Impact of COVID-19 vaccination on seminal and systemic inflammation in men.

Expedited development of SARS-CoV-2 vaccines led to public concerns regarding impacts of the novel vaccine on gametes in patients seeking assisted reproduction. In cases of an acute intermittent illness or fever in men, it is often advised to postpone ART treatments so that efforts can be made to enhance wellbeing and improve sperm parameters. However, it is unknown whether sperm parameters are altered in the acute (24-72 hour) phase following COVID-19 vaccination. We performed a longitudinal cohort study of 17 normospermic male patients attending a fertility clinic for semen analysis. Semen and matched peripheral blood samples were collected prior to vaccination, within 46 + 18.9 hours of vaccine course completion (acute) and at 88.4 + 12 days (3 months) post-vaccination. No overall change from baseline was seen in symptoms, mean volume, pH, sperm concentration, motility, morphology or DNA damage in the acute or long phase. Seminal plasma was found to be negative for anti-SARS-CoV2 Spike antibody detection, and MCP-1 levels showed an acute but transient elevation post-vaccine, while IL-8 was marginally increased 3 months after completion of vaccination. A modest, positive correlation was noted between serum levels of the anti-inflammatory cytokine IL-10 and self-reported symptoms post-vaccine. Our findings are reassuring in that no significant adverse effect of vaccination was noted and provide evidence to support the current recommendations of reproductive medicine organisations regarding timing of vaccination during fertility treatment.

A new class of 7-deazaguanine agents targeting autoimmune diseases: dramatic reduction of synovial fibroblast IL-6 production from human rheumatoid arthritis patients and improved performance against murine experimental autoimmune encephalomyelitis.

A simple in vitro assay involving the measurement of IL-6 production in human synovial fibroblasts from rheumatoid arthritis patients has been utilised to select candidates from a targeted library of queuine tRNA ribosyltransferase (QTRT) substrates for subsequent in vivo screening in murine experimental autoimmune encephalomyelitis (EAE - a model of multiple sclerosis). The in vitro activity assay discriminated between poor and excellent 7-deazaguanine QTRT substrates and allowed the identification of several structures which subsequently outperformed the previous lead in EAE. Two molecules were of significant promise: one rigidified analogue of the lead, and another considerably simpler structure incorporating an oxime motif which differs structurally from the lead to a considerable extent. These studies provide data from human cells for the first time and have expanded both the chemical space and current understanding of the structure-activity relationship underpinning the remarkable potential of 7-deazguanines in a Multiple Sclerosis disease model.

Physical Activity and Self-Determination towards Exercise among Esports Athletes.

BACKGROUND: Esports is competitive video gaming, performed within teams or individually, across multiple genres. Players are required to be sedentary for extended periods and require a high-level of cognitive skills for successful competitive performance. There are conflicting findings within the physical activity research in the esports industry. The aim of this research is to explore self-reported physical activity through accelerometer-assessed physical activity, to gain a better insight into the physical activity behaviours of international e'athletes. METHOD: Participants (n = 796) across multiple popular esports games, holding any in-game rank, competing at any level, were recruited. The survey consisted of demographic details, esports experience, the International Physical Activity Questionnaire-Long Form (IPAQ-LF), and Behavioural Regulations towards Exercise Questionnaire (BREQ-3). Within a convenience sample, local intervarsity e'athletes (n = 18) were recruited to wear a wrist-worn accelerometer to measure physical activity for 7-days and then complete the survey. Results from the accelerometers were compared to the survey results to explore physical activity reporting within this population. RESULTS: When comparing IPAQ-LF to accelerometer data, players significantly over-report moderate-to-vigorous physical activity and weekly MET-min- 1 (p = .018, r = .63 and p ≤ .001, r = .92). The BREQ-3 showed that e'athletes categorised as high physical activity displayed significantly higher levels of intrinsic motivation, when compared to players categorised as low and moderate physical activity. CONCLUSIONS: E'athletes significantly over report physical activity time when measured through the IPAQ-LF, suggesting previous surveys may overestimate physical activity and further research is needed. Given the exponential growth of the industry and the level of physical inactivity, esports may contribute to global physical inactivity levels.

