Epithelial damage and tissue γδ T cells promote a unique tumor-protective IgE response.

IgE is an ancient and conserved immunoglobulin isotype with potent immunological function. Nevertheless, the regulation of IgE responses remains an enigma, and evidence of a role for IgE in host defense is limited. Here we report that topical exposure to a common environmental DNA-damaging xenobiotic initiated stress surveillance by γδTCR+ intraepithelial lymphocytes that resulted in class switching to IgE in B cells and the accumulation of autoreactive IgE. High-throughput antibody sequencing revealed that γδ T cells shaped the IgE repertoire by supporting specific variable-diversity-joining (VDJ) rearrangements with unique characteristics of the complementarity-determining region CDRH3. This endogenous IgE response, via the IgE receptor FcεRI, provided protection against epithelial carcinogenesis, and expression of the gene encoding FcεRI in human squamous-cell carcinoma correlated with good disease prognosis. These data indicate a joint role for immunosurveillance by T cells and by B cells in epithelial tissues and suggest that IgE is part of the host defense against epithelial damage and tumor development.

Peptide nucleic acid-dependent artifact can lead to false-positive triplex gene editing signals.

Triplex gene editing relies on binding a stable peptide nucleic acid (PNA) sequence to a chromosomal target, which alters the helical structure of DNA to stimulate site-specific recombination with a single-strand DNA (ssDNA) donor template and elicits gene correction. Here, we assessed whether the codelivery of PNA and donor template encapsulated in Poly Lactic-co-Glycolic Acid (PLGA)-based nanoparticles can correct sickle cell disease and x-linked severe combined immunodeficiency. However, through this process we have identified a false-positive PCR artifact due to the intrinsic capability of PNAs to aggregate with ssDNA donor templates. Here, we show that the combination of PNA and donor templates but not either agent alone results in different degrees of aggregation that result in varying but highly reproducible levels of false-positive signal. We have identified this phenomenon in vitro and confirmed that the PNA sequences producing the highest supposed correction in vitro are not active in vivo in both disease models, which highlights the importance of interrogating and eliminating carryover of ssDNA donor templates in assessing various gene editing technologies such as PNA-mediated gene editing.

A numerical study on microfluidic devices to maintain the concentration and purity of dielectrophoresis-induced separated fractions of analyte.

Determining the physical and chemical properties of biologically important particles such as cells, organelles, viruses, exosomes, complexes, nucleotides, and proteins is needed to understand their function. These properties are determined with common analytical tools (mass spectrometry, cryo-EM, NMR, various spectroscopies, nucleotide sequencing, etc.) whose function can be improved when samples are pure and concentrated. Separations science plays a central role in conditioning samples, ranging from low-resolution benchtop operations like precipitations or extractions to higher-resolution chromatography and electrophoresis. In the last two decades, gradient insulator-based dielectrophoresis (g-iDEP) has emerged as a high-resolution separation technique capable of highly selective enrichment of cells, viruses, exosomes, and proteins. Specific evidence has been shown that pure homogeneous and concentrated fractions of cells and exosomes can be generated from complex mixtures. However, recovering those fractions for analysis has not been developed, limiting the technique to an analytical rather than a preparative one. Here, a finite element analysis was undertaken to identify geometries and operational parameters to efficiently remove the enriched fraction while retaining maximum concentration and providing total mass transfer. Geometric factors (e.g., side channel width and distance from the gradient-inducing gap) were studied, along with the addition of a second inlet side channel. Two flow-generating mechanisms-electroosmosis and hydrostatic pressure-were evaluated for semi-optimized device designs, including a comparison of the one- and two-inlet designs. Simulations indicate effectively one hundred percent mass transfer and a concentration increase by an order of magnitude for several device configurations and operational parameters.

Interfacing microfluidics with information-rich detection systems for cells, bioparticles, and molecules.

