Cryo-EM of soft-landed ß-galactosidase: Gas-phase and native structures are remarkably similar.

Native mass spectrometry (MS) has become widely accepted in structural biology, providing information on stoichiometry, interactions, homogeneity, and shape of protein complexes. Yet, the fundamental assumption that proteins inside the mass spectrometer retain a structure faithful to native proteins in solution remains a matter of intense debate. Here, we reveal the gas-phase structure of ß-galactosidase using single-particle cryo-electron microscopy (cryo-EM) down to 2.6-Å resolution, enabled by soft landing of mass-selected protein complexes onto cold transmission electron microscopy (TEM) grids followed by in situ ice coating. We find that large parts of the secondary and tertiary structure are retained from the solution. Dehydration-driven subunit reorientation leads to consistent compaction in the gas phase. By providing a direct link between high-resolution imaging and the capability to handle and select protein complexes that behave problematically in conventional sample preparation, the approach has the potential to expand the scope of both native mass spectrometry and cryo-EM.

Accuracy of a smart bottle in measuring fluid intake by American football players during pre-season training.

Rehydration is important for athlete performance and recovery. However, it can be challenging to follow appropriate fluid replacement practices due in part to difficulties in tracking fluid intake in real time. The purpose of this study was to determine the accuracy of a smart bottle in measuring fluid intake during exercise. Thirty male American football athletes drank from bottles equipped with a smart cap during outdoor pre-season practices (110 ± 30 min; 29.3 ± 3.0 °C; 75 ± 11% rh). The cap technology included optical sensors, microprocessors, batteries, and wireless connectivity that transmitted fluid volume data to a smartphone application in real-time. Reference measurements of fluid intake from the smart bottle were determined by gravimetry followed by conversion to volume using the density of the fluid consumed. There was no significant mean difference in fluid intake between the smart bottle and reference method (1220 ± 371 ml vs. 1236 ± 389 ml, p = 0.39 paired t test). Bland-Altman 95% limits of agreement between methods was - 212 to 180 ml. The smart bottle provided accurate measurements of fluid intake during exercise in real-world field conditions on a group level and within limits of agreement of - 212 to 180 ml (or approximately ± 15% of overall fluid intake) on an individual level.

The roles of child temperament, parent stress, and parenting style in family mealtimes.

Family mealtimes are associated with benefits for children, including healthy eating, fewer behavior problems, and healthy psychological well-being. However, the interactions during family mealtimes, and the parent and child characteristics, which may affect both the family mealtime environment and the associated benefits in children are not fully understood. The goal of this study was to examine the role of child and parent characteristics on the family mealtime environment. We tested several mediation models to explain how child temperament (negative affectivity), parent stress, and the dimensions of parent feeding style (responsiveness and demandingness) interact and influence each other to impact the structure and quality of the mealtime environment. Parents (68 mothers; 82 fathers) of children between 2 and 6 years completed an online survey. Measures included the Children's Behavior Questionnaire, Perceived Stress Scale, Caregiver's Feeding Styles Questionnaire, and The Meals in Our Household Questionnaire. Child negative affectivity was associated with poorer mealtime quality and structure. These associations were mediated through parent responsiveness, but not demandingness. The role of demandingness in family mealtimes may depend on parent responsiveness. When examined together in a serial mediation model, child negative affectivity increased parent stress, which reduced responsiveness, and led to poorer mealtime quality and structure. These results emphasize the complex relationships between child temperament, parent stress, and the dimensions of parenting styles that occur within the mealtime context. This line of research is essential for understanding family mealtime dynamics and informing future studies aimed at creating positive interactions between parents and children during mealtimes.

Regional and time course differences in sweat cortisol, glucose, and select cytokine concentrations during exercise.

