Optimizing Biocompatibility and Gene Delivery with DMAEA and DMAEAm: A Niacin-Derived Copolymer Approach.

Gene therapy is pivotal in nanomedicine, offering a versatile approach to disease treatment. This study aims to achieve an optimal balance between biocompatibility and efficacy, which is a common challenge in the field. A copolymer library is synthesized, incorporating niacin-derived monomers 2-acrylamidoethyl nicotinate (AAEN) or 2-(acryloyloxy)ethyl nicotinate (AEN) with N,N-(dimethylamino)ethyl acrylamide (DMAEAm) or hydrolysis-labile N,N-(dimethylamino)ethyl acrylate (DMAEA). Evaluation of the polymers' cytotoxicity profiles reveals that an increase in AAEN or DMAEA molar ratios correlates with improved biocompatibility. Remarkably, an increase in AAEN in both DMAEA and DMAEAm copolymers demonstrated enhanced transfection efficiencies of plasmid DNA in HEK293T cells. Additionally, the top-performing polymers demonstrate promising gene expression in challenging-to-transfect cells (THP-1 and Jurkat cells) and show no significant effect on modulating immune response induction in ex vivo treated murine monocytes. Overall, the best performing candidates exhibit an optimal balance between biocompatibility and efficacy, showcasing potential advancements in gene therapy.

Benefits of semiology taught using near-peer tutoring are sustainable.

BACKGROUND: Near-peer tutoring appears to be an efficient approach for teaching clinical skills. However, the clinical experience gained in the form of student medical internships may offset any interest in such tutoring programme. We then investigated the long-term benefits of this programme. METHODS: This study was conducted in a medical school that experimented in near-peer tutoring for semiology intended for undergraduate medical students. Objective Structured Clinical Examinations and a written semiology test were used to assess students' clinical skills immediately on its conclusion and repeated one and 2 years after the tutoring was completed. RESULTS: 116 students were evaluated initially (80 tutored and 36 untutored), 38 at 1 year (16 tutored and 22 untutored), 42 at 2 years (21 tutored and 21 untutored). In the global score for Objective Structured Clinical Examinations: at 1 year, the tutored group scored 14.0 ± 1.05 and the untutored group scored 11.3 ± 2.3 (p < 0.001), at 2 years, the tutored group scored 15.1 ± 1.5 and the untutored group scored 12.4 ± 2.2 (p < 0.001). We found a similar but smaller difference for the written semiology test. The difference for Objective Structured Clinical Examinations between tutored and untutored students vanished over time for cross-cutting skills. CONCLUSIONS: Near-peer tutoring in semiology for undergraduate medical students led to better results that remained with the passing of time. Though internships do allow an improvement in the clinical skills of untutored students, they did not reach the level of tutored students.

Effects of Hypohydration on Muscular Strength, Endurance, and Power in Women.

ABSTRACT: Gann, JJ, Andre, TL, Gallucci, AR, and Willoughby, DS. Effects of hypohydration on muscular strength, endurance, and power in women. J Strength Cond Res 35(2S): S102-S106, 2021-The purpose of this study was to determine the effects of dehydration on muscular strength, endurance, power, and perceptual measures in resistance-trained women. Ten resistance-trained women completed 2 bouts of exercise (1 repetition maximum [1RM] for bench press and angled leg press followed by 5 sets to failure of 75% of 1RM and vertical jump), either dehydrated (∼3% body mass) (DT) or heat-exposed with fluid replacement (HT). Paired t-tests revealed bench press 1RM was significantly lower for DT (42.7 ± 14.5 kg) compared with HT (44.1 ± 13.9 kg). No significant difference was found for leg press 1RM (DT = 216.1 ± 55.0 kg; HT = 223.4 ± 55.7 kg). There was also no difference in total reps completed for bench press (DT = 33.5 ± 5.0; HT = 33.0 ± 5.5) or leg press (DT = 42.6 ± 20.3; HT = 45.8 ± 19.7). There was no significant difference for vertical jump height (DT: 45.8 ± 5.2 cm, HT: 46.9 ± 6.0 cm). Ratings of perceived exertion (RPE) and session RPE were not significantly different between trials. Significant differences for perceived recovery status (DT: 5.1 ± 2.2, HT: 7.2 ± 1.1) and perceived readiness (DT: 4.2 ± 1.0, HT: 2.5 ± 0.5) indicate subjects expected impaired performance during DT. The current results suggest that previous night dehydration may have a negative impact on both bench press 1RM performance and perceptual feelings of recovery in resistance-trained women.

