Stimulated myotube contractions regulate membrane-bound and soluble TLR4 to prevent LPS-induced signaling and myotube atrophy in skeletal muscle cells.

Toll-like receptor 4 (TLR4) activation by lipopolysaccharides (LPS) increases proinflammatory cytokine production and upregulation of muscle atrophy signaling pathways. Muscle contractions can suppress LPS/TLR4 axis activation by reducing the protein expression of TLR4 on immune cells. However, the mechanism by which muscle contractions decrease TLR4 remains undefined. Moreover, it is not clear whether muscle contractions affect TLR4 expressed on skeletal muscle cells. The purpose of this study was to uncover the nature and mechanisms by which stimulated myotube contractions using electrical pulse stimulation (EPS) as an in vitro model of skeletal muscle contractions affect TLR4 expression and intracellular signaling to combat LPS-induced muscle atrophy. C2C12 myotubes were stimulated to contract via EPS with and without subsequent LPS exposure. We then examined the isolated effects of conditioned media (CM) collected following EPS and soluble TLR4 (sTLR4) alone on LPS-induced myotube atrophy. Exposure to LPS decreased membrane-bound and sTLR4, increased TLR4 signaling (decreased inhibitor of κBα), and induced myotube atrophy. However, EPS decreased membrane-bound TLR4, increased sTLR4, and prevented LPS-induced signaling and myotube atrophy. CM, which contained elevated levels of sTLR4, prevented LPS-induced upregulation of atrophy-related gene transcripts muscle ring finger 1 (MuRF1) and atrogin-1 and reduced myotube atrophy. Recombinant sTLR4 added to media prevented LPS-induced myotube atrophy. In summary, our study provides the first evidence that sTLR4 has anticatabolic effects by reducing TLR4-mediated signaling and atrophy. In addition, the study reveals a novel finding, by demonstrating that stimulated myotube contractions decrease membrane-bound TLR4 and increase the secretion of sTLR4 by myotubes.NEW & NOTEWORTHY Excessive Toll-like receptor 4 (TLR4) activation causes muscle atrophy. Muscle contractions can limit TLR4 activation on immune cells, but its impact on TLR4 expressed on skeletal muscle cells remains unclear. Here, we demonstrate in C2C12 myotubes for the first time that stimulated myotube contractions reduce membrane-bound TLR4 and increase soluble TLR4, preventing TLR4-mediated signaling and myotube atrophy. Further analyses revealed soluble TLR4 independently prevents myotube atrophy, supporting a potential therapeutic role in combating TLR4-mediated atrophy.

Effect of heat stress on heat shock protein expression and hypertrophy-related signaling in the skeletal muscle of trained individuals.

Muscle mass is balanced between hypertrophy and atrophy by cellular processes, including activation of the protein kinase B-mechanistic target of rapamycin (Akt-mTOR) signaling cascade. Stressors apart from exercise and nutrition, such as heat stress, can stimulate the heat shock protein A (HSPA) and C (HSPC) families alongside hypertrophic signaling factors and muscle growth. The effects of heat stress on HSP expression and Akt-mTOR activation in human skeletal muscle and their magnitude of activation compared with known hypertrophic stimuli are unclear. Here, we show a single session of whole body heat stress following resistance exercise increases the expression of HSPA and activation of the Akt-mTOR cascade in skeletal muscle compared with resistance exercise in a healthy, resistance-trained population. Heat stress alone may also exert similar effects, though the responses are notably variable and require further investigation. In addition, acute heat stress in C2C12 muscle cells enhanced myotube growth and myogenic fusion, albeit to a lesser degree than growth factor-mediated hypertrophy. Though the mechanisms by which heat stress stimulates hypertrophy-related signaling and the potential mechanistic role of HSPs remain unclear, these findings provide additional evidence implicating heat stress as a novel growth stimulus when combined with resistance exercise in human skeletal muscle and alone in isolated murine muscle cells. We believe these findings will help drive further applied and mechanistic investigation into how heat stress influences muscular hypertrophy and atrophy.NEW & NOTEWORTHY We show that acute resistance exercise followed by whole body heat stress increases the expression of HSPA and increases activation of the Akt-mTOR cascade in a physically active and resistance-trained population.

Exercise mitigates the Toll of muscle atrophy: a narrative review of the effects of exercise on Toll-like receptor-4 in leukocytes and skeletal muscle.

