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There is tantalizing evidence that proteins can be accurately and selectively manipulated by higher order electric field effects within microfluidic devices. The accurate and precise manipulation of proteins in these platforms promises to disrupt and revolutionize many fields, most notably analytical biochemistry. Several lines of experimental evidence suggest much higher forces are generated compared to those calculated from traditional theories and those higher forces arise from subtle structural features of the proteins providing selectivity. New theories reflect some of the experimental evidence in the magnitude of the force predicted and inclusion of subtle structural features absent in traditional continuum theory. Unfortunately, the experimental evidence is largely exploratory in nature and lacks one or more important elements that prevents a clear interpretation and comparison to not only the other existing data, but also quantitative comparison to the evolving theoretical descriptions. Here, a clear and interpretable experimental system is presented that quantitatively determines the dielectrophoretic susceptibility of unlabeled, unaggregated native-structure protein molecules that are exposed to modest electric fields (10 5 -10 6  V/m) for short periods of time (∼5 ms) without significant increases in local concentration. The platform uses sub-nanogram quantities of protein, the probed volume upon determination is a few picoliters, and the total analysis time is 10 s., (© 2024 Wiley‐VCH GmbH.)

Introduction: To date, there is no valid single test or battery of tests for informing return-to-play (RTP) decisions following an acute shoulder injury. The purpose of this exploratory study is to evaluate a diagnostic test battery based on a Delphi consensus at the time of unrestricted return to team training after acute shoulder injury., Methods and Analysis: Data for this prospective multicentre cohort study are collected at two measurement time points: when the respective physician clears the patient for RTP (t1) and 12 months after RTP (t2). The study participants are 18-35 years old athletes participating at a professional level in the following team sports: handball, basketball, ice hockey, soccer, volleyball and American football. Maximum comparability will be ensured via uninjured matched pair teammates. To assess the subjective assessment of shoulder functioning and the athlete's readiness to RTP, patient-reported outcome measures (Western Ontario Shoulder Instability Index, Quick-Disabilities of the Arm, Shoulder and Hand, Psychological Readiness of Injured Athlete to Return to Sport and Shoulder Instability-Return to Sport after Injury) will be completed. After a medical check-up with a range of motion and anthropometric measurements as well as clinical tests, the participants will perform a structured warm-up protocol. The functional tests comprise handgrip strength, upper quarter Y-balance test, isometric strength, closed kinetic chain upper extremity stability test, wall hop test, functional throwing performance index and the unilateral seated shot put test and isokinetic tests., Ethics and Dissemination: The results of this study will be disseminated through peer-reviewed publications and scientific presentations at national and international conferences. Ethical approval was obtained through the Institutional Review Board of Martin-Luther-University Halle-Wittenberg (reference number: 2022-016)., Trial Registration Number: DRKS00028265., Competing Interests: Competing interests: HB works for the sponsor of the study as an advisor in the prevention field of sport (prevention department) and IB as an advisor in the field of insurance and benefits (rehabilitation department). Both supervise research projects in their departments and accompany the project participants until successful completion of the study. This is usually defined as successful implementation and transfer of the study results into practice., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

To evaluate the extent and characteristics of COVID-19 cases in relation to environmental COVID-19 incidences in the four best European soccer leagues (Bundesliga, Premier League, Serie A and La Liga) from the first of January 2020 until the end of January 2022. Methods: A retrospective evaluation of all publicly available COVID-19 cases in the studied cohorts was performed. The 14-day case incidences from epidemiological national data were used as reference values. The leagues studied are the Bundesliga (Germany), Premier League (Great Britain), Serie A (Italy) and La Liga (Spain). For all cases, the duration of time loss and date of case notification were recorded. Results: League-specific mean time loss due to disease or quarantine per COVID-19 case differs significantly between La Liga (11.45; ±5.21 days) and the other leagues studied (Bundesliga 20.41; ±33.87; p 0.0242; Premier League 17.12; ±10.39; p 0.0001; Serie A 17.61; ±12.71; p < 0.0001). A positive correlation between 14-day national incidence with COVID-19 disease occurrence in soccer leagues was found for all leagues studied. The correlations were strong in the Bundesliga (r 0.5911; CI 0.4249−0.7187; p < 0.0001), Serie A (r 0.5979; CI 0.4336−0.7238; p < 0.0001) and La Liga (r 0.5251; CI 0.3432−0.6690; p < 0.0001). A moderate correlation was found for the Premier League (r 0.3308; CI 0.1147−0.5169; p 0.0026). Odds ratios for altered environmental case risk in the cohorts studied could be calculated for four different national COVID-19 incidence levels (<50/100.000 to >500/100.000). A trend towards shorter COVID-19 case duration in the second half of 2021 was shown for all leagues studied. Conclusions: There was a significantly lower mean time-loss caused by a COVID-19 infection for cases occurred in La Liga compared with the other three leagues studied. For all four leagues studied, a positive, significant correlation of national environmental COVID-19 incidence level and the incidence of COVID-19 cases in the cohort of a football league was found.