The development of elegant and numerous microfluidic manipulations has enabled significant advances in the processing of small volume samples and the detection of minute amounts of biomaterials. Effective isolation of single cells in a defined volume as well as manipulations of complex bioparticle or biomolecule mixtures allows for the utilization of information-rich detection methods including mass spectrometry, electron microscopy imaging, and amplification/sequencing. The art and science of translating biosamples from microfluidic platforms to highly advanced, information-rich detection system is the focus of this review, where we term the translation between the microfluidics elements to the external world "off-chipping." When presented with the challenge of presenting sub-nanoliter volumes of manipulated sample to a detection scheme, several delivery techniques have been developed for effective analysis. These techniques include spraying (electrospray, nano-electrospray, pneumatic), meniscus-defined volumes (droplets, plugs), constrained volumes (narrow channels, containers), and phase changes (deposition, freezing). Each technique has been proven effective in delivering highly defined samples from microfluidic systems to the detection elements. This review organizes and presents selective publications that illustrate the advancements of these delivery techniques with respect to the type of sample analyzed, while introducing each strategy and providing historical perspective. The publications highlighted in this review were chosen due to their significance and relevance in the development of their respective off-chip technique. This review highlights advancements of delivery methods off a microfluidic chip for additional information rich detection schemes.

Orders-of-Magnitude Larger Force Demonstrated for Dielectrophoresis of Proteins Enabling High-Resolution Separations Based on New Mechanisms.

Proteins are perhaps the most important yet frustratingly complicated and difficult class of compounds to analyze, manipulate, and use. One very attractive option to characterize and differentially concentrate proteins is dielectrophoresis, but according to accepted theory, the force on smaller particles the size of proteins is too low to overcome diffusive action. Here, three model proteins, immunoglobulin G, α-chymotrypsinogen A, and lysozyme, are shown to generate forces much larger than predicted by established theory are more consistent with new theoretical constructs, which include the dipole moment and interfacial polarizability. The forces exerted on the proteins are quantitatively measured against well-established electrophoretic and diffusive processes and differ for each. These forces are orders of magnitude larger than previously predicted and enable the selective isolation and concentration of proteins consistent with an extremely high-resolution separation and concentration system based on the higher-order electric properties. The separations occur over a small footprint, happen quickly, and can be made in series or parallel (and in any order) on simple devices.

Extreme occupational heat exposure is associated with elevated haematological and inflammatory markers in Fire Service Instructors.

NEW FINDINGS: What is the central question of this study? Fire service instructors are frequently exposed to live fire scenarios, representing the most extreme chronic occupational heat exposure. These individuals report a series of unique health issues. We sought to identify whether the number of exposures completed was associated with inflammatory and immunological markers and symptoms of ill health. What is the main finding and its importance? Fire service instructors exhibit greater levels of inflammatory markers in comparison to firefighters. The number of exposures to fire is positively related to the prevalence of ill health and inflammation. Implementation of a proposed limit of nine exposures per month might be appropriate to minimize health issues. ABSTRACT: Fire Service Instructors (FSIs) experience ∼10 times more fire exposures than firefighters (FFs), and the increased physiological stress from this potentially puts them at risk of ill health and future cardiac events. The aim of the study was to establish whether FSIs exhibit elevated biomarkers associated with cardiac event risk, identify whether FSIs experience systemic inflammation linked to the frequency of fire exposure and evaluate a proposed exposure limit of nine exposures per month. Blood samples were collected from 110 Fire Service personnel (mean ± SD, age,44 ± 7 years; height, 178.1 ± 7.1 cm; and body mass, 84.3 ± 12.0 kg; FSIs n = 53 and FFs n = 57) for biomarker analysis. Work history details were collected from all participants. Participants with biomarker concentrations above healthy reference ranges were classified as being 'at risk'. The neutrophil-to-lymphocyte ratio, platelet count, cardiac troponin T, interleukin (IL)-6, IL-1ß, C-reactive protein and immunoglobulin G were greater in FSIs than in FFs (P < 0.05). Multiple regression analysis revealed that 18.8% of IL-6, 24.9% of IL-1ß, 29.2% of C-reactive protein and 10.9% of immunoglobulin G variance could be explained by the number of exposures to heat per month. Odds ratios revealed that those FSIs above the nine per month exposure limit were six to 12 times more likely to be classified as 'at risk' and were 16 times more likely to experience symptoms of ill health. Increased cytokine levels suggest that FSIs experience systemic inflammation, which is related to symptoms of ill health. We propose that an exposure limit could reduce the prevalence of these biomarker risk factors and ill health.