Conditions characterized by muscle wasting such as cachexia and sarcopenia are devastating at the individual level, and they place a profound burden on public health. Evidence suggests that inflammation is likely a mechanistic contributor to the pathogenesis of these conditions. One specific molecule, lipopolysaccharide, has gained attention due to its role in initiating inflammation. Toll-like receptor-4 is the primary receptor for lipopolysaccharide and has been shown to be implicit in the downstream proinflammatory response associated with lipopolysaccharide. Importantly, Toll-like receptor-4 is expressed in various cell types throughout the human body such as leukocytes and skeletal muscle fibers and may have site-specific effects that contribute to muscle wasting conditions based on the location in which activation occurs. Accordingly, reducing proinflammatory signaling at these locations may be an effective strategy at mitigating muscle wasting. Regular exercise training is believed to elicit anti-inflammatory adaptations, but the mechanisms by which this occurs are yet to be fully understood. Understanding the mechanisms by which Toll-like receptor-4 activation contributes to muscle wasting and how exercise affects this may allow for the development of a nonpharmacological therapeutic intervention. Therefore, in this review, we summarize the current understanding of the lipopolysaccharide/Toll-like receptor-4 axis in leukocytes and skeletal muscle fibers on the pathogenesis of muscle wasting conditions and we critically examine the current evidence regarding the effects of exercise on this axis.

Can linear regression confirm VO2max was attained in middle-aged and older adults?

PURPOSE: Evaluate the efficacy of a regression method for identifying a VO2 plateau to confirm the attainment of VO2max compared to a verification trial in middle-aged and older adults. METHODS: Eleven men and ten women (age 61.0 ± 8.1, VO2max 21.8-50.3 ml/kg/min, n = 21) completed an individualized ramp graded exercise test (GXT) on the cycle ergometer, and one hour later, a verification trial at 105% of their maximal work rate (WR) achieved during the GXT. A plateau in VO2 was used to confirm VO2max was attained. VO2 plateau was identified using the difference between the highest VO2 between the two trials and a linear regression analysis of the VO2-WR relationship during the GXT. McNemar's test of marginal homogeneity was used to detect differences in the proportion of paired data of individuals' attainment of VO2max criteria. RESULTS: Of the 21 participants, 15 (71.4%) met the verification criterion while 6 (28.6%) did not, compared to the regression method where 16 (76.2%) achieved the regression criterion while 5 (23.8%) did not. McNemar's test revealed no significant difference between participants' ability to achieve the regression and verification criteria (p = 0.999). CONCLUSION: The regression method is an effective strategy for confirming VO2max was attained with middle-aged and older adults on a cycle ergometer. This time-efficient regression method is comparable with the verification criterion but does not require a second maximal test, which may be advantageous for those where the verification trial may not be practical.

The effect of interval and continuous work on markers of acute kidney injury in a hot environment.

PURPOSE: To examine the effect of high-intensity interval work (HIIW) and moderate-intensity continuous work (MICW) on markers of acute kidney injury (AKI) and kidney function in a hot environment. METHODS: Nine males completed 2 h of work (2 × 60 min with 10 min passive rest) in a hot environment (40 °C and 15% relative humidity) as either HIIW [2 min at 80% peak oxygen consumption (VO2peak) and 3 min at 30% VO2peak] or MICW (matched for total work of HIIW). Blood and urine samples were collected immediately before (Pre), after (Post), 1 h (1 h Post), and 24 h after (24 h Post) the trials. Urine flow rate (UFR), creatinine clearance, insulin-like growth factor binding protein 7 (IGFBP7), urinary neutrophil gelatinase-associated lipocalin (uNGAL), and urinary kidney injury marker 1 (uKIM-1) were measured to assess kidney function and injury. RESULTS: Log IGFBP7 (p < 0.01), log uNGAL (p < 0.01), and log uKIM-1 (p = 0.01) all displayed a main effect for time after both HIIW and MICW. IGFBP7 (p = 0.01) and uKIM-1 (p < 0.01), corrected for Uosm, were higher after HIIW compared to MICW at Post, while IGFBP7 was also higher 1 h Post after HIIW compared to MICW (p = 0.02). UFR significantly decreasing from Pre to Post (p < 0.01) and 1 h Post (p < 0.01), but no main effect for condition (p = 0.53). CONCLUSION: Both HIIW and MICW in a hot environment caused an increase in biomarkers of kidney injury (IGFBP7, KIM-1, and NGAL), but HIIW may have a greater impact on biomarkers related to AKI.