Ultrastructure and functional morphology of the appendages in the reef-building sedentary polychaete Sabellaria alveolata (Annelida, Sedentaria, Sabellida).

BACKGROUND: The sedentary polychaete Sabellaria alveolata, the sandcastle or honeycomb worm, possesses four different kinds of appendages besides the parapodia: opercular papillae, tentacular filaments, palps, and branchiae. It exhibits a highly specialized anterior end, the operculum, formed by the prostomium, peristomium, and two anterior segments. The operculum comprises opercular papillae, tentacular filaments, and palps. Paired branchiae are present from the second thoracic chaetiger onwards on the posteriorly following segments except for the last ones. Ultrastructural data on these appendages are either scanty, incomplete, or even lacking in Sabellariidae. In order to analyze their functional morphology, to bridge the data gap, and providing data for future phylogenetic and evolutionary analyses, we investigated the appendages of S. alveolata by applying light microscopy, confocal laser scanning microscopy, scanning, and transmission electron microscopy. RESULTS: In S. alveolata the entire body is covered by a thin cuticle characterized by the absence of layers of parallel collagen fibers with no differentiation between the various body regions including the branchiae. The opercular papillae bear numerous tufts of receptor cells and lack motile cilia. The tentacular filaments show a distinctive pattern of motile cilia. Their most conspicuous morphological feature is a cell-free cartilaginous endoskeletal structure enclosed by ECM. Besides musculature the filaments include a single coelomic cavity but blood vessels are absent. The palps are ciliated and possess two coelomic cavities and a single blind-ending internal blood vessel. Besides external ciliation and receptor cells, the coelomate branchiae are highly vascularized and equipped with numerous blood spaces extending deep between the epidermal cells resulting in low diffusion distances. CONCLUSIONS: All appendages, including the branchiae, bear receptor cells and, as such, are sensory. The opercular papillae resemble typical parapodial cirri. In contrast, the tentacular filaments have a triple function: sensing, collecting and transporting particles. A similarity to branchiae can be excluded. The palps are typical grooved palps. A revised classification of polychaete branchiae is suggested; thereby, the branchiae of S. alveolata belong to the most common type comprising coelom, musculature, and blood vessels. The results indicate that diffusion distances between blood and environment have been underestimated in many cases.

Esport: Fortnite Acutely Increases Heart Rate of Young Men.

Esports has rapidly increased in online play and viewing. A myriad of literature focuses on heart rate (HR) during traditional athletic competition, little research has addressed the HR responses of individuals playing esports and none in the esport: Fortnite. The purpose of this study was to compare the HR response during a 3-hour seated session of the esport: Fortnite to resting HR. Male college students (n = 23; age = 21 ± 1.8 years; BMI = 25.7 ± 3.9; Esport mean hours per week = 13 ± 8.7) were recruited to participate in the study. Mean and peak resting HR were collected during two HR measurements in a controlled laboratory setting. Additionally, participants wore the HR monitor to record mean and peak HR during their regular 3-hour esport: Fortnite session at home. Participants were also instructed to record their match statistics from each solo round. Separate paired sample t-tests were conducted to compare mean resting heart rate (rHR) vs. mean seated esport heart rate (eHR) and peak rHR vs. peak eHR. Pearson's r correlations were used to investigate relationships between variables. There was a statistically significant increase in mean eHR compared to mean rHR (76 ± 10 bpm vs. 70 ± 11 bpm; p < 0.05) and peak eHR compared to peak rHR (120 ± 16 bpm vs. 81 ± 11 bpm; p < 0.05). The average number of kills was 29 ± 18.6. This initial study suggests esport: Fortnite increases individuals seated HR, suggesting a physiological stressful event.

