Myths and methodologies: Considerations for evaluating the time course of thermoregulatory adaptation during heat acclimation.

Since the early 1900s, repeated heat exposure has been used as a method to induce physiological adaptations that enhance our ability to tolerate heat stress during athletic and occupational pursuits. Much of this work has been dedicated to quantifying the time course of adaptation and identifying the minimum duration of acclimation required to optimise performance or enhance safety. To achieve this, investigators have typically applied classical (constant load) heat acclimation, whereby 60-90 min exercise is performed at the same absolute or relative intensity in a hot environment for 3-24 days, with adaptations evaluated using an identical forcing function test before and after. This approach has provided a foundation from which to develop our understanding of changes in thermoregulatory function, but it has several, frequently overlooked shortcomings, which have resulted in misconceptions concerning the time course of adaptation. It is frequently suggested that most of the thermoregulatory adaptations during heat acclimation occur within a week, but this is an oversimplification and a predictable artefact of the experimental designs used. Consequently, the time course of complete human adaptation to heat remains poorly understood and appears to vary considerably due to numerous individual factors. The purpose of this communication is to highlight the key methodological considerations required when interpreting the existing literature documenting adaptation over time. We also propose potential means by which to improve the way we induce and quantify the magnitude of adaptation to expedite discovery.

Short-term isothermic heat acclimation elicits beneficial adaptations but medium-term elicits a more complete adaptation.

PURPOSE: To investigate the effects of 60 min daily, short-term (STHA) and medium-term (MTHA) isothermic heat acclimation (HA) on the physiological and perceptual responses to exercise heat stress. METHODS: Sixteen, ultra-endurance runners (female = 3) visited the laboratory on 13 occasions. A 45 min sub-maximal (40% Wmax) cycling heat stress test (HST) was completed in the heat (40 °C, 50% relative humidity) on the first (HSTPRE), seventh (HSTSTHA) and thirteenth (HSTMTHA) visit. Participants completed 5 consecutive days of a 60 min isothermic HA protocol (target Tre 38.5 °C) between HSTPRE and HSTSTHA and 5 more between HSTSTHA and HSTMTHA. Heart rate (HR), rectal (Tre), skin (Tsk) and mean body temperature (Tbody), perceived exertion (RPE), thermal comfort (TC) and sensation (TS) were recorded every 5 min. During HSTs, cortisol was measured pre and post and expired air was collected at 15, 30 and 45 min. RESULTS: At rest, Tre and Tbody were lower in HSTSTHA and HSTMTHA compared to HSTPRE, but resting HR was not different between trials. Mean exercising Tre, Tsk, Tbody, and HR were lower in both HSTSTHA and HSTMTHA compared to HSTPRE. There were no differences between HSTSTHA and HSTMTHA. Perceptual measurements were lowered by HA and further reduced during HSTMTHA. CONCLUSION: A 60 min a day isothermic STHA was successful at reducing physiological and perceptual strain experienced when exercising in the heat; however, MTHA offered a more complete adaptation.

Antigen-Presenting Human γδ T Cells Promote Intestinal CD4+ T Cell Expression of IL-22 and Mucosal Release of Calprotectin.

The cytokine IL-22 plays a critical role in mucosal barrier defense, but the mechanisms that promote IL-22 expression in the human intestine remain poorly understood. As human microbe-responsive Vγ9/Vδ2 T cells are abundant in the gut and recognize microbiota-associated metabolites, we assessed their potential to induce IL-22 expression by intestinal CD4+ T cells. Vγ9/Vδ2 T cells with characteristics of APCs were generated from human blood and intestinal organ cultures, then cocultured with naive and memory CD4+ T cells obtained from human blood or the colon. The potency of blood and intestinal γδ T-APCs was compared with that of monocytes and dendritic cells, by assessing CD4+ T cell phenotypes and proliferation as well as cytokine and transcription factor profiles. Vγ9/Vδ2 T cells in human blood, colon, and terminal ileum acquired APC functions upon microbial activation in the presence of microenvironmental signals including IL-15, and were capable of polarizing both blood and colonic CD4+ T cells toward distinct effector fates. Unlike monocytes or dendritic cells, gut-homing γδ T-APCs employed an IL-6 independent mechanism to stimulate CD4+ T cell expression of IL-22 without upregulating IL-17. In human intestinal organ cultures, microbial activation of Vγ9/Vδ2 T cells promoted mucosal secretion of IL-22 and ICOSL/TNF-α-dependent release of the IL-22 inducible antimicrobial protein calprotectin without modulating IL-17 expression. In conclusion, human γδ T-APCs stimulate CD4+ T cell responses distinct from those induced by myeloid APCs to promote local barrier defense via mucosal release of IL-22 and calprotectin. Targeting of γδ T-APC functions may lead to the development of novel gut-directed immunotherapies and vaccines.