The effect of prolonged interval and continuous exercise in the heat on circulatory markers of intestinal barrier integrity.

PURPOSE: The purpose of this study was to determine the effect of prolonged high-intensity interval (INT) and moderate-intensity continuous (CONT) treadmill exercise in the heat on markers of enterocyte injury and bacterial endotoxin translocation. METHODS: Nine males completed 2 h of work-matched exercise in the heat (40 °C and 15% RH) as either INT (2 min at 80% VO2max and 3 min at 30% VO2max) or CONT (~ 50% of VO2max). Blood samples collected pre- and post-exercise were assayed for intestinal fatty acid-binding protein (I-FABP), claudin-3 (CLDN-3), and lipopolysaccharide-binding protein (LBP). RESULTS: I-FABP was significantly increased from pre- to post-exercise in CONT (913.96 ± 625.13 to 1477.26 ± 760.99 pg•mL-1; p = 0.014, d = 0.766) and INT (714.59 ± 470.27 to 1547.93 ± 760.99 pg•mL-1; p = 0.001, d = 1.160). Pre- to post-exercise changes in I-FABP were not different between CONT and INT (p = 0.088, d = 0.414). LBP was significantly increased from pre- to post-exercise in INT (15.94 ± 2.90 to 17.35 ± 3.26 µg•mL-1; p = 0.028, d = 0.459) but not CONT (18.11 ± 5.35 to 16.93 ± 5.39 µg•mL-1; p = 0.070, d = 0.226), and pre- to post-exercise changes in LBP were higher in the INT compared to CONT (p < 0.001, d = 1.160). No significant changes were detected from pre- to post-exercise for CLDN-3 in CONT (14.90 ± 2.21 to 15.30 ± 3.07 µg•mL-1) or INT (15.55 ± 1.63 to 16.41 ± 2.11 µg•mL-1) (p > 0.05). CONCLUSIONS: We conclude that prolonged exercise in the heat induces enterocyte injury, but interval (or intermittent) exercise may cause greater bacterial endotoxin translocation which may increase the risk for local and systemic inflammation.

Repeated sprint exercise in hypoxia stimulates HIF-1-dependent gene expression in skeletal muscle.

PURPOSE: Our aim was to determine the effect of repeated sprint exercise in hypoxia on HIF-1 and HIF-1-regulated genes involved in glycolysis, mitochondrial turnover and oxygen transport. We also determined whether genes upregulated by exercise in hypoxia were dependent on the activation of HIF-1 in an in vitro model of exercise in hypoxia. METHODS: Eight endurance athletes performed bouts of repeated sprint exercise in control and hypoxic conditions. Skeletal muscle was sampled pre, post and 3 h post-exercise. HIF-1α protein and HIF1A, PDK1, GLUT4, VEGFA, BNIP3, PINK1 and PGC1A mRNA were measured. C2C12 myotubes were exposed to hypoxia and muscle contraction following treatment with a HIF-1α inhibitor to determine whether hypoxia-sensitive gene expression was dependent on HIF-1α. RESULTS: Sprint exercise in hypoxia increased HIF-1α protein expression immediately post-exercise [fold change (FC) = 3.5 ± 2.0]. Gene expression of PDK1 (FC = 2.1 ± 1.2), BNIP3 (FC = 2.4 ± 1.4) and VEGFA (FC = 2.7 ± 1.7) increased 3 h post-exercise in hypoxia but not control. PGC1A mRNA increased 3 h post-exercise in control (FC = 5.16) and hypoxia (FC = 5.7 ± 4.1) but there was no difference between the trials. Results from the in vitro experiment showed that hypoxia plus contraction also increased PDK1, BNIP3, and VEGFA gene expression. These responses were inhibited when HIF-1 protein activity was suppressed. CONCLUSION: Repeated sprint exercise in hypoxia upregulates some genes involved in glycolytic metabolism, mitochondrial turnover, and oxygen transport. HIF-1α is necessary for the expression of these genes in skeletal muscle cells.

The effect of repetition tempo on cardiovascular and metabolic stress when time under tension is matched during lower body exercise.