Physiological and Perceptual Response to a Live Collegiate Esports Tournament.

Competitive esports has grown rapidly across the globe justifying a need to quantify the physiological stress response to this environment. The purpose of this study was to describe the physiological and perceptual responses in a live collegiate esports tournament. Male members of the University of Mississippi Esports team (n = 14; age = 19.8 ± 1.0 years; BMI = 24.1 ± 5.5) completed the study during the esports Egg Bowl. Heart rate (HR) and heart rate variability (HrV) were collected pre-, during, and post-competition. Rating of perceived exertion for the session (S-RPE) and mental fatigue were collected post competition. Mean HR during competition were significantly elevated compared to mean pre- and post- (131.4 ± 19.0 bpm vs. 97.1 ± 19.9 bpm and 101.9 ± 17.4 bpm; p = 0.000) and peak HR during competition were significantly elevated compared to peak pre- and post-(188.1 ± 32.9 bpm vs. 119.6 ± 20.1 bpm and 119.9 ± 16.3 bpm; p = 0.000). R-R intervals were significantly lower in-competition (465.71 ± 68.99) compared to pre- (643.64 ± 138.54) or post- competition (616.07 ± 109.98; p = .000). No significant differences were found in rMSSD, (ln) rMSSD, SDNN, or NN50 across the three measurements. LF was lower post- competition than pre-competition (d = 0.278). Participants indicated moderate mental fatigue (3.7 ± 1.2; on a scale of 1-7). These findings demonstrate competing in esports causes a physiological stress response. Given the elevated HR, further understanding of the chronic physiological stress to competitive esports is warranted.

Restrictive Breathing Mask Reduces Repetitions to Failure During a Session of Lower-Body Resistance Exercise.

Andre, TL, Gann, JJ, Hwang, PS, Ziperman, E, Magnussen, MJ, and Willoughby, DS. Restrictive breathing mask reduces repetitions to failure during a session of lower-body resistance exercise. J Strength Cond Res 32(8): 2103-2108, 2018-The purpose of this study was to determine the effect of restrictive breathing mask (RBM) on muscle performance, hemodynamic, and perceived stress variables during a session of lower-body resistance exercise. In a crossover design, 10 participants performed 2 separate testing sessions, RBM and no mask, consisting of squat, leg press, and leg extension. The paired-samples t-test was used for session rating of perceived exertion (S-RPE), perceived stress before and after, heart rate (HR), pulse oximetry, and a 2 × 4 (session [mask, no mask] × time [squat exercise, leg press exercise, leg extension exercise, total resistance exercise session]) factorial analysis of variance with repeated measures (p ≤ 0.05). A significant decrease was found in total repetitions during the RBM condition (p < 0.01). A majority of the decrease in repetitions to failure occurred in the squat (p < 0.05) and in the leg press (p < 0.01), whereas no difference was observed in leg extension (p = 0.214). A significant increase was observed in S-RPE during the RBM session (p < 0.01). A significant increase was found in prestress (p < 0.01) and poststress (p = 0.01) in the RBM session. No significant difference existed for HR between exercise sessions (p = 0.08). A significant decrease existed in pulse oximetry during the RBM session (p < 0.01). The use of an RBM had a negative effect on the number of repetitions completed during an acute session of lower-body resistance training.

Skeletal Muscle Estrogen Receptor Activation in Response to Eccentric Exercise Up-Regulates Myogenic-Related Gene Expression Independent of Differing Serum Estradiol Levels Occurring during the Human Menstrual Cycle.

