Search Results (1,582)

Gene editing, particularly CRISPR/Cas technology, represents a promising approach for the treatment of rare genetic diseases, including inherited retinal dystrophies, for which effective therapies are largely unavailable. Despite extensive research investigating gene editing across a wide range of cell types, transient delivery of CRISPR/Cas components and efficient homology-directed repair (HDR) in differentiated cells remain challenging. In this study, we employed hiPSCs derived from patients with Stargardt disease or Best disease, carrying pathogenic variants in ABCA4 or BEST1, respectively, to explore gene editing in human models. CRISPR/Cas9 and Cas12 nucleases were delivered into hiPS-derived retinal pigment epithelium (RPE) and retinal organoids using lipoplexes and compared with electroporation. We evaluated transfection efficiency, sgRNA-mediated DNA cleavage, and HDR-based correction. Precise repair of the pathogenic BEST1 variant was successfully achieved in hiPS-derived RPE cells using both nucleases, with Cas12 yielding the highest efficiency, exceeding 10% of HDR correction. Edited RPE cells preserved normal morphology and expressed specific maturity markers. In contrast, retinal organoids exhibited moderate transfection efficiency but showed no detectable CRISPR/Cas-induced DNA cleavage, highlighting the need for further optimization of gene editing in more complex cellular tissues. This study demonstrates, for the first time, precise correction of a single-nucleotide mutation in patient-derived RPE using CRISPR/Cas9 and Cas12 delivered using lipoplexes. These findings underscore the therapeutic potential of CRISPR/Cas-based strategies for inherited retinal dystrophies and provide a proof of concept for future clinical approximations. Full article

Background: Sodium bicarbonate (SB) supplementation can enhance performance in short, high-intensity movements. However, its effectiveness in team sports such as rugby remains insufficiently explored. Methods: In this double-blind, parallel, controlled trial, 17 male professional rugby players ingested SB (0.3 g/kg) or a placebo 90 min before a high-intensity, rugby-specific training session monitored via GPS. The training session was conducted under real-world conditions to enhance ecological validity. Physical performance (countermovement jump, CMJ), fatigue markers (capillary lactate and ratings of perceived exertion, RPE), and gastrointestinal (GI) symptoms were assessed pre- and post-exercise. Results: No significant pre–post changes were observed in CMJ performance in either group. Lactate concentrations increased from pre- to post-exercise in both groups (both p < 0.001). The SB group showed higher GI symptom severity before, during and after exercise versus placebo, with several symptoms increasing over time solely in the SB group (p < 0.05). RPE increased similarly in both groups (SB: p = 0.012; PLA: p = 0.008). Due to the small sample size, only moderate-to-large within-group effects and very large between-group differences could be detected; therefore, the study was powered to detect moderate-to-large within-group effects but underpowered for detecting between-group differences. Conclusions: Acute SB ingestion at 0.3 g/kg did not result in detectable improvements in performance or fatigue markers during rugby-specific high-intensity training and was associated with a greater incidence of GI discomfort; however, the study was underpowered to detect small between-group differences. This study was registered on 23 May 2025 on ClinicalTrials.gov (NCT07017582). Full article

Background/Objectives: SETD2 is a dual-function methyltransferase important for methylation of histone H3 at lysine 36 and α-tubulin in spindle microtubules. Genetic inactivation of SETD2 during oncogenesis drives loss of H3K36me3, genomic instability, and cancer progression. This study asked if disruption of genomic stability was a canonical feature of SETD2 inactivation via different pathways. Methods: We evaluated the impact of EPZ-719, a pharmacologic SETD2 inhibitor, and an H3.3K36M mutant histone (“oncohistone”) that binds and sequesters SETD2, on methylation activity and genomic stability in human cell lines. SETD2 activity was measured using in vitro methylation assays, H3K36me3 loss confirmed by Western analysis, and mitotic defects, specifically micronuclei and chromatin bridges, quantified with cytogenetic analysis. Results: EPZ-719 caused a dose- and time-dependent reduction in SETD2 activity on both histone and tubulin substrates, accompanied by significant increases in chromatin bridges and micronuclei in retinal pigmented epithelial (RPE-1) and 786-O ccRCC cells. Similarly, oncohistone expression markedly decreased SETD2 function, as determined by H3K36me3 levels, and induced comparable mitotic defects in 786-O cells, and aneuploidy in two chondrocyte cell lines expressing the H3.3K36M oncohistone. Combining EPZ-719 with H3.3K36M expression did not exacerbate mitotic defects beyond either oncohistone or pharmacologic inhibition alone, consistent with inhibition of SETD2 as their shared underlying mechanism of action. Conclusions: Pharmacologic inhibition and oncohistone-mediated sequestration of SETD2 converge on the induction of mitotic defects, underscoring SETD2’s essential role in maintaining genomic stability. Identification of loss of genomic stability as a canonical feature of SETD2 inactivation points to a potential therapeutic liability associated with targeting SETD2 in cancers where it is overexpressed and reveals a mechanism that could contribute to the progression of cancers expressing oncohistone mutations. Full article