PURPOSE: To investigate the effect of repetition tempo on cardiovascular and metabolic stress when time under tension (TUT) and effort are matched during sessions of lower body resistance training (RT). METHODS: In a repeated-measures, cross-over design, 11 recreationally trained females (n = 5) and males (n = 6) performed 5 sets of belt squats under the following conditions: slow-repetition tempo (SLOW; 10 reps with 4-s eccentric and 2-s concentric) and traditional-repetition tempo (TRAD; 20 reps with 2-s eccentric and 1-s concentric). TUT (60 s) was matched between conditions and external load was adjusted so that lifters were close to concentric muscular failure at the end of each set. External load, total volume load (TVL), impulse (IMP), blood lactate, ratings of perceived exertion (RPE), HR, and muscle oxygenation were measured. RESULTS: Data indicated that TVL (p < 0.001), blood lactate (p = 0.017), RPE (p = 0.015), and HR (p < 0.001) were significantly greater during TRAD while external load (p = 0.030) and IMP (p = 0.002) were significantly greater during SLOW. Whether it was expressed as minimal values or change scores, muscle oxygenation was not different between protocols. CONCLUSION: When TUT is matched, TVL, cardiovascular stress, metabolic stress, and perceived exertion are greater when faster repetition tempos are used. In contrast, IMP and external load are greater when slower repetition tempos are used.

The combined effects of exercise-induced muscle damage and heat stress on acute kidney stress and heat strain during subsequent endurance exercise.

PURPOSE: The purpose of the study was to investigate the combined effect of downhill running and heat stress on muscle damage, as well as on heat strain and kidney stress during subsequent running in the heat. METHODS: In a randomized cross-over study, ten non-heat-acclimated, physically active males completed downhill running in temperate (EIMD in Temp) and hot (EIMD in Hot) conditions followed by an exercise-heat stress (HS) test after 3-h seated rest. Blood and urine samples were collected immediately pre- and post-EIMD and HS, and 24 h post-EIMD (post-24 h). Core temperature and thermal sensation were measured to evaluate heat strain. Serum creatine kinase (CK), maximal voluntary isometric contraction of the quadriceps (MVC) and perceived muscle soreness were measured to evaluate muscle damage. Urinary neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1) levels were measured to indicate acute kidney stress. RESULTS: CK, MVC and perceived soreness were not different between conditions at any timepoints. In the EIMD in Hot condition, urinary NGAL was significantly elevated from pre- to post-HS (pre-HS: 6.56 {1.53-12.24} ng/min, post-HS: 13.72 {7.67-21.46} ng/min, p = 0.034). Such elevation of NGAL or KIM-1 was not found in the EIMD in Temp condition. CONCLUSIONS: As compared with downhill running in a temperate environment, downhill running in a hot environment does not appear to aggravate muscle damage. However, elevated NGAL levels following EIMD in a hot environment suggest such exercise may increase risk of mild acute kidney injury during subsequent endurance exercise in the heat.

Effect of Cardiorespiratory Fitness on Verifying VO2max in Middle-aged and Older Adults.

We investigated the effect of cardiorespiratory fitness (CRF) on the probability of achieving the verification criterion to confirm that VO2max was obtained in a sample of middle-aged to older adults. Data from twelve men and nine women (60.7±8.5 years, VO2max: 34.8±9.4 mL/kg/min) were used for analysis. Participants had their VO2max measured via a maximal graded exercise test and confirmed using a verification bout on a cycle ergometer. Logistic regression was used to evaluate the effect of CRF (VO2max) on the probability a participant would successfully achieve the verification criterion. Odds ratios are reported to quantify the effect size. No statistically significant relationship was observed between CRF and achieving the verification criterion (ß=.081, SE=.0619, Wald=1.420, p=.156). Estimated odds ratio for the effect of CRF on the verification criterion indicated an increase of 8% [Exp(ß)=1.08, 95% CI (0.96, 1.22)] in the probability of achieving the verification criterion given a one unit increase in VO2max. Each 1 mL/kg/min increase in VO2max results in an 8% increase in the chance that an individual achieves the verification criterion confirming that VO2max was obtained. Therefore, CRF is likely of practical significance and should be considered when deciding to use a verification trial.

Aerobic Adaptations to Resistance Training: The Role of Time under Tension.