This study sought to determine if the differences in serum estradiol we have previously observed to occur during the mid-follicular (MF) and mid-luteal (ML) phases of the female menstrual cycle could be attributed to estrogen-induced receptor activation and subsequent effects on myogenic-related genes which may otherwise impact muscle regeneration in response to eccentric exercise. Twenty-two physically-active females (20.9 ± 1.4 years, 63.5 ± 9.0 kg, 1.65 ± 0.08 m) underwent an eccentric exercise bout of the knee extensors during the MF and ML phases of their 28-day menstrual cycle. Prior to (PRE), at 6 (6HRPOST), and 24 (24HRPOST) hours post-exercise for each session, participants had muscle biopsies obtained. Skeletal muscle estradiol and estrogen receptor-α (ER-α) content and ER-DNA binding were determined with ELISA. Real-time PCR was used to assess ER-α, Myo-D, and cyclin D1 mRNA expression. Data were analyzed utilizing a 2 x 3 repeated measures univariate analyses of variance (ANOVA) for each criterion variable (p ≤ .05). Skeletal muscle estradiol levels were not significantly impacted by either menstrual phase (p > 0.05); however, both ER-α mRNA and protein were significantly increased during MF (p < 0.05). ER-DNA binding and Myo-D mRNA expression increased significantly in both menstrual phases in response to exercise but were not different from one another; however, cyclin D1 mRNA expression was significantly greater during MF. This study demonstrates that skeletal muscle ER-α activation in response to eccentric exercise up-regulates myogenic-related gene expression independent of serum estradiol levels occurring during the human menstrual cycle.

Electron and photon diagnostics for plasma acceleration-based FELs.

It is now well established that laser plasma acceleration (LPA) is an innovative and good candidate in the beam acceleration field. Relativistic beams are indeed produced up to several GeV but their quality remains to be demonstrated in the highly demanding case of free-electron lasers (FELs). Several experiments have already shown the feasibility of synchrotron radiation delivery based on LPA but free-electron lasing has still to be achieved. Since the quality of the LPA beam inside the undulator is the critical issue, any LPA-based FEL experiment requires a refined characterization of the beam properties along the transport line and of the photon beam at the undulator exit. This characterization relies on diagnostics which must be adapted to the LPA specificities. Here, the electron and photon diagnostics already used on LPAs and required for LPA-based FELs are reviewed, and the critical points are illustrated using recent experiments performed around the world.

Effectiveness of Fish Oil Supplementation in Attenuating Exercise-Induced Muscle Damage in Women During Midfollicular and Midluteal Menstrual Phases.

McKinley-Barnard, SK, Andre, TL, Gann, JJ, Hwang, PS, and Willoughby, DS. Effectiveness of fish oil supplementation in attenuating exercise-induced muscle damage in females during midfollicular and midluteal menstrual phases. J Strength Cond Res 32(6): 1601-1612, 2018-The purpose of this study was to determine whether the differences in estrogen levels during the female menstrual cycle and fish oil supplementation would attenuate eccentric exercise-induced muscle damage and delayed-onset muscle soreness (DOMS). In a double-blind fashion, 22 physically active females (20.9 ± 1.4 years, 63.5 ± 9.0 kg, 165.2 ± 7.5 cm) were randomly assigned to ingest either 6 g of fish oil (n = 11) or placebo (n = 11) daily for 21 days. Participants underwent an eccentric exercise bout of the knee extensors on 2 occasions during the midfollicular (MF) and midluteal (ML) phases of the 28-day menstrual cycle. Before (PRE), at 6 (6HRPOST), and at 24 hours postexercise (24HRPOST) for each session, participants underwent assessments of DOMS, muscle strength, and had venous blood samples and muscle biopsies obtained. Data were analyzed using a 2 × 2 × 3 repeated-measures multivariate analysis of variance for each criterion variable (p ≤ 0.05). Further analysis of the main effects for the test was performed using separate 1-way analyses of variance. Delayed-onset muscle soreness was significantly greater at the 6HRPOST and 24HRPOST timepoints compared with PRE (p < 0.001). Superoxide dismutase and tumor necrosis factor-alpha (TNF-α) concentrations were significantly higher at the MF phase compared with the ML phase (p < 0.001 and p = 0.05, respectively). There were no statistically significant differences observed for muscle strength, myoglobin, NF-Kß p50, or NF-Kß p65. This study demonstrates that higher levels of estrogen may exert a cytoprotective effect on the sarcolemma.