How Much Energy Do E'Athletes Use during Gameplay? Quantifying Energy Expenditure and Heart Rate Variability Within E'Athletes.

BACKGROUND: Research into esports suggests that e'athletes experience physiological stressors and demands during competition and training. The physiological demands of esports are poorly understood and need to be investigated further to inform future training guidelines, optimise performance outcomes, and manage e'athlete wellbeing. This research aimed to quantify the metabolic rate of esports gameplay and compare this outcome with heart rate variability within expert e'athletes. RESULTS: Thirteen healthy male participants ranked within the top 10% of their respective esports title participated in the study (age = 20.7 ± 2.69 years; BMI = 24.6 ± 5.89 kg·m- 2). Expired gas analysis indirect calorimetry measured gas exchange during rest and gaming. Compared to resting conditions, competitive esports gameplay significantly increased median energy expenditure (1.28 (IQR 1.16-1.49) kcal·min- 1 vs. 1.45 (IQR 1.20-1.77) kcal·min- 1, p = .02), oxygen consumption (0.27 (IQR 0.24-0.30) L·min- 1 vs. 0.29 (IQR 0.24-0.35) L·min- 1, p = .02) and carbon dioxide production (0.20 (IQR 0.19-0.27) L·min- 1vs. 0.27 (IQR 0.24-0.33) L·min- 1, p = .01). Competitive gameplay also resulted in a significant increase in heart rate (84.5 (IQR 74.1-96.1) bpm vs. 87.1 (IQR 80.3-104) bpm, p = .01) and decrease in R-R interval's (710 (IQR 624-810) ms vs. 689 (IQR 579-747) ms, p = .02) when compared to rest. However, there were no significant differences in time or frequency measures of heart rate variability. CONCLUSIONS: The data reveal increased physiological responses to metabolic rate, energy expenditure and cardiovascular function to esports game play within expert e'athletes. Further physiological research into the physical demands on e'athletes, the influence of different training programs to esport performance, and the added multivariate determinants to elite level esport performance are warranted.

Effects of Repeated Jump Testing and Diurnal Changes on Subsequent Countermovement Jump and Squat Jump Output and Force-Time Characteristics.

ABSTRACT: Harrison, PW, James, LP, Jenkins, DG, Holmberg, PM, and Kelly, VG. Effects of repeated jump testing and diurnal changes on subsequent countermovement jump and squat jump output and force-time characteristics. J Strength Cond Res 38(1): 174-179, 2024-The aim of this brief study was to investigate the effects of repeated jump testing on performance over 2 consecutive days while considering the possibility of diurnal changes. Fourteen male subjects and 14 recreationally active female subjects completed countermovement jump (CMJ) and squat jump (SJ) testing on 5 occasions (baseline [0,800], 5 minutes [0,820], 8 hours [1,600], 24 hours [0,800], and 32 hours [1,600]) over 32 hours. An additional rested baseline test was conducted on a separate day in the afternoon (1,600) to compare jump performance between morning and afternoon baseline values. Excluding small decreases in CMJ height at 24 hours (p = 0.292, Cliff's delta = -0.225) in male subjects and similar decreases in CMJ height at 5 minutes (p = 0.034, Cliff's delta = -0.245) in addition to SJ height:contraction time at 32 hours (p = 0.126, Cliff's delta = 0.153) in female subjects, findings generally showed no changes in jump performance over multiple assessments. Squat jump metrics may have showed small improvements between morning and afternoon baseline values in male subjects (SJ height:contraction time [p = 0.030, Cliff's delta = 0.225]) and female subjects (SJ height [p = 0.013, Cliff's delta = 0.173] and SJ height:contraction time [p = 0.091, Cliff's delta = 0.163)]. As jump performance was largely unaffected by repeated jump testing, the present findings support the use of monitoring practices and research designs that require multiple jump assessments within acute periods (∼32 hours).

The Effects of Low-Load Squat Jump and Maximal Isometric Priming Exercise on Muscular Performance and Perceptual State.

