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Objective: This project was aimed at evaluating the impact of combat armor on physiological and cognitive functions during low-intensity exercise in hot-humid conditions (36 degrees C and 60% relative humidity).Methods: Nine males participated in three trials (2.5 hours), walking at two speeds and wearing different protective equipment: control (combat uniform and cloth hat); torso armor with uniform and cloth hat; and full armor (uniform, torso armor, and helmet).Results: As time progressed, core temperatures increased and deviated significantly among trials, rising at 0.37 degrees C h(-1) (control), 0.41 degrees C h(-1) (torso armor), and 0.51 degrees C h(-1) (full armor). Heart rates also progressively diverged, and subjects lost significantly more sweat during the two armored trials. However, cognitive-function tests revealed neither significant main effects nor time by treatment interactions.Conclusion: The combat armor and helmet significantly increased thermal and cardiovascular strain, but these were unlikely to lead to either exertional heat illness or impaired cognitive function during uneventful urban, military patrols in hot-humid conditions. [ABSTRACT FROM AUTHOR]

Background: Older adults with cancer are a growing population requiring tailored care to achieve optimum treatment outcomes. Their care is complicated by under-recognised and under-treated wasting disorders: malnutrition, sarcopenia, and cachexia. We aimed to investigate the prevalence, overlap, and patients' views and experiences of malnutrition, sarcopenia, and cachexia, in older adults with cancer., Methods: Mixed-methods study with cross-sectional study and qualitative interviews. Interviews were thematically analysed through a phenomenological lens, with feedback loop analysis investigating relationships between themes and findings synthesised using modified critical interpretative synthesis., Findings: n = 30 were screened for malnutrition, sarcopenia, and cachexia, n = 8 completed semi-structured interviews. Eighteen (60.0%) were malnourished, 16 (53.3%) sarcopenic, and 17 (56.7%) cachexic. One or more condition was seen in 80%, and all three in 30%. In univariate analysis, Rockwood clinical frailty score (OR 2.94 [95% CI: 1.26-6.89, p = 0.013]) was associated with sarcopenia, reported percentage meal consumption (OR 2.28 [95% CI: 1.24-4.19, p = 0.008]), and visible wasting (OR 8.43 [95% CI: 1.9-37.3] p = 0.005) with malnutrition, and percentage monthly weight loss (OR 8.71 [95% CI: 1.87-40.60] p = 0.006) with cachexia. Screening tools identified established conditions rather than 'risk'. Nutritional and functional problems were often overlooked, overshadowed, and misunderstood by both patients and (in patients' perceptions) by clinicians; misattributed to ageing, cancer, or comorbidities. Patients viewed these conditions as both personal impossibilities, yet accepted inevitabilities., Conclusion: Perceptions, identification, and management of these conditions needs to improve, and their importance recognised by clinicians and patients so those truly 'at risk' are identified whilst conditions are more remediable to interventions., (© 2024. The Author(s).)

Objective: The aim of this study was to model the effect of body armor coverage on body core temperature elevation and wet-bulb globe temperature (WBGT) offset., Background: Heat stress is a critical factor influencing the health and safety of military populations. Work duration limits can be imposed to mitigate the risk of exertional heat illness and are derived based on the environmental conditions (WBGT). Traditionally a 3°C offset to WBGT is recommended when wearing body armor; however, modern body armor systems provide a range of coverage options, which may influence thermal strain imposed on the wearer., Method: The biophysical properties of four military clothing ensembles of increasing ballistic protection coverage were measured on a heated sweating manikin in accordance with standard international criteria. Body core temperature elevation during light, moderate, and heavy work was modeled in environmental conditions from 16°C to 34°C WBGT using the heat strain decision aid., Results: Increasing ballistic protection resulted in shorter work durations to reach a critical core temperature limit of 38.5°C. Environmental conditions, armor coverage, and work intensity had a significant influence on WBGT offset., Conclusion: Contrary to the traditional recommendation, the required WBGT offset was >3°C in temperate conditions (<27°C WBGT), particularly for moderate and heavy work. In contrast, a lower WBGT offset could be applied during light work and moderate work in low levels of coverage., Application: Correct WBGT offsets are important for enabling adequate risk management strategies for mitigating risks of exertional heat illness.

Objective: To investigate the effects of repeated work bouts in the heat on peak body core temperature and to explore sex-based differences in body core temperature responses., Methods: Fourteen males and fifteen females performed four work bouts (two heavy and two moderate, alternating) in 32.5 °C Wet Bulb Globe Temperature (WBGT), each separated by 30-min seated rest in 28.0 °C WBGT. Participants wore a military combat uniform with body armour and helmet (10 kg load) during the work bouts, removing the vest and helmet during recovery periods., Results: Body core temperature elevation over time was faster in the first compared with subsequent work bouts of each intensity. Body core temperature elevation was similar between males and females during the first heavy work bout, then remained significantly lower in females for the reminder of the trial., Conclusions: Contrary to the assumed progressive elevation in strain, but in agreement with recent literature, a gradual reduction in heat storage in subsequent exercise bouts prevented a cumulative increase in heat strain in the conditions tested., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Identifying major histocompatibility complex (MHC) class I immunopeptide antigens represents a key step in the development of immune-based targeted therapeutics and vaccines. However, the complete characterization of these antigens by tandem mass spectrometry remains challenging due to their short sequence length, high degree of hydrophobicity, and/or lack of sufficiently basic amino acids. This study seeks to address the potential for 193 nm ultraviolet photodissociation (UVPD) to improve the analysis of MHC class I immunopeptides by offering enhanced characterization of these sequences in lower charge states and differentiation of prominent isomeric leucine and isoleucine residues in the HLA-A*02:01 motif. Although electron transfer dissociation-higher energy collisional dissociation (EThcD) offered some success in the differentiation of leucine and isoleucine, 193 nm UVPD was able to confirm the identity of nearly 60% of leucine and isoleucine residues in a synthetic peptide mixture. Furthermore, 193 nm UVPD led to significantly more peptide identifications and higher scoring metrics than EThcD for peptides obtained from immunoprecipitation of MHC class I immunopeptides from in vitro cell culture. Additionally, 193 nm UVPD represents a promising complementary technique to higher-energy collisional dissociation (HCD), in which 424 of the 2593 peptides identified by 193 nm UVPD were not identified by HCD in HLA-A*02:01-specific immunoprecipitation and 804 of the 3300 peptides identified by 193 nm UVPD were not identified by HCD for pan HLA-A, -B, and -C immunoprecipitation. These results highlight that 193 nm UVPD offers an option for the characterization of immunopeptides, including differentiation of leucine and isoleucine residues.

Background: Despite recommendations for nutritional risk screening of all inpatients, outpatients and care home residents, as well as work to assess clinician's experiences and the validity of tools, little attention has been paid to the experiences of patients undergoing nutritional screening. This review aims to synthesise systematically the current evidence regarding nutritional risk screening with respect to the experiences and views of patients, their families and carers., Methods: A systematic search was performed in MEDLINE, Embase, PsychINFO, CINAHL, Web of Science and British Nursing Database (inception - July 2019); with screening terms related to malnutrition, screening tools and experience. Titles, abstracts and full-text papers were independently reviewed by two reviewers and then quality-appraised. Qualitative papers and quantitative surveys were included. A narrative review of surveys and a thematic framework synthesis of interviews were used to identify themes., Results: Nine studies, including five qualitative interview papers, were included. Qualitative and quantitative study results were combined using a matrix chart to allow comparison. Surveyed participants reported processes of nutritional screening as acceptable. Three key themes emerged from qualitative data: (i) experience of nutritional screening; (ii) misunderstanding of malnutrition: of causes, role of screening and poor self-perception of risk; and (iii) barriers to and opportunities for change., Conclusions: Although the screening process is acceptable, patients' misunderstanding and poor knowledge regarding causes and consequences of malnutrition result in reduced risk perception and disbelief or disregard of nutritional screening results. Findings should inform policy and clinical practice, as well as highlight the known paucity of data regarding the effectiveness of screening on clinical outcomes., (© 2020 The Authors. Journal of Human Nutrition and Dietetics published by John Wiley & Sons Ltd on behalf of British Dietetic Association.)