Resistance Training-Induced Elevations in Muscular Strength in Trained Men Are Maintained After 2 Weeks of Detraining and Not Differentially Affected by Whey Protein Supplementation.

Hwang, PS, Andre, TL, McKinley-Barnard, SK, Morales Marroquín, FE, Gann, JJ, Song, JJ, and Willoughby, DS. Resistance training-induced elevations in muscular strength in trained men are maintained after 2 weeks of detraining and not differentially affected by whey protein supplementation. J Strength Cond Res 31(4): 869-881, 2017-Resistance training (RT) with nutritional strategies incorporating whey protein intake postexercise can stimulate muscle protein synthesis and elicit hypertrophy. The early phases of training-induced anabolic responses can be attenuated with longer-term training. It is currently unknown if short-term detraining (DT) can restore these blunted anabolic responses during a subsequent retraining (ReT) period. Twenty resistance-trained men (age 20.95 ± 1.23 years; n = 20) were randomized into one of 2 groups (PRO or CHO; 25 g) in a double-blind manner. Participants followed a 4-day per week RT program (4-week RT; 2-week DT; 4-week ReT) while consuming their respective supplement only on workout days during RT and ReT, but every day during DT. At baseline, 4 weeks after RT (post-RT), 2 weeks after DT (post-2-week DT), and after 4 weeks of ReT after DT (post-ReT), leg press strength (LPS) was assessed and rectus femoris cross-sectional area and lean mass changes were assessed by ultrasonography and dual-energy x-ray absorptiometry, respectively. A factorial 2 × 4 (group by time) analyses of variance with repeated measures were used with a probability level at ≤0.05. LPS was elevated throughout the 10-week training study (p = 0.003) with no decrease in LPS after DT in both groups. Although not statistically significant, both groups retained lean mass after DT. A 2-week period of DT appeared to retain muscular strength in resistance-trained men. Therefore, a short-term period of DT can potentially retain lower-body strength in young resistance-trained men irrespective of supplementing with 25 g of whey protein postexercise.

BAIBA Does Not Regulate UCP-3 Expression in Human Skeletal Muscle as a Response to Aerobic Exercise.

OBJECTIVE: ß-Aminoisobutyric acid (BAIBA) has shown to modulate uncoupling protein (UCP)-1 expression, which is mainly expressed in white adipose tissue; however, no studies to date have analyzed its potential effect on the main uncoupling protein of skeletal muscle, UCP-3. The main goal of this study was to assess the potential effect of acute aerobic exercise on serum BAIBA and skeletal muscle UCP-3. The secondary goal was to assess the potential involvement of the transcription factors proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) and peroxisome proliferator-activated receptor alpha (PPARα), as well as free fatty acids (FFAs) in UCP-3 expression. A tertiary goal of the study was to evaluate the potential effect of consuming a preexercise meal on the outcome of the first 2 objectives. METHODS: In a randomized crossover design, untrained participants performed 2 acute cycling sessions (350 kcal at 70% of their VO2peak) after 2 experimental conditions: (1) consumption of a multi-macronutrient shake and (2) a fasting period of 8 hours. Blood samples were taken at baseline, preexercise, postexercise, 1 hour, and 4 hours postexercise, and muscle biopsies were taken at the last 4 time points. UCP-3 protein concentration and expression, as well as the mRNA expression of PGC-1α and PPARα, were measured in muscle, and BAIBA, glucose, and FFA were measured in serum. RESULTS: Aerobic exercise failed to induce a significant effect on serum BAIBA, PGC-1α, and PPARα regardless on the feeding condition. Despite the lack of effect of exercise on the previous variables, UCP-3 expression and protein concentration significantly increased in the shake condition. CONCLUSION: The expression of human skeletal muscle UCP-3 as a result of exercise might be controlled by factors other than BAIBA.