ABSTRACT: Harrison, PW, James, LP, Jenkins, DG, McGuigan, MR, Holmberg, PM, and Kelly, VG. The effects of low-load squat jump and maximal isometric priming exercise on muscular performance and perceptual state. J Strength Cond Res 38(1): 1-9, 2024-The aim of this study was to examine responses at 3 and 27 hours after low-load jump squat (LL) and maximal isometric half-squat (ISO) priming stimuli. Fifteen resistance-trained males performed LL (4 × 3 at 20% 1 repetition maximum [1RM]), ISO (4 × 3 seconds), and control (CON) activities (standardized warm-up) in a randomized and counterbalanced order. Countermovement jump (CMJ) and isometric midthigh pull tests were conducted to assess performance changes after priming and CON activities. No clear changes in CMJ measures were found after priming activities compared with CON. However, small effect size improvements were found after priming stimuli completed on the same day. A 2.9% decrease in concentric phase duration (CI = 0.3-5.9, p = 0.333, Cliff's delta = -0.156) and a 9.1% increase in RSImod (CI = 0.2-12.3, p = 0.151, Cliff's delta = -0.218) occurred at 3 hours after LL compared with CON. Braking phase duration (CI = 0.8-10.6, p = 0.333, Cliff's delta = -0.213) was 2.9% shorter at 3 hours after ISO compared with CON. No clear changes in isometric peak force occurred after priming activities compared with CON. Additionally, questionnaires were completed to assess perceptual state and perceived effectiveness of the priming stimulus to influence performance. An increase in the "effect of activity" was perceived at 3 hours after LL and ISO (p = 0.013-0.044, Cliff's delta = 0.578-0.6) and at 27 hours after ISO (p = 0.99, Cliff's delta = 0.173) compared with CON. An increase in "muscular heaviness" was also reported at 3 hours after ISO compared with CON (p = 0.199, Cliff's delta = 0.320). The collective findings suggest limited benefits over the day after LL and ISO priming stimuli. However, as there was substantial variation in individual responses, the relative nature of priming responses should be considered when prescribing similar strategies in practical environments.

Networking to Optimize Dmd exon 53 Skipping in the Brain of mdx52 Mouse Model.

Duchenne muscular dystrophy (DMD) is caused by mutations in the DMD gene that disrupt the open reading frame and thus prevent production of functional dystrophin proteins. Recent advances in DMD treatment, notably exon skipping and AAV gene therapy, have achieved some success aimed at alleviating the symptoms related to progressive muscle damage. However, they do not address the brain comorbidities associated with DMD, which remains a critical aspect of the disease. The mdx52 mouse model recapitulates one of the most frequent genetic pathogenic variants associated with brain involvement in DMD. Deletion of exon 52 impedes expression of two brain dystrophins, Dp427 and Dp140, expressed from distinct promoters. Interestingly, this mutation is eligible for exon skipping strategies aimed at excluding exon 51 or 53 from dystrophin mRNA. We previously showed that exon 51 skipping can restore partial expression of internally deleted yet functional Dp427 in the brain following intracerebroventricular (ICV) injection of antisense oligonucleotides (ASO). This was associated with a partial improvement of anxiety traits, unconditioned fear response, and Pavlovian fear learning and memory in the mdx52 mouse model. In the present study, we investigated in the same mouse model the skipping of exon 53 in order to restore expression of both Dp427 and Dp140. However, in contrast to exon 51, we found that exon 53 skipping was particularly difficult in mdx52 mice and a combination of multiple ASOs had to be used simultaneously to reach substantial levels of exon 53 skipping, regardless of their chemistry (tcDNA, PMO, or 2'MOE). Following ICV injection of a combination of ASO sequences, we measured up to 25% of exon 53 skipping in the hippocampus of treated mdx52 mice, but this did not elicit significant protein restoration. These findings indicate that skipping mouse dystrophin exon 53 is challenging. As such, it has not yet been possible to answer the pertinent question whether rescuing both Dp427 and Dp140 in the brain is imperative to more optimal treatment of neurological aspects of dystrophinopathy.