Aberrant gene activation driven by the histone acetyltransferases p300 and CREB binding protein (CBP) has been linked to several diseases, including cancers. Because of this, many efforts have been aimed toward the targeting of the closely related paralogues, p300 and CBP, but these endeavors have been exclusively directed toward noncovalent inhibitors. X-ray crystallography of A-485 revealed that both p300 and CBP possess a cysteine (C1450) near the active site, thus rendering covalent inhibition an attractive chemical approach. Herein we report the development of compound 2 , an acrylamide-based inhibitor of p300/CBP that forms a covalent adduct with C1450. We demonstrated using mass spectrometry that compound 2 selectively targets C1450, and we also validated covalent binding using kinetics experiments and cellular washout studies. The discovery of covalent inhibitor 2 gives us a unique tool for the study of p300/CBP biology., Competing Interests: The authors declare no competing financial interest., (© 2021 American Chemical Society.)

Peptides presented by the class-I major histocompatibility complex (MHC-I) are important targets for immunotherapy. The identification of these peptide targets greatly facilitates the generation of T-cell-based therapeutics. Herein, we report the capability of proteolysis targeting chimera (PROTAC) compounds to induce the presentation of specific MHC class-I peptides derived from endogenous cellular proteins. Using LC-MS/MS, we identified several BET-derived MHC-I peptides induced by treatment with three BET-directed PROTAC compounds. To understand our ability to tune this process, we measured the relative rate of presentation of these peptides under varying treatment conditions using label-free mass spectrometry quantification. We found that the rate of peptide presentation reflected the rate of protein degradation, indicating a direct relationship between PROTAC treatment and peptide presentation. We additionally analyzed the effect of PROTAC treatment on the entire immunopeptidome and found many new peptides that were displayed in a PROTAC-specific fashion: we determined that these identifications map to the BET pathway, as well as, potential off-target or unique-to-PROTAC pathways. This work represents the first evidence of the use of PROTAC compounds to induce the presentation of MHC-I peptides from endogenous cellular proteins, highlighting the capability of PROTAC compounds for the discovery and generation of new targets for immunotherapy.

Unlabelled: Following phagocytosis, microbes are exposed to an array of antimicrobial weapons that include reactive oxygen species (ROS) and cationic fluxes. This is significant as combinations of oxidative and cationic stresses are much more potent than the corresponding single stresses, triggering the synergistic killing of the fungal pathogenCandida albicansby "stress pathway interference." Previously we demonstrated that combinatorial oxidative plus cationic stress triggers a dramatic increase in intracellular ROS levels compared to oxidative stress alone. Here we show that activation of Cap1, the major regulator of antioxidant gene expression inC. albicans, is significantly delayed in response to combinatorial stress treatments and to high levels of H2O2 Cap1 is normally oxidized in response to H2O2; this masks the nuclear export sequence, resulting in the rapid nuclear accumulation of Cap1 and the induction of Cap1-dependent genes. Here we demonstrate that following exposure of cells to combinatorial stress or to high levels of H2O2, Cap1 becomes trapped in a partially oxidized form, Cap1(OX-1) Notably, Cap1-dependent gene expression is not induced when Cap1 is in this partially oxidized form. However, while Cap1(OX-1)readily accumulates in the nucleus and binds to target genes following high-H2O2stress, the nuclear accumulation of Cap1(OX-1)following combinatorial H2O2and NaCl stress is delayed due to a cationic stress-enhanced interaction with the Crm1 nuclear export factor. These findings define novel mechanisms that delay activation of the Cap1 transcription factor, thus preventing the rapid activation of the stress responses vital for the survival ofC. albicanswithin the host., Importance: Combinatorial stress-mediated synergistic killing represents a new unchartered area in the field of stress signaling. This phenomenon contrasts starkly with "stress cross-protection," where exposure to one stress protects against subsequent exposure to a different stress. Previously we demonstrated that the pathogenCandida albicansis acutely sensitive to combinations of cationic and oxidative stresses, because the induction of H2O2-responsive genes is blocked in the presence of cationic stress. We reveal that this is due to novel mechanisms that delay activation of the Cap1 AP-1-like transcription factor, the major regulator of the H2O2-induced regulon. Cap1 becomes trapped in a partially oxidized form following simultaneous exposure to oxidative and cationic stresses. In addition, cationic stress promotes the interaction of Cap1 with the Crm1 nuclear export factor, thus inhibiting its nuclear accumulation. These mechanisms probably explain the potency of neutrophils, which employ multiple stresses to kill fungal pathogens., (Copyright © 2016 Kos et al.)

Military activities in hot environments pose 2 competing demands: the requirement to perform realistic training to develop operational capability with the necessity to protect armed forces personnel against heat-related illness. To ascertain whether work duration limits for protection against heat-related illness restrict military activities, this study examined the heat strain and risks of heat-related illness when conducting a military activity above the prescribed work duration limits. Thirty-seven soldiers conducted a march (10 km; ∼5.5 km h -1 ) carrying 41.8 ± 3.6 kg of equipment in 23.1 ± 1.8°C wet-bulb globe temperature. Body core temperature was recorded throughout and upon completion, or withdrawal, participants rated their severity of heat-related symptoms. Twenty-three soldiers completed the march in 107 ± 6.4 min (Completers); 9 were symptomatic for heat exhaustion, withdrawing after 71.6 ± 10.1 min (Symptomatic); and five were removed for body core temperature above 39.0°C (Hyperthermic) after 58.4 ± 4.5 min. Body core temperature was significantly higher in the Hyperthermic (39.03 ± 0.26°C), than Symptomatic (38.34 ± 0.44°C; P = 0.007 ) and Completers (37.94 ± 0.37°C; P<0.001 ) after 50 min. Heat-related symptom severity was significantly higher among Symptomatic (28.4 ± 11.8) compared to Completers (15.0 ± 9.8, P = 0.006 ) and Hyperthermic (13.0 ± 9.6, P = 0.029 ). The force protection provided by work duration limits may be preventing the majority of personnel from conducting activities in hot environments, thereby constraining a commander's mandate to develop an optimised military force. The dissociation between heat-related symptoms and body core temperature elevation suggests that the physiological mechanisms underpinning exhaustion during exertional heat stress should be re-examined to determine the most appropriate physiological criteria for prescribing work duration limits.

This project was based on the premise that decisions concerning the ballistic protection provided to defence personnel should derive from an evaluation of the balance between protection level and its impact on physiological function, mobility, and operational capability. Civilians and soldiers participated in laboratory- and field-based studies in which ensembles providing five levels of ballistic protection were evaluated, each with progressive increases in protection, mass (3.4-11.0 kg), and surface-area coverage (0.25-0.52 m(2)). Physiological trials were conducted on volunteers (N = 8) in a laboratory, under hot-dry conditions simulating an urban patrol: walking at 4 km·h(-1) (90 min) and 6 km·h(-1) (30 min or to fatigue). Field-based trials were used to evaluate tactical battlefield movements (mobility) of soldiers (N = 31) under tropical conditions, and across functional tests of power, speed, agility, endurance, and balance. Finally, trials were conducted at a jungle training centre, with soldiers (N = 32) patrolling under tropical conditions (averaging 5 h). In the laboratory, work tolerance was reduced as protection increased, with deep-body temperature climbing relentlessly. However, the protective ensembles could be grouped into two equally stressful categories, each providing a different level of ballistic protection. This outcome was supported during the mobility trials, with the greatest performance decrement evident during fire and movement simulations, as the ensemble mass was increased (-2.12%·kg(-1)). The jungle patrol trials similarly supported this outcome. Therefore, although ballistic protection does increase physiological strain, this research has provided a basis on which to determine how that strain can be balanced against the mission-specific level of required personal protection.

Background and Objectives: Approximately 400 000 students participate in US high school cheerleading annually, including 123,386 involved in competitive spirit squads. The degree of athleticism and the difficulty of cheerleading skills have increased in recent decades, renewing safety concerns. This study describes the epidemiology of high school cheerleading injuries and compares cheerleading injury rates and patterns relative to other sports., Methods: Data collected by the longitudinal, National High School Sports-Related Injury Surveillance Study from 2009/2010 through 2013/2014 were analyzed., Results: Injury rates in cheerleading ranked 18th of 22 sports, with an overall injury rate of 0.71 per 1000 athlete-exposures (AEs). Competition (0.85) and practice (0.76) injury rates were similar, whereas performance rates were lower (0.49). Although 96.8% of injured cheerleaders were girls, the overall injury rate was higher in boys (1.33 vs 0.69, rate ratio [RR]: 1.93, 95% confidence interval [CI]: 1.30-2.88). Although concussions were the most common cheerleading injury (31.1% of injuries), concussion rates were significantly lower in cheerleading (2.21 per 10,000 athlete-exposures) than all other sports combined (3.78; RR: 0.58, 95% CI: 0.51-0.66) and all other girls' sports (2.70; RR: 0.82, 95% CI: 0.72-0.93). Over half of all injuries occurred during stunts (53.2%)., Conclusions: Although safety remains a concern among cheerleaders, overall injury rates are lower than most other high school sports. Although overall injury rates are relatively low, cheerleading injuries may be more severe when they do occur. A detailed knowledge of cheerleading injury patterns relative to other sports is needed to drive targeted, evidence-based prevention efforts., (Copyright © 2016 by the American Academy of Pediatrics.)

