SARS-CoV-2 Spike Protein Induces Time-Dependent CTSL Upregulation in HeLa Cells and Alveolarspheres.

Autophagy and lysosomal pathways are involved in the cell entry of SARS-CoV-2 virus. To infect the host cell, the spike protein of SARS-CoV-2 binds to the cell surface receptor angiotensin-converting enzyme 2 (ACE2). To allow the fusion of the viral envelope with the host cell membrane, the spike protein has to be cleaved. One possible mechanism is the endocytosis of the SARS-CoV-2-ACE2 complex and subsequent cleavage of the spike protein, mainly by the lysosomal protease cathepsin L. However, detailed molecular and dynamic insights into the role of cathepsin L in viral cell entry remain elusive. To address this, HeLa cells and iPSC-derived alveolarspheres were treated with recombinant SARS-CoV-2 spike protein, and the changes in mRNA and protein levels of cathepsins L, B, and D were monitored. Additionally, we studied the effect of cathepsin L deficiency on spike protein internalization and investigated the influence of the spike protein on cathepsin L promoters in vitro. Furthermore, we analyzed variants in the genes coding for cathepsin L, B, D, and ACE2 possibly associated with disease progression using data from Regeneron's COVID Results Browser and our own cohort of 173 patients with COVID-19, exhibiting a variant of ACE2 showing significant association with COVID-19 disease progression. Our in vitro studies revealed a significant increase in cathepsin L mRNA and protein levels following exposure to the SARS-CoV-2 spike protein in HeLa cells, accompanied by elevated mRNA levels of cathepsin B and D in alveolarspheres. Moreover, an increase in cathepsin L promoter activity was detected in vitro upon spike protein treatment. Notably, the knockout of cathepsin L resulted in reduced internalization of the spike protein. The study highlights the importance of cathepsin L and lysosomal proteases in the SARS-CoV-2 spike protein internalization and suggests the potential of lysosomal proteases as possible therapeutic targets against COVID-19 and other viral infections.

Effectiveness of instrument-assisted soft tissue mobilization for the management of upper body, lower body, and spinal conditions. An updated systematic review with meta-analyses.

PURPOSE: To critically appraise randomized controlled trials (RCTs) on Instrument-Assisted Soft Tissue Mobilisation (IASTM) and quantify the effects of IASTM compared with other treatment in individuals with or without pathologies on function, pain, and range of motion. MATERIALS AND METHODS: We search four electronic databases from January 1999 to January 2022 and included RCTs of healthy participants/athletes and people with upper, lower, or spinal conditions, who received IASTM versus other active treatment for clinical outcomes (function, pain, and range of motion). RESULTS: Forty-six RCTs were considered eligible for data analysis. Effects of IASTM plus other treatment versus other treatment on function and pain intensity were not statistically significant or clinically meaningful (very low quality, SMD -0.28, 95% CI -0.66 to 0.09) and (very low quality, SMD -0.05, 95% CI -0.53 to 0.43) at up to one-year follow-up respectively. No clinically meaningful improvements were found on range of motion outcomes. Out of the 46 included RCTs, only 10 assessed and reported IASTM-related adverse events. CONCLUSION: Evidence of very low-quality certainty does not support the efficacy of IASTM in individuals with or without various pathologies on function, pain, and range of motion in the management of upper body, lower body, or spinal conditions. IMPLICATIONS FOR REHABILITATIONThe included RCTs had a high risk of bias and were assessed as very-low quality evidence for all the included outcomes.IASTM does not lead to clinically meaningful improvements in function, pain, or range of motion in individuals with upper body, lower body, and spinal conditions.The publication of IASTM trials in suspected predatory journals is increasing.The available evidence on IASTM does not support its use to improve function, pain, or range of motion in individuals with upper body, lower body, and spinal conditions.Health care practitioners should consider other evidence-based management strategies (physical activity and exercise) to improve function, pain, or range of motion in individuals with musculoskeletal injuries and disorders.Given the rise of publications on IASTM in suspected predatory journals, health care practitioners should be judicious to examine the legitimacy of a journal when searching for evidence on IASTM treatment technique.

An alternative splicing modulator decreases mutant HTT and improves the molecular fingerprint in Huntington's disease patient neurons.