Does Moderate-Load Priming Activity Influence Maximal Upper-Body Performance and Perceptual State?

ABSTRACT: Harrison, PW, Kelly, VG, Jenkins, DG, McGuigan, MR, Holmberg, PM, and James, LP. Does moderate-load priming activity influence maximal upper-body performance and perceptual state?. J Strength Cond Res 37(11): e581-e587, 2023-The results of previous research indicate that resistance exercise "priming" may improve strength-power measures within 48 hours after their completion. Although researchers have primarily examined performance responses after lower-body priming stimuli, investigations examining the effects of upper-body resistance priming exercises are presently limited. Therefore, the aim of this study was to examine upper-body pushing and pulling performance in addition to perceptual responses 3 and 27 hours after moderate-load (ML) upper-body resistance priming exercise. Fourteen resistance-trained men were assigned to complete ML priming (4 × 3 bench press and bench pull at 65% 1RM [repetition maximum]) and control (rest) protocols in a randomized and counterbalanced order. Peak velocity during the bench throw and bench pull tests involving different loads (25, 50, and 75% 1RM) showed no practical change at 3 and 27 hours after the priming session (p = 0.216-0.99, Cliff's d = -0.041 to 0.225). Small effect size increases in perceptual measures ("physical feeling," "physical performance," "aggression" [p = 0.400-0.553, Cliff's d = 0.183-0.201], and "muscular heaviness" [p = 0.178, Cliff's d = 0.231]) were found at 3 hours postpriming. A moderate practical increase was observed in perceived "physical feeling" compared with control (p = 0.385, Cliff's d = 0.349) in addition to small effect size increases in perceived "physical performance" and "aggression" (Cliff's d = 0.243-0.290) at 27 hours after priming activities. These results indicate that upper-body strength-power changes within 27 hours after ML upper-body resistance exercise priming are not practically meaningful.

Thinking About Elite Performance: The Experience and Impact of Mental Fatigue in Elite Sport Coaching.

PURPOSE: Mental fatigue causes decreases in aspects of athletes' performance. Elite coaches commonly undertake cognitively demanding tasks and are seemingly at similar risk of subsequent performance impairment. However, elite sport coaches' experiences of mental fatigue, alongside other markers of psychobiological stress, have yet to be quantified. METHODS: Three elite coaching and performance staff (2 women and 1 man) provided 100-mm visual analog scale ratings of mental fatigue, physical fatigue, readiness to perform, and salivary samples for later cortisol (sCort) and alpha-amylase (sAA) analysis. Data were obtained on the same morning each week across a 16-week preseason. Data were subset by individual coach for descriptive and repeated-measures correlational analyses. RESULTS: Fluctuating mental fatigue was observed over the 16 weeks (min-max; coach 1 = 25-86 AU; coach 2 = 0-51 AU; and coach 3 = 15 - 76 AU). Elevated levels of mental fatigue were reported at multiple time points, with individual variability observed. sCort (in nanomoles per liter), sAA (in micromoles per liter), and sAA:sCort indicated that coaches experienced psychophysiological stress (min-max; coach 1 sCort = 8.42-17.31, sAA = 52.40-113.06, sAA:sCort = 3.20-12.80; coach 2 sCort = 4.20-9.70, sAA = 158.80-307.20, sAA:sCort = 21.10-61.70; and coach 3 sCort = 6.81-19.66, sAA = 86.55-495.85, sAA:sCort = 4.90-35.50). A significant inverse relationship between mental fatigue and readiness to perform (r = -.44 [-.64 to -.17], P = .002) was identified. CONCLUSIONS: Elite sport coaches report elevated instances of mental fatigue during a preseason training period. Those involved in elite sports should act to understand the presence and potential subsequent impacts of staff mental fatigue and consider management or mitigation strategies. Optimization of the cognitive performance of coaches and performance staff presents as a potential source of competitive advantage.