Effect of Cold (14° C) vs. Ice (5° C) Water Immersion on Recovery From Intermittent Running Exercise.

Anderson, D, Nunn, J, and Tyler, CJ. Effect of cold (14° C) vs. ice (5° C) water immersion on recovery from intermittent running exercise. J Strength Cond Res 32(3): 764-771, 2018-The purpose was to compare 14° C (CWI14° C) and 5° C (CWI5° C) cold water immersion after intermittent running. On 3 occasions, 9 male team-sport players undertook 12 minutes of CWI14° C, CWI5° C, or nonimmersed seated recovery (CON) after 45 minutes of intermittent running exercise. Maximal cycling performance and markers of recovery were measured before and in the 0-72 hours after exercise. Peak power output (PPO) was immediately reduced after all interventions (d = 1.8). CWI5° C was more effective at restoring PPO than CWI14° C (d = 0.38) and CON (d = 0.28) 24 hours after exercise, whereas both CON (d = 0.20) and CWI5 (d = 0.37) were more effective than CWI14° C after 48 hours. Cold water immersion (CWI) was more effective than CON at restoring PPO 72 hours after exercise (d = 0.28-0.30). Mean power output (MPO) was higher in CON compared with CWI5° C (d = 0.30) and CWI14° C (d = 0.21), but there was no difference between CWI5° C and CWI14° C (d = 0.08). CWI5° C was more effective than CWI14° C for restoring MPO to baseline levels 24 hours (d = 0.28) and 72 hours (d = 0.28) after exercise; however, CON was more, or equally, effective as CWI5° C and CWI14° C throughout. Lactate and creatine kinase concentrations were unaffected. Perceived muscle soreness remained elevated in CWI5 and CON throughout but was similar to baseline in CWI14° C after 72 hours. In conclusion, repeated bouts of exercise are initially impaired after 5 and 14° C CWI, but PPO may be improved 72 hours after exercise. Cold water immersion is not recommended for acute recovery based on these data. Athletes and coaches should use the time currently allocated to CWI for more effective and alternative recovery modalities.

The Antigen-Presenting Potential of Vγ9Vδ2 T Cells During Plasmodium falciparum Blood-Stage Infection.

During Plasmodium falciparum infections, erythrocyte-stage parasites inhibit dendritic cell maturation and function, compromising effective antimalarial adaptive immunity. Human Vγ9Vδ2 T cells can act in vitro as antigen-presenting cells (APCs) and induce αß T-cell activation. However, the relevance of this activity in vivo has remained elusive. Because Vγ9Vδ2 T cells are activated during the early immune response against P. falciparum infection, we investigated whether they could contribute to the instruction of adaptive immune responses toward malaria parasites. In P. falciparum-infected patients, Vγ9Vδ2 T cells presented increased surface expression of APC-associated markers HLA-DR and CD86. In response to infected red blood cells in vitro, Vγ9Vδ2 T cells upregulated surface expression of HLA-DR, HLA-ABC, CD40, CD80, CD83, and CD86, induced naive αß T-cell responses, and cross- presented soluble prototypical protein to antigen-specific CD8+ T cells. Our findings qualify Vγ9Vδ2 T cells as alternative APCs, which could be harnessed for therapeutic interventions and vaccine design.

Microbe-specific unconventional T cells induce human neutrophil differentiation into antigen cross-presenting cells.

