Characterization and Modification of Light-Sensitive Phosphodiesterases from Choanoflagellates

Enzyme rhodopsins, including cyclase opsins (Cyclops) and rhodopsin phosphodiesterases (RhoPDEs), were recently discovered in fungi, algae and protists. In contrast to the well-developed light-gated guanylyl/adenylyl cyclases as optogenetic tools, ideal light-regulated phosphodiesterases are still in demand. Here, we investigated and engineered the RhoPDEs from Salpingoeca rosetta, Choanoeca flexa and three other protists. All the RhoPDEs (fused with a cytosolic N-terminal YFP tag) can be expressed in Xenopus oocytes, except the AsRhoPDE that lacks the retinal-binding lysine residue in the last (8th) transmembrane helix. An N296K mutation of YFP::AsRhoPDE enabled its expression in oocytes, but this mutant still has no cGMP hydrolysis activity. Among the RhoPDEs tested, SrRhoPDE, CfRhoPDE1, 4 and MrRhoPDE exhibited light-enhanced cGMP hydrolysis activity. Engineering SrRhoPDE, we obtained two single point mutants, L623F and E657Q, in the C-terminal catalytic domain, which showed ~40 times decreased cGMP hydrolysis activity without affecting the light activation ratio. The molecular characterization and modification will aid in developing ideal light-regulated phosphodiesterase tools in the future.

Effect of annealing time and pressure on electrical activation and surface morphology of Mg-implanted GaN annealed at 1300 °C in ultra-high-pressure nitrogen ambient

We performed an isothermal annealing study on Mg-implanted GaN at 1300 °C in an ultra-high-pressure (1 GPa) nitrogen ambient. Annealing for more than 30 min resulted in a high acceptor activation ratio and a low compensation ratio that were comparable to those obtained with annealing at 1400 °C for 5 min. We also performed annealing at 1300 °C in a reduced nitrogen pressure of 300 MPa which makes us possible to expand inner diameter of annealing equipment in the future. High electrical activation, similar to one obtained by annealing at 1 GPa, was successfully obtained.

Effects of Whole-Body Vibration Therapy on Quadriceps Function in Patients With Anterior Cruciate Ligament Reconstruction: A Systematic Review

Context: Quadriceps dysfunction is common for patients after anterior cruciate ligament reconstruction (ACLR). Whole-body vibration (WBV) could effectively treat quadriceps dysfunction. Objective: To summarize WBV protocols for patients with ACLR and to evaluate the effects of WBV on quadriceps function. Data Sources: PubMed, CINAHL, SportDiscus, Web of Science, Medline, and Embase were searched from inception to January 2020. Study Selection: Randomized controlled trials recruiting patients with ACLR, using WBV as intervention, and reporting at least 1 of the following outcomes, strength, rate of torque development (RTD), and voluntary activation ratio of quadriceps, were included. Study Design: Systematic review. Evidence Level: Level 3. Methods: This systematic review was reported according to PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines. Quality of evidence was determined by PEDro criteria and GRADE system. Participant characteristics, interventions, and the relevant results of the included studies were extracted and synthesized in a narrative way. Results: In total, 8 studies were included. Of these, 2 studies had serious risk of bias. Five of 8 studies implemented a series of WBV program ranging from 2 to 10 weeks in duration, while the other 3 studies implemented a single session of WBV. Eight WBV protocols were reported. The reported outcomes consisted of quadriceps strength, RTD, and central activation ratio. WBV protocols were heterogeneous. Low quality of evidence supported that exclusive conventional rehabilitation was more effective than exclusive WBV therapy in increasing quadriceps strength. Low quality of evidence supported that WBV combined with conventional rehabilitation was more beneficial in increasing quadriceps strength when compared with conventional rehabilitation alone. Very low quality of evidence supported the efficacy of a single session of WBV on quadriceps function. Conclusions: There is no standardized WBV protocol for patients with ACLR, and the effectiveness of WBV in rehabilitation on quadriceps function remains inconclusive.