Queuine Analogues Incorporating the 7-Aminomethyl-7-deazaguanine Core: Structure-Activity Relationships in the Treatment of Experimental Autoimmune Encephalomyelitis.

A library of queuine analogues targeting the modification of tRNA isoacceptors for Asp, Asn, His and Tyr catalysed by queuine tRNA ribosyltransferase (QTRT, also known as TGT) was evaluated in the treatment of a chronic multiple sclerosis model: murine experimental autoimmune encephalomyelitis. Several active 7-deazaguanines emerged, together with a structure-activity relationship involving the necessity for a flexible alkyl chain of fixed length.

The Perceptions of Elite Professional Rugby League Players and Staff on the National Rugby League Annual Calendar: A Mixed-Methods Study.

BACKGROUND: In recent years, the length of elite sporting competitions has raised concerns regarding player well-being, highlighting a need to review current match calendars. Therefore, this study aimed to explore the perceptions of elite National Rugby League (NRL) players and staff on the annual training and competition calendar from a player workload and well-being perspective. METHODS: This study adopted a mixed-methods approach, using a sequential explanatory design. Phase one implemented a cross-sectional survey, and phase two utilised semi-structured interviews. Four hundred and thirty-nine elite rugby league players and 46 staff completed the survey. Eighteen elite professional NRL players and six football staff were interviewed, and verbal data were analysed into pre-defined topic summaries using qualitative coding reliability methods. Topics included in-season, off-season, pre-season and well-being. RESULTS: Data analysis suggests that elite NRL players and staff believe players appear particularly comfortable with the current number of games; however, they are at their maximum capacity. Importantly, this study identified several minority groups that may require support to enhance player well-being. Players believe reducing the pre-season would negate fatigue experienced later in the subsequent season. Players and staff believe this timeframe still provides sufficient time to prepare for the upcoming season. Further, players were open to extending the off-season to 8-10 weeks and believed that extra time would allow for greater recovery from the previous season. Mid-season congested scheduling affects players following the intensified period and requires attention to alleviate fatigue. CONCLUSION: The results of this study convey important implications for the NRL, emphasising a need to review their annual training and competitive calendar, or to implement specific strategies to enhance the well-being of minority groups. The findings from this study should be considered when discussing the ideal length and structure of the match calendar to support players' physical and mental welfare.

The Effectiveness of Online Mental Health First Aid Training in Community Rugby: A Mixed-Methods Approach.

Mental Health First Aid (MHFA) training exists to improve supportive behaviours towards peers, increase mental health literacy, and reduce stigma. Community sport clubs have potential to successfully deliver mental health programs. This study investigated the effectiveness of online MHFA training undertaken by members of the rugby community and evaluated the feasibility and usefulness of the online delivery mode and users' engagement with it. A mixed-methods approach was used to provide depth of understanding through qualitative analysis, combined with quantitative outcomes. Online surveys examining participants' knowledge and perceptions were administered pre- and post-MHFA training. Significant improvements (p < 0.05) across all assessed domains were observed post- compared to pre-MHFA training. A large effect size was identified in relation to advice giving and sign and symptom identification. A moderate effect size pre- to post-improvement was identified for users' perceptions of therapy's effectiveness, the ability of people with severe mental health conditions to recover, and benefit of a healthcare professional. Participants endorsed the MHFA program to improve mental health literacy, advance non-technical skills, and improve confidence. MHFA training can increase the awareness and knowledge of mental health issues in key individuals in community sport clubs and enable them to aid people with mental health concerns. Online MHFA training is associated with improved mental health literacy and may be a suitable and economically sustainable model for community sport.

Investigating the Impact of Delivery Routes for Exon Skipping Therapies in the CNS of DMD Mouse Models.