The early immune response to microbes is dominated by the recruitment of neutrophils whose primary function is to clear invading pathogens. However, there is emerging evidence that neutrophils play additional effector and regulatory roles. The present study demonstrates that human neutrophils assume Ag cross-presenting functions and suggests a plausible scenario for the local generation of APC-like neutrophils through the mobilization of unconventional T cells in response to microbial metabolites. Vγ9/Vδ2 T cells and mucosal-associated invariant T cells are abundant in blood, inflamed tissues, and mucosal barriers. In this study, both human cell types responded rapidly to neutrophils after phagocytosis of Gram-positive and Gram-negative bacteria producing the corresponding ligands, and in turn mediated the differentiation of neutrophils into APCs for both CD4(+) and CD8(+) T cells through secretion of GM-CSF, IFN-γ, and TNF-α. In patients with acute sepsis, circulating neutrophils displayed a similar APC-like phenotype and readily processed soluble proteins for cross-presentation of antigenic peptides to CD8(+) T cells, at a time when peripheral Vγ9/Vδ2 T cells were highly activated. Our findings indicate that unconventional T cells represent key controllers of neutrophil-driven innate and adaptive responses to a broad range of pathogens.

Acute L-arginine supplementation has no effect on cardiovascular or thermoregulatory responses to rest, exercise, and recovery in the heat.

PURPOSE: To investigate the effect of acute L-arginine (L-ARG) supplementation on cardiovascular and thermoregulatory responses to rest, exercise, and recovery in the heat. METHODS: Eight healthy men (age 27 ± 6 years; stature 176 ± 6 cm; body mass 76 ± 4 kg; maximal power output 237 ± 39 W) participated in a double-blind, crossover study, attending the laboratory for two experimental trials. On each occasion, participants consumed 500 ml of a black currant-flavoured cordial beverage 30 min before completing a 90 min experiment in the heat (35 °C and 50% rh). The experiment consisted of 30 min of seated rest, followed by 30 min submaximal cycling (60% maximal power output) and 30 min passive seated recovery. On one visit the drink contained 10 g of dissolved L-ARG while on the other visit it did not. RESULTS: L-ARG supplementation increased plasma L-ARG concentrations (peak +223 ± 80% after 60 min of the 90 min experiment); however, supplementation had no effect on rectal temperature, mean skin temperature, heart rate, arterial pressure, forearm skin vascular conductance, oxygen consumption or sweat loss at rest, during exercise, or during recovery in the heat (p > 0.05). CONCLUSION: Acute ingestion of 10 g L-ARG supplementation failed to elicit any changes in the cardiovascular or thermoregulatory responses to active or passive heat exposure in young, healthy males.

Human Vγ9/Vδ2 T cells: Innate adaptors of the immune system.

Unconventional T cells are gaining center stage as important effector and regulatory cells that orchestrate innate and adaptive immune responses. Human Vγ9/Vδ2 T cells are amongst the best understood unconventional T cells, as they are easily accessible in peripheral blood, can readily be expanded and manipulated in vitro, respond to microbial infections in vivo and can be exploited for novel tumor immunotherapies. We here review findings that suggest that Vγ9/Vδ2 T cells, and possibly other unconventional human T cells, play an important role in bridging innate and adaptive immunity by promoting the activation and differentiation of various types of antigen-presenting cells (APCs) and even turning into APCs themselves, and thereby pave the way for antigen-specific effector responses and long-term immunological memory. Although the direct physiological relevance for most of these mechanisms still needs to be demonstrated in vivo, these findings may have implications for novel therapies, diagnostic tests and vaccines.

IL-21 conditions antigen-presenting human γδ T-cells to promote IL-10 expression in naïve and memory CD4+ T-cells.

Direct interaction between T-cells exerts a major influence on tissue immunity and inflammation across multiple body sites including the human gut, which is highly enriched in 'unconventional' lymphocytes such as γδ T-cells. We previously reported that microbial activation of human Vγ9/Vδ2+ γδ T-cells in the presence of the mucosal damage-associated cytokine IL-15 confers the ability to promote epithelial barrier defence, specifically via induction of IL-22 expression in conventional CD4+ T-cells. In the current report, we assessed whether other cytokines enriched in the gut milieu also functionally influence microbe-responsive Vγ9/Vδ2 T-cells. When cultured in the presence of IL-21, Vγ9/Vδ2 T-cells acquired the ability to induce expression of the immunoregulatory cytokine IL-10 in both naïve and memory CD4+ T-cells, at levels surpassing those induced by monocytes or monocyte-derived DCs. These findings identify an unexpected influence of IL-21 on Vγ9/Vδ2 T-cell modulation of CD4+ T-cell responses. Further analyses suggested a possible role for CD30L and/or CD40L reverse signalling in mediating IL-10 induction by IL-21 conditioned Vγ9/Vδ2 T-cells. Our findings indicate that the local microenvironment exerts a profound influence on Vγ9/Vδ2 T-cell responses to microbial challenge, leading to induction of distinct functional profiles among CD4+ T-cells that may influence inflammatory events at mucosal surfaces. Targeting these novel pathways may offer therapeutic benefit in disorders such as inflammatory bowel disease.