Biophysical differentiation of susceptibility and chemical differences in Staphylococcus aureus.

Differentiating bacteria strains using biophysical forces has been the focus of recent studies using dielectrophoresis (DEP). The refinement of these studies has created high-resolution separations such that very subtle properties of the cells are enough to induce significant differences in measurable biophysical properties. These high-resolution capabilities build upon the advantages of DEP which include small sample sizes and fast analysis times. Studies focusing on differentiating antimicrobial resistant and susceptible bacteria potentially have significant impact on human health and medical care. A prime example is Staphylococcus aureus, which commonly colonizes adults without ill effects. However, the pathogen is an important cause of infections, including surgical site infections. Treatment of S. aureus infections is generally possible with antimicrobials, but antimicrobial resistance has emerged. Of special importance is resistance to methicillin, an antimicrobial created in response to resistance to penicillin. Here, dielectrophoresis is used to study methicillin-resistant (MRSA) and -susceptible S. aureus (MSSA) strains, both with and without the addition of a fluorescent label. The capture onset potential of fluorescently-labeled MRSA (865 ± 71 V) and thus the ratio of electrokinetic to dielectrophoretic mobility, was found to be higher than that of fluorescently-labeled MSSA (685 ± 61 V). This may be attributable to the PBP2a enzyme present in the MRSA strain and not in the MSSA bacteria. Further, unlabeled MRSA was found to have a capture onset potential of 732 ± 44 V, while unlabeled MSSA was found to have a capture onset potential of 562 ± 59 V. This shows that the fluorescently-labeled bacteria require a higher applied potential, and thus ratio of mobilities, to capture than the unlabeled bacteria.

Dielectrophoresis of proteins: experimental data and evolving theory.

The ability to selectively move and trap proteins is core to their effective use as building blocks and for their characterization. Analytical and preparative strategies for proteins have been pursued and modeled for nearly a hundred years, with great advances and success. Core to all of these studies is the separation, isolation, purification, and concentration of pure homogeneous fractions of a specific protein in solution. Processes to accomplish this useful solution include biphasic equilibrium (chromatographies, extractions), mechanical, bulk property, chemical equilibria, and molecular recognition. Ultimately, the goal of all of these is to physically remove all non-like protein molecules-to the finest detail: all atoms in the full three-dimensional structure being identical down the chemical bond and bulk structure chirality. One strategy which has not been effectively pursued is exploiting the higher order subtle electrical properties of the protein-solvent system. The advent of microfluidic systems has enabled the use of very high electric fields and well-defined gradients such that extremely high resolution separations of protein mixtures are possible. These advances and recognition of these capabilities have caused a re-evaluation of the underlying theoretical models and they were found to be inadequate. New theoretical descriptions are being considered which align more closely to the total forces present and the subtlety of differences between similar proteins. These are focused on the interfacial area between the protein and hydrating solvent molecules, as opposed to the macroscale assumptions of homogeneous solutions and particles. This critical review examines all data which has been published that place proteins in electric field gradients which induce collection of those proteins, demonstrating a force greater than dispersive effects or countering forces. Evolving theoretical constructs are presented and discussed, and a general estimate of future capabilities using the higher order effects and the high fields and precise gradients of microfluidic systems is discussed. Graphical abstract.

Electrochemically Modulated Liquid Chromatography in Fused Silica Capillary Columns.