In chronic lymphocytic leukemia (CLL), mutation and loss of p53 and ATM abrogate DNA damage signalling and predict poorer response and shorter survival. We hypothesised that poly (ADP-ribose) polymerase (PARP) activity, which is crucial for repair of DNA breaks induced by oxidative stress or chemotherapy, may be an additional predictive biomarker and a target for therapy with PARP inhibitors.We measured PARP activity in 109 patient-derived CLL samples, which varied widely (192 - 190052 pmol PAR/10⁶ cells) compared to that seen in healthy volunteer lymphocytes (2451 - 7519 pmol PAR/10⁶ cells). PARP activity was associated with PARP1 protein expression and endogenous PAR levels. PARP activity was not associated with p53 or ATM loss, Binet stage, IGHV mutational status or survival, but correlated with Bcl-2 and Rel A (an NF-kB subunit). Levels of 8-hydroxy-2'-deoxyguanosine in DNA (a marker of oxidative damage) were not associated with PAR levels or PARP activity. The potent PARP inhibitor, talazoparib (BMN 673), inhibited CD40L-stimulated proliferation of CLL cells at nM concentrations, independently of Binet stage or p53/ATM function.PARP activity is highly variable in CLL and correlates with stress-induced proteins. Proliferating CLL cells (including those with p53 or ATM loss) are highly sensitive to the PARP inhibitor talazoparib.

ATR is an attractive target in cancer therapy because it signals replication stress and DNA lesions for repair and to S/G2 checkpoints. Cancer-specific defects in the DNA damage response (DDR) may render cancer cells vulnerable to ATR inhibition alone. We determined the cytotoxicity of the ATR inhibitor VE-821 in isogenically matched cells with DDR imbalance. Cell cycle arrest, DNA damage accumulation and repair were determined following VE-821 exposure.Defects in homologous recombination repair (HRR: ATM, BRCA2 and XRCC3) and base excision repair (BER: XRCC1) conferred sensitivity to VE-821. Surprisingly, the loss of different components of the trimeric non-homologous end-joining (NHEJ) protein DNA-PK had opposing effects. Loss of the DNA-binding component, Ku80, caused hypersensitivity to VE-821, but loss of its partner catalytic subunit, DNA-PKcs, did not. Unexpectedly, VE-821 was particularly cytotoxic to human and hamster cells expressing high levels of DNA-PKcs. High DNA-PKcs was associated with replicative stress and activation of the DDR. VE-821 suppressed HRR, determined by RAD51 focus formation, to a greater extent in cells with high DNA-PKcs.Defects in HRR and BER and high DNA-PKcs expression, that are common in cancer, confer sensitivity to ATR inhibitor monotherapy and may be developed as predictive biomarkers for personalised medicine.

Background: Patients with malignant pleural effusions (MPEs) generally have advanced disease with poor survival and few therapeutic options. Cells within MPEs may be used to stratify patients for targeted therapy. Targeted therapy with poly(ADP ribose) polymerase inhibitors (PARPi) depends on identifying homologous recombination DNA repair (HRR)-defective cancer cells. We aimed to determine the feasibility of assaying HRR status in MPE cells., Methods: A total of 15 MPE samples were collected from consenting patients with non-small-cell lung cancer (NSCLC), mesothelioma and ovarian and breast cancer. Primary cultures were confirmed as epithelial by pancytokeratin, and HRR status was determined by the detection of γH2AX and RAD51 foci following a 24-h exposure to rucaparib, by immunofluorescence microscopy. Massively parallel next-generation sequencing of DNA repair genes was performed on cultured MPE cells., Results: From 15 MPE samples, 13 cultures were successfully established, with HRR function successfully determined in 12 cultures. Four samples - three NSCLC and one mesothelioma - were HRR defective and eight samples - one NSCLC, one mesothelioma, one sarcomatoid, one breast and four ovarian cancers - were HRR functional. No mutations in DNA repair genes were associated with HRR status, but there was probable loss of heterozygosity of FANCG, RPA1 and PARP1., Conclusions: HRR function can be successfully detected in MPE cells demonstrating the potential to stratify patients for targeted therapy with PARPi.

Unlabelled: Most heat acclimation data are from regimes longer than 1 week, and acclimation advice is to prevent dehydration., Objectives: We hypothesized that (i) short-term (5-day) heat acclimation would substantially improve physiological strain and exercise tolerance under heat stress, and (ii) dehydration would provide a thermally independent stimulus for adaptation., Methods: Nine aerobically fit males heat acclimated using controlled-hyperthermia (rectal temperature 38.5°C) for 90 min on 5 days; once euhydrated (EUH) and once dehydrated (DEH) during acclimation bouts. Exercising heat stress tests (HSTs) were completed before and after acclimations (90-min cycling in Ta 35°C, 60% RH)., Results: During acclimation bouts, [aldosterone]plasma rose more across DEH than EUH (95%CI for difference between regimes: 40-411 pg ml(-1); P = 0.03; n = 5) and was positively related to plasma volume expansion (r = 0.65; P = 0.05), which tended to be larger in DEH (CI: -1 to 10%; P = 0.06; n = 9). In HSTs, resting forearm perfusion increased more in DEH (by 5.9 ml 100 tissue ml(-1) min(-1): -11.5 to -1.0; P = 0.04) and end-exercise cardiac frequency fell to a greater extent (by 11 b min(-1): -1 to 22; P = 0.05). Hydration-related effects on other endocrine, cardiovascular, and psychophysical responses to HSTs were unclear. Rectal temperature was unchanged at rest but was 0.3°C lower at end exercise (P < 0.01; interaction: P = 0.52)., Conclusions: Short-term (5-day) heat acclimation induced effective adaptations, some of which were more pronounced after fluid-regulatory strain from permissive dehydration, and not attributable to dehydration effects on body temperature., (Copyright © 2014 Wiley Periodicals, Inc.)

Aim: This experiment was designed to test the hypothesis that the plasma volume is not selectively defended during exercise- and heat-induced dehydration following humid-heat acclimation., Methods: Eight physically active males were heat acclimated (39.8 °C, relative humidity 59.2%) using 17 days of controlled hyperthermia (core temperature: 38.5 °C). Inter-compartmental fluid losses and movements were tracked (radioisotopes and Evans blue dye) during progressive dehydration (cycling) in these same conditions and also during a resting recovery without fluid replacement (28 °C), before (day 1), during (day 8) and after heat acclimation (day 22)., Results: On days 8 and 22, there were significant increases in total body water, interstitial fluid and plasma volume (P < 0.05), but the intracellular compartments did not change (P > 0.05). The baseline plasma volume remained expanded throughout: 43.4 [±2.6 (day 1)], 49.1 [±2.4 (day 8); P < 0.05] and 48.9 mL kg(-1) [±3.0 (day 22); P < 0.05]. During progressive dehydration, plasma reductions of 9.0% (±0.9: day 1), 12.4% (±1.6: day 8) and 13.6% (±1.2: day 22) were observed, with day 8 and 22 losses significantly exceeding day 1 (P < 0.05). During recovery, plasma volume restoration commenced, with the intracellular fluid contribution becoming more pronounced as acclimation progressed., Conclusion: It is concluded that the plasma volume was not defended more vigorously following humid-heat acclimation. Indeed, a greater fluid loss may well underlie the mechanisms for enhancing plasma volume recovery when heat acclimation is induced using the controlled-hyperthermia technique., (© 2013 Scandinavian Physiological Society. Published by John Wiley & Sons Ltd.)

