Isolation and characterization of IgG3 glycan-targeting antibodies with exceptional cross-reactivity for diverse viral families.

Broadly reactive antibodies that target sequence-diverse antigens are of interest for vaccine design and monoclonal antibody therapeutic development because they can protect against multiple strains of a virus and provide a barrier to evolution of escape mutants. Using LIBRA-seq (linking B cell receptor to antigen specificity through sequencing) data for the B cell repertoire of an individual chronically infected with human immunodeficiency virus type 1 (HIV-1), we identified a lineage of IgG3 antibodies predicted to bind to HIV-1 Envelope (Env) and influenza A Hemagglutinin (HA). Two lineage members, antibodies 2526 and 546, were confirmed to bind to a large panel of diverse antigens, including several strains of HIV-1 Env, influenza HA, coronavirus (CoV) spike, hepatitis C virus (HCV) E protein, Nipah virus (NiV) F protein, and Langya virus (LayV) F protein. We found that both antibodies bind to complex glycans on the antigenic surfaces. Antibody 2526 targets the stem region of influenza HA and the N-terminal domain (NTD) region of SARS-CoV-2 spike. A crystal structure of 2526 Fab bound to mannose revealed the presence of a glycan-binding pocket on the light chain. Antibody 2526 cross-reacted with antigens from multiple pathogens and displayed no signs of autoreactivity. These features distinguish antibody 2526 from previously described glycan-reactive antibodies. Further study of this antibody class may aid in the selection and engineering of broadly reactive antibody therapeutics and can inform the development of effective vaccines with exceptional breadth of pathogen coverage.

Development of LIBRA-seq for the guinea pig model system as a tool for the evaluation of antibody responses to multivalent HIV-1 vaccines.

Consistent elicitation of serum antibody responses that neutralize diverse clades of HIV-1 remains a primary goal of HIV-1 vaccine research. Prior work has defined key features of soluble HIV-1 Envelope (Env) immunogen cocktails that influence the neutralization breadth and potency of multivalent vaccine-elicited antibody responses including the number of Env strains in the regimen. We designed immunization groups that consisted of different numbers of SOSIP Env strains to be used in a cocktail immunization strategy: the smallest cocktail (group 2) consisted of a set of two Env strains, which were a subset of the three Env strains that made up group 3, which, in turn, were a subset of the six Env strains that made up group 4. Serum neutralizing titers were modestly broader in guinea pigs that were immunized with a cocktail of three Envs compared to cocktails of two and six, suggesting that multivalent Env immunization could provide a benefit but may be detrimental when the cocktail size is too large. We then adapted the LIBRA-seq platform for antibody discovery to be compatible with guinea pigs, and isolated several tier 2 neutralizing monoclonal antibodies. Three antibodies isolated from two separate guinea pigs were similar in their gene usage and CDR3s, establishing evidence for a guinea pig public clonotype elicited through vaccination. Taken together, this work investigated multivalent HIV-1 Env immunization strategies and provides a novel methodology for screening guinea pig B cell receptor antigen specificity at a high-throughput level using LIBRA-seq.IMPORTANCEMultivalent vaccination with soluble Env immunogens is at the forefront of HIV-1 vaccination strategies but little is known about the influence of the number of Env strains included in vaccine cocktails. Our results suggest that adding more strains is sometimes beneficial but may be detrimental when the number of strains is too high. In addition, we adapted the LIBRA-seq platform to be compatible with guinea pig samples and isolated several tier 2 neutralizing monoclonal antibodies, some of which share V and J gene usage and >70% CDR3 identity, thus establishing the existence of public clonotypes in guinea pigs elicited through vaccination.

Development of an Aptamer-Based Electrochemical Microfluidic Device for Viral Vaccine Quantitation.