Central activation deficits contribute to post stroke lingual weakness in a rat model

Lingual weakness frequently occurs after stroke and is associated with deficits in speaking and swallowing. Chronic weakness after stroke has been attributed to both impaired central activation of target muscles and reduced force generating capacity within muscles. How these factors contribute to lingual weakness is not known. We hypothesized that lingual weakness due to middle cerebral artery occlusion (MCAO) would manifest as reduced muscle force capacity and reduced muscle activation. Rats were randomized into MCAO or sham surgery groups. Maximum volitional tongue forces were quantified 8 weeks after surgery. Hypoglossal nerve stimulation was used to assess maximum stimulated force, muscle twitch properties, and force-frequency response. The central activation ratio was determined by maximum volitional/maximum stimulated force. Genioglossus muscle fiber type properties and neuromuscular junction innervation were assessed. Maximum volitional force and the central activation ratio were significantly reduced with MCAO. Maximum stimulated force was not significantly different. No significant differences were found for muscle twitch properties, unilateral contractile properties, muscle fiber type percentages, or fiber size. However, the twitch/tetanus ratio was significantly increased in the MCAO group relative to sham. A small but significant increase in denervated NMJs and fiber-type grouping occurred in the contralesional genioglossus. Results suggest the primary cause of chronic lingual weakness after stroke is impaired muscle activation rather than a deficit of force generating capacity in lingual muscles. Increased fiber type grouping and denervated NMJs in the contralesional genioglossus suggest partial reinnervation of muscle fibers may have preserved force generating capacity, but not optimal activation patterns.

Reproducibility and Concurrent Validity of Manual Palpation with Rehabilitative Ultrasound Imaging for Assessing Deep Abdominal Muscle Activity: Analysis with Preferential Ratios

The abdominal drawing-in maneuver (ADIM) is a clinical tool used for identifying preferential activity of deep abdominal muscles. However, concurrent validity and reproducibility of palpation during the ADIM has not been formally investigated. The aims of this study were (1) to assess intra- and interrater reliability of manual palpation during the ADIM, and (2) to determine the concurrent validity of manual palpation during the ADIM by calculating preferential activation ratio cut-off as assessed with ultrasound imaging (RUSI). Thirty-two subjects (n = 16 patients with nonspecific low back pain and 16 comparable healthy individuals) performed the ADIM in a supine hook-lying position. Two experienced assessors evaluated the presence or absence of preferential contraction of the deep abdominal muscles by palpation during the ADIM on 2 different days. Intrarater (test-retest) and interrater reliability of palpation were calculated using Cohen’s kappa coefficients. Muscle thickness of the transverse abdominis (TrA), internal oblique (IO), and external oblique (EO) muscles at rest and during the ADIM were also measured. TrA-Contraction Ratio (TrA-CR), TrA-Preferential Activation Ratio (TrA-PAR), and Modified-TrA-PR (M-TrA-PAR) were calculated. The concurrent validity of manual palpation was determined using the correlation between manual palpation and imaging and by calculating ROC curve (operating characteristics curve), Youden index, and sensitivity and specificity. Intra- and interrater reliability of manual palpation during the ADIM was excellent (k: 0.82–1.00) and good to excellent (k: 0.71–1.00), respectively. Interrater reliability for muscle thickness ranged from good to excellent (ICC3,1 0.79–0.91). Manual palpation and TrA ratio showed low to moderate correlations (r: 0.36–0.60). When evaluating the diagnostic accuracy of manual palpation, the best predictive model (ROC value: 0.89; p < 0.001) for correct a preferential contraction of TrA was obtained when the M-TrA-PAR was ≥0.08 (sensitivity: 0.95–1.00; specificity: 0.62). Good to excellent intra- and interrater reliability of manual palpation was found during the ADIM in both patients and healthy groups. Manual palpation showed concurrent validity for identifying the preferential activity of the TrA muscle supporting its use in clinical practice.