Aims: As Candida albicans is the major fungal pathogen of humans, there is an urgent need to understand how this pathogen evades toxic reactive oxygen species (ROS) generated by the host immune system. A key regulator of antioxidant gene expression, and thus ROS resistance, in C. albicans is the AP-1-like transcription factor Cap1. Despite this, little is known regarding the intracellular signaling mechanisms that underlie the oxidation and activation of Cap1. Therefore, the aims of this study were; (i) to identify the regulatory proteins that govern Cap1 oxidation, and (ii) to investigate the importance of Cap1 oxidation in C. albicans pathogenesis., Results: In response to hydrogen peroxide (H2O2), but not glutathione-depleting/modifying oxidants, Cap1 oxidation, nuclear accumulation, phosphorylation, and Cap1-dependent gene expression, is mediated by a glutathione peroxidase-like enzyme, which we name Gpx3, and an orthologue of the Saccharomyces cerevisiae Yap1 binding protein, Ybp1. In addition, Ybp1 also functions to stabilise Cap1 and this novel function is conserved in S. cerevisiae. C. albicans cells lacking Cap1, Ybp1, or Gpx3, are unable to filament and thus, escape from murine macrophages after phagocytosis, and also display defective virulence in the Galleria mellonella infection model., Innovation: Ybp1 is required to promote the stability of fungal AP-1-like transcription factors, and Ybp1 and Gpx3 mediated Cap1-dependent oxidative stress responses are essential for the effective killing of macrophages by C. albicans., Conclusion: Activation of Cap1, specifically by H2O2, is a prerequisite for the subsequent filamentation and escape of this fungal pathogen from the macrophage.

Individuals exposed to extreme heat may experience reduced physiological and cognitive performance, even during very light work. This can have disastrous effects on the operational capability of aircrew, but such impairment could be prevented by auxiliary cooling devices. This hypothesis was tested under very hot-dry conditions, in which eight males performed 2 h of low-intensity exercise (~30 W) in three trials, whilst wearing biological and chemical protective clothing: temperate (control: 20°C, 30% relative humidity) and two hot-dry trials (48°C, 20% relative humidity), one without (experimental) and one with liquid cooling (water at 15°C). Physiological strain and six cognitive functions were evaluated (MiniCog Rapid Assessment Battery), and participants drank to sustain hydration state. Maximal core temperatures averaged 37.0°C (±0.1) in the control trial, and were significantly elevated in the experimental trial (38.9°C ± 0.3; P < 0.05). Similarly, heart rates peaked at 92 beats min(-1) (±7) and 133 beats min(-1) (±4; P < 0.05), respectively. Liquid cooling reduced maximal core temperatures (37.3°C ± 0.1; P < 0.05) and heart rates 87 beats min(-1) (±3; P < 0.05) in the heat, such that neither now differed significantly from the control trial (P > 0.05). However, despite inducing profound hyperthermia and volitional fatigue, no cognitive degradation was evident in the heat (P > 0.05). Since extensive dehydration was prevented, it appears that thermal strain in the absence of dehydration may have minimal impact upon cognitive function, at least as evaluated within this experiment.

A need exists to identify dehydrated individuals under stressful settings beyond the laboratory. A predictive index based on changes in saliva osmolality has been proposed, and its efficacy and sensitivity was appraised across mass (water) losses from 1 to 7%. Twelve euhydrated males [serum osmolality: 286.1 mOsm kg(-1) H(2)O (SD 4.3)] completed three exercise- and heat-induced dehydration trials (35.6°C, 56% relative humidity): 7% dehydration (6.15 h), 3% dehydration (with 60% fluid replacement: 2.37 h), repeat 7% dehydration (5.27 h). Expectorated saliva osmolality, measured at baseline and at each 1% mass change, was used to predict instantaneous hydration state relative to mass losses of 3 and 6%. Saliva osmolality increased linearly with dehydration, although its basal osmolality and its rate of change varied among and within subjects across trials. Receiver operating characteristic curves indicated a good predictive power for saliva osmolality when used with two, single-threshold cutoffs to differentiate between hydrated and dehydrated individuals (area under curve: 3% cutoff = 0.868, 6% cutoff = 0.831). However, when analysed using a double-threshold detection technique (3 and 6%), as might be used in a field-based monitor, <50% of the osmolality data could correctly identify individuals who exceeded 3% dehydration. Indeed, within the 3-6% dehydration range, its sensitivity was 64%, while beyond 6% dehydration, this fell to 42%. Therefore, while expectorated saliva osmolality tracked mass losses within individuals, its large intra- and inter-individual variability limited its predictive power and sensitivity, rendering its utility questionable within a universal dehydration monitor.

Junin virus (JUNV) is the aetiological agent of Argentine haemorrhagic fever. The pathogenesis of the infection is not well understood, no licensed vaccines exist and no specific antiviral therapy is available. Previous studies have demonstrated the ability of ribavirin to delay and reduce JUNV disease and virus burden in guinea pigs without preventing death. Based on available data, we performed three different studies to determine the efficacy of ribavirin against JUNV in the guinea pig model with a focus on survival. Different doses and treatment schedules of ribavirin were tested in a lethal model of JUNV infection. Our results show that prolonged treatment with high doses of ribavirin significantly reduces the mortality in guinea pigs infected with JUNV. These results may be useful in future experimental studies or clinical testing., (© 2011 Blackwell Verlag GmbH.)

Effectiveness of short-term acclimation has generally been undertaken using untrained and moderately-trained participants. The purpose of this study was to determine the impact of short-term (5-day) heat acclimation on highly trained athletes. Eight males (mean ± SD age 21.8 ± 2.1 years, mass 75.2 ± 4.6 kg, VO(2peak) 4.9 ± 0.2 L min(-1) and power output 400 ± 27 W) were heat acclimated under controlled hyperthermia (rectal temperature 38.5 °C), for 90-min on five consecutive days (T(a) = 39.5 °C, 60% relative humidity). Acclimation was undertaken with dehydration (no fluid-intake) during daily bouts. Participants completed a rowing-specific, heat stress test (HST) 1 day before and after acclimation (T(a) = 35 °C, 60% relative humidity). HST consisted 10-min rowing at 30% peak power output (PPO), 10 min at 60% PPO and 5-min rest before a 2-km performance test, without feedback cues. Participants received 250 mL fluid (4% carbohydrate; osmolality 240-270 mmol kg(-1)) before the HST. Body mass loss during acclimation bouts was 1.6 ± 0.3 kg (2.1%) on day 1 and 2.3 ± 0.4 kg (3.0%) on day 5. In contrast, resting plasma volume increased by 4.5 ± 4.5% from day 1 to 5 (estimated from [Hb] & Hct). Plasma aldosterone increased at rest (52.6 pg mL(-1); p = 0.03) and end-exercise (162.4 pg mL(-1); p = 0.00) from day 1 to 5 acclimation. During the HST T(re) and f(c) were lowered 0.3 °C (p = 0.00) and 14 b min(-1) (p = 0.00) after 20-min exercise. The 2-km performance time (6.52.7 min) improved by 4 s (p = 0.00). Meaningful physiological and performance improvements occurred for highly trained athletes using a short-term (5-day) heat acclimation under hyperthermia control, with dehydration.

Purpose: PARP inhibitors are being developed as therapeutic agents for cancer. More than six compounds have entered clinical trials. The majority of these compounds are β-nicotinamide adenine dinucleotide (NAD(+))-competitive inhibitors. One exception is iniparib, which has been proposed to be a noncompetitive PARP inhibitor. In this study, we compare the biologic activities of two different structural classes of NAD(+)-competitive compounds with iniparib and its C-nitroso metabolite., Experimental Design: Two chemical series of NAD(+)-competitive PARP inhibitors, iniparib and its C-nitroso metabolite, were analyzed in enzymatic and cellular assays. Viability assays were carried out in MDA-MB-436 (BRCA1-deficient) and DLD1(-/-) (BRCA2-deficient) cells together with BRCA-proficient MDA-MB-231 and DLD1(+/+) cells. Capan-1 and B16F10 xenograft models were used to compare iniparib and veliparib in vivo. Mass spectrometry and the (3)H-labeling method were used to monitor the covalent modification of proteins., Results: All NAD(+)-competitive inhibitors show robust activity in a PARP cellular assay, strongly potentiate the activity of temozolomide, and elicit robust cell killing in BRCA-deficient tumor cells in vitro and in vivo. Cell killing was associated with an induction of DNA damage. In contrast, neither iniparib nor its C-nitroso metabolite inhibited PARP enzymatic or cellular activity, potentiated temozolomide, or showed activity in a BRCA-deficient setting. We find that the nitroso metabolite of iniparib forms adducts with many cysteine-containing proteins. Furthermore, both iniparib and its nitroso metabolite form protein adducts nonspecifically in tumor cells., Conclusions: Iniparib nonselectively modifies cysteine-containing proteins in tumor cells, and the primary mechanism of action for iniparib is likely not via inhibition of PARP activity., (©2011 AACR.)

