAPK/JNK produced by burn injury, may be responsible for these changes in phosphorylation.

Confocal Raman spectroscopy is introduced as a noninvasive in vivo optical method to measure molecular concentration profiles in the skin. It is shown how it can be applied to determine the water concentration in the stratum corneum as a function of distance to the skin surface, with a depth resolution of 5 microm. The resulting in vivo concentration profiles are in qualitative and quantitative agreement with published data, obtained by in vitro X-ray microanalysis of skin samples. Semi-quantitative concentration profiles were determined for the major constituents of natural moisturizing factor (serine, glycine, pyrrolidone-5-carboxylic acid, arginine, ornithine, citrulline, alanine, histidine, urocanic acid) and for the sweat constituents lactate and urea. A detailed description is given of the signal analysis methodology that enables the extraction of this information from the skin Raman spectra. No other noninvasive in vivo method exists that enables an analysis of skin molecular composition as a function of distance to the skin surface with similar detail and spatial resolution. Therefore, it may be expected that in vivo confocal Raman spectroscopy will find many applications in basic and applied dermatologic research.

The hypermetabolic inflammatory state that occurs after major trauma has been extensively studied at the whole body level, however, there is only limited information on metabolic changes in individual tissues. In this study, the effect of thermal injury on metabolic function of uninjured hind limb muscle of rabbits was measured noninvasively by positron emission tomography (PET). Rabbits were subjected to full thickness burn on 25% of their body surface area. Two to three weeks after injury, PET and arterial blood sampling was performed during inhalation of 15O2, C15O2 and 11CO and after injection of 18FDG. The tissue and blood data were analyzed by standard kinetic models for blood flow, oxygen extraction fraction (OEF), oxygen utilization and glucose metabolism. A total of seven injured and five sham animals were studied. Total body oxygen consumption was measured by indirect calorimetry and plasma concentrations of glucose, insulin and IGF-1 were measured with standard assays. Compared to sham rabbits, blood flow to muscle of injured animals was unchanged. However, OEF, oxygen utilization and glucose metabolism were significantly reduced (p<0.01) in uninjured muscle of burned rabbits. These data demonstrate that thermal injury is associated with alterations in muscle metabolism, which are not related to change in blood flow.

In response to severe injury, extraordinary quantities of lactate that are released from the peripheral tissues serve as substrate for hepatic gluconeogenesis. It is possible that as a result of burn injury, reaction kinetics involving lactate could be directly changed within the liver. The metabolic fate of [U-14C]lactate was examined in vitro in fresh liver slices after a 20 per cent total body surface area injury. Glucose and CO2 were produced in vitro by the liver tissues after the injury and no differences were seen in the metabolism of these substrates between the injured and control animals. These findings suggest that the intrinsic enzymatic processes within the liver are not directly altered by injury itself or by any of the associated inflammatory mediators which appear early after burns.

The biodistribution of the plasma expander colloidal solution, heta-starch (HES), has been examined in rats, subjected to thermal injury or sepsis. The ability of these solutions to alter RES phagocytic function of [99mTc]-sulphur colloid ([99mTc]SC) uptake in vivo has been examined. The biodistribution of radiolabelled HES has been determined in normal rats. The HES colloidal solution has no deleterious effect upon RES function in vivo in the thermally injured animals or the septic animals as compared to sham controls. In addition, the HES colloidal solution seemed to be distributed primarily within the liver, spleen and kidney, with a small amount residing in the lung. Thermal injury did not increase the uptake of this material by the lung. These results suggest that the use of HES in thermally injured and septic individuals has no deleterious effects on RES function, nor does it accumulate in the lungs, and hence, should be advocated for use in these situations.

The effect of chronic interleukin-2 (IL-2) injection upon reticuloendothelial system (RES) function in the rat has been determined. Seven-day treatments with two doses of human recombinant IL-2 resulted in a dramatic reduction in the phagocytic uptake of the liver and spleen, while increasing the weight of both organs. There were dramatic histological changes in the intestine, liver and spleen as well. These results suggest that the chronic use of IL-2 can result in hepatic dysfunction, which is associated with altered RES phagocytic function.