Effects of Fish Oil Supplementation on Postresistance Exercise Muscle Soreness.

The aim of this study was to examine the effects of fish oil supplementation on the magnitude and time-course of postresistance exercise muscle soreness. This study was a randomized, placebo-controlled, double-blind trial. Nonresistance trained females were randomized into one of two groups: fish oil supplementation (6 g/day; 5:1 eicosapentaenoic acid to docosahexaenoic acid (EPA:DHA)) or placebo (6 g/day corn/soy oil). After consuming the supplements for one week, participants underwent a single bout of resistance exercise consisting of 10 sets to failure of elbow flexion and leg extension machines. Muscle soreness was measured daily over the next week via grounded visual analog scale while participants continued to consume their assigned supplement. At 48 hours and one week postexercise, soreness during functional movements and limb circumferences were measured. The fish oil group perceived less static and functional muscle soreness than placebo, although the differences were not statistically significant. Effect sizes for resistance exercise-induced static and functional soreness responses were 33 to 42% lower in fish oil versus placebo without changes in upper arm and thigh circumferences. Supplementing the diet with 6 g per day of fish oil may alleviate muscle soreness experienced after resistance training in young untrained females.

Eight Weeks of Phosphatidic Acid Supplementation in Conjunction with Resistance Training Does Not Differentially Affect Body Composition and Muscle Strength in Resistance-Trained Men.

This study attempted to determine the effects of eight weeks of resistance training (RT) combined with phosphatidic acid (PA) supplementation at a dose of either 250 mg or 375 mg on body composition and muscle size and strength. Twenty-eight resistance-trained men were randomly assigned to ingest 375 mg [PA375 (n = 9)] or 250 mg [PA250 (n = 9)] of PA or 375 mg of placebo [PLC (n = 10)] daily for eight weeks with RT. Supplements were ingested 60 minutes prior to RT and in the morning on non-RT days. Participants' body composition, muscle size, and lower-body muscle strength were determined before and after training/supplementation. Separate group x time ANOVAs for each criterion variable were used employing an alpha level of ≤ 0.05. Magnitude- based inferences were utilized to determine the likely or unlikely impact of PA on each criterion variable. A significant main effect for time was observed for improvements in total body mass (p = 0.003), lean mass (p = 0.008), rectus femoris cross-sectional area [RF CSA (p = 0.011)], and lower-body strength (p < 0.001), but no significant interactions were present (p > 0.05). Collectively, magnitude-based inferences determined both doses of PA to have a likely impact of increasing body mass (74.2%), lean mass (71.3%), RF CSA (92.2%), and very likely impact on increasing lower-body strength (98.1% beneficial). When combined with RT, it appears that PA has a more than likely impact on improving lower-body strength, whereas a likely impact exists for increasing muscle size and lean mass.

Effect of resistance exercise intensity on the expression of PGC-1α isoforms and the anabolic and catabolic signaling mediators, IGF-1 and myostatin, in human skeletal muscle.