A rethinking of current heat-acclimation strategies is required as most research and advice for improving physiological strain in the heat includes maintaining hydration using long-term acclimation protocols (>10 days). Furthermore, these strategies have tended to use untrained and moderately trained participants. Therefore, the aims of this review were to (i) investigate the effectiveness of short-term heat acclimation (STHA) with moderately and highly trained athletes; (ii) determine the importance of fluid regulatory strain, which has a thermally independent role in heat adaptation; (iii) assess the impact of STHA on a marker of thermotolerance (inducible heat-shock protein 70 [HSP70]); and (iv) provide further information on the decay of acclimation to heat. The review suggests that 5-day STHA is effective, and adaptations may be more pronounced after fluid regulatory strain from a dehydration-acclimation regimen. Furthermore, highly trained athletes may have similar physiological gains to those who are less trained using STHA. However, research has tended to focus on untrained or moderately trained participants and more information is required for highly trained populations. HSP70 response is upregulated across STHA. This indicates increased thermotolerance and protective adaptive change that may indicate HSP70 response as a useful marker of heat acclimation. Physiological adaptations after heat acclimation are relatively short term and may vanish only a few days or weeks after removal from heat exposure. From a practical perspective 5-day STHA may be the preferred acclimation regimen for moderately and highly trained athletes as it has been shown to be effective, less expensive and less likely to disrupt the tapering for competition in elite performers. Furthermore, updated information on the time course of acclimation decay may allow a reliable estimate of how long individuals can be free from heat exposure before reacclimation is required. This is particularly pertinent in present times as many athletes, civilians and military personnel increasingly have to relocate to different climates of the world, often within a short period of time.

Introduction: The aim of this study was to evaluate how the textile composition of torso undergarment fabrics may impact upon thermal strain, moisture transfer, and the thermal and clothing comfort of fully clothed, armored individuals working in a hot-dry environment (41.2 degrees C and 29.8% relative humidity)., Methods: Five undergarment configurations were assessed using eight men who walked for 120 min (4 km x h(-1)), then alternated running (2 min at 10 km x h(-1)) and walking (2 min at 4 km x h(-1)) for 20 min. Trials differed only in the torso undergarments worn: no t-shirt (Ensemble A); 100% cotton t-shirt (Ensemble B); 100% woolen t-shirt (Ensemble C); synthetic t-shirt (Ensemble D: nylon, polyethylene, elastane); hybrid shirt (Ensemble E)., Results: Thermal and cardiovascular strain progressively increased throughout each trial, with the average terminal core temperature being 38.5 degrees C and heart rate peaking at 170 bpm across all trials. However, no significant between-trial separations were evident for core or mean skin temperatures, or for heart rate, sweat production, evaporation, the within-ensemble water vapor pressures, or for thermal or clothing discomfort., Conclusion: Thus, under these conditions, neither the t-shirt textile compositions, nor the presence or absence of an undergarment, offered any significant thermal, central cardiac, or comfort advantages. Furthermore, there was no evidence that any of these fabrics created a significantly drier microclimate next to the skin.

We previously showed that the cell-cell junction protein plakoglobin (PG) not only suppresses motility of keratinocytes in contact with each other, but also, unexpectedly, of single cells. Here we show that PG deficiency results in extracellular matrix (ECM)-dependent disruption of mature focal adhesions and cortical actin organization. Plating PG⁻/⁻ cells onto ECM deposited by PG+/⁻ cells partially restored normal cell morphology and inhibited PG⁻/⁻ cell motility. In over 70 adhesion molecules whose expression we previously showed to be altered in PG⁻/⁻ cells, a substantial decrease in fibronectin (FN) in PG⁻/⁻ cells stood out. Re-introduction of PG into PG⁻/⁻ cells restored FN expression, and keratinocyte motility was reversed by plating PG⁻/⁻ cells onto FN. Somewhat surprisingly, based on previously reported roles for PG in regulating gene transcription, PG-null cells exhibited an increase, not a decrease, in FN promoter activity. Instead, PG was required for maintenance of FN mRNA stability. PG⁻/⁻ cells exhibited an increase in activated Src, one of the kinases controlled by FN, a phenotype reversed by plating PG⁻/⁻ cells on ECM deposited by PG+/⁻ keratinocytes. PG⁻/⁻ cells also exhibited Src-independent activation of the small GTPases Rac1 and RhoA. Both Src and RhoA inhibition attenuated PG⁻/⁻ keratinocyte motility. We propose a novel role for PG in regulating cell motility through distinct ECM-Src and RhoGTPase-dependent pathways, influenced in part by PG-dependent regulation of FN mRNA stability.

The ability of the major systemic fungal pathogen of humans, Candida albicans, to sense and respond to reactive oxygen species (ROS), such as H(2)O(2) generated by the host immune system, is required for survival in the host. However, the intracellular signaling mechanisms underlying such responses are poorly understood. Here, we show that thioredoxin (Trx1), in addition to its antioxidant activity, plays a central role in coordinating the response of C. albicans to ROS by regulating multiple pathways. In particular, Trx1 function is important for H(2)O(2)-induced phosphorylation of the Hog1 stress-activated protein kinase and to reverse H(2)O(2)-induced oxidation and activation of the AP-1 like transcription factor Cap1. Furthermore, Trx1 regulates H(2)O(2)-induced hyperpolarized bud growth in a mechanism that involves activation of the Rad53 checkpoint kinase. Consistent with its key roles in responses to ROS, cells lacking Trx1 displayed significantly attenuated virulence in a murine model of C. albicans systemic infection. Collectively, our data indicate that Trx1 has a multifaceted role in H(2)O(2) signaling and promotes C. albicans survival in the host.

The ability of cells to modulate interactions with each other and the substrate is essential for epithelial tissue remodeling during processes such as wound healing and tumor progression. However, despite strides made in the field of proteomics, proteins involved in adhesion have been difficult to study. Here, we report a method for the enrichment and analysis of proteins associated with the basal surface of the cell and its underlying matrix. The enrichment involves deroofing the cells with 20 mM ammonium hydroxide and the removal of cytosolic and organellar proteins by stringent water wash. Proteomic profiling was achieved by LC-FTMS, which allowed comparison of differentially expressed or shared proteins under different cell states. First, we analyzed and compared the basal cell components of mouse keratinocytes lacking the cell-cell junction molecule plakoglobin with their control counterparts. Changes in the molecules involved in motility and invasion were detected in plakoglobin-deficient cells, including decreased detection of fibronectin, integrin beta(4), and FAT tumor suppressor. Second, we assessed the differences in basal cell components between two human oral squamous cell carcinoma lines originating from different sites in the oral cavity (CAL33 and UM-SCC-1). The data show differences between the two lines in the type and abundance of proteins specific to cell adhesion, migration, and angiogenesis. Therefore, the method described here has the potential to serve as a platform to assess proteomic changes in basal cell components including extracellular and adhesion-specific proteins involved in wound healing, cancer, and chronic and acquired adhesion-related disorders.

The purpose of this work was to investigate adaptation and decay from short-term (5-day) heat acclimation (STHA). Ten moderately trained males (mean +/- SD age 28 +/- 7 years; body mass 74.6 +/- 4.4 kg; VO2peak 4.26 +/- 0.37 l min(-1)) underwent heat acclimation (Acc) for 90-min on 5-days consecutively (T (a) = 39.5 degrees C, 60% RH), under controlled hyperthermia (rectal temperature 38.5 degrees C). Participants completed a heat stress test (HST) 1 week before acclimation (Acc), then on the 2nd and 8th day (1 week) following Acc (T (a) = 35 degrees C, 60% RH). Seven participants completed HSTs 2 and 3 weeks after Acc. HST consisted of 90-min cycling at 40% peak power output before an incremental performance test. Rectal temperature at rest (37.1 +/- 0.4 degrees C) was not lowered by Acc (95% CI -0.3 to 0.2 degrees C), after 90-min exercise (38.6 +/- 0.5 degrees C) it reduced 0.3 degrees C (-0.5 to -0.1 degrees C) and remained at this level 1 week later (-0.5 to -0.1 degrees C), but not two (0.1 degrees C -0.4 to 0.5 degrees C; n = 7) or 3 weeks. Similarly, heart rate after 90-min exercise (146 +/- 21 b min(-1)) was reduced (-13: -6 to -20 b min(-1)) and remained at this level after 1 week (-13: -6 to -20 b min(-1)) but not two (-9: 6 to -23 b min(-1); n = 7) or 3 weeks. Performance (746 s) increased 106 s: 59 to 152 s after Acc and remained higher after one (76 s: 31 to 122) but not two (15 s: -88 to 142 s; n = 7) or 3 weeks. Therefore, STHA (5-day) induced adaptations permitting increased heat loss and this persisted 1 week but not 2 weeks following Acc.