Global deployment of vaccines poses significant challenges in the distribution and use of the accompanying immunoassays, one of the standard methods for quality control of vaccines, particularly when establishing assays in countries worldwide to support testing/release upon importation. This work describes our effort toward developing an integrated, portable device to carry out affinity assays for viral particles quantification in viral vaccines by incorporating (i) aptamers, (ii) microfluidic devices, and (iii) electrochemical detection. We generated and characterized more than eight aptamers against multiple membrane proteins of cytomegalovirus (CMV), which we used as a model system and designed and fabricated electrochemical microfluidic devices to measure CMV concentrations in a candidate vaccine under development. The aptamer-based assays provided a half maximal effective concentration, EC50, of 12 U/mL, comparable to that of an ELISA using a pair of antibodies (EC50 60 U/mL). The device measured relative CMV concentrations accurately (within ±10% bias) and precisely (11%, percent relative standard deviation). This work represents the critical first steps toward developing simple, affordable, and robust affinity assays for global deployment without the need for sensitive equipment and extensive analyst training.

Comparison of VO2peak from the Progressive Aerobic Cardiovascular Endurance Run (PACER) and treadmill in children.

Background/Objectives: The purpose of this study was to investigate the difference in peak oxygen consumption (VO2peak) during a graded treadmill test and the Progressive Aerobic Cardiovascular Endurance Run (PACER) in a sample of 7-14 year old children. Methods: Forty-four participants (25 boys, 19 girls) had VO2 assessed during a peak treadmill test and the PACER by a portable indirect calorimeter on non-consecutive days. Exercise parameters were compared between exercise tests by paired t-tests. Results: The PACER elicited a greater measured VO2peak (49.4 ± 9.4 vs. 46.7 ± 7.5 ml. kg-1·min-1) and maximum respiratory exchange ratio (1.14 ± 0.08 vs. 1.07 ± 0.08) than the treadmill test (p < 0.05). Rating of perceived exertion was higher (8.1 ± 3.5 vs 7.6 ± 3.8) during the treadmill test compared to the PACER test (p < 0.05). There was no difference in maximum heart rate between treadmill test and PACER test (196.9 ± 9.3 vs. 198.6 ± 8.8, p > 0.05). Conclusions: The PACER provides an acceptable measure of cardiorespiratory fitness in children but the finding that children elicit a higher measured VO2peak during the PACER compared to a graded treadmill test warrants continual refinement in future aerobic fitness prediction equations from the PACER.

Factors associated with nurses emotional distress during the COVID-19 pandemic.

BACKGROUND: Nurses are among the frontline healthcare workers directly impacted by the burden of the coronavirus disease of 2019 (COVID-19) pandemic. This study aimed to examine the prevalence of emotional distress and the associated factors among nurses practicing in South Dakota during the COVID-19 pandemic. METHODS: An online survey was conducted among practicing, licensed nurses in South Dakota during the pandemic (July 2020 - August 2020). Emotional distress was measured using the Depression, Anxiety, and Stress Scale (DASS-21). Logistic regression models were performed to examine the association of emotional distress and the three DASS-21 subscales with: sociodemographic and work environment factors (e.g., work setting, job satisfaction, number of COVID-19 cases seen at the facility, preparedness, concerns with worsening pre-exiting mental health conditions due to the pandemic, and contracting the illness). RESULTS: Among 1505 participants, overall emotional distress was reported by 22.2%, while anxiety, depression and stress were 15.8%, 14.5% and 11.9%, respectively. Factors associated with moderate to severe emotional distress, depression, anxiety, and stress were as follows: concerns for worsening of pre-existing mental health conditions, job dissatisfaction, encountering higher number of COVID-19 cases at one's work facility, feeling unprepared for the pandemic, and concern for contracting the illness (all p < 0.05). CONCLUSIONS: Our study suggests a high prevalence of emotional distress among nurses and highlights the factors associated with emotional distress during the COVID-19 pandemic. Promoting appropriate support is imperative to reduce nurses' emotional distress and promote psychological well-being during the COVID-19 world health crisis and in future pandemics.

Combined Sprint Interval, Plyometric, and Strength Training in Adolescent Soccer Players: Effects on Measures of Speed, Strength, Power, Change of Direction, and Anaerobic Capacity.