INTRODUCTION: The use of sweat as a biofluid for non-invasive sampling and diagnostics is a popular area of research. However, concentrations of cortisol, glucose, and cytokines have not been described across anatomical regions or as time progresses throughout exercise. PURPOSE: To determine regional and time course differences in sweat cortisol, glucose, and select cytokines (EGF, IFN-γ, IL-1ß, IL-1α, IL-1ra, TNF-α, IL-6, IL-8, and IL-10). METHODS: Sweat was collected with absorbent patches from eight subjects (24-44 y; 80.2 ± 10.2 kg) on the forehead (FH), right dorsal forearm (RDF), right scapula (RS), and right triceps (RT) at 0-25 min, 30-55 min, and 60-85 min during 90 min of cycling (~ 82% HRmax) in a heated chamber (32 °C, 50% rh). ANOVA was used to determine the effect of site and time on outcomes. Data are reported as LS means ± SE. RESULTS: There was a significant effect of location on sweat analyte concentrations with FH having higher values than most other regions for cortisol (FH: 1.15 ± 0.08 ng/mL > RDF: 0.62 ± 0.09 ng/mL and RT: 0.65 ± 0.12 ng/mL, P = 0.02), IL-1ra (P < 0.0001), and IL-8 (P < 0.0001), but lower concentrations for glucose (P = 0.01), IL-1α (P < 0.0001), and IL-10 (P = 0.02). Sweat IL-1ß concentration was higher on the RS than RT (P < 0.0001). Sweat cortisol concentration increased (25 min: 0.34 ± 0.10 ng/mL < 55 min: 0.89 ± 0.07 ng/mL < 85 min: 1.27 ± 0.07 ng/mL; P < 0.0001), while EGF (P < 0.0001), IL-1ra (P < 0.0001), and IL-6 (P = 0.02) concentrations decreased over time. CONCLUSION: Sweat analyte concentrations varied with time of sampling and anatomical region, which is essential information to consider when conducting future work in this area. CLINICAL TRIAL IDENTIFIER: NCT04240951 registered January 27, 2020.

Uncovering the Role of N-Glycan Occupancy on the Cooperative Assembly of Spike and Angiotensin Converting Enzyme 2 Complexes: Insights from Glycoengineering and Native Mass Spectrometry.

Interactions between the SARS-CoV-2 Spike protein and ACE2 are one of the most scrutinized reactions of our time. Yet, questions remain as to the impact of glycans on mediating ACE2 dimerization and downstream interactions with Spike. Here, we address these unanswered questions by combining a glycoengineering strategy with high-resolution native mass spectrometry (MS) to investigate the impact of N-glycan occupancy on the assembly of multiple Spike-ACE2 complexes. We confirmed that intact Spike trimers have all 66 N-linked sites occupied. For monomeric ACE2, all seven N-linked glycan sites are occupied to various degrees; six sites have >90% occupancy, while the seventh site (Asn690) is only partially occupied (∼30%). By resolving the glycoforms on ACE2, we deciphered the influence of each N-glycan on ACE2 dimerization. Unexpectedly, we found that Asn432 plays a role in mediating dimerization, a result confirmed by site-directed mutagenesis. We also found that glycosylated dimeric ACE2 and Spike trimers form complexes with multiple stoichiometries (Spike-ACE2 and Spike2-ACE2) with dissociation constants (Kds) of ∼500 and <100 nM, respectively. Comparing these values indicates that positive cooperativity may drive ACE2 dimers to complex with multiple Spike trimers. Overall, our results show that occupancy has a key regulatory role in mediating interactions between ACE2 dimers and Spike trimers. More generally, since soluble ACE2 (sACE2) retains an intact SARS-CoV-2 interaction site, the importance of glycosylation in ACE2 dimerization and the propensity for Spike and ACE2 to assemble into higher oligomers are molecular details important for developing strategies for neutralizing the virus.

Explaining variation in sweat sodium concentration: effect of individual characteristics and exercise, environmental, and dietary factors.