Purpose: To contrast the effects of three postcooling techniques in reducing body core temperature (Tc) in exercise-induced hyperthermic participants on the cessation of exercise., Methods: Eleven healthy active male volunteers were cooled during a 40-min period using three different methods: ice packs to the neck, axillae, and groin (ICE); water spray and fan (FAN); and 2 L of chilled (20 degrees C) intravenous saline administered during a 20-min period (IV). Rate of decrease in Tc, cardiovascular responses, and any incidence of reported adverse effects were investigated. Trials were presented in a counterbalanced order with the volunteers' body core temperature being elevated to 40.0 degrees C on three occasions via an intermittent walk-run (2 min at 6 km x h and 4 min at 10 km x h) protocol conducted within a climate-controlled chamber (34.2 +/- 0.5 degrees C and 62.3 +/- 3.1% relative humidity)., Results: Rate of Tc reduction during the first 20 min of cooling was greater for FAN compared with ICE (0.09 +/- 0.02 degrees C.min vs 0.07 +/- 0.02 degrees C.min, P < 0.05), whereas IV did not differ with the other trials (0.08 +/- 0.01 degrees C.min, P > 0.05). Three participants complained of numbness or paresthesia in their arm or hand during administration of the chilled saline, although these symptoms resolved within 5 min of ceasing the infusion., Conclusions: All three cooling techniques reduced Tc and would be suitable for first aid application in a field setting during transportation to adequate medical facilities. Chilled IV saline did not produce any contraindications, providing a suitable alternative for Tc cooling.

The reduction of core body temperature (T(C)) is vitally important in the treatment of hyperthermia; however, little is known regarding the impact of cooling treatments on the autonomic control of heart rate (HR). The aim of the present study was to examine the influence of three field-based hyperthermia treatments on the neural control of HR via heart rate variability (HRV). Following exercise-induced hyperthermia (T(C) approximately 40.0 degrees C) in a warm environment (34.2 +/- 0.5 degrees C), nine healthy, active men were treated during recovery, in a randomized order, with intravenous cold saline infusion (IV) or ice packs (ICE) or fan cooling with intermittent water spray (FAN) for 40 min. During each treatment, HR dynamics via power spectral (VLF, LF, HF), Poincare plot (SD1, SD2), approximate entropy (ApEn) and short- (alpha(1)) and long-term (alpha(2)) fractal scaling analyses were determined every 10 min. At recovery onset, HR and T(C) were similar between treatments and were significantly reduced over the 40 min recovery period. During recovery, HR and alpha(2) were significantly reduced from initial levels but were significantly greater for IV compared with ICE and FAN. In contrast, VLF, LF, HF, SD1, SD2 and ApEn increased during recovery, with all being significantly lower for IV compared with ICE and/or FAN. The present results demonstrated that IV, compared with ICE and FAN, resulted in significantly greater HR, reduced spectral and geometrical HRV, lower HR complexity and reduced long-term HR control, indicative of reduced vagal and/or increased sympathetic modulation. Specific treatments for exercise-induced hyperthermia may result in an altered sympathovagal balance that requires further examination.

Unlabelled: Amphotericin B (AmphoB) remains the preferred therapy for invasive fungal infections despite many side effects, such as nephrotoxicity and electrolyte imbalance. Our previous study suggested that high sodium (Na) intake >4 mEq/kg per day may be associated with lower nephrotoxicity in extremely premature infants treated with AmphoB. Subsequently, it became a standard of care in our unit to administer Na >4 mEq/kg per day to extremely premature infants treated with AmphoB. The purpose of this study was to evaluate the effect of high Na intake > 4 mEq/kg per day on the incidence of AmphoB-induced nephrotoxicity among extremely premature infants with birth weight <1250 gm. All extremely premature infants with birth weight <1250 gm born between 1992 and 2004 and treated with AmphoB for systemic fungal infections were included in the study. The study infants were divided into two groups: a control (CL) group (1/1992-12/1999, n = 21) consisting of extremely premature infants given a maintenance Na intake during AmphoB therapy, and a high sodium intake (High Na) group (1/2000-12/2004, n = 16) consisting of extremely premature infants given a high Na intake >4 mEq/kg per day during AmphoB therapy. Nephrotoxicity was defined as serum creatinine levels >1 mg/dl, urinary output (UOP) < 1 ml/kg per hour or a decrease in UOP of 50%, compared with the previous 2 days, and persisting for at least 2 days. Invasive fungal infection was diagnosed in 5.7% of the infants (44/763 infants). Thirty-seven infants were eligible for the study and seven were excluded. There were no differences between the two groups in gestational age, birth weight, age at fungal infection diagnosis, length of AmphoB therapy, daily fluid intake or hyponatremia. Nephrotoxicity was significantly higher in the CL group than in the High Na group (13/21 vs. 3/16; P = 0.02). In the CL group, nephrotoxicity occurred at (mean +/- SD) 1.9 +/- 3.2 days after the initiation of AmphoB treatment and lasted for 5.5 +/- 4.7 days. In this group, nephrotoxicity occurred in two of the 13 infants before the initiation of AmphoB therapy. In the High Na group, nephrotoxicity occurred before the start of AmphoB therapy in two of the three infants. In the third infant, nephrotoxicity lasted for 1 day. Mean Na intake was not different between the two groups during the 4-day period prior to AmphoB therapy. Mean Na intake during the first 10-day period of AmphoB therapy was significantly lower in the CL group (3.7 vs 6.2; P < 0.001)., Conclusion: High Na intake was associated with a reduction in the incidence of AmphoB-induced nephrotoxicity in extremely premature infants with birth weight <1250 gm. We recommend the use of a high Na intake of >4 mEq/kg per day for extremely premature infants during Amphotericin B therapy.

Background: Morbidity and mortality related to respiratory syncytial virus (RSV) in pediatric acute myeloid leukemia (AML) is not known., Procedure: We combined data from three Children's Oncology Group AML studies and determined the prevalence of RSV infection and RSV-related mortality in children treated for de novo AML., Results: We found that the prevalence of RSV infection ranged from 0% to 1% in induction and between 0.3 and 2.2% in consolidation. Four children died from RSV resulting in RSV-specific mortality of 0.2% among all children. However, the risk of RSV-related mortality among RSV infection episodes was high (4/40, 10%)., Conclusions: RSV infections and deaths are rare in pediatric AML.

A combination of experiment and density functional theory was used to investigate the energetics of CO adsorption onto several small M(x)S(y)(+) (M = Mo, W; x/y = 2/6, 3/7, 5/7, 6/8) clusters as a probe of their atomic and electronic structure. Experimentally, tandem mass spectrometry was used to measure the relative yields of M(x)S(y)(+)(CO)(n) cluster adducts formed by collisions between a beam of mass-selected M(x)S(y)(+) cluster ions and CO molecules in a high-pressure collision cell (hexapole ion guide). The most probable M(x)S(y)(+)(CO)(n) adducts observed are those with n < or = x, that is, only one CO molecule bound to each metal site. The notable exception is the M(5)S(7)(+) cluster, for which the n = 6 adduct is found to have nearly the same intensity as the n = x = 5 adduct. Density functional calculations were used to search for the lowest energy structures of the bare M(x)S(y)(+) clusters and to obtain their relative stability for sequential CO binding. The calculated trends in CO binding energies were then compared to the experimental adduct distributions for assigning the ground-state structures. In this way, it was possible to distinguish between two nearly isoenergetic ground-state isomers for the M(2)S(6)(+) and M(3)S(7)(+) clusters, as only one isomer gave a calculated CO stabilization energy trend that was consistent with the experimental data. Similar comparisons of predicted and observed CO adsorption trends also provide evidence for assigning the ground-state structures of the M(5)S(7)(+) and M(6)S(8)(+) clusters. The latter contain metallic cores with most of the sulfur atoms bonded along the edges or in the faces of the metal core structure. The n = 6 and 7 adducts of M(5)S(7)(+) are predicted to be more stable than the n = x = 5 adduct, but only the n = 6 adduct is observed experimentally. The DFT calculations show that the n = 7 adduct undergoes internal bond breaking whereas the n = 6 framework is stable, albeit highly distorted. For the M(6)S(8)(+) cluster, the calculations predict that the two lowest energy isomers can bind more than six CO molecules without fragmentation; however, the apparent binding energy drops significantly for adducts with n > 6. In general, the ability of these small M(x)S(y)(+) clusters to bind more CO molecules than the number of metal atoms is a balance between the gain in CO adsorption energy versus the strain introduced into the cluster structure caused by CO crowding, the consequences of which can be fragmentation of the M(x)S(y)(+)(CO)(n) cluster adduct (n > x).