The objective of this study was to investigate the effect of nitrate (NO₃⁻) supplementation on exercise performance in multiple sets of bench press (BP) and leg press (LP) at 80% of one-repetition maximum (1RM), to determine whether it would be beneficial towards the number of repetitions to failure (RTF). A total of 18 trained male subjects (25 ± 3 years old) completed two sessions of repeated number of maximum repetition (NMR) tests in BP and LP to assess RTF, power output, heart rate (HR), rating of perceived exertion (RPE) 2 h after NO₃⁻ or placebo intake. Comparisons between dependent variables were conducted using a two-factor analysis of variance (ANOVA) with repeated measures, examining the factors of condition and sets. The results for RTF showed only a main effect of set for BP and LP. No significant differences were found between conditions for total RTF. Our results showed that the NO₃⁻ supplementation had no significant effect on RTF, mean power, peak power, HR, and RPE when compared to placebo conditions. Results demonstrated that for physically active male individuals with experience in strength training, NO₃⁻ supplementation did not affect strength endurance performance. Full article

This study examined weekly internal load and neuromuscular performance in elite junior female boxers over 10 weeks. Internal load was quantified using session rating of perceived exertion (sRPE), from which weekly monotony and strain were derived. Neuromuscular performance was assessed weekly using wall-sit endurance and a repetitive jump test. Twenty elite junior female boxers (Mean ± SD: 18.9 ± 1.2) were monitored during regular training without experimental manipulation. Weekly sRPE-derived training load, monotony, and strain showed statistically significant week-to-week fluctuations (p < 0.001). Neuromuscular performance improved in week 2, declined during weeks 3–5, and partially recovered in week 6. The findings demonstrated consistent temporal alignment between internal-load indices and week-to-week neuromuscular performance changes within an observational monitoring framework. Inter-individual variability was observed across athletes. Overall, sRPE-derived indices reflected training stress patterns and were aligned with neuromuscular performance changes in elite female boxers, supporting their use for contextual monitoring of weekly training responses. These findings support the practical integration of internal-load and performance monitoring in elite female combat-sport settings. Future research incorporating boxing-specific external-load metrics, physiological markers, and longer monitoring periods may further refine individualized load-management strategies. Full article

Background: Caffeine is an ergogenic aid shown to delay fatigue, increase arousal, and improve performance. Recommended doses are 3–6 mg/kg BM, although evidence supports lower doses (<3 mg/kg). Some conflicting results have highlighted that lower doses may still be ergogenic, and with new pouch and gum products, further research is warranted. Method: This study investigated the effects of novel low-dose caffeine products on muscular endurance, strength, and power. A repeated-measure, crossover design (pouch 80 mg, gum 80 mg, control gum 0 mg) was employed, recruiting nineteen participants (age 22.4 + 4.8 yrs; weight 72.8 + 16.9 kg; relative caffeine dose 1.1 mg/kg). Participants completed a battery of tests, including 60% 1 RM single leg press (LP) and shoulder press (SP) to exhaustion, counter-movement jump, and isometric mid-thigh pull, in addition to providing ratings of perceived exertion (RPE) during endurance tests. One-way repeated measures ANOVA was conducted on all measures associated with physical tasks, with a two-way repeated measures ANOVA conducted for RPE. Results: No significance was observed among conditions for physical tests. However, effect sizes, employing Cohen’s D classification, identified a moderate (d = 0.55) and small (d = 0.45) effect for the caffeine pouch compared to the placebo and caffeine gum for the LP respectively. Further, small effects for the pouch compared to the placebo were observed (d = 0.33) for the SP. Significant differences were produced for RPE during the LP (p = 0.022), with post hoc analysis identifying significant differences between the placebo vs. caffeine pouch (p = 0.032). Conclusion: Low-dose caffeine has the potential to produce meaningful effects on strength endurance, likely linked to caffeine mechanisms reducing RPE. Full article