Electrochemically modulated liquid chromatography (EMLC) uses electrical potentials, applied to a conductive chromatographic stationary phase (e.g., porous graphitic carbon [PGC]), to manipulate analyte retention. This paper reports the design of a capillary EMLC column with a smaller internal diameter (ID; 250 µm) than that of the standard bore predecessor (3.3 mm ID). The new capillary EMLC columns are configured so that the PGC stationary phase serves as the working electrode in a two-electrode electrochemical cell and simplifies electrode placement by obviating the need for a counter electrode. This configuration also eliminates the internal Nafion sleeve that is critical to operation for the standard bore columns, thereby avoiding Nafion deformation as a source of chromatographic band broadening and rupturing as a mode of column failure. Indeed, values for chromatographic efficiency obtained on the capillary columns meet or exceed those measured for the standard columns (20 000-40 000 vs 14 000 plates/m, respectively) with near symmetric elution bands (asymmetry factors of 1.1-1.4 for well-packed capillaries) that surpass band symmetries observed in all prior studies. A test suite of aromatic sulfonates was used to characterize the chromatographic performance of the capillary EMLC columns. Separations of this test mixture showed that retention factors for individual analytes could be manipulated by as much as 21× by changing the applied potential at the PGC stationary phase. Changes in retention behavior at different potential ranges, hypothesized to result from differences in adsorption orientation, were also observed and are consistent with past work. Collectively, the retention behavior unique to EMLC is operative in this new capillary configuration and promises to open new avenues in tuning LC separations.

Simulation and experiment of asymmetric electrode placement for electrophoretic exclusion in a microdevice.

Electrophoretic exclusion (EE) is a counterflow gradient technique that exploits hydrodynamic flow and electrophoretic forces to exclude, enrich, and separate analytes. Resolution for this technique has been theoretically examined and the smallest difference in electrophoretic mobilities that can be completely separated is estimated to be 10-13  cm2 /Vs. Traditional and mesoscale systems have been used, whereas microfluidics offers a greater range of geometries and configurations towards approaching this theoretical limit. To begin to understand the impact of seemingly subtle changes to the entrance flow and the electric field configurations, three closely related microfluidic interfaces were modeled, fabricated, and tested. These interfaces consisted of systematically varying placement of an asymmetric electrode relative to a channel entrance: leading electrode placed outside the channel entrance, leading electrode aligned with the channel, and leading electrode placed within the channel. A charged fluorescent dye is used as a sensitive and accurate probe for the model and to test the concentration variation at these interfaces. Models and experiments focused on visualizing the concentration profile in areas of high temporal dynamics, thus providing a severe test of the models. Experimental data and simulation results showed strong qualitative agreement. The complexity of the electric and flow fields about this interface and the agreement between models and testing suggests the theoretical assessment capabilities can be used to faithfully design novel and more efficient interfaces.

Creatine supplementation improves performance above critical power but does not influence the magnitude of neuromuscular fatigue at task failure.

NEW FINDINGS: What is the central question of this study? Does the magnitude of neuromuscular fatigue depend on the amount of work done (W') at task failure when cycling above critical power (CP)? What is the main finding and its importance? Creatine supplementation increases W' and enhances supra-CP performance, but induces similar magnitudes of neuromuscular fatigue at task failure compared to placebo. Increased W' does not lead to higher levels of neuromuscular fatigue. This supports the notion of a critical level of neuromuscular fatigue at task failure and challenges a direct causative link between W' depletion and neuromuscular fatigue. ABSTRACT: The present study examined the effect of creatine supplementation on neuromuscular fatigue and exercise tolerance when cycling above critical power (CP). Eleven males performed an incremental cycling test with four to five constant-load trials to task failure (TTF) to obtain asymptote (CP) and curvature constant (W') of the power-duration relationship, followed by three constant-load supra-CP trials: (1) one TTF following placebo supplementation (PLA); (2) one TTF following creatine supplementation (CRE); and (3) one trial of equal duration to PLA following creatine supplementation (ISO). Neuromuscular assessment of the right knee extensors was performed pre- and post-exercise to measure maximal voluntary contraction (MVC), twitch forces evoked by single (Qpot ) and paired high- (PS100) and low- (PS10) frequency stimulations and voluntary activation. Creatine supplementation increased TTF in CRE vs. PLA by ∼11% (P = 0.017) and work done above CP by ∼10% (P = 0.015), with no difference (P > 0.05) in reductions in MVC (-24 ± 8% vs. -20 ± 9%), Qpot (-39 ± 13% vs. -32 ± 14%), PS10 (-42 ± 14% vs. -36 ± 13%), PS100 (-25 ± 10% vs. -18 ± 12%) and voluntary activation (-7 ± 8% vs. -5 ± 7%). No significant difference was found between ISO and either PLA or CRE (P > 0.05). These findings suggest similar levels of neuromuscular fatigue can be found following supra-CP cycling despite increases in performance time and amount of work done above CP, supporting the notion of a critical level of neuromuscular fatigue and challenging a direct causative link between W' depletion and neuromuscular fatigue.