The pathway which results in a loss of intestinal barrier function and transepithelial transfer of macromolecules after cutaneous thermal injury is unknown. To determine the enhanced absorption pathway, transepithelial transport of horseradish peroxidase (HRP) was examined ultrastructurally after a thermal injury. Within 6 h after the injury, increased HRP uptake was seen in the portal and systemic blood with the maximal increase in uptake measured at 18 h postinjury; permeability returned to normal by 72 h postinjury. Morphologically, the increased uptake was found to be transcellular through ultrastructurally normal intestinal absorptive cells. Occasional focal regions of enhanced HRP uptake were found and this enhanced uptake was attributed to focal intestinal epithelial disruptions. This increase in intestinal permeability represents a transient loss of intestinal barrier function and potentially allows absorption of macromolecules such as endotoxin from the intestinal lumen into the portal circulation early after thermal injury.

The effect of injection or chronic infusion of lipopolysaccharide (LPS) into unanaesthetized rats on the distribution of [99Tcm-]SC has been determined. At a dose of 2.5 mg/kg, LPS injection caused a marked alteration in the distribution of the radiolabelled material, with more uptake being achieved in the lung while less was taken up by the spleen. Kidney and liver uptake were also changed. Chronic infusion of LPS at a similar dose (3 mg/kg in 24 h) caused a marked increase in the uptake of the radioactive material by the lung only. These data are consistent with a working hypothesis that the alterations in RES phagocytic activity of the lung observed in rats subjected to burn trauma could be related in part to LPS, either coming as a bolus, or being continuously presented.

The synthesis of proteoglycans (PG) by normal human skin, and normal and hypertrophic scars were compared using tissue explants in culture. Newly synthesized PG were labelled with [35S]Na2SO4. Significant differences were found in the proportion of [35S]-radio-labelled incorporation of PG in the tissue and accumulation of [35S]PG in culture medium in the different tissues. The rate of PG biosynthesis in all three tissue types occurred in two phases. There was an initial phase of PG synthesis occurring at 0-3 h and a later phase that occurred at 3-18 h [35S]-labelled PG were isolated and characterized by Sepharose CL-6B chromatography and cellulose acetate electrophoresis. The results showed that the hypertrophic scar tissue and its culture medium contained higher proportions of dermatan sulphate (DS), chondroitin sulphate (CS) and DS' PG than the normal skin fractions. These results suggest that abnormal scarring is related to a change in the level of PG synthesis during the burn injury repair process.

Platelet-activating factor (PAF) was injected into conventional mice, endotoxin-resistant mice (C3H/HEJ), conventional rats, germ-free rats and specific-pathogen-free (SPF) mice. The PAF resulted in significant necrosis and damage to the small intestines of all the animals tested. In general, the frequency and severity of the lesions were similar in all groups. All the conventional rats and mice, as well as the endotoxin-resistant HEJ mice, were dead 18 h after the injection of the PAF, while all the germ-free rats and the SPF mice survived. These data demonstrate that development of massive intestinal lesions, in the absence of aerobic bacteria, is not sufficient to cause the death of the host from septic shock and endotoxaemia.

The role of haemolysis of blood in the alterations in the uptake of [99mTc]SC ([99mTc]-sulphur colloid) in vivo in the rat has been examined. When the haemolysed blood (produced by first freezing the blood in liquid nitrogen) was infused into synergenic Lewis rats via the tail vein, there was a significant reduction in the uptake of the [99mTc]SC by the spleen, but lung, liver and kidney uptake remained constant. These results suggest that haemolysis of the blood may play a role in the alterations in RES phagocytic activity observed in the spleen following thermal injury.

The effect of platelet activating factor (PAF) injections on the uptake of 99mTc-SC (99mTc-SC (99mTc-sulphur colloid) was determined in vivo. PAF (2 micrograms) injected intravenously into unanaesthetized, unrestrained rats was associated with the development of lesions in the small intestine and alteration of 99mTc-SC uptake in vivo. 99mTc-SC uptake into the lung was increased while spleen uptake was decreased. Pretreatment of the animals with a PAF antagonist, SRI-64-441, prevented the intestinal lesions and alterations of 99mTc-SC uptake. Macrophages, isolated from lung lavage of the PAF-treated rats, demonstrated a decreased generation of hydrogen peroxide in vitro. The present results suggest that, in addition to its other effects on the immune system, PAF can also alter the in vivo phagocytic activity of the reticuloendothelial system in the rat.

We have examined the role of neutrophils in the alterations observed in the intestines of rats subjected to 40 per cent surface area scald injury. Histologically, there was no evidence of neutrophilia in the intestinal tissue, and myeloperoxidase activity in mucosal scrapings was not elevated. The distribution of labelled human neutrophils injected into the burned rats showed no enhancement of uptake in the intestines, although there was increased uptake by the lung. The present data suggest that neutrophil migration may not play a role in the alterations in small intestinal function and morphology seen in burn trauma in the rat, but may be a factor in lung damage associated with thermal injury.