Nucleic acid-based therapies have demonstrated great potential for the treatment of monogenetic diseases, including neurologic disorders. To date, regulatory approval has been received for a dozen antisense oligonucleotides (ASOs); however, these chemistries cannot readily cross the blood-brain barrier when administered systemically. Therefore, an investigation of their potential effects within the central nervous system (CNS) requires local delivery. Here, we studied the brain distribution and exon-skipping efficacy of two ASO chemistries, PMO and tcDNA, when delivered to the cerebrospinal fluid (CSF) of mice carrying a deletion in exon 52 of the dystrophin gene, a model of Duchenne muscular dystrophy (DMD). Following intracerebroventricular (ICV) delivery (unilateral, bilateral, bolus vs. slow rate, repeated via cannula or very slow via osmotic pumps), ASO levels were quantified across brain regions and exon 51 skipping was evaluated, revealing that tcDNA treatment invariably generates comparable or more skipping relative to that with PMO, even when the PMO was administered at higher doses. We also performed intra-cisterna magna (ICM) delivery as an alternative route for CSF delivery and found a biased distribution of the ASOs towards posterior brain regions, including the cerebellum, hindbrain, and the cervical part of the spinal cord. Finally, we combined both ICV and ICM injection methods to assess the potential of an additive effect of this methodology in inducing efficient exon skipping across different brain regions. Our results provide useful insights into the local delivery and associated efficacy of ASOs in the CNS in mouse models of DMD. These findings pave the way for further ASO-based therapy application to the CNS for neurological disease.

Disruption to tRNA Modification by Queuine Contributes to Inflammatory Bowel Disease.

BACKGROUNDS AND AIMS: Transfer RNA (tRNA) is the most extensively modified RNA in cells. Queuosine modification is a fundamental process for ensuring the fidelity and efficiency of translation from RNA to protein. In eukaryotes, Queuosine tRNA (Q-tRNA) modification relies on the intestinal microbial product queuine. However, the roles and potential mechanisms of Q-containing tRNA (Q-tRNA) modifications in inflammatory bowel disease (IBD) are unknown. METHODS: We explored the Q-tRNA modifications and expression of QTRT1 (queuine tRNA-ribosyltransferase 1) in patients with IBD by investigating human biopsies and reanalyzing datasets. We used colitis models, QTRT1 knockout mice, organoids, and cultured cells to investigate the molecular mechanisms of Q-tRNA modifications in intestinal inflammation. RESULTS: QTRT1 expression was significantly downregulated in ulcerative colitis and Crohn's disease patients. The 4 Q-tRNA-related tRNA synthetases (asparaginyl-, aspartyl-, histidyl-, and tyrosyl-tRNA synthetase) were decreased in IBD patients. This reduction was further confirmed in a dextran sulfate sodium-induced colitis model and interleukin-10-deficient mice. Reduced QTRT1 was significantly correlated with cell proliferation and intestinal junctions, including downregulation of ß-catenin and claudin-5 and the upregulation of claudin-2. These alterations were confirmed in vitro by deleting the QTRT1 gene from cells and in vivo using QTRT1 knockout mice. Queuine treatment significantly enhanced cell proliferation and junction activity in cell lines and organoids. Queuine treatment also reduced inflammation in epithelial cells. Moreover, altered QTRT1-related metabolites were found in human IBD. CONCLUSIONS: tRNA modifications play an unexplored novel role in the pathogenesis of intestinal inflammation by altering epithelial proliferation and junction formation. Further investigation of the role of tRNA modifications will uncover novel molecular mechanisms for the prevention and treatment of IBD.

Structural basis of Qng1-mediated salvage of the micronutrient queuine from queuosine-5'-monophosphate as the biological substrate.

Eukaryotic life benefits from-and ofttimes critically relies upon-the de novo biosynthesis and supply of vitamins and micronutrients from bacteria. The micronutrient queuosine (Q), derived from diet and/or the gut microbiome, is used as a source of the nucleobase queuine, which once incorporated into the anticodon of tRNA contributes to translational efficiency and accuracy. Here, we report high-resolution, substrate-bound crystal structures of the Sphaerobacter thermophilus queuine salvage protein Qng1 (formerly DUF2419) and of its human ortholog QNG1 (C9orf64), which together with biochemical and genetic evidence demonstrate its function as the hydrolase releasing queuine from queuosine-5'-monophosphate as the biological substrate. We also show that QNG1 is highly expressed in the liver, with implications for Q salvage and recycling. The essential role of this family of hydrolases in supplying queuine in eukaryotes places it at the nexus of numerous (patho)physiological processes associated with queuine deficiency, including altered metabolism, proliferation, differentiation and cancer progression.