The magnitude of neuromuscular fatigue is not intensity dependent when cycling above critical power but relates to aerobic and anaerobic capacities.

NEW FINDINGS: What is the central question of this study? Is the magnitude of neuromuscular fatigue dependent upon exercise intensity above critical power (CP) when W' (the curvature constant of the power-duration relationship) is depleted? What is the main finding and its importance? The magnitude of neuromuscular fatigue is the same after two bouts of supra-CP cycling (3 versus 12 min) when controlling for W' depletion but is larger for individuals of greater anaerobic capacity after the shorter bout and smaller for individuals of greater aerobic capacity after the longer exercise bout. These findings provide new insight into the mechanisms underpinning exercise above CP. ABSTRACT: The aim of the present study was to test whether the development of neuromuscular fatigue within the severe-intensity domain could be linked to the depletion of the curvature constant (W') of the power-duration relationship. Twelve recreationally active men completed tests to determine peak oxygen consumption, critical power (CP) and W', followed by two randomly assigned constant-load supra-CP trials set to deplete W' fully in 3 (P-3) and 12 min (P-12). Pre- to postexercise changes in maximal voluntary contraction, potentiated quadriceps twitch force evoked by single (Qpot ) and paired high- (PS100) and low-frequency (PS10) stimulations and voluntary activation were determined. Cycling above CP reduced maximal voluntary contraction (P-3, -20 ± 10% versus P-12, -15 ± 7%), measures associated with peripheral fatigue (Qpot , -35 ± 13 versus -31 ± 14%; PS10, -38 ± 13 versus -37 ± 17%; PS100, -18 ± 9 versus -13 ± 8% for P-3 and P-12, respectively) and voluntary activation (P-3, -12 ± 3% versus P-12, -13 ± 3%; P < 0.05), with no significant difference between trials (P > 0.05). Changes in maximal voluntary contraction and evoked twitch forces were inversely correlated with CP and peak oxygen consumption after P-12, whereas W' was significantly correlated with changes in Qpot and PS10 after P-3 (P < 0.05). Therefore, the magnitude of neuromuscular fatigue does not depend on exercise intensity when W' is fully exhausted during severe-intensity exercise; nonetheless, exploration of inter-individual variations suggests that mechanisms underpinning exercise tolerance within this domain differ between short- and long-duration exercise.

A smart curtailment approach for reducing bat fatalities and curtailment time at wind energy facilities.

The development and expansion of wind energy is considered a key global threat to bat populations. Bat carcasses are being found underneath wind turbines across North and South America, Eurasia, Africa, and the Austro-Pacific. However, relatively little is known about the comparative impacts of techniques designed to modify turbine operations in ways that reduce bat fatalities associated with wind energy facilities. This study tests a novel approach for reducing bat fatalities and curtailment time at a wind energy facility in the United States, then compares these results to operational mitigation techniques used at other study sites in North America and Europe. The study was conducted in Wisconsin during 2015 using a new system of tools for analyzing bat activity and wind speed data to make near real-time curtailment decisions when bats are detected in the area at control turbines (N = 10) vs. treatment turbines (N = 10). The results show that this smart curtailment approach (referred to as Turbine Integrated Mortality Reduction, TIMR) significantly reduced fatality estimates for treatment turbines relative to control turbines for pooled species data, and for each of five species observed at the study site: pooled data (-84.5%); eastern red bat (Lasiurus borealis, -82.5%); hoary bat (Lasiurus cinereus, -81.4%); silver-haired bat (Lasionycteris noctivagans, -90.9%); big brown bat (Eptesicus fuscus, -74.2%); and little brown bat (Myotis lucifugus, -91.4%). The approach reduced power generation and estimated annual revenue at the wind energy facility by ≤ 3.2% for treatment turbines relative to control turbines, and we estimate that the approach would have reduced curtailment time by 48% relative to turbines operated under a standard curtailment rule used in North America. This approach significantly reduced fatalities associated with all species evaluated, each of which has broad distributions in North America and different ecological affinities, several of which represent species most affected by wind development in North America. While we recognize that this approach needs to be validated in other areas experiencing rapid wind energy development, we anticipate that this approach has the potential to significantly reduce bat fatalities in other ecoregions and with other bat species assemblages in North America and beyond.