Ferley, DD, Scholten, S, and Vukovich, MD. Combined sprint interval, plyometric, and strength training in adolescent soccer players: effects on measures of speed, strength, power, change of direction, and anaerobic capacity. J Strength Cond Res 34(4): 957-968, 2020-During winter, many soccer players train indoors to improve the aerobic and anaerobic demands of their sport. Sprint interval training (SIT) performed on a treadmill using level and graded conditions represents a viable alternative to traditional endurance conditioning. To date, little research exists contrasting the effects of these conditions. Therefore, the purpose of this investigation included examining the effects of 2 approaches combining SIT, plyometrics, and strength training on performance measures in soccer players aged 13-18 years over 8 weeks. Forty-six subjects were divided into 3 groups. Group 1 performed SIT using predominantly inclined treadmill conditions combined with resistance and plyometric training (INC, n = 17). Group 2 performed SIT using level treadmill grades and completed the same resistance and plyometric training (LEV, n = 14). Group 3 was a control group representing various sports who continued their normal training (CON, n = 15). Pre- and posttests assessed speed, strength, change of direction, and anaerobic capacity, including sprint speed (9.1 and 18.3 m sprint), unilateral triple hop for distance (3HOP_L and 3HOP_R), pro agility change of direction (PA); treadmill running to exhaustion on a 20% grade (CFMod), and hip flexor maximum strength (HF_1RM). After training, INC and LEV improved more in all measures compared with CON. Furthermore, INC improved significantly more compared with LEV in 9.1- and 18.3-m sprint, 3HOP_L and 3HOP_R, PA, CFMod, and HF_1RM (p < 0.05). We conclude that strength and plyometric training combined with incline-based SIT is more effective than a similar training approach using level-grade SIT.

Assessing the Reliability of Using a Horizontal Leg Press Equipped With a Force Plate to Report on Measures of Positive and Negative Neuromuscular Characteristics.

Ferley, DD and Vukovich, MD. Assessing the reliability of using a horizontal leg press equipped with a force plate to report on measures of positive and negative neuromuscular characteristics. J Strength Cond Res 33(2): 360-371, 2019-Individuals participating in sport or returning from a lower-extremity injury routinely perform assessments of lower-body anaerobic capacity and power to gauge fitness or readiness to return to competition. However, many commonly used assessments lack the specificity of movement and muscle contraction demonstrated in sprinting, jumping, and changing direction. Therefore, this investigation assessed the reliability of a novel lower-body power test called a Plyo Press Power Quotient (3PQ), which involves jumping on a horizontal leg press equipped with a force plate. Thirty participants completed 1 repetition maximum (1RM) strength testing for 1 and 2 legs and a countermovement vertical jump. Two trials of 1- and 2-leg jump tests were performed using 30, 40, 50, and 60% of 1RM for 20 and 30 seconds, respectively. Dependent variables were a variety of positive (concentric) and negative (eccentric) neuromuscular indices. Statistical significance was set to p ≤ 0.05. No significant differences existed between trial 1 and 2 for any measures. One-way analysis of variance between %1RM conditions revealed significant differences in peak force (right; both), average negative power (left; right), average negative work (left; right), rate of power development (both), percent positive and negative fatigue (left; right; both), and velocity of movement (left; right; both). Peak power of all 2-leg jump tests correlated highly with vertical jump (r ≥ 0.89) but was significantly different. We conclude 3PQ testing provides a reliable assessment of anaerobic capacity and power, with the added benefit of easy single-leg comparisons.

Predicting the Intensity for Performing Supramaximal Incline Treadmill Interval Training in Distance Runners.