Reliability of Tibialis Anterior Muscle Voluntary Activation Using the Interpolated Twitch Technique and the Central Activation Ratio in People with Stroke

Voluntary activation (VA) is measured by applying supramaximal electrical stimulation to a muscle during a maximal voluntary contraction (MVC). The amplitude of the evoked muscle twitch is used to determine any VA deficit, and indicates incomplete central neural drive to the motor units. People with stroke experience VA deficits and greater levels of central fatigue, which is the decrease in VA that occurs following exercise. This study investigated the between-session reliability of VA and central fatigue of the tibialis anterior muscle (TA) in people with chronic stroke (n = 12), using the interpolated twitch technique (ITT), adjusted-ITT, and central activation ratio (CAR) methods. On two separate sessions, supramaximal electrical stimulation was applied to the TA when it was at rest and maximally activated, at the start and end of a 30-s isometric dorsiflexor MVC. The most reliable measures of VA were obtained using the CAR calculation on transformed data, which produced an ICC of 0.92, and a lower bound confidence interval in the good range (95% CI 0.77 to 0.98). Reliability was lower for the CAR calculation on non-transformed data (ICC 0.82, 95% CI 0.63 to 0.91) and the ITT and adjusted-ITT calculations on transformed data (ICCs 0.82, 95% CIs 0.51 to 0.94), which had lower bound confidence intervals in the moderate range. The two ITT calculations on non-transformed data demonstrated the poorest reliability (ICCs 0.62, 95% CI 0.25 to 0.74). Central fatigue measures demonstrated very poor reliability. Thus, the reliability for VA in people with chronic stroke ranged from good to poor, depending on the calculation method and statistical analysis method, whereas the reliability for central fatigue was very poor.

Core Strength Training Can Alter Neuromuscular and Biomechanical Risk Factors for Anterior Cruciate Ligament Injury

Background: Core stability is influential in the incidence of lower extremity injuries, including anterior cruciate ligament (ACL) injuries, but the effects of core strength training on the risk for ACL injury remain unclear. Hypothesis: Core muscle strength training increases the knee flexion angle, hamstring to quadriceps (H:Q) coactivation ratio, and vastus medialis to vastus lateralis (VM:VL) muscle activation ratio, as well as decreases the hip adduction, knee valgus, and tibial internal rotation angles. Study Design: Controlled laboratory study. Methods: A total of 48 male participants were recruited and randomly assigned to either the intervention group (n = 32) or the control group (n = 16). Three-dimensional trunk, hip, knee, and ankle kinematic data and muscle activations of selected trunk and lower extremity muscles were obtained while the participants performed side-step cutting. The core endurance scores were measured before and after training. Two-way analyses of variance were conducted for each dependent variable to determine the effects of 10 weeks of core strength training. Results: The trunk endurance scores in the intervention group significantly increased after training ( P < .05 for all comparisons). The intervention group showed decreased knee valgus ( P = .038) and hip adduction angles ( P = .032) but increased trunk flexion angle ( P = .018), rectus abdominis to erector spinae coactivation ratio ( P = .047), H:Q coactivation ratio ( P = .021), and VM:VL activation ratio ( P = .016). In addition, the knee valgus angle at initial contact was negatively correlated with the VM:VL activation ratio in the precontact phase ( R2 = 0.188; P < .001) but was positively correlated with the hip adduction angle ( R2 = 0.120; P < .005). No statistically significant differences were observed in the trunk endurance scores, kinematics, and muscle activations for the control group. Conclusion: Core strength training altered the motor control strategies and joint kinematics for the trunk and the lower extremity by increasing the trunk flexion angle, VM:VL activation ratio, and H:Q activation ratio and reducing the knee valgus and hip adduction angles. Clinical Relevance: Training core muscles can modify the biomechanics associated with ACL injuries in a side-step cutting task; thus, core strength training might be considered in ACL injury prevention programs to alter the lower extremity alignment in the frontal plane and muscle activations during sports-related tasks.