Effects of surface treatments on trapping with DC insulator-based dielectrophoresis.

A central challenge in measuring the biophysical properties of cells with electrokinetic approaches is the assignment of these biophysical properties to specific biological characteristics. Changes in the electrokinetic behavior of cells may come from mutations, altered gene expression levels, post-translation modifications, or environmental effects. Here we assess the electrokinetic behavior of chemically surface-modified bacterial cells in order to gain insight into the biophysical properties that are specifically affected by changes in surface chemistry. Using E. coli as a scaffold, an amine coupling reaction was used to covalently attach glycine, spermine, bovine serum albumin (protein), or 7-amino-4-methyl-3-coumarinylacetic acid (fluorescent dye) to the free carboxylic acid groups on the surface of the cells. These populations, along with unlabeled control cells, were subject to electrokinetic and dielectrophoretic measurements to quantify any changes in the biophysical properties upon alteration. The properties associated with each electrokinetic force are discussed relative to the specific reactant used. We conclude that relatively modest and superficial changes to cell surfaces can cause measurable changes in their biophysical properties.

Identification of neural stem and progenitor cell subpopulations using DC insulator-based dielectrophoresis.

Neural stem and progenitor cells (NSPCs) are an extremely important group of cells that form the central nervous system during development and have the potential to repair damage in conditions such as stroke impairment, spinal cord injury and Parkinson's disease degradation. Current schemes for separation of NSPCs are inadequate due to the complexity and diversity of cells in the population and lack sufficient markers to distinguish diverse cell types. This study presents an unbiased high-resolution separation and characterization of NSPC subpopulations using direct current insulator-based dielectrophoresis (DC-iDEP). The properties of the cells were identified by the ratio of electrokinetic (EK) to dielectrophoretic (DEP) mobilities. The ratio factor of NSPCs showed more heterogeneity variance (SD = 3.4-3.9) than the controlled more homogeneous human embryonic kidney cells (SD = 1.1), supporting the presence of distinct subpopulations of cells in NSPC cultures. This measure reflected NSPC fate potential since the ratio factor distribution of more neurogenic populations of NSPCs was distinct from the distribution of astrogenic NSPC populations (confidence level >99.9%). The abundance of NSPCs captured with different ranges of ratio of EK to DEP mobilities also exhibit final fate trends consistent with established final fates of the chosen samples. DC-iDEP is a novel, label-free and non-destructive method for differentiating and characterizing, and potentially separating, neural stem cell subpopulations that differ in fate.

A mathematical model of dielectrophoretic data to connect measurements with cell properties.

Dielectrophoresis (DEP) brings about the high-resolution separations of cells and other bioparticles arising from very subtle differences in their properties. However, an unanticipated limitation has arisen: difficulty in assignment of specific biological features which vary between two cell populations. This hampers the ability to interpret the significance of the variations. To realize the opportunities made possible by dielectrophoresis, the data and the diversity of structures found in cells and bioparticles must be linked. While the crossover frequency in DEP has been studied in-depth and exploited in applications using AC fields, less attention has been given when a DC field is present. Here, a new mathematical model of dielectrophoretic data is introduced which connects the physical properties of cells to specific elements of the data from potential- or time-varied DEP experiments. The slope of the data in either analysis is related to the electrokinetic mobility, while the potential at which capture initiates in potential-based analysis is related to both the electrokinetic and dielectrophoretic mobilities. These mobilities can be assigned to cellular properties for which values appear in the literature. Representative examples of high and low values of properties such as conductivity, zeta potential, and surface charge density for bacteria including Streptococcus mutans, Rhodococcus erythropolis, Pasteurella multocida, Escherichia coli, and Staphylococcus aureus are considered. While the many properties of a cell collapse into one or two features of data, for a well-vetted system the model can indicate the extent of dissimilarity. The influence of individual properties on the features of dielectrophoretic data is summarized, allowing for further interpretation of data. Graphical abstract.