The Hog1 mitogen-activated protein kinase (MAPK) plays a central role in stress responses in the human pathogen Candida albicans. Here, we have investigated the MAPK kinase kinase (MAPKKK)-dependent regulation of the pathway. In contrast to the Hog1 pathway in Saccharomyces cerevisiae, which is regulated by three MAPKKKs (Ssk2, Ssk22, and Ste11), our results demonstrate that Hog1 in C. albicans is regulated by a single MAPKKK Ssk2. Deletion of SSK2 results in comparable stress and morphological phenotypes exhibited by hog1Delta cells, and Ssk2 is required for the stress-induced phosphorylation and nuclear accumulation of Hog1, and for Hog1-dependent gene expression. Furthermore, phenotypes associated with deletion of SSK2 can be circumvented by expression of a phosphomimetic mutant of the MAPKK Pbs2, indicating that Ssk2 regulates Hog1 via activation of Pbs2. In S. cerevisiae, the Hog1 pathway is also regulated by the MAPKKK Ste11. However, we can find no connection between Ste11 and the regulation of Hog1 in C. albicans. Furthermore, expression of a chimeric Pbs2 protein containing the Ste11-dependent regulatory region of S. cerevisiae Pbs2, fails to stimulate Ste11-dependent stress signaling in C. albicans. Collectively, our data show that Ssk2 is the sole MAPKKK to relay stress signals to Hog1 in C. albicans and that the MAPK signaling network in C. albicans has diverged significantly from the corresponding network in S. cerevisiae.

The reactivity of the Ti(8)C(12)(+) met-car cation toward thiophene was investigated using density functional theory (DFT) and mass selective ion chemistry. It is shown that the experimentally observed mass spectrum can be well described by the DFT calculations. In contrast to the weak bonding interactions seen for thiophene on a TiC(001) surface, the Ti(8)C(12)(+) met-car cation is able to interact strongly with up to four thiophene molecules with the cluster staying intact. In the most stable conformation, the thiophene molecules bond to the four low-coordinated Ti(0) sites of Ti(8)C(12)(+) via a eta(5)-C,S coordination. The stability and the activity of the Ti(8)C(12)(+) met-car is observed to increase with an increasing number of attached thiophene molecules at the Ti(0) sites, which is associated with a significant transfer of electron density from thiophene to the cluster. The additional electron density on the Ti(8)C(12)(+) cation cluster, however, is not sufficient to cleave the C-S bonds of thiophene and the dissociation reaction of thiophene is predicted to be a highly activated process. By contrast, DFT calculations for the neutral Ti(8)C(12) met-car predict that the dissociation reaction leading to adsorbed S and C(4)H(4) fragments is energetically favorable for the first thiophene molecule. The binding behavior for subsequent addition of thiophene molecules to the neutral met-car is also presented and compared to that of the cation.

Gas-phase Ti(x)C(y)+ clusters (x/y = 3/5, 4/7, 5/9, 6/9, 7/12, 8/12, 9/12) including the magic Ti8C12+ (met-car) have been produced by reactive sputtering with a magnetron cluster source. The gas-phase reactivity of the met-car with SCO, CS2, and SO2 was investigated in a hexapole collision cell by way of tandem mass spectrometry. Results indicate an increase in activity as the oxygen-to-sulfur ratio increases (SO2 > SCO > CS2) with products ranging from association to break down of the met-car cluster. Trends in the mass spectra also indicate SCO and CS2 may bond to the met-car in a unique way not observed in previous reactivity studies on Ti8C12+. To investigate this, several possible single molecule-cluster bonding configurations were calculated with density functional theory. The results indicate that bridge bonding of the intact molecules is energetically preferred. In addition, the energy barriers and transition states leading to dissociation products were calculated and the trends are found to be in qualitative agreement with experiment. The effects of the different types of bonding and number of adsorbed species on the reactivity of the met-car along with proposed reaction mechanisms for product formation are also discussed.

Aim: In this study, we quantified acute changes in the intracellular and extracellular fluid compartments during upright neutral- and cold-water immersion. We hypothesized that, during short-term cold immersion, fluid shifts would be wholly restricted to the extracellular space., Methods: Seven males were immersed 30 days apart: control (33.3 degrees SD 0.6 degrees C); and cold (18.1 degrees SD 0.3 degrees C). Posture was controlled for 4 h prior to a 60-min seated immersion., Results: Significant reductions in terminal oesophageal (36.9 degrees +/- 0.1 degrees -36.3 degrees +/- 0.1 degrees C) and mean skin temperatures (30.3 degrees +/- 0.3 degrees -23.0 degrees +/- 0.3 degrees C) were observed during the cold, but not the control immersion. Both immersions elicited a reduction in intracellular fluid [20.17 +/- 6.02 mL kg(-1) (control) vs. 22.72 +/- 9.90 mL kg(-1)], while total body water (TBW) remained stable. However, significant plasma volume (PV) divergence was apparent between the trials at 60 min [12.5 +/- 1.0% (control) vs. 6.1 +/- 3.1%; P < 0.05], along with a significant haemodilution in the control state (P < 0.05). Plasma atrial natriuretic peptide concentration increased from 18.0 +/- 1.6 to 58.7 +/- 15.1 ng L(-1) (P < 0.05) during cold immersion, consistent with its role in PV regulation. We observed that, regardless of the direction of the PV change, both upright immersions elicited reductions in intracellular fluid., Conclusion: These observations have two implications. First, one cannot assume that PV changes reflect those of the entire extracellular compartment. Second, since immersion also increases interstitial fluid pressure, fluid leaving the interstitium must have been rapidly replaced by intracellular water.

Aim: To determine whether acute exercise-heat-induced dehydration affects sweat composition, eight males cycled for 2 h at 39.5 +/- 1.6% VO2peak on two separate occasions in a hot-humid environment (38.0 +/- 0.0 degrees C, 60.0 +/- 0.1% relative humidity)., Methods: During exercise, subjects ingested either no fluid (dehydration) or a 20 mmol L(-1) sodium chloride solution (euhydration). The volume of solution, calculated from whole-body sweat loss and determined in a familiarization trial, was ingested at 0 min and every 15 min thereafter. Venous blood was collected at 0, 60 and 120 min of exercise and sweat was aspirated from a patch located on the dominant forearm at 120 min., Results: Following the 2-h cycling exercise, sweat [Na+] and [Cl-] was greater (P < 0.05) in the dehydration trial (Na+ 91.1 +/- 6.8 mmol L(-1); Cl- 73.3 +/- 3.5 mmol L(-1)) compared with the euhydration trial (Na+ 81.1 +/- 5.9 mmol L(-1); Cl- 68.5 +/- 3.3 mmol L(-1)). In addition, dehydration invoked a greater serum [Na+] (142.2 +/- 0.7 mmol L(-1); P < 0.05), [Cl-] (105.8 +/- 0.6 mmol L(-1); P < 0.05) and [K+] (5.27 +/- 0.2 mmol L(-1); P < 0.05) over the euhydration values for [Na+], [Cl-] and [K+], respectively (138.9 +/- 0.6, 102.9 +/- 0.5 and 4.88 +/- 0.1 mmol L(-1)). Plasma aldosterone was also significantly higher during exercise in the dehydration trial compared with the euhydration trial (53.8 +/- 3.8 vs. 40.0 +/- 4.3 ng dL(-1); P < 0.05)., Conclusions: Acute exercise-heat stress without fluid replacement resulted in a greater sweat [Na+] and [Cl-] which was potentially related to greater extracellular fluid [Na+], plasma aldosterone or sympathetic nervous activity.