The validity of the SmartJump contact mat.

The aim of this study was to establish the criterion validity of the SmartJump contact mat in assessing vertical jump height (VJH) and peak power (PP). Twenty-three participants (15 men, age = 26 ± 6 years; 8 women, age = 26 ± 9 years) completed a maximal effort vertical jump using 3 different jump types (countermovement jump [CMJ], countermovement with arms [CMJA], and squat jump [SJ]). Data were simultaneously collected on both the contact mat and force platform. Vertical jump height was calculated using the time in air (TIA) method with both force platform (TIA(platform)) and contact mat (TIA(mat)) data and the takeoff velocity (TOV) method using the force platform (TOV(platform)) data. Peak power was calculated using a validated equation. The results showed that VJH and PP calculated using the TIA(mat) method were significantly greater than that calculated from the TIA(platform) and TOV(platform) methods for all jump types (VJH: p < 0.001, PP: p < 0.001). The results from this study show clear discrepancies between apparatus and calculation methods that may have implications for practitioners and should be considered when assessing VJH and PP in the field.

The Influence of a Competitive Field Hockey Match on Cognitive Function.

Despite the known positive effects of acute exercise on cognition, the effects of a competitive team sport match are unknown. In a randomized crossover design, 20 female and 17 male field hockey players (19.7 ± 1.2 years) completed a battery of cognitive tests (Visual Search, Stroop, Corsi Blocks, and Rapid Visual Information Processing) prior to, at half-time, and immediately following a competitive match (or control trial of seated rest); with effect sizes (ES) presented as raw ES from mixed effect models. Blood samples were collected prior to and following the match and control trial, and analyzed for adrenaline, noradrenaline, brain derived neurotrophic factor (BDNF), cathepsin B, and cortisol. The match improved response times for a simple perception task at full-time (ES = -14 ms; P < 0.01) and response times on the complex executive function task improved at half-time (ES = -44 ms; P < 0.01). Working memory declined at full-time on the match (ES = -0.6 blocks; P < 0.01). The change in working memory was negatively correlated with increases in cortisol (r = -0.314, P = 0.01; medium), as was the change in simple perception response time and the change in noradrenaline concentration (r = -0.284, P = 0.01; small to medium). This study is the first to highlight the effects a competitive hockey match can have on cognition. These findings have implications for performance optimization, as understanding the influence on specific cognitive domains across a match allows for the investigation into strategies to improve these aspects.

Antibody secreting cells are critically dependent on integrin α4ß7/MAdCAM-1 for intestinal recruitment and control of the microbiota during chronic colitis.

T and B cells employ integrin α4ß7 to migrate to intestine under homeostatic conditions. Whether those cells differentially rely on α4ß7 for homing during inflammatory conditions has not been fully examined. This may have implications for our understanding of the mode of action of anti-integrin therapies in inflammatory bowel disease (IBD). Here, we examined the role of α4ß7 integrin during chronic colitis using IL-10-/- mice, ß7-deficient IL-10-/-, IgA-deficient IL-10-/- mice, and antibody blockade of MAdCAM-1. We found that α4ß7 was predominantly expressed by B cells. ß7 deficiency and MAdCAM-1 blockade specifically depleted antibody secreting cells (ASC) (not T cells) from the colonic LP, leading to a fecal pan-immunoglobulin deficit, severe colitis, and alterations of microbiota composition. Colitis was not due to defective regulation, as dendritic cells (DC), regulatory T cells, retinaldehyde dehydrogenase (RALDH) expression, activity, and regulatory T/B-cell cytokines were all comparable between the strains/treatment. Finally, an IgA deficit closely recapitulated the clinical phenotype and altered microbiota composition of ß7-deficient IL-10-/- mice. Thus, a luminal IgA deficit contributes to accelerated colitis in the ß7-deficient state. Given the critical/nonredundant dependence of IgA ASC on α4ß7:MAdCAM-1 for intestinal homing, B cells may represent unappreciated targets of anti-integrin therapies.