Generally, skeletal muscle adaptations to exercise are perceived through a dichotomous lens where the metabolic stress imposed by aerobic training leads to increased mitochondrial adaptations while the mechanical tension from resistance training leads to myofibrillar adaptations. However, there is emerging evidence for cross over between modalities where aerobic training stimulates traditional adaptations to resistance training (e.g., hypertrophy) and resistance training stimulates traditional adaptations to aerobic training (e.g., mitochondrial biogenesis). The latter is the focus of the current review in which we propose high-volume resistance training (i.e., high time under tension) leads to aerobic adaptations such as angiogenesis, mitochondrial biogenesis, and increased oxidative capacity. As time under tension increases, skeletal muscle energy turnover, metabolic stress, and ischemia also increase, which act as signals to activate the peroxisome proliferator-activated receptor gamma coactivator 1-alpha, which is the master regulator of mitochondrial biogenesis. For practical application, the acute stress and chronic adaptations to three specific forms of high-time under tension are also discussed: Slow-tempo, low-intensity resistance training, and drop-set resistance training. These modalities of high-time under tension lead to hallmark adaptations to resistance training such as muscle endurance, hypertrophy, and strength, but little is known about their effect on traditional aerobic training adaptations.

Efficacy of estimating VO2max with the Heart Rate Ratio Method in middle-aged and older adults.

PURPOSE: Investigate difference, agreement, and bias between maximal oxygen consumption (VO2max) measured by indirect calorimetry and the Heart Rate Ratio Method (HRRM) for estimating VO2max using measured and age-predicted maximum heart rate (HRmax) in a sample of middle-aged and older adults. METHODS: Eleven men and nine women (age: 61.8 ± 8.5 years, VO2max: 36.6 ± 9.5 mL/kg/min, N = 20) had their VO2max measured via a maximal graded exercise test and confirmed using a verification bout on a treadmill ergometer. Participants' VO2max was estimated via the HRRM using measured and age-predicted HRmax. Difference, agreement, and bias between methods were assessed using a one-way repeated measures Analysis of Variance (ANOVA), Lin's concordance correlation coefficient, and Bland-Altman plots, respectively. RESULTS: There were no statistical differences between methods of assessing VO2max, F(1.13, 21.47) = 0.583, p = 0.473, generalized eta-squared = 0.012. The standard error of the estimate (SEE) for the HRRM using measured or predicted HRmax was 5.2% and 5.6% of the mean VO2max, respectively. Both HRRM predictions had poor agreement with VO2max (rc ≤ 0.40). There was a systematic bias to underestimate as VO2max increased for HRRM using measured HRmax, F(1, 18) = 17.40, p < 0.001, and when using an age-predicted HRmax, F(1, 18) = 13.39, p = 0.002. CONCLUSION: Both versions of the HRRM have poor agreement, relatively large SEEs, and a bias to misrepresent VO2max. When possible, VO2max should be measured via a maximal graded exercise test with indirect calorimetry for middle-aged and older adults.

Does heart rate response confirm the attainment of maximal oxygen uptake in adults 45 years and older?

PURPOSE: To test the efficacy of a plateau in heart rate (HRplat) as an effective indicator for confirming [Formula: see text]max attainment in a middle-aged to older sample. METHODS: Nine men and eleven women (age 60 ± 8.5 years, [Formula: see text]max 35.9 ± 9.4 ml/kg/min, N = 20) completed a single [Formula: see text]max test on both the treadmill and cycle ergometer.[Formula: see text]max was confirmed using a plateau in [Formula: see text] ([Formula: see text]plat) of ≤ 150 ml/min, a verification bout, and HRplat (≤ 4 bpm). RESULTS: [Formula: see text]plat occurred in 100% and 95% of participants on the treadmill and cycle ergometer, respectively. Verification criteria ([Formula: see text]max during verification ≤ 2% of [Formula: see text]max during incremental test) were met by 80% of participants on both modalities. HRplat was achieved by 90% and 70% of participants on the treadmill and cycle ergometer, respectively. CONCLUSION: These results suggest that a verification bout is reliable for confirming [Formula: see text]max in older adults on both modalities. In our sample of middle-aged and older adults, [Formula: see text]plat was the most robust method to assess [Formula: see text]max when indirect calorimetry is available. Although more research is warranted, when indirect calorimetry is not available, a HRplat of ≤ 4 bpm may be a useful alternative to get an accurate representation of maximal effort in middle-aged and older adults.

Effect of Predicted Versus Measured Thoracic Gas Volume on Body Fat Percentage in Young Adults.