This study determined the relative importance of several individual characteristics and dietary, environmental, and exercise factors in determining sweat [Na+] during exercise. Data from 1944 sweat tests were compiled for a retrospective analysis. Stepwise multiple regression (P < 0.05 threshold for inclusion) and T values were used to express the relative importance of each factor in a model. Three separate models were developed based on available independent variables: model 1 (1,944 sweat tests from 1,304 subjects); model 2 (subset with energy expenditure: 1,003 sweat tests from 607 subjects); model 3 (subset with energy expenditure, dietary sodium, and V̇o2max: n = 48). Whole body sweat [Na+] was predicted from forearm sweat patches in models 1 and 2 and directly measured using whole body washdown in model 3. There were no significant effects of age group, race/ethnicity, relative humidity, exercise duration, pre-exercise urine specific gravity, exercise fluid balance, or dietary or exercise sodium intake on any model. Significant predictors in model 1 (adjusted r2 = 0.17, P < 0.001) were season of the year (warm, T = -6.8), exercise mode (cycling, T = 6.8), sex (male, T = 4.9), whole body sweating rate (T = 4.5), and body mass (T = -3.0). Significant predictors in model 2 (adjusted r2 = 0.19, P < 0.001) were season of the year (warm, T = -5.2), energy expenditure (T = 4.7), exercise mode (cycling, T = 3.6), air temperature (T = 3.0), and sex (male, T = 2.7). The only significant predictor in model 3 (r2 = 0.23, P < 0.001) was energy expenditure (T = 3.8). In summary, the models accounted for 17%-23% of the variation in whole body sweat [Na+] and energy expenditure and season of the year (proxy for heat acclimatization) were the most important factors.NEW & NOTEWORTHY This comprehensive analysis of a large, diverse data set contributes to our overall understanding of the factors that influence whole body sweat [Na+]. The main finding was that energy expenditure was directly associated with whole body sweat [Na+], potentially via the relation between energy expenditure and whole body sweating rate (WBSR). Warmer months (proxy for heat acclimatization) were associated with lower whole body sweat [Na+]. Exercise mode, air temperature, and sex may also have small effects, but other variables (age group, race/ethnicity, fluid balance, sodium intake, relative V̇o2max) had no association with whole body sweat [Na+]. Taken together, the models explained 17%-23% of the variation in whole body sweat [Na+].

Multiple regression analyses to determine the effect of sweating rate and tattoo characteristics on sweat outcome measures during exercise.

PURPOSE: To compare local sweating rate (LSR) and local sweat sodium ([Na+]), chloride ([Cl-]), and potassium ([K+]) concentrations of tattooed skin and contralateral non-tattooed skin during exercise. METHODS: Thirty-three recreational exercisers (17 men, 16 women) with ≥ 1 unilateral permanent tattoo on the torso/arms were tested during cycling, running, or fitness sessions (26 ± 4 °C and 54 ± 13% relative humidity). Forty-eight tattoos with a range of ink colors, ages (3 weeks to 20 years), and densities (10-100%) were included. Before exercise, the skin was cleaned with alcohol and patches (3 M Tegaderm + Pad) were placed on the tattooed and contralateral non-tattooed skin. LSR was calculated from sweat mass (0.80 ± 0.31 g), patch surface area (11.9 cm2), and duration (62 ± 14 min). Sweat [Na+], [Cl-], and [K+] were measured via ion chromatography. RESULTS: Based on the analysis of variance results, there were no differences between tattooed and non-tattooed skin for LSR (1.16 ± 0.52 vs. 1.12 ± 0.53 mg/cm2/min; p = 0.51), sweat [Na+] (60.2 ± 23.5 vs. 58.5 ± 22.7 mmol/L; p = 0.27), sweat [Cl-] (52.1 ± 22.4 vs. 50.6 ± 22.0 mmol/L; p = 0.31), or sweat [K+] (5.8 ± 1.6 vs. 5.9 ± 1.4 mmol/L; p = 0.31). Multiple regression analyses suggested that younger tattoos were associated with higher sweat [Na+] (p = 0.045) and colorful tattoos were associated with higher sweat [Cl-] (p = 0.04) compared with contralateral non-tattooed skin. Otherwise, there were no effects of LSR or tattoo characteristics on regression models for LSR or sweat electrolyte concentrations. CONCLUSION: There were no effects of tattoos on LSR and sweat [K+] during exercise-induced sweating, but tattoo age and color had small effects on sweat [Na+] and sweat [Cl-], respectively. CLINICAL TRIAL IDENTIFIERS: NCT04240951 was registered on January 27, 2020 and NCT04920266 was registered on June 9, 2021.

Sweating Rate and Sweat Chloride Concentration of Elite Male Basketball Players Measured With a Wearable Microfluidic Device Versus the Standard Absorbent Patch Method.