We measured intra- and extravascular body-fluid compartments in 12 resting males before (day 1; control), during (day 8) and after (day 22) a 3-week, exercise-heat acclimation protocol to investigate plasma volume (PV) changes. Our specific focus was upon the selective nature of the acclimation-induced PV expansion, and the possibility that this expansion could be sustained during prolonged acclimation. Acclimation was induced by cycling in the heat, and involved 16 treatment days (controlled hyperthermia (90 min); core temperature = 38.5 degrees C) and three experimental exposures (40 min rest, 96.9 min (s.d. 9.5 min) cycling), each preceded by a rest day. The environmental conditions were a temperature of 39.8 degrees C (s.d. 0.5 degrees C) and relative humidity of 59.2% (s.d. 0.8%). On days 8 and 22, PV was expanded and maintained relative to control values (day 1: 44.0 +/- 1.8; day 8: 48.8 +/- 1.7; day 22: 48.8 +/- 2.0 ml kg(-1); P < 0.05). The extracellular fluid compartment (ECF) was equivalently expanded from control values on days 8 (279.6 +/- 14.2 versus 318.6 +/- 14.3 ml kg(-1); n= 8; P < 0.05) and 22 (287.5 +/- 10.6 versus 308.4 +/- 14.8 ml kg(-1); n= 12; P < 0.05). Plasma electrolyte, total protein and albumin concentrations were unaltered following heat acclimation (P > 0.05), although the total plasma content of these constituents was elevated (P < 0.05). The PV and interstitial fluid (ISF) compartments exhibited similar relative expansions on days 8 (15.0 +/- 2.2%versus 14.7 +/- 4.1%; P > 0.05) and 22 (14.4 +/- 3.6% versus 6.4 +/- 2.2%; P= 0.10). It is concluded that the acclimation-induced PV expansion can be maintained following prolonged heat acclimation. In addition, this PV expansion was not selective, but represented a ubiquitous expansion of the extracellular compartment.

We investigated the impact of cold-water acclimation on whole-body fluid regulation using tracer-dilution methods to differentiate between the intracellular and extracellular fluid compartments. Seven euhydrated males [age 24.7 (8.7) years, mass 74.4 (6.4) kg, height 176.8 (7.8) cm, sum of eight skinfolds 107.4 (20.4) mm; mean (SD)] participated in a 14-day cold-water acclimation protocol, with 60-min resting cold-water stress tests [CWST; 18.1 (0.1) degrees C] on days 1, 8 and 15, and 90-min resting cold-water immersions [18.4 (0.4) degrees C] on intervening days. Subjects were immersed to the 4th intercostal space. Intracellular and extracellular fluid compartments, and plasma protein, electrolyte and hormone concentrations were investigated. During the first CWST, the intracellular fluid (5.5%) and plasma volumes were reduced (6.1%), while the interstitial fluid volume was simultaneously expanded (5.4%). This pattern was replicated on days 8 and 15, but did not differ significantly among test days. Acclimation did not produce significant changes in the pre-immersion distribution of total body water, or changes in plasma osmolality, total protein, electrolyte, atrial natriuretic peptide or aldosterone concentrations. Furthermore, a 14-day cold-water acclimation regimen did not elicit significant changes in body-fluid distribution, urine production, or the concentrations of plasma protein, electrolytes or the fluid-regulatory hormones. While acclimation trends were not evident, we have confirmed that fluid from extravascular cells is displaced into the interstitium during acute cold-water immersion, both before and after cold acclimation.

Seven active male subjects cycled for 60 min at 29.5 (0.8)% peak work rate on three separate occasions in a hot environmental condition [36.0 (0.1) degrees C, 60 (1)% relative humidity] in order to determine the reproducibility of a closed-pouch sweat collection technique for sweat composition at the scapula, forearm and thigh. To confirm that sweat composition was not influenced by between-trial variations in sudomotor drive, local sweat rate, whole-body sweat rate, heart rate (HR), rectal temperature (T(re)) and mean skin temperature (T(sk)) responses were also measured, consequently reproducibility was also established for these variables. Sweat composition did not differ among trials, with the mean coefficients of variation (CVs) for sweat [Na(+)], [K(+)] and pH being 10.4 (7.4)%, 8.1 (6.5)% and 1.3 (1.1)%, respectively. Local sweat rates did not differ among the three trials (P>0.05) although whole-body sweat rate was reduced in the third trial (P<0.05). The mean CVs were 11.0 (7.8)% and 4.7 (1.6)% for local and whole-body sweat rates, respectively. Between-trial differences were not evident for T(re), T(sk) or HR with mean CVs of 0.3 (0.2)%, 0.7 (0.6)% and 3.9 (1.7)%, respectively, although HR tended to be greater in the first trial ( P=0.08). It is proposed that moderate variations in sweat composition were influenced by variations in the local sweat rate, which were induced by application of the pouch.

We tested the hypothesis that local sweat rates would not display a systematic postadaptation redistribution toward the limbs after humid heat acclimation. Eleven nonadapted males were acclimated over 3 wk (16 exposures), cycling 90 min/day, 6 days/wk (40 degrees C, 60% relative humidity), using the controlled-hyperthermia acclimation technique, in which work rate was modified to achieve and maintain a target core temperature (38.5 degrees C). Local sudomotor adaptation (forehead, chest, scapula, forearm, thigh) and onset thresholds were studied during constant work intensity heat stress tests (39.8 degrees C, 59.2% relative humidity) conducted on days 1, 8, and 22 of acclimation. The mean body temperature (Tb) at which sweating commenced (threshold) was reduced on days 8 and 22 (P < 0.05), and these displacements paralleled the resting thermoneutral Tb shift, such that the Tb change to elicit sweating remained constant from days 1 to 22. Whole body sweat rate increased significantly from 0.87 +/- 0.06 l/h on day 1 to 1.09 +/- 0.08 and 1.16 +/- 0.11 l/h on days 8 and 22, respectively. However, not all skin regions exhibited equivalent relative sweat rate elevations from day 1 to day 22. The relative increase in forearm sweat rate (117 +/- 31%) exceeded that at the forehead (47 +/- 18%; P < 0.05) and thigh (42 +/- 16%; P < 0.05), while the chest sweat rate elevation (106 +/- 29%) also exceeded the thigh (P < 0.05). Two unique postacclimation observations arose from this project. First, reduced sweat thresholds appeared to be primarily related to a lower resting Tb, and more dependent on Tb change. Second, our data did not support the hypothesis of a generalized and preferential trunk-to-limb sweat redistribution after heat acclimation.

The colonic epithelium undergoes a continuous cycle of proliferation, differentiation, and apoptosis. To characterize factors important for colonic homeostasis and its dysregulation, human fetal colonic epithelial cells were isolated and seeded on a collagen type I matrix with embedded colonic fibroblasts. The epithelial cells rapidly spread from clusters and proliferated, and within 3 days, a columnar layer of polarized epithelium surrounded the surface of the constricted collagen matrix. The polarized enterocytes developed brush borders, tight junctions and desmosomes, and goblet and enteroendocrine cells were present. A balance of growth and differentiation was maintained for several weeks in the presence of collagen-embedded fibroblasts and a complex mixture of growth factors. Leukemia inhibitory factor (LIF) was critical for proliferation of enterocytes and inhibited expression of the differentiation marker carbonic anhydrase II. In the presence of LIF, the relative number of goblet cells remained stable, whereas enteroendocrine relative cell number declined. LIF-stimulated epithelial cells remained dependent on the presence of fibroblasts in the matrix. In combination with stem cell factor and endothelin 3, LIF induced formation of disorganized structures of stratified and semi-stratified cells, suggesting that the homeostatic balance in the normal human colon requires cooperation with differentiation-inducing factors.

Participants generated lists of exemplars from the categories of animals, tools, and fruit, and their lists were used to determine the relative accessibility of individual exemplars. Measures of accessibility included output dominance (the number of participants who listed an exemplar), rank (how early instances were listed), and two scores that reflect their combination-output precedence and dominance/rank. Other participants drew and described novel exemplars of those categories that might exist on an imaginary planet and reported on the factors that influenced their creations. References to Earth animals, tools, or fruit were used to determine imagination frequency (the number of participants who mentioned relying on particular Earth exemplars). Items high in accessibility were also high in imagination frequency, implying that those items that come to mind most readily are the ones most likely to serve as starting points for the development of novel ideas. This result held even when task constraints weighed against the use of such items (Experiment 2) and when participants were encouraged to be as creative as possible (Experiment 4), suggesting that it is difficult to avoid the influence of highly accessible category exemplars. Other measures of category structure, including the rated typicality, familiarity, and frequency of exemplars, did not predict imagination frequency as well. The results are discussed in terms of expanding concept boundaries and the inadvertent application of knowledge that is readily accessible.