Heat tolerance of Fire Service Instructors.

OBJECTIVES: Fire Service Instructors (FSI) experience repeated fire exposures a median of 13 ±â€¯8 times a month; consequently they may develop an acclimatised state. However, the chronic immunological implications of heat acclimation are yet to be understood. This study aimed to establish whether FSI exhibit an increased heat tolerance and altered immunological response to heat exposures, compared to non-exposed individuals. The study also aimed to identify if heat tolerance is related to symptoms of ill health. METHODS: Twenty-two participants were recruited: 11 FSI (age: 41 ±â€¯7 yrs, body mass: 77.4 ±â€¯12.2 kg, height: 174.1 ±â€¯8.2 cm) and 11 non-exposed controls (CON) (age: 41 ±â€¯7 yrs, body mass: 75.9 ±â€¯12.2 kg, height: 177.0 ±â€¯8.1 cm). Participants completed a 40 min heat occupational tolerance test (HOTT) exercising at 6 W kg-1 (50.0 ±â€¯1.0 °C, 12.3 ±â€¯3.3% relative humidity) on two occasions, separated by 2 months. Physiological and perceptual measures were collected throughout and venous blood samples were collected prior to and post exposure. RESULTS: FSI displayed significantly reduced peak rectal temperature (Tre) (-0.42 °C), change in Tre (-0.33 °C), and thermal sensation (-1.0) and increased sweat rate (+0.25 L h-1) at the end of the HOTT compared to CON (p < 0.05). FSI exhibited similar responses to the HOTT as CON for all haematological variables. However, resting interleukin-6, interleukin-1ß, and immunoglobulin G were significantly greater in FSI than CON. There was no difference in responses following the 2 month working period. FSI peak Tre was negatively correlated with symptoms of ill health (rpb = -0.473, p = 0.026) and the number of fire exposures in the previous 2 weeks (rs = -0.589, p = 0.004). CONCLUSION: Despite increased heat tolerance compared to non-exposed individuals, FSI may develop a maladaptation to repeated fire exposures, with elevated resting cytokine levels and an increased prevalence of ill health symptoms.

The acute effect of training fire exercises on fire service instructors.

Fire service instructors (FSI) regularly experience different types of fire exercises, however the strain experienced from these scenarios is not well understood. This study aims to identify the physiological and perceptual strain of Fire Service Instructors (FSI) to three training exercises: DEMO, ATTACK, COMPARTMENT, and the different roles performed: SETTER, INSTRUCTOR. The study also aims to assess the effect that different exercise patterns over a day (BOX, MULTI, COMBINATION) have on immunological responses. Sixteen FSI (age: 41 ± 8 years, body mass: 83.7 ± 6.7 kg, height: 177.0 ± 6.7 cm) were recruited, with 10 FSI completing the three exercises. Physiological and perceptual measures were collected prior to and immediately post each exercise. Venous blood samples were collected at the beginning and end of each day. One-way analysis of variance (ANOVA) were conducted to assess differences in physiological variables between exercise types, independent samples t-tests were conducted between roles. Day changes in hematological variables were assessed by paired sample t-tests and analyzed by one-way ANOVAs to identify differences between exercise patterns. The COMPARTMENT exercise resulted in a greater change in rectal temperature (ΔTre) (0.49 ± 0.28 °C) than both the DEMO (0.23 ± 0.19 °C, p = 0.045) and ATTACK (0.27 ± 0.22 °C, p = 0.016). Within the COMPARTMENT exercise, the SETTER resulted in a greater ΔTre and rating of perceived exertion than the INSTRUCTOR (0.67 ± 0.29 °C vs. 0.43 ± 0.18 °C, p = 0.027 and 14 ± 2 vs. 11 ± 2, p = 0.001, respectively). Following a day of fire exercises white blood cells (WBC), neutrophils, lymphocytes (LYMPH), monocytes (MONO), platelets (PLT), mean platelet volume (MPV), Interleukin (IL)-6, and cardiac troponin T (cTnT) all increased (p < 0.05). Exercise patterns containing a COMPARTMENT exercise resulted in greater PLT, MPV, and IL-6. Total daily variation in ΔTre was correlated with post-exercise WBC, MONO, and LYMPH. COMPARTMENT exercises produce the greatest physiological strain, with the SETTER role within this exercise causing the greatest ΔTre. Although predominately physiological responses remain within safe limits. Exercise patterns that include a COMPARTMENT exercise also generate a greater inflammatory response.