The purpose of this study was to compare a wearable microfluidic device and standard absorbent patch in measuring local sweating rate (LSR) and sweat chloride concentration ([Cl-]) in elite basketball players. Participants were 53 male basketball players (25 ± 3 years, 92.2 ± 10.4 kg) in the National Basketball Association's development league. Players were tested during a moderate-intensity, coach-led practice (98 ± 30 min, 21.0 ± 1.2 °C). From the right ventral forearm, sweat was collected using an absorbent patch (3M Tegaderm™ + Pad). Subsequently, LSR and local sweat [Cl-] were determined via gravimetry and ion chromatography. From the left ventral forearm, LSR and local sweat [Cl-] were measured using a wearable microfluidic device and associated smartphone application-based algorithms. Whole-body sweating rate (WBSR) was determined from pre- to postexercise change in body mass corrected for fluid/food intake (ad libitum), urine loss, and estimated respiratory water and metabolic mass loss. The WBSR values predicted by the algorithms in the smartphone application were also recorded. There were no differences between the absorbent patch and microfluidic patch for LSR (1.25 ± 0.91 mg·cm-2·min-1 vs. 1.14 ±0.78 mg·cm-2·min-1, p = .34) or local sweat [Cl-] (30.6 ± 17.3 mmol/L vs. 29.6 ± 19.4 mmol/L, p = .55). There was no difference between measured and predicted WBSR (0.97 ± 0.41 L/hr vs. 0.89 ± 0.35 L/hr, p = .22; 95% limits of agreement = 0.61 L/hr). The wearable microfluidic device provides similar LSR, local sweat [Cl-], and WBSR results compared with standard field-based methods in elite male basketball players during moderate-intensity practices.

Mass-selective and ice-free electron cryomicroscopy protein sample preparation via native electrospray ion-beam deposition.

Despite tremendous advances in sample preparation and classification algorithms for electron cryomicroscopy (cryo-EM) and single-particle analysis (SPA), sample heterogeneity remains a major challenge and can prevent access to high-resolution structures. In addition, optimization of preparation conditions for a given sample can be time-consuming. In the current work, it is demonstrated that native electrospray ion-beam deposition (native ES-IBD) is an alternative, reliable approach for the preparation of extremely high-purity samples, based on mass selection in vacuum. Folded protein ions are generated by native electrospray ionization, separated from other proteins, contaminants, aggregates, and fragments, gently deposited on cryo-EM grids, frozen in liquid nitrogen, and subsequently imaged by cryo-EM. We demonstrate homogeneous coverage of ice-free cryo-EM grids with mass-selected protein complexes. SPA reveals that the complexes remain folded and assembled, but variations in secondary and tertiary structures are currently limiting information in 2D classes and 3D EM density maps. We identify and discuss challenges that need to be addressed to obtain a resolution comparable to that of the established cryo-EM workflow. Our results show the potential of native ES-IBD to increase the scope and throughput of cryo-EM for protein structure determination and provide an essential link between gas-phase and solution-phase protein structures.

In-Season Nutrition Strategies and Recovery Modalities to Enhance Recovery for Basketball Players: A Narrative Review.

Basketball players face multiple challenges to in-season recovery. The purpose of this article is to review the literature on recovery modalities and nutritional strategies for basketball players and practical applications that can be incorporated throughout the season at various levels of competition. Sleep, protein, carbohydrate, and fluids should be the foundational components emphasized throughout the season for home and away games to promote recovery. Travel, whether by air or bus, poses nutritional and sleep challenges, therefore teams should be strategic about packing snacks and fluid options while on the road. Practitioners should also plan for meals at hotels and during air travel for their players. Basketball players should aim for a minimum of 8 h of sleep per night and be encouraged to get extra sleep during congested schedules since back-to back games, high workloads, and travel may negatively influence night-time sleep. Regular sleep monitoring, education, and feedback may aid in optimizing sleep in basketball players. In addition, incorporating consistent training times may be beneficial to reduce bed and wake time variability. Hydrotherapy, compression garments, and massage may also provide an effective recovery modality to incorporate post-competition. Future research, however, is warranted to understand the influence these modalities have on enhancing recovery in basketball players. Overall, a strategic well-rounded approach, encompassing both nutrition and recovery modality strategies, should be carefully considered and implemented with teams to support basketball players' recovery for training and competition throughout the season.

Predicted sweat rates for group water planning in sport: accuracy and application.