Ferley, DD and Vukovich, MD. Predicting the intensity for performing supramaximal incline treadmill interval training in distance runners. J Strength Cond Res 33(5): 1354-1361, 2019-Recent evidence highlights the effectiveness of 30-second bouts paired with level-grade supramaximal interval training (SMIT) and incline treadmill training (INC), respectively, in distance runners. Although INC has been described as a form of SMIT, no investigation of INC involving a supramaximal intensity and 30-second bouts has occurred; hence, no established recommendation for prescribing a supramaximal intensity with 30-second bouts for INC exists. Therefore, the purpose of this investigation included reporting on the time-to-exhaustion (Tmax) response and test-retest reliability of running on a 5% grade using supramaximal intensities of 110, 115, 120, 125, and 130% of the velocity associated with maximum oxygen consumption (Vmax). Additionally, these measures were assessed during 140% Vmax and 1% grade. A second aim included determining the %Vmax associated with a 30-second effort via bivariate analysis. Twelve distance runners (age, 26.9 ± 4.8 years; body mass, 69.2 ± 11.7 kg; height, 177.3 ± 10.2 cm; and V[Combining Dot Above]O2max, 61.4 ± 6.3 ml·kg·min) completed 2 Tmax trials at each intensity for measures of reliability. The dependent variable was the Tmax of each condition. Statistical significance was set to p ≤ 0.05. Student's t-test revealed no significant differences between trials for all intensities. One-way analysis of variance revealed (a) that Tmax during INC at 110% Vmax was significantly different than all conditions except 115% Vmax and (b) no significant difference in Tmax between 120, 125, 130, and 140% Vmax conditions. In conclusion, Tmax of all conditions proved reliable, and bivariate analysis revealed running at 125% Vmax on a 5% grade yielded a 30-second effort.

Protein Supplementation During a 6-Month Concurrent Training Program: Effect on Body Composition and Muscular Strength in Sedentary Individuals.

We examined the effect of a protein supplement on muscular strength and body composition during 6 months of a 5 days/week concurrent strength and endurance training program. Sedentary males (n = 26) and females (n = 25), 18-25 years, were randomly assigned to receive a protein (PRO, 42 g/serving) or carbohydrate (CON) supplement twice daily. Strength and body composition (dual-energy X-ray absorptiometry) were assessed at baseline, 3 (3M), and 6 (6M) months. Protein intake was higher in PRO (PRO: 2.2 g/kg; CON: 1.1 g/kg; p < .001). Females in both groups gained similar strength at 3M and 6M in bench press and hip sled. Males in PRO gained more bench press strength at 3M (PRO: 24.6 ± 3.2 kg; CON: 14.3 ± 3.8 kg; p = .06) and 6M (PRO: 34.4 ± 4.3 kg; CON: 18.7 ± 5.1 kg; p = .03) and hip sled strength at 3M (PRO: 67.7 ± 9.2 kg; CON: 40.8 ± 10.8 kg, p = .07) and 6M (PRO: 94.0 ± 10.6 kg; CON: 65.1 ± 12.4 kg; p = .09) compared with CON. Females in PRO experienced a greater reduction in fat mass over the course of the study (6M) than CON (PRO: -1.7 ± 0.5 kg; CON: 0.1 ± 0.5 kg; p = .06). Changes in lean mass were similar for females in PRO and CON. Loss in fat mass was similar for males in PRO and CON at 3M and 6M. Males in PRO gained more lean mass at 3M compared with CON (PRO: 3.2 ± 0.3 kg; CON: 2.2 ± 0.4 kg; p = .1) but similar gains at 6M (PRO: 2.6 ± 0.4 kg; CON: 2.2 ± 0.5 kg; p = .6). The results of this study demonstrate that PRO used during a concurrent training program may augment positive changes in body composition in young sedentary males and females, and strength gains in males.

The Effect of Nitrate Supplementation on Exercise Tolerance and Performance: A Systematic Review and Meta-Analysis.

