Anthropometrical Determinants of Deadlift Variant Performance.

The barbell deadlift is a popular exercise and one of the three lifts in competitive powerlifting. While muscle activation has been tested between the sumo (SDL) and conventional deadlift (CDL), the relationships between anthropometrics and deadlift performance in the two styles is not yet known. The purpose of this study was to investigate the relationships between anthropometrics and SDL versus CDL performance (SDL:CDL strength ratio). Forty-seven (n = 28 male, n = 19 female) deadlift naïve subjects participated in this study. Anthropometric measurements were arm and hand length, wrist and ankle girth, seated height, thigh length, and lower leg length. Deadlift instructions for the two styles were provided on day 1 and 2. On day 3 and 4, deadlift 1RM was tested for the SDL or CDL in random order, and then deadlift repetitions to volitional fatigue with 60% of 1RM were measured. No significant differences between CDL 1RM and SDL 1RM were found. The only significant correlation found between the anthropometric predictors and the SDL:CDL strength ratio was an inverse relationship with the sitting height to total height ratio (r = 0.297, p = 0.043). Total repetitions to volitional fatigue was higher in females compared to males for both lifts (p = 0.041). Our findings suggest that the sumo deadlift may be slightly mechanically advantageous for deadlift naïve individuals with longer torsos, while the conventional deadlift may be better suited for those with shorter torsos.

One in a million: flow cytometric sorting of single cell-lysate assays in monodisperse picolitre double emulsion droplets for directed evolution.

Directed evolution relies on iterative cycles of randomization and selection. The outcome of an artificial evolution experiment is crucially dependent on (i) the numbers of variants that can be screened and (ii) the quality of the assessment of each clone that forms the basis for selection. Compartmentalization of screening assays in water-in-oil emulsion droplets provides an opportunity to screen vast numbers of individual assays with good signal quality. Microfluidic systems have been developed to make and sort droplets, but the operator skill required precludes their ready implementation in nonspecialist settings. We now establish a protocol for the creation of monodisperse double-emulsion droplets in two steps in microfluidic devices with different surface characteristics (first hydrophobic, then hydrophilic). The resulting double-emulsion droplets are suitable for quantitative analysis and sorting in a commercial flow cytometer. The power of this approach is demonstrated in a series of enrichment experiments, culminating in the successful recovery of catalytically active clones from a sea of 1 000 000-fold as many low-activity variants. The modular workflow allows integration of additional steps: the encapsulated lysate assay reactions can be stopped by heat inactivation (enabling ready control of selection stringency), the droplet size can be contracted (to concentrate its contents), and storage (at -80 °C) is possible for discontinuous workflows. The control that can be thus exerted on screening conditions will facilitate exploitation of the potential of protein libraries compartmentalized in droplets in a straightforward protocol that can be readily implemented and used by protein engineers.

Correction: Roberts et al. "Satiating Effect of High Protein Diets on Resistance-Trained Individuals in Energy Deficit" Nutrients 2019, 11(1), 56.

The authors wish to make a correction to the published version of their paper [...].

High-Throughput, Lysis-Free Screening for Sulfatase Activity Using Escherichia coli Autodisplay in Microdroplets.

Directed evolution of enzymes toward improved catalytic performance has become a powerful tool in protein engineering. To be effective, a directed evolution campaign requires the use of high-throughput screening. In this study we describe the development of an ultra high-throughput lysis-free procedure to screen for improved sulfatase activity by combining microdroplet-based single-variant activity sorting with E. coli autodisplay. For the first step in a 4-step screening procedure, we quantitatively screened >105 variants of the homodimeric arylsulfatase from Silicibacter pomeroyi (SpAS1), displayed on the E. coli cell surface, for improved sulfatase activity using fluorescence activated droplet sorting. Compartmentalization of the fluorescent reaction product with living E. coli cells autodisplaying the sulfatase variants ensured the continuous linkage of genotype and phenotype during droplet sorting and allowed for direct recovery by simple regrowth of the sorted cells. The use of autodisplay on living cells simplified and reduced the degree of liquid handling during all steps in the screening procedure to the single event of simply mixing substrate and cells. The percentage of apparent improved variants was enriched >10-fold as a result of droplet sorting. We ultimately identified 25 SpAS1 variants with improved performance toward 4-nitrophenyl sulfate (up to 6.2-fold) and/or fluorescein disulfate (up to 30-fold). In SpAS1 variants with improved performance toward the bulky fluorescein disulfate, many of the beneficial mutations occur in residues that form hydrogen bonds between α-helices in the C-terminal oligomerization region, suggesting a previously unknown role for the dimer interface in shaping the substrate binding site of SpAS1.

Satiating Effect of High Protein Diets on Resistance-Trained Subjects in Energy Deficit.