The uptake of radiolabelled colloid or bacteria was compared in normal rats and animals subjected to acute burn trauma. The uptake of colloid by the liver was unaffected by burn trauma, but uptake of the labelled bacteria was reduced. Spleen uptake of both colloid and bacteria was reduced by burn trauma while lung uptake was increased. These data are consistent with the hypothesis that acute burn trauma alters reticuloendothelial system phagocytic activity in the rat towards both inert particles and live bacteria.

Acute burn trauma has been demonstrated to depress reticuloendothelial system (RES) phagocytic activity, which could partially explain the development of septicaemia in burn patients. In the present study we have attempted to determine the role that skin plays in the depression of the RES. One group of Lewis rats was subjected to a 100 degrees C scald burn for 10 s. Eschar was then removed and implanted onto the backs of a second group of normal controls. A third group of Lewis rats were subjected to sham treatment; the eschar of these animals was removed and implanted onto the back of a fourth group of normal rats. The excision sites of the donor animals were immediately covered with Biobrane. Twenty-four hours later the technetium-99m sulphur colloid [( 99mTS]SC) method described earlier (Trop et al., 1989) was used to determine phagocytosis in these four groups of animals. Acute burn trauma produced a marked reduction in colloid uptake by the spleen and a marked increase in the uptake of colloid material by the lung, although no effect was observed on liver or kidney uptake. Implantation of the burn eschar into normal control rats had no statistically significant effect upon colloid uptake. These data suggest that alterations in phagocytic activity of the spleen and lung occur within minutes after burn injury and may be unrelated to the presence of the burn eschar itself.

Insulin, glucagon, and epidermal growth factor (EGF) addition stimulated DNA synthesis in primary hepatocyte cell cultures prepared from adult rat liver. The addition of ethanol (20-200mM) to the culture medium resulted in a substantial reduction in DNA synthesis as measured by 3H-thymidine incorporation and autoradiography. This effect was specific for differentiated hepatocytes compared to fibroblasts and two other human hepatoma cell lines. These studies demonstrate in a cell culture system that one of the major properties of ethanol is the inhibition of hepatocyte DNA synthesis.

Hepatic repair and regeneration which is extremely important after thermal injuries can be inhibited by the acute inflammatory reaction. Since thermal injury initiates this acute inflammatory reaction, DNA synthesis was studied in the regenerating liver following this injury. In vivo incorporation of [3H]-thymidine into hepatic DNA, autoradiographic determination of a labeling index, and thymidine kinase activity were determined. Incorporation of [3H]-thymidine into hepatic DNA and labeling indices were markedly diminished at 24 hours if partial hepatectomy and thermal injury were carried out concurrently. After partial hepatectomy, the expected elevations in thymidine kinase activity were inhibited by the thermal injury (p less than 0.01) and elevation of serum fibrinogen, a marker of the acute phase reaction that normally follows thermal injury, was blunted by the partial hepatectomy (p less than 0.05). The combination of thermal injury and partial hepatectomy resulted in a greatly diminished DNA replicative response as compared to partial hepatectomy alone and suggests that multiplicative injury is more likely to result in multi-system failure.

The acute phase protein response following inflammation is associated with an increased total protein-bound carbohydrate content in plasma in the form of glycoproteins. Glycosyltransferases in liver may serve as a regulator of this increased glycosylation activity in the plasma and may also serve as a marker for the acute phase response. Sialyltransferase is an example of a glycosyltransferases in which sialic acid is transferred to oligosaccharides of glycopeptides in the Golgi prior to glycopeptide secretion. In this study, sialyltransferase activities were determined in plasma, liver, and intestinal mucosa following a standardized 25% body surface area thermal injury in the rat. A statistically significant increase in sialyltransferase activity was found in liver and small intestine which were maximal at 24 hours after the injury. These increased sialyltransferase activities were accompanied by a statistically significant 2 to 4 fold elevation in plasma sialyltransferase activity at 24 hours. The plasma and liver elevations in these activities were similar to elevations seen in other models of acute inflammation using turpentine injections and bacterial infections. The increased sialyltransferase activity within the rat intestine was comparable to increases in intestinal sialyltransferase activity following colchicine treatment and may represent a similar mechanism(s).