Van De Walle, GP and Vukovich, MD. The effect of nitrate supplementation on exercise tolerance and performance: a systematic review and meta-analysis. J Strength Cond Res 32(6): 1796-1808, 2018-The purpose of this article was to systematically review the current literature and evaluate the overall efficacy of nitrate supplementation on exercise tolerance and performance by meta-analysis. Studies were eligible for inclusion if they met the following criteria: (a) were an experimental trial published in an English peer-reviewed journal; (b) compared the effects of inorganic nitrate consumption with a non-bioactive supplement control or placebo; (c) used a quantifiable measure of exercise performance; and (d) was carried out in apparently healthy participants without disease. A total of 29 studies were identified that investigated the effects of nitrate supplementation on exercise tolerance or performance in accordance with the criteria outlined. Analysis using time to exhaustion as the outcome variable revealed a significant effect of nitrate supplementation on exercise tolerance (ES = 0.28; 95% confidence interval [CI]: 0.08-0.47; p = 0.006) compared with placebo. Analysis using time to complete a specific distance as the outcome variable revealed no significant effect of nitrate supplementation on exercise performance (ES = -0.05; 95% CI: -0.28 to 0.17; p = 0.64) compared with placebo. Nitrate supplementation is likely to improve exercise tolerance and capacity that may improve exercise performance. More research is required to determine the optimal dose and duration of nitrate supplementation. It would also be important to consider the type of athlete performing the exercise and the duration, intensity, and mode of the exercise performed because these factors are likely to influence the efficacy of nitrate supplementation.

Multiparameter Particle Display (MPPD): A Quantitative Screening Method for the Discovery of Highly Specific Aptamers.

Aptamers are a promising class of affinity reagents because they are chemically synthesized, thus making them highly reproducible and distributable as sequence information rather than a physical entity. Although many high-quality aptamers have been previously reported, it is difficult to routinely generate aptamers that possess both high affinity and specificity. One of the reasons is that conventional aptamer selection can only be performed either for affinity (positive selection) or for specificity (negative selection), but not both simultaneously. In this work, we harness the capacity of fluorescence activated cell sorting (FACS) for multicolor sorting to simultaneously screen for affinity and specificity at a throughput of 107 aptamers per hour. As a proof of principle, we generated DNA aptamers that exhibit picomolar to low nanomolar affinity in human serum for three diverse proteins, and show that these aptamers are capable of outperforming high-quality monoclonal antibodies in a standard ELISA detection assay.

Time-to-Fatigue During Incline Treadmill Running: Implications for Individualized Training Prescription.

Uphill running has been touted as a key interval training tactic for distance runners despite few scientifically derived recommendations for individualized training prescription. To date, a majority of uphill training research has focused on shorter, faster training bouts; however, longer, slower bouts based on an individual's velocity at maximum oxygen consumption (Vmax) may prove more effective. One potential longer bout length may be associated with the time Vmax can be maintained (Tmax), an approach proven effective in level-grade interval training. Therefore, the purpose of this investigation included examining the Tmax, heart rate, and test-retest reliability of incline treadmill running (INC) on a 10% grade at 65, 70, 75, 80, and 85%Vmax compared with level-grade running at Vmax. Twelve moderately trained distance runners (age, 26.4 ± 4.8; body mass, 64.3 ± 12.9 kg; height, 171.2 ± 9.3 cm; and V̇O2max, 56.6 ± 7.6 ml·min⁻¹·kg⁻¹) completed 2 Tmax INC trials at each submaximal Vmax and a level-grade Tmax at Vmax. The dependent variables were Tmax, heart rate plateau (HR(Plateau)), and half-time to heart rate plateau (½HR(Plateau)) of each condition. Statistical significance was set to p ≤ 0.05. Student's t-test revealed no significant differences in Tmax, HR(Plateau), and ½HR(Plateau) between trials 1 and 2 at any INC condition. One-way analysis of variance revealed significant differences in (a) Tmax during INC at 75, 80, and 85%Vmax and level-grade at Vmax and (b) ½HR(Plateau) during INC at 80 and 85%Vmax and all other conditions. In conclusion, Tmax and heart rate dynamics during INC proved reliable, and simple regression analysis revealed ∼68%Vmax during INC yields the same level-grade Tmax at Vmax.

Changes in Body Composition in Division I Football Players Over a Competitive Season and Recovery in Off-Season.