Knockout of ABCC1 in NCI-H441 cells reveals CF to be a suboptimal substrate to study MRP1 activity in organotypic in vitro models.

Multidrug resistance-associated protein 1 (MRP1/ABCC1) is an efflux transporter responsible for the extrusion of endogenous substances as well as xenobiotics and their respective metabolites. Its high expression levels in lung tissue imply a key role in pulmonary drug disposition. Moreover, its association with inflammatory lung diseases underline MRP1's relevance in drug development and precision-medicine. With the aim to develop a tool to better understand MRP1's role in drug disposition and lung disease, we generated an ABCC1-/- clone based on the human distal lung epithelial cell line NCI-H441 via a targeted CRISPR/Cas9 approach. Successful knockout (KO) of MRP1 was confirmed by qPCR, immunoblot and Sanger sequencing. To assess potential compensatory upregulation of transporters with a similar substrate recognition pattern as MRP1, expression levels of MRP2-9 as well as OAT1-4, 6, 7 and 10 were measured. Functional transporter activity was determined via release studies with two prodrug/substrate pairs, i.e. 5(6)-carboxyfluorescein (CF; formed from its diacetate prodrug) and S-(6-(7-methylpurinyl))glutathione (MPG; formed from its prodrug 6-bromo-7-methylpurine, BMP), respectively. Lastly, transepithelial electrical resistance (TEER) of monolayers of the KO clone were compared with wildtype (WT) NCI-H441 cells. Of eight initially generated clones, the M2 titled clone showed complete absence of mRNA and protein in accordance with the designed genome edit. In transport studies using the substrate CF, however, no differences between the KO clone and WT NCI-H441 cells were observed, whilst no differences in expression of potential compensatory transporters was noted. On the other hand, when using BMP/MPG, the release of MPG was reduced to 11.5% in the KO clone. Based on these results, CF appears to be a suboptimal probe for the study of MRP1 function, particularly in organotypic in vitro and ex vivo models. Our ABCC1-/- NCI-H441 clone further retained the ability to form electrically tight barriers, making it a useful model to study MRP1 function in vitro.

Relationship Between Training Workloads, Match Workloads, and Match Performance in Elite Netball.

PURPOSE: This study examined the relationships between training workloads, game workloads, and match performance in an elite netball team. METHODS: Ten elite female netball athletes were monitored over a complete season. Training and game external workloads were determined through inertial movement units and expressed as absolute PlayerLoad (PL) and change of direction (COD). Monthly workload and training efficiency index were also calculated, which used internal workloads (session rating of perceived exertion and summated heart-rate zones). Game performance was assessed through a performance analysis statistic algorithm called NetPoints. To account for the influence of team game workloads on each other, the average workload for midcourt positions (avgMC) was calculated for each game. Data for each athlete were transformed into z scores, and linear mixed modeling was used to build models to examine the relationships between workloads and game performance. RESULTS: Monthly PL, training efficiency index PL, and avgMC PL were statistically significant (P < .05) and positively related to game PL (z = 0.20-0.35, P < .001-.02). For game COD, statistically significant positive relationships were found between monthly COD (z = 0.29 [0.11], P = .01) and avgMC COD (z = 0.21 [0.09], P = .03). The models for NetPoints found significant negative relationships with monthly PL (z = 0.46 [0.12], P < .001) and COD (z = -0.36 [0.11], P = .01). CONCLUSIONS: Higher monthly workloads are related to higher game workload; however, they are also related to decreases in match performance. Therefore, netball practitioners should consider that increases to training workload in a 4-week period prior to a game can influence game workloads and performance.

Altered countermovement jump force profile and muscle-tendon unit kinematics following combined ballistic training.