The BodPod® (COSMED, Concord, CA) uses predicted (pTGV) or measured thoracic gas volume (mTGV) during estimations of percentage body fat (%BF). In young adults, there is inconsistent evidence on the variation between pTGV and mTGV, and the effect of sex as a potential covariate on this relationship is unknown. This study examined the difference between TGV assessments and its effect on %BF and potential sex differences that may impact this relationship. A retrospective analysis of BodPod® pTGV and mTGV for 95 men and 86 women ages 18-30 years was performed. Predicted TGV was lower than mTGV for men (-0.49 ± 0.7 L; p < .0001). For men, %BF derived by pTGV was lower than that by mTGV (-1.3 ± 1.8%; p < .0001). For women, no differences were found between pTGV and mTGV (-0.08 ± 0.6 L; p > .05) or %BF (-0.03 ± 0.2%; p > .05). The two-predictor model of sex and height was able to account for 57.9% of the variance in mTGV, F(2, 178) = 122.5, p < .0001. Sex corrected for the effect of height was a significant predictor of mTGV (ß = 0.483 L, p < .0001). There is bias for pTGV to underestimate mTGV in individuals with a large mTGV, which can lead to significant underestimations of %BF in young adults; this was especially evident for men in this study. Sex is an important covariate that should be considered when deciding to use pTGV. The results indicate that TGV should be measured whenever possible for both men and women ages 18-30 years.

Divergent mechanisms regulate TLR4 expression on peripheral blood mononuclear cells following workload-matched exercise in normoxic and hypoxic environments.

Exercise in hypoxia increases immune responses compared with normoxic exercise, and while Toll-like receptor 4 (TLR4) is implicated in these responses, its regulation remains undefined. The purpose of this study was to 1) investigate TLR4 regulation during workload-matched endurance exercise in normoxic and hypoxic conditions in vivo and 2) determine the independent effects of hypoxia and muscle contractions on TLR4 expression in vitro. Eight recreationally active men cycled for 1 h at 65% of their V̇o2max in normoxia (630 mmHg) and in hypobaric hypoxia (440 mmHg). Exercise in normoxia decreased TLR4 expressed on peripheral blood mononuclear cells (PBMCs), had no effect on the expression of inhibitor of κBα (IκBα), and increased the concentration of soluble TLR4 (sTLR4) in circulation. In contrast, exercise in hypoxia decreased the expression of TLR4 and IκBα in PBMCs, and sTLR4 in circulation. Markers of physiological stress were higher during exercise in hypoxia, correlating with markers of intestinal barrier damage, circulating lipopolysaccharides (LPS), and a concurrent decrease in circulating sTLR4, suggesting heightened TLR4 activation, internalization, and degradation in response to escalating physiological strain. In vitro, both hypoxia and myotube contractions independently, and in combination, reduced TLR4 expressed on C2C12 myotubes, and these effects were dependent on hypoxia-inducible factor 1 (HIF-1). In summary, the regulation of TLR4 varies depending on the physiological stress during exercise. To our knowledge, our study provides the first evidence of exercise-induced effects on sTLR4 in vivo and highlights the essential role of HIF-1 in the reduction of TLR4 during contraction and hypoxia in vitro.NEW & NOTEWORTHY We provide the first evidence of exercise affecting soluble Toll-like receptor 4 (sTLR4), a TLR4 ligand decoy receptor. We found that the degree of exercise-induced physiological stress influences TLR4 regulation on peripheral blood mononuclear cells (PBMCs). Moderate-intensity exercise reduces PBMC TLR4 and increases circulating sTLR4. Conversely, workload-matched exercise in hypoxia induces greater physiological stress, intestinal barrier damage, circulating lipopolysaccharides, and reduces both TLR4 and sTLR4, suggesting heightened TLR4 activation, internalization, and degradation under increased strain.

Could Orthostatic Stress Responses Predict Acute Mountain Sickness Susceptibility Prior to High Altitude Travel? A Pilot Study.