This study investigated changes in body composition over 1 competitive football season in D-I collegiate football players (N = 53; by position, 21 linemen vs. 32 nonline; or by seniority, 30 upperclassmen vs. 23 underclassmen) and additional changes by the following spring season (N = 46; 20 linemen vs. 26 nonline; 27 upperclassmen vs. 19 underclassmen). Body composition by dual-energy x-ray absorptiometry (DXA) was completed pre- and post-season and the following spring. For the team as a whole, player weight decreased 1.3 kg (1.2%) and lean mass decreased 1.4 kg (1.6%) over the season. Absolute fat mass showed no change; however, percent body fat showed a 0.5% increase. There was an interaction between player position and seniority for changes in lean mass (p < 0.01). In nonline positions upperclassmen lost more lean mass than underclassmen, whereas in line positions underclassmen lost more lean mass than upperclassmen. Spring measures indicate that weight did not increase during the off-season, but improvement in body composition was noted. Lean mass increased by 2.2 kg (2.6%), whereas absolute fat mass decreased by 1.4 kg (6.7%). Although weight and lean mass losses during the competitive season were recovered in the off-season, changes in collegiate football programs that include nutrition counseling, dietary recommendations, monitoring of weight, and skin-fold testing as an estimate of body fat change would be beneficial to players. Strength and conditioning coaches and staff need to consider strategies to incorporate these practices into their programs.

The effects of incline and level-grade high-intensity interval treadmill training on running economy and muscle power in well-trained distance runners.

Despite a paucity of evidence, uphill running has been touted as a sport-specific resistance-to-movement training tactic capable of enhancing metabolic, muscular, and neuromuscular processes in distance runners in ways similar to previously established resistance-to-movement training methods, such as heavy and/or explosive strength training and plyometric training. Therefore, the purpose of this investigation included documenting the effects of incline and level-grade interval treadmill training on indices of running economy (RE) (i.e., oxygen consumption [VO2] and blood lactate [BLa] responses of submaximal running) and muscle power. Thirty-two well-trained distance runners (age, 27.4 ± 3.8 years; body mass, 64.8 ± 8.9 kg; height, 173.6 ± 6.4 cm; and VO2max, 60.9 ± 8.5 ml·min(-1)·kg(-1)) received assignment to an uphill (GHill = 12), level-grade (GFlat = 12), or control (GCon = 8) group. GHill and GFlat completed 12 interval and 12 continuous run sessions over 6 weeks, whereas GCon maintained their normal training. Dependent variables measured before and after training were VO2 and BLa at 2 separate velocities associated with lactate threshold (VLT) (VO2-60% and VO2-80%; and BLa-60% and BLa-80%, respectively); percentage of VO2max at lactate threshold (%VO2max at VLT); muscle power as assessed through a horizontal 5-jump test (5Jmax); and isokinetic knee extension and flexion at 3 angular velocities (90, 180, and 300°·s(-1)). Statistical significance was set to p ≤ 0.05. All groups significantly improved 5Jmax, VO2-60%, VO2-80%, BLa-60%, and BLa-80%. Additionally, GHill and GFlat significantly improved %VO2max at VLT. Other indices of RE and muscle power did not improve. We conclude incline treadmill training effective for improving the components of RE, but insufficient as a resistance-to-movement exercise for enhancing muscle power output.

Antibody sequence determinants of viral antigen specificity.