Combined heavy- and light-load ballistic training is often employed in high-performance sport to improve athletic performance and is accompanied by adaptations in muscle architecture. However, little is known about how training affects muscle-tendon unit (MTU) kinematics during the execution of a sport-specific skill (e.g., jumping), which could improve our understanding of how training improves athletic performance. The aim of this study was to investigate vastus lateralis (VL) MTU kinematics during a countermovement jump (CMJ) following combined ballistic training. Eighteen young, healthy males completed a 10-week program consisting of weightlifting derivatives, plyometrics, and ballistic tasks under a range of loads. Ultrasonography of VL and force plate measurements during a CMJ were taken at baseline, mid-test, and post-test. The training program improved CMJ height by 11 ± 13%. During the CMJ, VL's MTU and series elastic element (SEE) length changes and velocities increased from baseline to post-test, but VL's fascicle length change and velocity did not significantly change. It is speculated that altered lower limb coordination and increased force output of the lower limb muscles during the CMJ allowed more energy to be stored within VL's SEE. This may have contributed to enhanced VL MTU work during the propulsion phase and an improved CMJ performance following combined ballistic training.

Design and synthesis of multifunctional microtubule targeting agents endowed with dual pro-apoptotic and anti-autophagic efficacy.

Autophagy is a lysosome dependent cell survival mechanism and is central to the maintenance of organismal homeostasis in both physiological and pathological situations. Targeting autophagy in cancer therapy attracted considerable attention in the past as stress-induced autophagy has been demonstrated to contribute to both drug resistance and malignant progression and recently interest in this area has re-emerged. Unlocking the therapeutic potential of autophagy modulation could be a valuable strategy for designing innovative tools for cancer treatment. Microtubule-targeting agents (MTAs) are some of the most successful anti-cancer drugs used in the clinic to date. Scaling up our efforts to develop new anti-cancer agents, we rationally designed multifunctional agents 5a-l with improved potency and safety that combine tubulin depolymerising efficacy with autophagic flux inhibitory activity. Through a combination of computational, biological, biochemical, pharmacokinetic-safety, metabolic studies and SAR analyses we identified the hits 5i,k. These MTAs were characterised as potent pro-apoptotic agents and also demonstrated autophagy inhibition efficacy. To measure their efficacy at inhibiting autophagy, we investigated their effects on basal and starvation-mediated autophagic flux by quantifying the expression of LC3II/LC3I and p62 proteins in oral squamous cell carcinoma and human leukaemia through western blotting and by immunofluorescence study of LC3 and LAMP1 in a cervical carcinoma cell line. Analogues 5i and 5k, endowed with pro-apoptotic activity on a range of hematological cancer cells (including ex-vivo chronic lymphocytic leukaemia (CLL) cells) and several solid tumor cell lines, also behaved as late-stage autophagy inhibitors by impairing autophagosome-lysosome fusion.

Effects of the Off-Season on Muscular Power in Professional Rugby League.

PURPOSE: Periods away from training and competition are necessary for physical and mental restoration in sport. There is limited research investigating changes to physical qualities in rugby league following prolonged breaks. This study aimed to evaluate the effects of the off-season on muscular power in rugby league. In addition, this study aimed to determine whether the type and volume of training players chose to complete had any relationship to feelings of restoration and/or readiness to return to training. METHODS: Twenty professional rugby league players participated in this study. Lower-body muscular power was evaluated using a countermovement jump prior to the off-season and at the recommencement of training. Players completed a questionnaire to identify training and activities completed during the break and to assess feelings of restoration and readiness to commence training. Linear regression was used to estimate the effects of the number of days off on muscular power. One-way repeated-measures analysis of variance was conducted to examine differences in lower-body power throughout the study. Spearman rank order correlation was calculated to determine the relationship between off-season activities and feelings of restoration and readiness. RESULTS: Peak velocity and peak force significantly increased following the off-season break. There were significant relationships between mental restoration and upper-body resistance training, as well as physical restoration and full-body resistance training. CONCLUSIONS: The off-season has a positive effect on a player's ability to generate muscular power. Coaches may want to encourage players to complete resistance training sessions with the aim simply to maintain training load and potentially help players to feel rested.