Myofibroblasts derived from retinal pigment epithelial (RPE) cells play a key role in the pathogenesis of retinal fibrotic conditions such as proliferative vitreoretinopathy (PVR). Upon exposure to growth factors and cytokines such as TNF-α and TGF-β (TNT), RPE cells undergo epithelial-mesenchymal transition and subsequent transdifferentiation to contractile myofibroblasts. In this study, the effects of JD5037, a peripherally restricted CB1 antagonist, on myofibroblast transdifferentiation of primary cultures of human RPE cells were assessed. JD5037 significantly reduced TNT-induced, RPE cell-mediated collagen gel contraction, an indicator of myofibroblast function, in a concentration-dependent manner. Western blot analysis showed that JD5037 attenuated TNT-induced expression of α-SMA and fibronectin, two molecular markers of myofibroblasts. Furthermore, siRNA knockdown of CB1 cannabinoid receptor partially inhibited TNT-induced myofibroblast transdifferentation of human RPE cells and eliminated the inhibitory effects of JD5037 on myofibroblast transdifferentiation. These data demonstrate, for the first time, that peripherally restricted antagonists, such as JD5037, targeting the CB1 cannabinoid receptor have therapeutic potential for PVR and other retinal fibrotic conditions. Full article

Brown macroalgae are a valuable source of bioactive polysaccharides, particularly alginates and fucoidans, with significant potential for cosmetic and pharmaceutical applications. In this study, polysaccharides were extracted from four species (Padina pavonica, Sargassum ilicifolium, S. latifolium, and Turbinaria decurrens) collected along the Djibouti coastline. Structural characterization by FT-IR and ¹H-NMR revealed pronounced interspecific variability. Alginates displayed distinct mannuronate/guluronate (M/G) ratios, with Sargassum latifolium showing the highest and T. decurrens the lowest, reflecting differences in polymer composition and structural flexibility. Fucoidan spectra exhibited characteristic sulfate bands at 1217–1220 and 840 cm⁻¹, with lower transmission values for T. decurrens indicating a relatively higher degree of sulfation. Biological assays demonstrated that alginate extracts exhibited moderate antioxidant activity, whereas fucoidans showed significantly stronger radical scavenging (DPPH) and ferric reducing (FRAP) capacities, in some cases comparable to vitamin C. Fucoidans also displayed potent inhibition of skin-aging enzymes, with elastase inhibition reaching 62.1% for P. pavonica and tyrosinase inhibition peaking at 63% for S. ilicifolium at 0.5 mg/mL. These results highlight the critical role of structural features, particularly sulfation patterns, in determining biological activity. Overall, Djiboutian brown algal polysaccharides combine antioxidant and enzyme inhibitory properties, confirming their potential as multifunctional and sustainable marine-derived ingredients for cosmetic formulations. Full article

This study investigated the acute effects of anodal transcranial direct current stimulation (a-tDCS) applied over the dorsolateral prefrontal cortex (DLPFC) and primary motor cortex (M1) on neuromuscular performance during a velocity-based back squat exercise. Fifteen recreationally trained men participated in a randomized, double-blind, crossover design, completing three experimental conditions (SHAM, DLPFC, and M1 stimulation) consisting of 20 min of 2 mA a-tDCS followed by a squat protocol performed to a 15% velocity loss threshold. Total repetitions, repetitions per set, mean concentric velocity, and rating of perceived exertion (RPE) were recorded. No significant differences between stimulation conditions were observed for any outcome variable. However, two individuals showed reversed responses, consistent with previously reported inter-individual variability in response to tDCS. Given the high inter-individual variability in response to a-tDCS, we additionally performed a post hoc sensitivity analysis based on response direction relative to SHAM. This analysis indicated that a-tDCS over M1 and DLPFC resulted in a significantly greater total number of repetitions compared with SHAM, whereas repetitions per set, mean velocity, and RPE were not different between conditions. Accordingly, a systematic and individualized approach may be needed to address inter-individual variability in response to tDCS to optimize its effect on fatigue tolerance. Full article