Bellovary, Bryanne N., Andrew D. Wells, Zachary J. Fennel, Jeremy B. Ducharme, Jonathan M. Houck, Trevor J. Mayschak, Ann L. Gibson, Scott N. Drum, and Christine M. Mermier. Could orthostatic stress responses predict acute mountain sickness susceptibility before high altitude travel? A pilot study. High Alt Med Biol. 24:19-26, 2023. Purpose: This study assessed head-up tilt (HUT) responses in relation to acute mountain sickness (AMS)-susceptibility during hypoxic exposure. Materials and Methods: Fifteen participants completed three lab visits: (1) protocol familiarization and cycle maximal oxygen consumption (VO2max) test; (2) HUT test consisting of supine rest for 20 minutes followed by 70° tilting for ≤40 minutes; and (3) 6 hours of hypobaric hypoxic exposure (4,572 m) where participants performed two 30-minute cycling bouts separated by 1 hour at a 50% VO2max workload within the first 3 hours and rested when not exercising. During HUT, systolic blood pressure (SBP), diastolic blood pressure, heart rate (HR), and variability (blood pressure variability [BPV] and HR variability [HRV]) were measured continuously. The AMS scores were determined after 6 hours of exposure. Correlations determined relationships between HUT cardiovascular responses and AMS scores. Repeated-measures analysis of variance (ANOVA) assessed differences between those with and without AMS symptoms during HUT. Results: Higher AMS scores correlated with greater change in SBP variability (r = 0.52, p = 0.048) and blunted changes in HRV (root mean square of successive differences between normal heartbeats r = 0.81, p = 0.001, percentage of adjacent normal sinus intervals that differ by more than 50 milliseconds [pNN50] r = 0.87, p < 0.001) during HUT. A pNN50 interaction (p = 0.02) suggested elevated cardiac sympathetic activity at baseline and a blunted increase in cardiac sympathetic influence throughout HUT in those with AMS (pNN50 baseline: AMS = 26.2% ± 15.3%, no AMS = 51.0% ± 13.5%; first 3 minutes into HUT: AMS = 17.2% ± 19.1%, no AMS = 17.1% ± 10.9%; end of HUT: AMS = 6.2% ± 9.1%, no AMS 11.0% ± 10.0%). Conclusions: The results suggest autonomic responses via HUT differ in AMS-susceptible individuals. Changes in HRV and BPV during HUT may be a promising predictive measurement for AMS-susceptibility, but further research is needed for confirmation.

Ibuprofen Increases Markers of Intestinal Barrier Injury But Suppresses Inflammation at Rest and After Exercise in Hypoxia.

PURPOSE: The purpose of this study was to evaluate the effects of acute ibuprofen consumption (2 × 600-mg doses) on markers of enterocyte injury, intestinal barrier dysfunction, inflammation, and symptoms of gastrointestinal (GI) distress at rest and after exercise in hypobaric hypoxia. METHODS: Using a randomized double-blind placebo-controlled crossover design, nine men (age, 28 ± 3 yr; weight, 75.4 ± 10.5 kg; height, 175 ± 7 cm; body fat, 12.9% ± 5%; V̇O 2 peak at 440 torr, 3.11 ± 0.65 L·min -1 ) completed a total of three visits including baseline testing and two experimental trials (placebo and ibuprofen) in a hypobaric chamber simulating an altitude of 4300 m. Preexercise and postexercise blood samples were assayed for intestinal fatty acid binding protein (I-FABP), ileal bile acid binding protein, soluble cluster of differentiation 14, lipopolysaccharide binding protein, monocyte chemoattractant protein-1, tumor necrosis factor α (TNF-α), interleukin-1ß, and interleukin-10. Intestinal permeability was assessed using a dual sugar absorption test (urine lactulose-to-rhamnose ratio). RESULTS: Resting I-FABP (906 ± 395 vs 1168 ± 581 pg·mL -1 ; P = 0.008) and soluble cluster of differentiation 14 (1512 ± 297 vs 1642 ± 313 ng·mL -1 ; P = 0.014) were elevated in the ibuprofen trial. Likewise, the urine lactulose-to-rhamnose ratio (0.217 vs 0.295; P = 0.047) and the preexercise to postexercise change in I-FABP (277 ± 308 vs 498 ± 479 pg·mL -1 ; P = 0.021) were greater in the ibuprofen trial. Participants also reported greater upper GI symptoms in the ibuprofen trial ( P = 0.031). However, monocyte chemoattractant protein-1 ( P = 0.007) and TNF-α ( P = 0.047) were lower throughout the ibuprofen trial compared with placebo (main effect of condition). CONCLUSIONS: These data demonstrate that acute ibuprofen ingestion aggravates markers of enterocyte injury and intestinal barrier dysfunction at rest and after exercise in hypoxia. However, ibuprofen seems to suppress circulating markers of inflammation.

Circulating markers of intestinal barrier injury and inflammation following exertion in hypobaric hypoxia.