This study tested the accuracy of a novel, limited-availability web application (H2Q™) for predicting sweat rates in a variety of sports using estimates of energy expenditure and air temperature only. The application of predictions for group water planning was investigated for soccer match play. Fourteen open literature studies were identified where group sweat rates were reported (n = 20 group means comprising 230 individual observations from 179 athletes) with fidelity. Sports represented included: walking, cycling, swimming, and soccer match play. The accuracy of H2Q™ sweat rates was tested by comparing to measured group sweat rates using the concordance correlation coefficient (CCC) with 95% confidence interval [CI]. The relative absolute error (RAE) with 95% [CI] was also assessed, whereby the mean absolute error was expressed relative to an acceptance limit of 0.250 L/h. The CCC was 0.98 [0.95, 0.99] and the RAE was 0.449 [0.279, 0.620], indicating that the prediction error was on average 0.112 L/h. The RAE was < 1.0 for 19/20 observations (95%). Drink volumes modeled as a proxy for sweat losses during soccer match play prevented dehydration (< 1% loss of body mass). The H2Q™ web application demonstrated high group sweat prediction accuracy for the variety of sports activities tested. Water planning for soccer match play suggests the feasibility of easily and accurately predicting sweat rates to plan group water needs and promote optimal hydration in training and/or competition.

Nutritional considerations to counteract gastrointestinal permeability during exertional heat stress.

Intestinal barrier integrity and function are compromised during exertional heat stress (EHS) potentially leading to consequences that range from minor gastrointestinal (GI) disturbances to fatal outcomes in exertional heat stroke or septic shock. This mini-review provides a concise discussion of nutritional interventions that may protect against intestinal permeability during EHS and suggests physiological mechanisms responsible for this protection. Although diverse nutritional interventions have been suggested to be protective against EHS-induced GI permeability, the ingestion of certain amino acids, carbohydrates, and fluid per se is potentially effective strategy, whereas evidence for various polyphenols and pre/probiotics is developing. Plausible physiological mechanisms of protection include increased blood flow, epithelial cell proliferation, upregulation of intracellular heat shock proteins, modulation of inflammatory signaling, alteration of the GI microbiota, and increased expression of tight junction (TJ) proteins. Further clinical research is needed to propose specific nutritional candidates and recommendations for their application to prevent intestinal barrier disruption and elucidate mechanisms during EHS.

Re-envisioning paradigms of education: towards awareness, alignment, and pluralism.

In this article we introduce a synthesis of education "paradigms," adapted from a multi-disciplinary body of literature and tailored to health professions education (HPE). Each paradigm involves a particular perspective on the purpose of education, the nature of knowledge, what knowledge is valued and included in the curriculum, what it means to learn and how learning is assessed, and the roles of teachers and learners in the learning process. We aim to foster awareness of how these different paradigms look in practice and to illustrate the importance of alignment between teaching, learning and assessment practices with paradigmatic values and assumptions. Finally, we advocate for a pluralistic approach that purposefully and meaningfully integrates paradigms of education, enhancing our ability to drive quality in HPE.

DNA origami signposts for identifying proteins on cell membranes by electron cryotomography.

Electron cryotomography (cryoET), an electron cryomicroscopy (cryoEM) modality, has changed our understanding of biological function by revealing the native molecular details of membranes, viruses, and cells. However, identification of individual molecules within tomograms from cryoET is challenging because of sample crowding and low signal-to-noise ratios. Here, we present a tagging strategy for cryoET that precisely identifies individual protein complexes in tomograms without relying on metal clusters. Our method makes use of DNA origami to produce "molecular signposts" that target molecules of interest, here via fluorescent fusion proteins, providing a platform generally applicable to biological surfaces. We demonstrate the specificity of signpost origami tags (SPOTs) in vitro as well as their suitability for cryoET of membrane vesicles, enveloped viruses, and the exterior of intact mammalian cells.

Fluid Balance, Sweat Na+ Losses, and Carbohydrate Intake of Elite Male Soccer Players in Response to Low and High Training Intensities in Cool and Hot Environments.