The rapid integration of generative AI in education often frames teachers as technology users who primarily need technical training. Existing prompt engineering frameworks offer technical guidance but have limited grounding in theories of teacher professional development or reflective practice. This misses a key feature of prompt engineering: prompting can externalize pedagogical thinking, making AI interaction a process of knowledge externalization. Through systematic conceptual analysis, this paper proposes a reconceptualization of prompt engineering from a technical competency to a reflective professional practice. The methodology integrates three theoretical traditions: Schön’s reflective practice theory (for externalizing tacit knowledge), Wiggins and McTighe’s backward design (for structuring instructional decisions), and Celik’s AI-TPACK framework (as integrated knowledge base). This synthesis suggests that effective prompting can be understood as an act of pedagogical externalization requiring integrated professional knowledge. The paper develops a seven-strategy framework (RPE framework) as an analytic lens for examining prompt engineering sophistication. This theoretical framework offers theory-derived hypotheses that require future empirical validation rather than presenting verified outcomes. Ultimately, the RPE framework offers a conceptual basis for potentially shifting the focus from technical training to teacher professional development by repositioning educators as AI-assisted instructional designers rather than mere AI users. Full article

Background/Objectives: While CRISPR/Cas9 technology offers a revolutionary approach for correcting genetic ocular blindness, efficient and safe delivery remains the primary bottleneck. Traditional viral vectors, despite their efficacy, face challenges regarding cargo size limitations and potential genomic integration risks. Non-viral vectors offer distinct comparative advantages, including large cargo capacity for diverse CRISPR tools and transient expression to minimize off-target effects, but must overcome the eye’s formidable static and dynamic barriers, specifically the corneal epithelium, vitreous humor, and the inner limiting membrane. In this review, we present an anatomically guided framework for non-viral CRISPR/Cas9 delivery, mapping engineering strategies to specific ocular tissue targets. We first delineate the mechanisms of key physiological barriers, including the corneal stroma, aqueous humor circulation, and the vitreous–retina interface. Subsequently, we critically evaluate the latest advancements in non-viral platforms, such as pH-responsive lipid nanoparticles and engineered virus-like particles. The core focus of this review is on site-specific breakthrough strategies: from utilizing mucoadhesive polymers to counteract tear clearance in the cornea to exploiting specialized administration routes, such as suprachoroidal space and subretinal injection, to bypass retinal barriers, and deep-penetrating intravitreal carriers for targeting the photoreceptor-RPE complex. By integrating material science with precise administration routes, this review highlights feasible translational pathways for next-generation, carrier-free, or biomimetic ocular gene editing therapies. Full article

Background: Water inertia load training using equipment such as water vests provides unstable resistance that enhances trunk muscle activation. However, practical methods for prescribing exercise intensity without expensive electromyography (EMG) equipment remain limited. This pilot study aimed to develop prediction models for estimating trunk muscle activation using rating of perceived exertion (RPE) during water inertia load exercises. Methods: Seventeen healthy adults (20.45 ± 2.02 years) performed lateral trunk flexion exercises wearing a water vest at five progressive loads (8–16 kg in 2 kg increments). Surface EMG was recorded from four trunk muscles (rectus abdominis, external oblique, internal oblique, erector spinae) and normalized to maximal voluntary isometric contraction (%MVIC). Rating of perceived exertion (RPE) was assessed using the Borg CR-10 scale. Load-dependent changes in muscle activation were examined using repeated-measures ANOVA, and relationships between RPE and EMG were analyzed using regression and linear mixed-effects models. Results: All trunk muscles showed significant increases in activation with increasing load (all p < 0.001, ηp² = 0.381). RPE demonstrated significant positive correlations with all abdominal muscles (r = 0.37–0.46, p < 0.001). Simple regression analyses indicated predictive accuracy (R² = 0.267), representing a 29% increase compared with the strongest individual muscle model. Linear mixed-effects modeling confirmed RPE as a significant predictor after accounting for inter-individual variability. Conclusions: This pilot study provides preliminary evidence that RPE can be used to estimate trunk muscle activation during water inertia load exercise. The proposed composite activation index enhances prescription when EMG measurement is not feasible. Full article