Females exposed to 24 h of sleep deprivation do not experience greater physiological strain, but do perceive heat illness symptoms more severely, during exercise-heat stress.

There is limited and inconclusive evidence surrounding the physiological and perceptual responses to heat stress while sleep deprived, especially for females. This study aimed to quantify the effect of 24 h sleep deprivation on physiological strain and perceptual markers of heat-related illness in females. Nine females completed two 30-min heat stress tests (HST) separated by 48 h in 39°C, 41% relative humidity at a metabolic heat production of 10 W · kg-1. The non-sleep deprived HST was followed by the sleep deprivation (SDHST) trial for all participants during the follicular phase of the menstrual cycle. Physiological and perceptual measures were recorded at 5 min intervals during the HSTs. On the cessation of the HSTs, heat illness symptom index (HISI) was completed. HISI scores increased after sleep deprivation by 28 ± 16 versus 20 ± 16 (P = 0.01). Peak (39.40 ± 0.35°C vs. 39.35 ± 0.33°C) and change in rectal temperature (1.91 ± 0.21 vs. 1.93 ± 0.34°C), and whole body sweat rate (1.08 ± 0.31 vs. 1.15 ± 0.36 L · h-1) did not differ (P > 0.05) between tests. No difference was observed in peak, nor rise in: heart rate, mean skin temperature, perceived exertion or thermal sensation during the HSTs. Twenty-four hours sleep deprivation increased perceptual symptoms associated with heat-related illness; however, no thermoregulatory alterations were observed.

A new occupational heat tolerance test: A feasibility study.

Heat tolerance tests identify those susceptible to heat illnesses and monitor heat adaptations. Currently, tolerance tests do not replicate the uncompensable heat strain environments experienced in some occupations. In addition, tests can take up to 2 h to complete, and cannot offer intra and inter individual comparisons, due to the use of a fixed exercise intensity. This study aimed to assess the validity and reliability of a new heat occupational tolerance test (HOTT: 40 min at 6 W kg-1 metabolic heat production, 50 °C 10% RH, in protective clothing) to the standard heat tolerance test (HTT: 2 h walk at 5 km h-1 1% gradient, 40 °C 40% RH, in shorts and t-shirt). Eighteen participants (age: 21 ±â€¯3 yrs, body mass: 81.3 ±â€¯5.9 kg) completed trials to assess the validity and/or reliability of the HOTT. Peak rectal temperature (Tre) displayed strong agreement and low measurement error (0.19 °C) between HTT (38.7 ±â€¯0.4 °C) and HOTT (38.6 ±â€¯0.4 °C). Strong agreement was also displayed for physiological and perceptual measures between the two HOTT trials, including peak Tre (38.5 ±â€¯0.4 °C vs. 38.5 ±â€¯0.4 °C) and peak heart rate (182 ±â€¯20 b min-1 vs. 182 ±â€¯21 b min-1). The HOTT is the first tolerance test that assesses individuals' responses whilst wearing protective clothing in high temperatures. It can consistently identify individuals' levels of heat tolerance within a reduced time frame. In addition, it allows for participant monitoring over time and comparisons between individuals to be made. A continuum based approach is recommended when assessing individuals' responses to the HOTT.