Hypohydration increases physiological strain and reduces physical and technical soccer performance, but there are limited data on how fluid balance responses change between different types of sessions in professional players. This study investigated sweat and fluid/carbohydrate intake responses in elite male professional soccer players training at low and high intensities in cool and hot environments. Fluid/sodium (Na+) losses and ad-libitum carbohydrate/fluid intake of fourteen elite male soccer players were measured on four occasions: cool (wet bulb globe temperature (WBGT): 15 ± 7 °C, 66 ± 6% relative humidity (RH)) low intensity (rating of perceived exertion (RPE) 2-4, m·min-1 40-46) (CL); cool high intensity (RPE 6-8, m·min-1 82-86) (CH); hot (29 ± 1 °C, 52 ± 7% RH) low intensity (HL); hot high intensity (HH). Exercise involved 65 ± 5 min of soccer-specific training. Before and after exercise, players were weighed in minimal clothing. During training, players had ad libitum access to carbohydrate beverages and water. Sweat [Na+] (mmol·L-1), which was measured by absorbent patches positioned on the thigh, was no different between conditions, CL: 35 ± 9, CH: 38 ± 8, HL: 34 ± 70.17, HH: 38 ± 8 (p = 0.475). Exercise intensity and environmental condition significantly influenced sweat rates (L·h-1), CL: 0.55 ± 0.20, CH: 0.98 ± 0.21, HL: 0.81 ± 0.17, HH: 1.43 ± 0.23 (p =0.001), and percentage dehydration (p < 0.001). Fluid intake was significantly associated with sweat rate (p = 0.019), with no players experiencing hypohydration > 2% of pre-exercise body mass. Carbohydrate intake varied between players (range 0-38 g·h-1), with no difference between conditions. These descriptive data gathered on elite professional players highlight the variation in the hydration status, sweat rate, sweat Na+ losses, and carbohydrate intake in response to training in cool and hot environments and at low and high exercise intensities.

Skin-interfaced microfluidic system with personalized sweating rate and sweat chloride analytics for sports science applications.

Advanced capabilities in noninvasive, in situ monitoring of sweating rate and sweat electrolyte losses could enable real-time personalized fluid-electrolyte intake recommendations. Established sweat analysis techniques using absorbent patches require post-collection harvesting and benchtop analysis of sweat and are thus impractical for ambulatory use. Here, we introduce a skin-interfaced wearable microfluidic device and smartphone image processing platform that enable analysis of regional sweating rate and sweat chloride concentration ([Cl-]). Systematic studies (n = 312 athletes) establish significant correlations for regional sweating rate and sweat [Cl-] in a controlled environment and during competitive sports under varying environmental conditions. The regional sweating rate and sweat [Cl-] results serve as inputs to algorithms implemented on a smartphone software application that predicts whole-body sweating rate and sweat [Cl-]. This low-cost wearable sensing approach could improve the accessibility of physiological insights available to sports scientists, practitioners, and athletes to inform hydration strategies in real-world ambulatory settings.

Cross-validation of equations to predict whole-body sweat sodium concentration from regional measures during exercise.

We have previously published equations to estimate whole-body (WB) sweat sodium concentration ([Na+ ]) from regional (REG) measures; however, a cross-validation is needed to corroborate the applicability of these prediction equations between studies. The purpose of this study was to determine the validity of published equations in predicting WB sweat [Na+ ] from REG measures when applied to a new data set. Forty-nine participants (34 men, 15 women; 75 ± 12 kg) cycled for 90 min while WB sweat [Na+ ] was measured using the washdown technique. REG sweat [Na+ ] was measured from seven regions using absorbent patches (3M Tegaderm + Pad). Published equations were applied to REG sweat [Na+ ] to determine predicted WB sweat [Na+ ]. Bland-Altman analysis of mean bias (raw and predicted minus measured) and 95% limits of agreement (LOA) were used to compare raw (uncorrected) REG sweat [Na+ ] and predicted WB sweat [Na+ ] to measured WB sweat [Na+ ]. Mean bias (±95% LOA) between raw REG sweat [Na+ ] and measured WB sweat [Na+ ] was 10(±20), 0(±19), 9(±20), 22(±25), 23(±24), 0(±15), -4(±18) mmol/L for the dorsal forearm, ventral forearm, upper arm, chest, upper back, thigh, and calf, respectively. The mean bias (±95% LOA) between predicted WB sweat [Na+ ] and measured WB sweat [Na+ ] was 3(±14), 4(±12), 0(±14), 2(±17), -2(±16), 5(±13), 4(±15) mmol/L for the dorsal forearm, ventral forearm, upper arm, chest, upper back, thigh, and calf, respectively. Prediction equations improve the accuracy of estimating WB sweat [Na+ ] from REG and are therefore recommended for use when determining individualized sweat electrolyte losses.