Short-term energy deficit strategies are practiced by weight class and physique athletes, often involving high protein intakes to maximize satiety and maintain lean mass despite a paucity of research. This study compared the satiating effect of two protein diets on resistance-trained individuals during short-term energy deficit. Following ethical approval, 16 participants (age: 28 ± 2 years; height: 1.72 ± 0.03 m; body-mass: 88.83 ± 5.54 kg; body-fat: 21.85 ± 1.82%) were randomly assigned to 7-days moderate (PROMOD: 1.8 g·kg-1·d-1) or high protein (PROHIGH: 2.9 g·kg-1·d-1) matched calorie-deficit diets in a cross-over design. Daily satiety responses were recorded throughout interventions. Pre-post diet, plasma ghrelin and peptide tyrosine tyrosine (PYY), and satiety ratings were assessed in response to a protein-rich meal. Only perceived satisfaction was significantly greater following PROHIGH (67.29 ± 4.28 v 58.96 ± 4.51 mm, p = 0.04). Perceived cravings increased following PROMOD only (46.25 ± 4.96 to 57.60 ± 4.41 mm, p = 0.01). Absolute ghrelin concentration significantly reduced post-meal following PROMOD (972.8 ± 130.4 to 613.6 ± 114.3 pg·mL-1; p = 0.003), remaining lower than PROHIGH at 2 h (-0.40 ± 0.06 v -0.26 ± 0.06 pg·mL-1 normalized relative change; p = 0.015). Absolute PYY concentration increased to a similar extent post-meal (PROMOD: 84.9 ± 8.9 to 147.1 ± 11.9 pg·mL-1, PROHIGH: 100.6 ± 9.5 to 143.3 ± 12.0 pg·mL-1; p < 0.001), but expressed as relative change difference was significantly greater for PROMOD at 2 h (+0.39 ± 0.20 pg·mL-1 v -0.28 ± 0.12 pg·mL-1; p = 0.001). Perceived hunger, fullness and satisfaction post-meal were comparable between diets (p > 0.05). However, desire to eat remained significantly blunted for PROMOD (p = 0.048). PROHIGH does not confer additional satiating benefits in resistance-trained individuals during short-term energy deficit. Ghrelin and PYY responses to a test-meal support the contention that satiety was maintained following PROMOD, although athletes experiencing negative symptoms (i.e., cravings) may benefit from protein-rich meals as opposed to over-consumption of protein.

The short-term effect of high versus moderate protein intake on recovery after strength training in resistance-trained individuals.

BACKGROUND: Dietary protein intakes up to 2.9 g.kg-1.d-1 and protein consumption before and after resistance training may enhance recovery, resulting in hypertrophy and strength gains. However, it remains unclear whether protein quantity or nutrient timing is central to positive adaptations. This study investigated the effect of total dietary protein content, whilst controlling for protein timing, on recovery in resistance trainees. METHODS: Fourteen resistance-trained individuals underwent two 10-day isocaloric dietary regimes with a protein content of 1.8 g.kg-1.d-1 (PROMOD) or 2.9 g.kg-1.d-1 (PROHIGH) in a randomised, counterbalanced, crossover design. On days 8-10 (T1-T3), participants undertook resistance exercise under controlled conditions, performing 3 sets of squat, bench press and bent-over rows at 80% 1 repetition maximum until volitional exhaustion. Additionally, participants consumed a 0.4 g.kg-1 whey protein concentrate/isolate mix 30 min before and after exercise sessions to standardise protein timing specific to training. Recovery was assessed via daily repetition performance, muscle soreness, bioelectrical impedance phase angle, plasma creatine kinase (CK) and tumor necrosis factor-α (TNF-α). RESULTS: No significant differences were reported between conditions for any of the performance repetition count variables (p > 0.05). However, within PROMOD only, squat performance total repetition count was significantly lower at T3 (19.7 ± 6.8) compared to T1 (23.0 ± 7.5; p = 0.006). Pre and post-exercise CK concentrations significantly increased across test days (p ≤ 0.003), although no differences were reported between conditions. No differences for TNF-α or muscle soreness were reported between dietary conditions. Phase angle was significantly greater at T3 for PROHIGH (8.26 ± 0.82°) compared with PROMOD (8.08 ± 0.80°; p = 0.012). CONCLUSIONS: When energy intake and peri-exercise protein intake was controlled for, a short term PROHIGH diet did not improve markers of muscle damage or soreness in comparison to a PROMOD approach following repeated days of intensive training. Whilst it is therefore likely that moderate protein intakes (1.8 g.kg-1.d-1) may be sufficient for resistance-trained individuals, it is noteworthy that both lower body exercise performance and bioelectrical phase angle were maintained with PROHIGH. Longer term interventions are warranted to determine whether PROMOD intakes are sufficient during prolonged training periods or when extensive exercise (e.g. training twice daily) is undertaken.

Enzyme engineering in biomimetic compartments.

The success of a directed evolution approach to creating custom-made enzymes relies in no small part on screening as many clones as possible. The miniaturisation of assays into pico to femtoliter compartments (emulsion droplets, vesicles or gel-shell beads) makes directed evolution campaigns practically more straightforward than current large scale industrial screening that requires liquid handling equipment and much manpower. Several recent experimental formats have established protocols to screen more than 10 million compartments per day, representing unprecedented throughput at low cost. This review introduces the emerging approaches towards making biomimetic man-made compartments that are poised to be adapted by a wider circle of researchers. In addition to cost and time saving, control of selection pressures and conditions, the quantitative readout that reports on every library members and the ability to develop strategies based on these data will increase the degrees of freedom in designing and testing strategies for directed evolution experiments.