Huntington's disease (HD) is a neurodegenerative disorder caused by poly-Q expansion in the Huntingtin (HTT) protein. Here, we delineate elevated mutant HTT (mHTT) levels in patient-derived cells including fibroblasts and iPSC derived cortical neurons using mesoscale discovery (MSD) HTT assays. HD patients' fibroblasts and cortical neurons recapitulate aberrant alternative splicing as a molecular fingerprint of HD. Branaplam is a splicing modulator currently tested in a phase II study in HD (NCT05111249). The drug lowers total HTT (tHTT) and mHTT levels in fibroblasts, iPSC, cortical progenitors, and neurons in a dose dependent manner at an IC50 consistently below 10 nM without inducing cellular toxicity. Branaplam promotes inclusion of non-annotated novel exons. Among these Branaplam-induced exons, there is a 115 bp frameshift-inducing exon in the HTT transcript. This exon is observed upon Branaplam treatment in Ctrl and HD patients leading to a profound reduction of HTT RNA and protein levels. Importantly, Branaplam ameliorates aberrant alternative splicing in HD patients' fibroblasts and cortical neurons. These findings highlight the applicability of splicing modulators in the treatment of CAG repeat disorders and decipher their molecular effects associated with the pharmacokinetic and -dynamic properties in patient-derived cellular models.

Aberrant NOVA1 function disrupts alternative splicing in early stages of amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a fatal disease characterized by aberrant alternative splicing (AS). Nuclear loss and cytoplasmic accumulation of the splicing factor TDP-43 in motor neurons (MN) are hallmarks of ALS at late stages of the disease. However, it is unknown if altered AS is present before TDP-43 pathology occurs. Here, we investigate altered AS and its origins in early stages of ALS using human induced pluripotent stem cell-derived motor neurons (MNs) from sporadic and familial ALS patients. We find high levels of the RNA-binding proteins NOVA1, NOVA2, and RBFOX2 in the insoluble protein fractions and observe that AS events in ALS-associated MNs are enriched for binding sites of these proteins. Our study points to an early disrupted function of NOVA1 that drives AS changes in a complex fashion, including events caused by a consistent loss of NOVA1 function. NOVA1 exhibits increased cytoplasmic protein levels in early stage MNs without TDP-43 pathology in ALS postmortem tissue. As nuclear TDP-43 protein level depletes, NOVA1 is reduced. Potential indications for a reduction of NOVA1 also came from mice over-expressing TDP-43 lacking its nuclear localization signal and iPSC-MN stressed with puromycin. This study highlights that additional RBP-RNA perturbations in ALS occur in parallel to TDP-43.

Psychometric Properties of the Patient-Specific Functional Scale in Patients with Low Back Pathology: A Systematic Review and Meta-Analysis.

Purpose: This systematic review and meta-analysis identifies, critically appraises, synthesizes, and meta-analyses the reported psychometric properties of the Patient-Specific Functional Scale (PSFS) in patients with low back pain or pathology. Method: The MEDLINE, Embase, PubMed, and Google Scholar databases were searched from their inception to September 2019. We included prospective measurement studies that reported on the psychometric properties (reliability, validity, responsiveness) of the PSFS in people with low back pain or pathology. We followed the COnsensus-based Standards for the selection of health Measurement INstruments 2018 guideline for systematic reviews. We performed both quantitative and qualitative syntheses in which the results were summarized on the basis of the reported measurement properties and study quality. Results: Ten eligible studies were included. The pooled PSFS reliability measure was excellent (intra-class correlation coefficient = 0.89; 95% CI: 0.75, 0.95). Validity measures displayed correlations that ranged from -0.47 to 0.69 when compared with other patient-reported outcome measures (PROMs) or other tests. Eight studies had assessed the responsiveness of the PSFS. Effect sizes reported were large (≥ 0.91). Conclusions: The PSFS is a reliable, valid, and responsive PROM for patients with low back pain or pathology.

Objectif : procéder à la détermination, à l'évaluation critique, à la synthétisation et à la méta-analyse des propriétés psychométriques déclarées de l'échelle fonctionnelle propre aux patients (PSFS) ayant des douleurs ou une pathologie dorsales. Méthodologie : les chercheurs ont consulté les bases de données MEDLINE, Embase, PubMed et Google Scholar depuis leur création jusqu'en septembre 2019. Ils ont inclus les études de mesures prospectives sur les propriétés psychométriques (fiabilité, viabilité, réactivité) du PSFS chez les personnes souffrant de douleurs ou d'une pathologie dorsales. Ils ont respecté les directives COSMIN 2018 sur les normes consensuelles pour la sélection d'instruments de mesure de la santé en vue d'analyses systématiques. Ils ont effectué à la fois une synthèse quantitative et une synthèse qualitative dans lesquelles ils ont résumé les résultats en fonction des propriétés métriques déclarées et de la qualité des études. Résultats : dix études admissibles ont été retenues. La mesure de fiabilité regroupée de la PSFS était excellente, avec un coefficient de corrélation intraclasse de 0,89 (IC à 95 % : 0,75, 0,95). Les mesures de validité ont révélé des corrélations entre −0,47 et 0,69 par rapport à d'autres mesures de résultats déclarées par les patients (PROM) ou d'autres tests. Huit études ont évalué les mesures de réactivité du PSFS. Les ampleurs de l'effet déclarées étaient importantes, à 0,91 ou plus. Conclusion : la PSFS est une PROM fiable, valide et réactive chez les patients ayant des douleurs ou des pathologies lombaires.