The effects of combined action observation and motor imagery on corticospinal excitability and movement outcomes: Two meta-analyses.

Motor simulation interventions involving motor imagery (MI) and action observation (AO) have received considerable interest in the behavioral sciences. A growing body of research has focused on using AO and MI simultaneously, termed 'combined action observation and motor imagery' (AOMI). The current paper includes two meta-analyses that quantify changes in corticospinal excitability and motor skill performance for AOMI compared to AO, MI and control conditions. Specifically, the first meta-analysis collated and synthesized existing motor evoked potential (MEP) amplitude data from transcranial magnetic stimulation studies and the second meta-analysis collated and synthesized existing movement outcome data from behavioral studies. AOMI had a positive effect compared to control and AO but not MI conditions for both MEP amplitudes and movement outcomes. No methodological factors moderated the effects of AOMI, indicating a robust effect of AOMI across the two outcome variables. The results of the meta-analyses are discussed in relation to existing literature on motor simulation and skill acquisition, before providing viable directions for future research on this topic.

The effects of pre- and per-cooling interventions used in isolation and combination on subsequent 15-minute time-trial cycling performance in the heat.

OBJECTIVES: To investigate the effects of pre- and per-cooling interventions on subsequent 15-min time-trial (TT) cycling performance in the heat. DESIGN: Randomized cross-over design. METHODS: Nine male athletes completed four experimental trials in the heat (40 °C, 50% rh): no-cooling (CON); warm-up per-cooling (PER: neck-cooling collar applied during the preload); pre-cooling (PRE: 30 min of cold water (22 °C) immersion [CWI]); and pre- and per-cooling combined (PRE + PER). In each trial, participants completed a 45-min preload exercise (50% V̇O2peak), followed by a 15-min TT. Physiological (rectal [Tre], skin [Tsk], and neck [Tneck] temperature, and heart rate [HR]) and perceptual data (ratings of perceived exertion [RPE], thermal comfort [TC] and thermal sensation [TS]) were measured throughout. RESULTS: Tre and Tsk were lower in PRE and PRE + PER at the start of the preload (p < 0.001). Tre remained lower throughout the preload following CWI although these differences were no longer present at the start of the TT (p = 0.22). Tneck was lowered throughout in PER and PRE + PER (p < 0.001). No other physiological or perceptual differences were observed at the start or end of the preload or TT. Participants covered a similar TT distance in all trials (15.7-15.9 km, p = 0.77). CONCLUSIONS: Pre-cooling induced thermoregulatory benefits for ~45 min and perceptual benefits for the same duration when supplemented with per-cooling. Neck per-cooling offered no such benefits when used in isolation. Neither pre- nor per-cooling, in isolation or combination, improved subsequent 15-min cycling time-trial performance in well-trained participants in the heat (40 °C).

Progressive hyperthermia elicits distinct responses in maximum and rapid torque production.

OBJECTIVES: To investigate the effect of progressive whole-body hyperthermia on maximal, and rapid voluntary torque production, and their neuromuscular determinants. DESIGN: Repeated measures, randomised. METHODS: Nine participants performed sets of neuromuscular assessments in HOT conditions (∼50°C, ∼35% relative humidity) at rectal temperatures (Tre) of 37, 38.5 and 39.5°C and in CON conditions (∼22°C, ∼35% relative humidity) at a Tre of ∼37°C and pre-determined comparative time-points. Electrically evoked twitch (single impulse) and octet (8 impulses at 300Hz) responses were measured at rest. Maximum voluntary torque (MVT), surface electromyography (EMG) normalised to maximal M-wave, and voluntary activation (VA) were measured during 3-5s isometric maximal voluntary contractions. Rate of torque development (RTD) and normalised EMG were measured during rapid voluntary isometric contractions from rest. RESULTS: All neuromuscular variables were unaffected by time in CON. In HOT, MVT, normalised EMG at MVT and VA were lower at 39.5°C compared to 37°C (p<0.05). Early- (0-50ms) and middle- (50-100ms) phase voluntary RTD were unaffected by increased Tre (p>0.05), despite lower normalised EMG at Tre 39.5°C (p<0.05) in rapid contractions. In contrast, late-phase (100-150ms) voluntary RTD was lower at 38.5°C and 39.5°C compared to 37°C (p<0.05) in HOT. Evoked twitch and octet RTD increased with increased Tre (p<0.05). CONCLUSIONS: Hyperthermia reduced late-phase voluntary RTD, likely due to reduced neural drive and the reduction in MVT. In contrast, early- and middle-phase voluntary RTD were unaffected by hyperthermia, likely due to the conflicting effects of reduced neural drive but faster intrinsic contractile properties.