Cellular Electron Cryo-Tomography to Study Virus-Host Interactions.

Viruses are obligatory intracellular parasites that reprogram host cells upon infection to produce viral progeny. Here, we review recent structural insights into virus-host interactions in bacteria, archaea, and eukaryotes unveiled by cellular electron cryo-tomography (cryoET). This advanced three-dimensional imaging technique of vitreous samples in near-native state has matured over the past two decades and proven powerful in revealing molecular mechanisms underlying viral replication. Initial studies were restricted to cell peripheries and typically focused on early infection steps, analyzing surface proteins and viral entry. Recent developments including cryo-thinning techniques, phase-plate imaging, and correlative approaches have been instrumental in also targeting rare events inside infected cells. When combined with advances in dedicated image analyses and processing methods, details of virus assembly and egress at (sub)nanometer resolution were uncovered. Altogether, we provide a historical and technical perspective and discuss future directions and impacts of cryoET for integrative structural cell biology analyses of viruses.

Reducing Pediatric Tracheostomy Wound Complications: An Evidence-Based Literature Review.

OBJECTIVE: To discuss prevention strategies that can mitigate the frequency of tracheostomy-related wound complications. DATA SOURCES: A systematic literature review of PubMed between 2010 and 2019. STUDY SELECTION: Full-text articles written in English language and studying human participants younger than 18 years. DATA EXTRACTION: The primary outcome was the rate of tracheostomy-related skin complications after implementation of a given intervention. Secondary outcomes included rates of accidental decannulation or other complications. DATA SYNTHESIS: A total of 348 studies were identified and 6 met inclusion criteria. There were 1,607 children included with interventions designed to reduce peristomal and cervical wound complications in 1,174 (73.1%). Strategies considered safe and effective included protective skin barriers, reducing prolonged pressure, and early wound identification protocols. CONCLUSIONS: Wound complications after pediatric tracheostomy can be reduced using a multifaceted approach by providers committed to making skin care a priority.

Permanent tattooing has no impact on local sweat rate, sweat sodium concentration and skin temperature or prediction of whole-body sweat sodium concentration during moderate-intensity cycling in a warm environment.

PURPOSES: This study investigated the impact of permanently tattooed skin on local sweat rate, sweat sodium concentration and skin temperature and determined whether tattoos alter the relationship between local and whole-body sweat sodium concentration. METHODS: Thirteen tattooed men (27 ± 6 years) completed a 1 h (66 ± 4% of [Formula: see text]) cycling trial at 32 °C, 35% relative humidity. Sweat rate and sweat sodium concentration were measured using the whole-body washdown and local absorbent patch techniques. Patches and skin-temperature probes were applied over the right/left thighs and tattooed/non-tattooed (contralateral) regions. RESULTS: Local sweat rates did not differ (p > 0.05) between the right (1.11 ± 0.38) and left (1.21 ± 0.37) thighs and the permanently tattooed (1.93 ± 0.82) and non-tattooed (1.72 ± 0.81 mg cm-2 min-1) regions. There were no differences in local sweat sodium concentration between the right (58.2 ± 19.4) and left (55.4 ± 20.3) thighs and the permanently tattooed (73.0 ± 22.9) and non-tattooed (70.2 ± 18.9 mmol L-1) regions. Difference in local skin temperature between the right and left thighs (- 0.043) was similar to that between the permanently tattooed and non-tattooed (- 0.023 °C) regions. Prediction of whole-body sweat sodium concentration for the permanently tattooed (41.0 ± 6.7) and the non-tattooed (40.2 ± 5.3 mmol L-1) regions did not differ. CONCLUSION: Permanent tattoos do not alter local sweat rate, sweat sodium concentration or local skin temperature during moderate-intensity cycling exercise in a warm environment. Results from a patch placed over a tattooed surface correctly predicts whole-body sweat sodium concentration from an equation developed from a non-tattooed region.