Psychometric properties of Patient-Specific Functional Scale in patients with upper extremity disorders. A systematic review.

PURPOSE: To identify, critically appraise, and synthesise the measurement properties of Patient-Specific Functional Scale (PSFS) in patients with upper extremity musculoskeletal disorders. METHODS: Medline, Embase, PubMed, and Google Scholar databases from January 1999 to November 2020 were searched. Prospective measurement studies that included patients with upper extremity musculoskeletal disorders, that reported on the psychometric properties of PSFS were included. We used the COnsensus-based Standards for the selection of health Measurement INstruments (COSMIN) 2018 guideline for systematic reviews to appraise the studies. RESULTS: Fourteen eligible studies were included. Three studies with adequate-very good quality and sufficient properties indicated excellent intra-class correlation coefficients (ICC) (≥0.75) in patients with shoulder pain (mean age 48 ± 11 years), multiple shoulder disorders (mean age 55 ± 16 years), and hand osteoarthritis (mean age 64 ± 9 years). The construct validity estimates of PSFS were moderate, when compared with Upper Extremity Functional Index (UEFI) (r = 0.50) and Numeric Pain Rating Scale (NPRS) (r = 0.51) in patients with combined upper extremity musculoskeletal disorders (shoulder/upper arm, wrist/hand and elbow/forearm). CONCLUSIONS: The patient-specific functional scale can be considered as a reliable, valid, and responsive tool in assessing functional change in patients with shoulder disorders/pain.Implications for rehabilitationThe Patient-Specific Functional Scale (PSFS) can be considered as: • a reliable outcome measure in assessing functional change in patients with shoulder pain and hand osteoarthritis; • a valid measure in assessing functional limitation in patients with upper extremity disorders; • a measure that is sensitive to change (displays longitudinal validity) in assessing functional change in patients with upper extremity disorders and in patients with shoulder pain; and • a responsive outcome measure in assessing functional change in patients with upper extremity disorders.

Examining changes in corticospinal excitability and balance performance in response to social-comparative feedback.

BACKGROUND: Social-comparative feedback informs an individual that their performance was better or worse than the group. Previous studies have found that compared to knowledge of results alone, social-comparative feedback produces a valence response that results in larger improvements in balance performance. However, the neural processes contributing to these motor improvements have not yet been examined. RESEARCH QUESTION: Does social-comparative feedback alter corticospinal excitability and consequently, balance performance? METHODS: Thirty-six healthy young adults stood and maintained their balance on a stabiliometer for eight trials. After three of the trials, the neutral (i.e., only knowledge of results) group received their performance feedback (i.e., time on balance) while the other two groups also received positive (i.e., performed better than the group) or negative (i.e., performed worse than the group) social-comparative feedback. To measure corticospinal excitability, soleus motor-evoked potentials were elicited using transcranial magnetic stimulation at the beginning of the experiment, after the presentation of feedback, and at the end of the experiment. Pre- and post- ratings of confidence, perceived skill, motivation, and anxiety were also collected. RESULTS: The negative feedback group reported decreases in perceived skill (43 ± 29%) and balance confidence (26 ± 28%), while the positive group reported a 13 ± 17% increase in perceived skill. Despite these group differences in feedback perception, all three groups improved their balance performance by ≈35% (p < 0.001) by the eighth trial. However, this improvement in balance performance was not matched by any changes in corticospinal excitability over time (19.2 ± 55.9% change; p = 0.340) or between groups (p = 0.734). SIGNIFICANCE: Our findings suggest that social-comparative feedback, as presented in this study, does not affect corticospinal excitability and balance performance differently than knowledge of results (neutral feedback) alone. More arousing and more frequent forms of social-comparative feedback may be necessary for observing larger changes in the functional or neural control of balance.