The purpose of this study was to investigate the acute messenger (mRNA) expression of the peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) isoforms, insulin-like growth factor-1Ea (IGF-1Ea), and myostatin in response to 2 resistance exercise intensities. In a uniform-balanced, crossover design, 10 participants performed 2 separate testing sessions involving a lower body resistance exercise component consisting of a lower intensity (50% of 1-repetition maximum; 1RM) protocol and a higher intensity (80% of 1RM) protocol of equal volumes. Muscle samples were obtained at before exercise, 45 min, 3 h, 24 h, and 48 h postexercise. Resistance exercise did not alter total PGC-1α mRNA expression; however, distinct responses of each PGC-1α isoform were observed. The response of each isoform was consistent between sessions, suggesting no effect of resistance exercise intensity on the complex transcriptional expression of the PGC-1α gene. IGF-1Ea mRNA expression significantly increased following the higher intensity session compared with pre-exercise and the lower intensity session. Myostatin mRNA expression was significantly reduced compared with pre-exercise values at all time points with no difference between exercise intensity. Further research is needed to determine the effects of the various isoforms of PGC-1α in human skeletal muscle on the translational level as well as their relation to the expression of IGF-1 and myostatin.

Effects of Hypohydration on Repeated 40-yd Sprint Performance.

This study examined the effects of hypohydration on repeated 40-yd sprint performance. Anaerobically fit current and former Division II male athletes (n = 12) completed 2 bouts of 10 × 40-yd sprints followed by an agility test, dehydrated (∼3% body weight [DT]), or hydrated trial (HT). Statistical analysis of group means indicated that hypohydration had little effect on sprint times for either the first (DT= 5.38 ± 0.37; HT = 5.35 ± 0.34) or second (DT = 5.47 ± 0.39; HT = 5.42 ± 0.39) bout of 10 sprints with only sprint number 2, 5, and 6 of bout 2 reaching statistical significance. However, when individual sprint performance was considered, a greater effect was seen. In all, 83% (10 of 12) of subjects experienced a meaningful change (≥0.1 seconds) (positive or negative) in mean sprint time (DT vs. HT) for one or more bout of 10 sprints. Ratings of perceived exertion was significantly higher (∼1 unit on a 10 point scale) for DT in all sprints during bout 1 and the first 2 sprints of bout 2. These results indicate that the effect of hypohydration on repeated sprint performance varies among individuals. Some improved performance with hypohydration, while others experienced detrimental effects. Hypohydration also resulted in a particularly notable negative impact on perceptual measures of exertion even when performance was similar.

Acute Myosin Heavy Chain Isoform mRNA Expression in Response to Two Resistance Exercise Intensities With Equal Volume Load in Resistance-Trained Men.

The purpose of this study was to determine if resistance exercise intensity, in the context of equal volume load, differentially affected myosin heavy chain (MHC) isoform messenger RNA (mRNA) expression in resistance-trained men. In a crossover, uniform-balanced design, 10 male participants (23.7 ± 2.8 years, 178.8 ± 5.9 cm, 85.9 ± 9.2 kg) completed 2 lower-body resistance exercise sessions of different intensities with equal volume load. For the higher-intensity exercise session, participants performed 5 sets of 6 repetitions at 80% of 1 repetition maximum (1RM). For the lower-intensity exercise session, participants performed 3 sets of 16 repetitions at 50% of 1RM. Muscle samples from the vastus lateralis were acquired before exercise (PRE), 45 minutes postexercise (45MINPE), 3 hours postexercise (3HRPE), 24 hours postexercise (24HRPE), and 48 hours postexercise (48HRPE). Statistical analyses of mRNA expression were performed using separate 2 × 5 two-way repeated-measures analyses of variance for each criterion variable (p ≤ 0.05). There were no statistically significant interactions between intensity and time. Likewise, there were no significant differences between exercise intensity in MHC expression. Expression of mRNA for all MHC isoforms decreased at all postexercise time points, except 3HRPE (p = 0.051), compared with PRE following both exercise bouts (p ≤ 0.05). The results of this study found no difference in mRNA expression of MHC isoforms as a function of resistance exercise intensity. In addition, in contrast to results found in previous studies of untrained men, MHC mRNA expression seems to decrease in response to acute resistance exercise in previously resistance-trained men.