Background: This study aimed to compare perceived exertion and changes in selected salivary biomarkers in young swimmers and basketball players. Methods: A total of 50 athletes participated in the study, including 23 swimmers (mean age: 13.8 ± 1.2 years; height: 172.4 ± 10.7 cm; body mass: 62.1 ± 11.4 kg) and 27 basketball players (mean age: 13.5 ± 0.6 years; height: 176.1 ± 11.0 cm; body mass: 61.2 ± 11.4 kg). All participants completed a single training session conducted at the same time of day. Saliva samples were collected from all athletes before and after the training session, and post-exercise changes were expressed as percentage differences. Participants also reported a rating of perceived exertion (RPE) using a 1–10 scale. Results: No significant between-group differences were observed in post-exercise changes in salivary flow rate, saliva pH, total protein concentration, α-amylase activity, or cortisol concentration. RPE did not differ significantly between groups, but moderate associations were found between salivary flow rate in both groups (r = 0.32, r = 0.34). Conclusions: Although perceived exertion was comparable, biochemical responses differed depending on the specific training environment. In turn, salivary flow rate appears to influence subjective perceptions of exercise intensity independently of exercise environment. Full article

Background/Objectives: The aim of this study was to compare the different forms of caffeine (CAF) administration in CrossFit^® participants. The countermovement jump (CMJ), the rate of perceived exertion (RPE), the total number of repetitions, and the maximum (HR_max) and mean heart rate (HR_mean) were evaluated. Methods: Fourteen males with more than six months of continuous CrossFit^® training (30.9 [5.62] years, 179 [1.33] cm, 78 [5.75] kg, 24.3 [1.33] kg·m⁻²) participated in this randomized, placebo-controlled, crossover study. Participants were randomized in a repeated measures design using caffeine capsule (CC), caffeine chewing gum (CCG), and caffeine mouth rinse (CMR) protocols, along with a placebo group (PG). Participants were unaware of whether any of the delivery methods contained caffeine. A 7-day washout period before each crossover was used. To ensure ecological validity, we replicated the real-world practice of CAF ingestion 30 min prior to training, mirroring typical athlete pre-workout routines. The participants of CrossFit^® performed the ‘Cindy’ protocol, and the CMJ as a primary outcome was measured pre- and post-intervention, while the RPE, HR, and the number of repetitions were tracked at the end of the workout for comparisons. Results: No significant differences were found between CAF forms in internal load measures (RPE, HR_max, HR_mean) or the number of repetitions. While no changes were observed with other CAF forms, CMR significantly improved the CMJ performance compared to the baseline (Δ: +3.5; Cohen’s d_unb: 0.51], which exceeded the estimated SWC by approximately three-fold. However, the inferential analysis revealed no significant main effects of the caffeine administration method on any measured outcomes. Conclusions: While CAF delivery forms did not improve internal load measures or performance parameters, such as the RPE, HR, or the total number of repetitions, the caffeine mouth rinse (CMR) showed a potentially meaningful improvement in CMJ performance after CrossFit^® training in participants with a certain level of experience. Full article

Background: Basketball performance depends strongly on physical preparation. A novel approach is accentuated eccentric loading within contrast training, though its acute effects using dumbbells remain underexplored. Methods: Twelve youth basketball players (age = 16.0 ± 0.3 years; body mass = 81.5 ± 7.6 kg) completed three sessions with dumbbell loads equivalent to 15%, 30% and 45% BW. CMJ performance was measured using dual wireless dual force plates. Assessments were conducted before the protocol and at 3, 9, and 15 min post intervention. Subjective responses were collected via wellness, RPE and readiness questionnaires. A two-way repeated measures ANOVA with Bonferroni corrections was applied, and the significance level was set to α = 0.05. Results: Significant decreases in jump height (p = 0.010) and average propulsive power (p = 0.005) were observed in the 45% BW condition at 3 and 9 min. Jump momentum decreased significantly at 30% and 45% BW at 3 and 9 min (p = 0.010; p = 0.033). No significant differences were detected in other CMJ force–time metrics (p > 0.05). Conclusions: Dumbbell-loaded CMJs as an accentuated eccentric loading contrast exercise did not produce generalized improvements but induced acute decreases at higher loads. However, they may still be useful in individual cases for athletes with favorable responses after monitoring. Full article