Hypoxia induced intestinal barrier injury, microbial translocation, and local/systemic inflammation may contribute to high-altitude associated gastrointestinal complications or symptoms of acute mountain sickness (AMS). Therefore, we tested the hypothesis that six-hours of hypobaric hypoxia increases circulating markers of intestinal barrier injury and inflammation. A secondary aim was to determine if the changes in these markers were different between those with and without AMS. Thirteen participants were exposed to six hours of hypobaric hypoxia, simulating an altitude of 4572 m. Participants completed two 30-minute bouts of exercise during the early hours of hypoxic exposure to mimic typical activity required by those at high altitude. Pre- and post-exposure blood samples were assessed for circulating markers of intestinal barrier injury and inflammation. Data below are presented as mean ± standard deviation or median [interquartile range]. Intestinal fatty acid binding protein (Δ251 [103-410] pg•mL-1; p = 0.002, d = 0.32), lipopolysaccharide binding protein (Δ2 ± 2.4 µg•mL-1; p = 0.011; d = 0.48), tumor necrosis factor-α (Δ10.2 [3-42.2] pg•mL-1; p = 0.005; d = 0.25), interleukin-1ß (Δ1.5 [0-6.7] pg•mL-1 p = 0.042; d = 0.18), and interleukin-1 receptor agonist (Δ3.4 [0.4-5.2] pg•mL-1p = 0.002; d = 0.23) increased from pre- to post-hypoxia. Six of the 13 participants developed AMS; however, the pre- to post-hypoxia changes for each marker were not different between those with and without AMS (p > 0.05 for all indices). These data provide evidence that high altitude exposures can lead to intestinal barrier injury, which may be an important consideration for mountaineers, military personnel, wildland firefighters, and athletes who travel to high altitudes to perform physical work or exercise.

Predictive model specific to young adults for estimating thoracic gas volume for air-displacement plethysmography.

BACKGROUND: Thoracic gas volume either measured (mTGV) or predicted by the BodPod® (bpTGV) is used during air-displacement plethysmography to obtain a better estimate of percent body fat. Evidence suggests that bpTGV underestimates mTGV for young adults and this is especially evident for young males. AIMS: We developed, validated, and cross-validated a TGV prediction model (pTGV) for males and females 18-30 years of age to address this underestimation. MATERIALS & METHODS: Participants (N = 181; 18-30 years) that had their body composition assessed with the BodPod® were retrospectively randomly assigned to one of two independent subgroups, a validation (n = 145) or cross-validation (n = 36) sample. Ten iterations of the k-fold validation procedure were performed to assess the internal replicability of pTGV within the validation sample. External replicability of pTGV was evaluated by assessing the difference and standard error of the estimate (SEE) compared to mTGV in the cross-validation group. RESULTS: The model using height, sex and body mass yielded the highest adjusted R2 (0.627) and the lowest SEE (0.56 L): pTGV = 0.615338 × Sex (0 = Female, 1 = Male) + 0.056267 × Height (cm) - 0.011006 × Body Mass (kg) - 5.358839. R2 remained stable across 10 iterations of the k-fold procedure (average R2 = 0.64). Differences between pTGV and mTGV were not significantly different than zero for the total cross-validation sample (-0.06 ± 0.7 L; SEE = 3.0%), for males (-0.11 ± 0.7 L; SEE = 3.7%), or for females (-0.02 ± 0.7 L; SEE = 5.3%). CONCLUSION: We recommend that when it is impractical to obtain mTGV, the strong internal and external replicability of the new prediction model supports its use for males and females ages 18-30 years old during air-displacement plethysmography.

Worth the Wait? Time Course of Supine Shifts in Body Water Compartments on Variables of Bioelectrical Impedance Analysis.

Bioelectrical impedance analysis (BIA) reference values are based on supine assessments. Little is known regarding the effects of time course shifts in body water compartments after assuming a supine position. The aim of this study was to characterize these effects and provide recommendations regarding the optimal waiting time to perform BIA. Thirty-eight healthy adults underwent BIA via the RJL Quantum Legacy analyzer immediately upon lying down and every 5 minutes for 15 minutes. Differences in resistance (R), reactance (Xc), intracellular (ICW), extracellular (ECW), total body water (TBW), body fat percentage (%BF), and phase angle (PhA) were assessed. There were small but significant increases in R, Xc, and %BF (all p<0.001), as well as small but significant decreases in ICW, ECW, and TBW (all p<0.001) over 15 minutes. No difference was observed for PhA (p=0.065). Average values changed over 15 minutes by +7.14Ω, +1.36Ω, -0.2L, -0.2L, -0.4L, +0.05° and +0.1% for R, Xc, ICW, ECW, TBW, PhA and %BF, respectively. BIA measurements are affected by shifts in body water compartments after assuming a supine position, but these differences lack clinical significance in healthy adults. Technicians working with healthy adults can perform BIA within 15 minutes after participants assume a supine position.