Purpose: Choroidal neovascularization in age-related macular degeneration is caused, to a large extent, by increased secretion of vascular endothelial growth factor (VEGF)-A by the retinal pigment epithelium (RPE). The purpose of the study was to identify pathways that lead to increased VEGF secretion by the RPE., Methods: Ca(2+) signaling was studied in ARPE-19 and human RPE cells in primary culture by means of Ca(2+) imaging. Membrane conductance was measured in the whole-cell configuration of the patch-clamp technique. VEGF-A secretion was measured by using ELISA., Results: Freshly isolated RPE cells or ARPE-19 cells were shown to express TRPV1, -2, -3, and -4 channels. Increasing the temperature or stimulation by IGF-1 increased the VEGF-A secretion rate in both cell types. These effects were both sensitive to the TRPV channel blocker ruthenium red (20 μM). The heat-inducible Ca(2+) signals were blocked by the TRPV channel blockers La(3+) and ruthenium red by 68% and 52%, respectively. In contrast, high concentrations of 2-APB (3 mM) increased [Ca(2+)](i), whereas the TRPV1 channel opener capsaicin and the TRPV3 channel opener camphor had no effect. Reduction of TRPV2 expression by siRNA attenuated the heat-evoked Ca(2+) response. In addition, a heat-activated inwardly rectifying current was measured that was completely blocked by ruthenium red. IGF-1 also increased whole-cell current with a corresponding increase in [Ca(2+)](i), which was blocked by the PI3-kinase blocker LY294002., Conclusions: The data strongly suggest that TRPV2 channels expressed by the RPE are involved in the Ca(2+) signaling that mediates both heat-dependent and IGF-1 (via PI3-kinase activation)-induced VEGF secretion.

Purpose: In different tissues the activation of large conductance Ca2+-activated (BK) potassium channels has been shown to be coupled to voltage-gated Ca2+ channels as well as ryanodine receptors. As activation of BK channels leads to hyperpolarization of the cell, these channels provide a negative feedback mechanism for Ca2+-induced functions. Many cellular functions of the retinal pigment epithelium (RPE) are coupled to changes in [Ca2+]i. The aim of this study was to identify which Ca2+-entry pathway leads to the activation of BK channels in the RPE., Methods: We used freshly isolated human RPE cells and the ARPE-19 cell line for the detection of transcripts of BK channel alpha subunits. Patch-Clamp measurements were used to characterize BK channels in ARPE-19 cells electrophysiologically. To monitor changes in [Ca2+]i ARPE-19 cells were loaded with Fura-2., Results: Freshly isolated human RPE cells and ARPE-19 cells were shown to express BK channels. In ARPE-19 cells these channels were shown to be functionally active. Application of iberiotoxin led to a block of outward currents by 28.15%. At +50 mV ARPE-19 cells had a BK channel-mediated current density of 2.42 pA/pF. Activation of ryanodine receptors by caffeine led to a significant increase in [Ca2+]i by 34.16%. Nevertheless, caffeine-induced Ca2+ signals were not sufficient to activate BK channels. Instead, the activation of L-type Ca2+ channels by BayK 8644 caused a dramatic increase in BK channel activity and a shift of the reversal potential of the ARPE-19 cells by -22.6 mV., Conclusions: We have shown here for the first time that human RPE cells express BK channels. These channels are activated in RPE cells by increases in [Ca2+]i that are mediated by the opening of voltage gated L-type Ca2+ channels. As Ca2+ entering the RPE cells through these Ca2+ channels are known to be important for growth factor secretion and light-induced transepithelial transport, we speculate that BK channels coupled directly to these Ca2+ channels may provide a good tool for negative feedback control of the L-type Ca2+ channels.