Throughout life, humans experience repeated exposure to viral antigens through infection and vaccination, resulting in the generation of diverse, antigen-specific antibody repertoires. A paramount feature of antibodies that enables their critical contributions in counteracting recurrent and novel pathogens, and consequently fostering their utility as valuable targets for therapeutic and vaccine development, is the exquisite specificity displayed against their target antigens. Yet, there is still limited understanding of the determinants of antibody-antigen specificity, particularly as a function of antibody sequence. In recent years, experimental characterization of antibody repertoires has led to novel insights into fundamental properties of antibody sequences but has been largely decoupled from at-scale antigen specificity analysis. Here, using the LIBRA-seq technology, we generated a large data set mapping antibody sequence to antigen specificity for thousands of B cells, by screening the repertoires of a set of healthy individuals against 20 viral antigens representing diverse pathogens of biomedical significance. Analysis uncovered virus-specific patterns in variable gene usage, gene pairing, somatic hypermutation, as well as the presence of convergent antiviral signatures across multiple individuals, including the presence of public antibody clonotypes. Notably, our results showed that, for B-cell receptors originating from different individuals but leveraging an identical combination of heavy and light chain variable genes, there is a specific CDRH3 identity threshold above which B cells appear to exclusively share the same antigen specificity. This finding provides a quantifiable measure of the relationship between antibody sequence and antigen specificity and further defines experimentally grounded criteria for defining public antibody clonality.IMPORTANCEThe B-cell compartment of the humoral immune system plays a critical role in the generation of antibodies upon new and repeated pathogen exposure. This study provides an unprecedented level of detail on the molecular characteristics of antibody repertoires that are specific to each of the different target pathogens studied here and provides empirical evidence in support of a 70% CDRH3 amino acid identity threshold in pairs of B cells encoded by identical IGHV:IGL(K)V genes, as a means of defining public clonality and therefore predicting B-cell antigen specificity in different individuals. This is of exceptional importance when leveraging public clonality as a method to annotate B-cell receptor data otherwise lacking antigen specificity information. Understanding the fundamental rules of antibody-antigen interactions can lead to transformative new approaches for the development of antibody therapeutics and vaccines against current and emerging viruses.

The effects of uphill vs. level-grade high-intensity interval training on VO2max, Vmax, V(LT), and Tmax in well-trained distance runners.

Uphill running represents a frequently used and often prescribed training tactic in the development of competitive distance runners but remains largely uninvestigated and unsubstantiated as a training modality. The purpose of this investigation included documenting the effects of uphill interval training compared with level-grade interval training on maximal oxygen consumption (VO2max), the running speed associated with VO2max (Vmax), the running speed associated with lactate threshold (V(LT)), and the duration for which Vmax can be sustained (Tmax) in well-trained distance runners. Thirty-two well-trained distance runners (age, 27.4 ± 3.8 years; body mass, 64.8 ± 8.9 kg; height, 173.6 ± 6.4 cm; and VO2max, 60.9 ± 8.5 ml·min(-1)·kg(-1)) received assignment to an uphill interval training group (G(Hill) = 12), level-grade interval training group (G(Flat) = 12), or control group (G(Con) = 8). G(Hill) and G(Flat) completed 12 interval and 12 continuous running sessions over 6 weeks, whereas G(Con) maintained their normal training routine. Pre- and posttest measures of VO2max, Vmax, V(LT), and Tmax were used to assess performance. A 3 × 2 repeated measures analysis of variance was performed for each dependent variable and revealed a significant difference in Tmax in both G(Hill) and G(Flat) (p < 0.05). With regard to running performance, the results indicate that both uphill and level-grade interval training can induce significant improvements in a run-to-exhaustion test in well-trained runners at the speed associated with VO2max but that traditional level-grade training produces greater gains.

Negative Binomial Mixture Model for Identification of Noise in Antigen-Specificity Predictions by LIBRA-seq.

Motivation: LIBRA-seq (linking B cell receptor to antigen specificity by sequencing) provides a powerful tool for interrogating the antigen-specific B cell compartment and identifying antibodies against antigen targets of interest. Identification of noise in LIBRA-seq antigen count data is critical for improving antigen binding predictions for downstream applications including antibody discovery and machine learning technologies. Results: In this study, we present a method for denoising LIBRA-seq data by clustering antigen counts into signal and noise components with a negative binomial mixture model. This approach leverages the VRC01 negative control cells included in a recent LIBRA-seq study(Abu-Shmais et al.) to provide a data-driven means for identification of technical noise. We apply this method to a dataset of nine donors representing separate LIBRA-seq experiments and show that our approach provides improved predictions for in vitro antibody-antigen binding when compared to the standard scoring method used in LIBRA-seq, despite variance in data size and noise structure across samples. This development will improve the ability of LIBRA-seq to identify antigen-specific B cells and contribute to providing more reliable datasets for future machine learning based approaches to predicting antibody-antigen binding as the corpus of LIBRA-seq data continues to grow.