Frequency scaling with miniature COmpact MIcrowave and Coaxial ion sources.

We will present recent basic developments about possible extension of the COMIC (for COmpact MIcrowave and Coaxial) devices up to 5.8 GHz in place of the present 2.45 GHz operation [P. Sortais, T. Lamy, J. Médard, J. Angot, L. Latrasse, and T. Thuillier, Rev. Sci. Instrum. 81, 02B314 (2010)]. New applications associating multiple COMIC devices for thin film deposition will be described and we will explain why an increase of the current density delivered by each individual ion source could lead to the increase of the deposition rate. For this purpose, we will present results of about two devices working at 5.8 GHz. The first one is a tiny ion source, the world smallest microwave ion source, exactly similar to COMIC but operating at 5.8 GHz with a quarter wave cavity structure and a few watts microwave power consumption. We will show that the frequency scaling effect is effective inside such small machines. The second one is a more ambitious ion source designed around a three quarter wave structure that works with a few tens of watts at 5.8 GHz.

Peptide microarrays for the profiling of cytotoxic T-lymphocyte activity using minimum numbers of cells.

The identification of epitopes that elicit cytotoxic T-lymphocyte activity is a prerequisite for the development of cancer-specific immunotherapies. However, especially the parallel characterization of several epitopes is limited by the availability of T cells. Microarrays have enabled an unprecedented miniaturization and parallelization in biological assays. Here, we developed peptide microarrays for the detection of CTL activity. MHC class I-binding peptide epitopes were pipetted onto polymer-coated glass slides. Target cells, loaded with the cell-impermeant dye calcein, were incubated on these arrays, followed by incubation with antigen-expanded CTLs. Cytotoxic activity was detected by release of calcein and detachment of target cells. With only 200,000 cells per microarray, CTLs could be detected at a frequency of 0.5% corresponding to 1,000 antigen-specific T cells. Target cells and CTLs only settled on peptide spots enabling a clear separation of individual epitopes. Even though no physical boundaries were present between the individual spots, peptide loading only occurred locally and cytolytic activity was confined to the spots carrying the specific epitope. The peptide microarrays provide a robust platform that implements the whole process from antigen presentation to the detection of CTL activity in a miniaturized format. The method surpasses all established methods in the minimum numbers of cells required. With antigen uptake occurring on the microarray, further applications are foreseen in the testing of antigen precursors that require uptake and processing prior to presentation.

Selectivity of competitive multivalent interactions at interfaces.

The development of synthetic, low-molecular-weight ligand receptor systems for the selective control of biomolecular interactions remains a major challenge. Binding of oligohistidine peptides to chelators containing Ni2+-loaded nitrilotriacetic acid (NTA) moieties is one of the most widely used and best-characterised recognition systems. Recognition units containing multiple NTA moieties (multivalent chelator headgroups, MCHs) recognise oligohistidines with substantially increased binding affinities. Different multivalencies both at the level of the MCH and at that of the oligohistidine ligand provide a powerful means to vary the affinity of the interaction systematically. Here we have explored the selectivity for the binding of different oligohistidines to immobilised MCH. Using microarrays of mono-, bis-, tris- and tetrakis-NTA chelators spotted at different surface densities, we explored the ability of these binders to discriminate fluorescently labelled hexa- and decahistidine peptides. When hexa- and decahistidine were tested alone, the discrimination of ligands showed little dependence either on the nature or on the density of the chelator. In contrast, coincubation of both peptides decreased the affinity of hexahistidine, increased the affinity of decahistidine, and made the binding of decahistidine highly dependent on MCH density. Kinetic binding assays by dual-colour total internal reflection fluorescence spectroscopy revealed active exchange of His6 by His10 and confirmed the high selectivity towards His10. Our results establish the key role of surface multivalency for the selectivity of multivalent interactions at interfaces.