An integrin αEß7-dependent mechanism of IgA transcytosis requires direct plasma cell contact with intestinal epithelium.

Efficient IgA transcytosis is critical for the maintenance of a homeostatic microbiota. In the canonical model, locally-secreted dimeric (d)IgA reaches the polymeric immunoglobulin receptor (pIgR) on intestinal epithelium via simple diffusion. A role for integrin αE(CD103)ß7 during transcytosis has not been described, nor its expression by intestinal B cell lineage cells. We found that αE-deficient (αE-/-) mice have a luminal IgA deficit, despite normal antibody-secreting cells (ASC) recruitment, local IgA production and increased pIgR expression. This deficit was not due to dendritic cell (DC)-derived retinoic acid (RA) nor class-switching defects, as stool from RAG-/- mice reconstituted with αE-/- B cells was also IgA deficient. Flow cytometric, ultrastructural and transcriptional profiling showed that αEß7-expressing ASC represent an undescribed subset of terminally-differentiated intestinal plasma cells (PC) that establishes direct cell to cell contact with intestinal epithelium. We propose that IgA not only reaches pIgR through diffusion, but that αEß7+ PC dock with E-cadherin-expressing intestinal epithelium to directly relay IgA for transcytosis into the intestinal lumen.

Inherent Immune Cell Variation Within Colonic Segments Presents Challenges for Clinical Trial Design.

BACKGROUND AND AIMS: Intestinal biopsy sampling during IBD trials represents a valuable adjunct strategy for understanding drug responses at the tissue level. Given the length and distinctive embryonic origins of the proximal and distal colon, we investigated whether inherent regional differences of immune cell composition could introduce confounders when sampling different disease stages, or pre/post drug administration. Here, we capitalise on novel mass cytometry technology to perform deep immunophenotyping of distinct healthy colonic segments, using the limited numbers of biopsies that can be harvested from patients. METHODS: Biopsies [2.8 mm] were collected from the caecum, transverse colon, descending colon, and rectum of normal volunteers. Intestinal leukocytes were isolated, stained with a panel of 37 antibodies, and mass cytometry data acquired. RESULTS: Site-specific patterns of leukocyte localisation were observed. The proximal colon featured increased CD8+ T cells [particularly resident memory], monocytes, and CD19+ B cells. Conversely, the distal colon and rectum tissues exhibited enrichment for CD4+ T cells and antibody-secreting cells. The transverse colon displayed increased abundance of both γδ T cells and NK cells. Subsets of leukocyte lineages also displayed gradients of expression along the colon length. CONCLUSIONS: Our results show an inherent regional immune cell variation within colonic segments, indicating that regional mucosal signatures must be considered when assessing disease stages or the prospective effects of trial drugs on leukocyte subsets. Precise protocols for intestinal sampling must be implemented to allow for the proper interpretation of potential differences observed within leukocyte lineages present in the colonic lamina propria.

Sphingosine-1-Phosphate Lyase Inhibition Alters the S1P Gradient and Ameliorates Crohn's-Like Ileitis by Suppressing Thymocyte Maturation.