Pre-sleep feeding, sleep quality, and markers of recovery in division I NCAA female soccer players.

Pre-sleep nutrition habits in elite female athletes have yet to be evaluated. A retrospective analysis was performed with 14 NCAA Division I female soccer players who wore a WHOOP, Inc. band - a wearable device that quantifies recovery by measuring sleep, activity, and heart rate metrics through actigraphy and photoplethysmography, respectively - 24 h a day for an entire competitive season to measure sleep and recovery. Pre-sleep food consumption data were collected via surveys every 3 days. Average pre-sleep nutritional intake (mean ± sd: kcals 330 ± 284; cho 46.2 ± 40.5 g; pro 7.6 ± 7.3 g; fat 12 ± 10.5 g) was recorded. Macronutrients and kcals were grouped into high and low categories based upon the 50th percentile of the mean to compare the impact of a high versus low pre-sleep intake on sleep and recovery variables. Sleep duration (p = 0.10, 0.69, 0.16, 0.17) and sleep disturbances (p = 0.42, 0.65, 0.81, 0.81) were not affected by high versus low kcal, PRO, fat, CHO intake, respectively. Recovery (p = 0.81, 0.06, 0.81, 0.92), RHR (p = 0.84, 0.64, 0.26, 0.66), or HRV (p = 0.84, 0.70, 0.76, 0.93) were also not affected by high versus low kcal, PRO, fat, or CHO consumption, respectively. Consuming a small meal before bed may have no impact on sleep or recovery.

Moderate protein intake improves total and regional body composition and insulin sensitivity in overweight adults.

A high protein intake (approximately 40% of energy intake) combined with aerobic and resistance exercise training is more closely associated with improved body composition and cardiovascular risk profile than a traditional protein intake (approximately 15% of intake) combined with moderate-intensity aerobic exercise. However, there is concern that such high-protein diets may adversely affect health. We therefore tested the hypothesis that moderate protein intake (approximately 25% of energy intake) would elicit similar benefits on body composition and metabolic profile as high protein intake. Twenty-four overweight/obese men and women (body mass index [BMI] = 32.2 +/- 3.4, percentage of body fat [%BF] = 37.3 +/- 8.0) were matched for BMI and %BF and randomly assigned to one of 3 groups for a 3-month nutrition/exercise training intervention: (1) high-protein diet (approximately 40% of energy intake) and combined high-intensity resistance and cardiovascular training (HPEx, n = 8, 5 female and 3 male), (2) moderate-protein diet (approximately 25% of energy intake) and combined high-intensity resistance and cardiovascular training (MPEx, n = 8, 5 female and 3 male), or (3) high-protein diet only (HPNx, n = 8, 5 female and 3 male). Total and regional body composition (dual-energy x-ray absorptiometry), insulin sensitivity (insulin sensitivity index to the oral glucose tolerance test), insulin-like growth factor-1 (IGF-1), IGF binding protein-1 (IGFBP-1), IGF binding protein-3 (IGFBP-3), and blood lipids were measured at baseline and after the intervention. All groups experienced significant (P < .05) and similar losses of body weight, BMI, and total and abdominal %BF, and similar improvements in insulin sensitivity (HPEx, 6.3 +/- 1.2 vs 9.5 +/- 0.98; MPEx, 6.2 +/- 1.4 vs 8.4 +/- 1.6; HPNx, 3.7 +/- 1.1 vs 7.0 +/- 1.1; insulin sensitivity index to the oral glucose tolerance test; P < .05) and leptin levels. Furthermore, the HPEx group demonstrated decreases in total cholesterol (TC) and triglycerides, and increases in IGF-1 and IGFBP-1. The MPEx group experienced decreases in TC, whereas the HPNx group had increases in high-density lipoprotein cholesterol, TC to high-density lipoprotein, IGF-1, and IGFBP-1. In conclusion, moderate protein intake elicits similar benefits in body composition and insulin sensitivity as a high-protein diet. These findings may have practical implications for individuals interested in diets containing elevated dietary protein.