BACKGROUND: Lymphocytes recirculate from tissues to blood following the sphingosine-1-phosphate (S1P) gradient (low in tissues, high in blood), maintained by synthetic and degradative enzymes, among which the S1P lyase (SPL) irreversibly degrades S1P. The role of SPL in the intestine, both during homeostasis and IBD, is poorly understood. We hypothesized that modulation of tissue S1P levels might be advantageous over S1P receptor (S1PR) agonists (eg, fingolimod, ozanimod, etrasimod), as without S1PR engagement there might be less likelihood of potential off-target effects. METHODS: First we examined SPL mRNA transcripts and SPL localization in tissues by quantitative reverse transcription polymerase chain reaction and immunohistochemistry. The in vivo effects of the SPL inhibitors 4-deoxypyridoxine hydrochloride (30 mg/L) and 2-acetyl-4 (tetrahydroxybutyl)imidazole (50 mg/L) were assessed through their oral administration to adult TNF∆ARE mice, which spontaneously develop Crohn's-like chronic ileitis. The effect of SPL inhibition on circulating and tissue lymphocytes, transcriptional regulation of proinflammatory cytokines, and on the histological severity of ileitis was additionally examined. Tissue S1P levels were determined by liquid chromatography-mass spectrometry. Mechanistically, the potential effects of high S1P tissue levels on intestinal leukocyte apoptosis were assessed via terminal deoxynucleotidyl transferase dUTP nick end-labeling assay and annexin 5 staining. Finally, we examined the ability of T cells to home to the intestine, along with the effects of SPL inhibition on cellular subsets within immune compartments via flow and mass cytometry. RESULTS: S1P lyase was ubiquitously expressed. In the gut, immunohistochemistry predominantly localized it to small intestinal epithelia, although the lamina propria leukocyte fraction had higher mRNA transcripts. Inhibition of SPL markedly increased local intestinal S1P levels, induced peripheral lymphopenia, downregulated proinflammatory cytokines, and attenuated chronic ileitis in mice. SPL inhibition reduced T and myeloid cells in secondary lymphoid tissues and the intestine and decreased naïve T-cell recruitment. The anti-inflammatory activity of SPL inhibition was not mediated by leukocyte apoptosis, nor by interference with the homing of lymphocytes to the intestine, and was independent of its peripheral lymphopenic effect. However, SPL inhibition promoted thymic atrophy and depleted late immature T cells (CD4+CD8+ double positive), with accumulation of mature CD4+CD8- and CD4-CD8+ single-positive cells. CONCLUSIONS: Inhibition of the S1P lyase alters the S1P gradient and attenuates chronic ileitis via central immunosuppression. SPL inhibition could represent a potential way to tame an overactive immune response during IBD and other T-cell-mediated chronic inflammatory diseases.

Impairment of Cycling Capacity in the Heat in Well-Trained Endurance Athletes After High-Intensity Short-Term Heat Acclimation.

PURPOSE: To investigate the effects of short-term, high-intensity interval-training (HIIT) heat acclimation (HA). METHODS: Male cyclists/triathletes were assigned into either an HA (n = 13) or a comparison (COMP, n = 10) group. HA completed 3 cycling heat stress tests (HSTs) to exhaustion (60% Wmax; HST1, pre-HA; HST2, post-HA; HST3, 7 d post-HA). HA consisted of 30-min bouts of HIIT cycling (6 min at 50% Wmax, then 12 × 1-min 100%-Wmax bouts with 1-min rests between bouts) on 5 consecutive days. COMP completed HST1 and HST2 only. HST and HA trials were conducted in 35°C/50% relative humidity. Cycling capacity and physiological and perceptual data were recorded. RESULTS: Cycling capacity was impaired after HIIT HA (77.2 [34.2] min vs 56.2 [24.4] min, P = .03) and did not return to baseline after 7 d of no HA (59.2 [37.4] min). Capacity in HST1 and HST2 was similar in COMP (43.5 [8.3] min vs 46.8 [15.7] min, P = .54). HIIT HA lowered resting rectal (37.0°C [0.3°C] vs 36.8°C [0.2°C], P = .05) and body temperature (36.0°C [0.3°C] vs 35.8°C [0.3°C], P = .03) in HST2 compared with HST1 and lowered mean skin temperature (35.4°C [0.5°C] vs 35.1°C [0.3°C], P = .02) and perceived strain on day 5 compared with day 1 of HA. All other data were unaffected. CONCLUSIONS: Cycling capacity was impaired in the heat after 5 d of consecutive HIIT HA despite some heat adaptation. Based on data, this approach is not recommended for athletes preparing to compete in the heat; however, it is possible that it may be beneficial if a state of overreaching is avoided.