Analyzing the Eye Gaze Behaviour of Students and Experienced Physiotherapists during Observational Movement Analysis.

Purpose: Physiotherapists use observational movement analysis (OMA) to inform clinical reasoning. This study aimed to (1) determine the feasibility of characterizing eye gaze behaviour during OMA with eye-tracking technology, (2) characterize experienced neurological physiotherapists' and physiotherapy students' eye gaze behaviour during OMA, and (3) investigate differences in eye gaze behaviour during OMA between physiotherapy students and experienced physiotherapists. Method: Eight students and eight physiotherapists wore an eye-tracking device while watching a video of a person with a history of stroke and subsequent concussion perform sit to stand. Feasibility criteria were (1) successful calibration of the eye tracker, and successful collection of data, for 80% of the participants and (2) moderate interrater reliability of the investigators, measured by intra-class correlation coefficients (ICCs). Three investigators independently recorded the participants' foveal fixations. Differences between physiotherapists and students in number of fixations, duration per fixation, and total duration of fixations were evaluated using unpaired t-tests, mean differences, and 95% CIs. Results: Data were collected for all participants. ICCs ranged from 0.64 to 0.78. Fixations by physiotherapists were shorter (mean 368.5 [SD 80.8] ms) and greater in number (mean 18.9 [SD 2.2]) than those by students (mean 459.0 [SD 64.2] ms, p = 0.03, and mean 15.9 [SD 2.7], p = 0.03), respectively. Conclusions: Measuring eye gaze behaviour during OMA using eye tracker technology is feasible. Physiotherapists made more fixations of shorter duration than students. Further investigation of how experienced therapists perform OMA and apply it to clinical reasoning may inform the instruction of OMA.

Objectif : les physiothérapeutes font appel à l'analyse observationnelle des mouvements (AOM) pour éclairer leur raisonnement clinique. La présente étude visait à 1) déterminer la faisabilité de caractériser le comportement du regard pendant l'AOM par la technologie de l'oculométrie, 2) caractériser le comportement du regard des physiothérapeutes neurologiques expérimentés et des étudiants en physiothérapie pendant l'AOM et 3) examiner les différences de comportement du regard des élèves physiothérapeutes par rapport aux physiothérapeutes d'expérience pendant l'AOM. Méthodologie : huit étudiants et huit physiothérapeutes ont porté un oculomètre en regardant la vidéo d'une personne ayant des antécédents d'accident vasculaire cérébral (AVC) et de commotion subséquente pour effectuer l'exercice assis-debout. Les critères de faisabilité s'établissaient comme suit : 1) la calibration réussie de l'oculomètre et la collecte réussie des données pour 80 % des participants et 2) une variabilité interévaluateurs modérée des chercheurs, mesurée par les coefficients de corrélation intraclasse (CCI). Trois chercheurs ont enregistré la fixation fovéale des participants. Ils ont évalué les différences entre les physiothérapeutes et les étudiants pour ce qui est du nombre de fixations, de la durée de chaque fixation et de la durée totale des fixations à l'aide des tests de Student non appariés, des différences moyennes et des intervalles de confiance à 95 %. Résultats : les données ont été colligées pour tous les participants. Les CCI se situaient entre 0,64 et 0,78. Les fixations des physiothérapeutes étaient plus courtes (368,5 [ÉT 80,8] ms) et plus fréquentes (18,9 [ÉT 2,2]) que celles des étudiants (459,0 [ÉT 64,2] ms, p = 0,03; 15,9 [ÉT 2,7], p = 0,03), respectivement. Conclusion : il est possible de mesurer le comportement du regard par oculométrie pendant l'AOM. Les physiothérapeutes avaient plus de fixations de courte durée que les étudiants. D'autres recherches sur la manière dont les thérapeutes d'expérience procèdent à l'AOM et l'appliquent à leur raisonnement clinique pourront éclairer les directives sur l'AOM.

Training to Improve Walking after Pediatric Spinal Cord Injury: A Systematic Review of Parameters and Walking Outcomes.

Walking or locomotor training is often initiated following pediatric spinal cord injury (SCI). There is no synthesis of the literature on interventions targeting walking for pediatric SCI, although this would assist future clinical trials and interventions. To address this need, we completed a systematic review to summarize the who, what, when, and how of walking interventions in children with SCI. Participant characteristics, training parameters, and walking outcomes with training in pediatric SCI were identified and compared with training parameters and outcomes in adults with SCI. The PubMed, Medline, AMED, Embase, PsycInfo, Cochrane Central Register of Controlled Trials, Cochrane Database of Systematic Reviews, and CINAHL databases were searched for studies that included participants aged 1-17 years with a SCI acquired post-birth, physical interventions, and pre- and post-training walking measures. Two researchers evaluated each study's risk of bias using a domain-based approach. Training parameters and walking outcomes were extracted. Total training duration (duration × frequency × number of weeks) was calculated. Thirteen pediatric studies (n = 43 children) were included; all but one were case series/reports. Risk of bias was high in the pediatric studies. A 2012 adult review was updated (11 studies added). As with adults, the training durations, frequencies, and modes used with the children varied; however, overground walking practice was included in 10/13 pediatric studies. Improvements in walking capacity, speed, and distance were comparable between children and adults. There was a trend for greater gains with greater total training durations. There is a paucity of high-quality research examining interventions targeting walking after pediatric SCI; however, intensive training, including practice overground, results in notable improvements.

Leucine-rich repeat kinase 2 (LRRK2): a key player in the pathogenesis of Parkinson's disease.

Parkinson's disease (PD) is the most common neurodegenerative movement disorder, with a prevalence of more than 1% after the age of 65 years. Mutations in the gene encoding leucine-rich repeat kinase-2 (LRRK2) have recently been linked to autosomal dominant, late-onset PD that is clinically indistinguishable from typical, idiopathic disease. LRRK2 is a multidomain protein containing several protein interaction motifs as well as dual enzymatic domains of GTPase and protein kinase activities. Disease-associated mutations are found throughout the multidomain structure of the protein. LRRK2, however, is unique among the PD-causing genes, because a missense mutation, G2019S, is a frequent determinant of not only familial but also sporadic PD. Thus, LRRK2 has emerged as a promising therapeutic target for combating PD. In this Mini-Review, we look at the current state of knowledge regarding the domain structure, amino acid substitutions, and potential functional roles of LRRK2.

The Roc domain of leucine-rich repeat kinase 2 is sufficient for interaction with microtubules.

Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the leading cause of genetically inherited Parkinson's disease (PD). Although this multidomain protein has been shown to have both GTPase and kinase activities through the Roc and MAPKKK domains, respectively, the protein-protein interactions and pathways involved in LRRK2-mediated signaling remain elusive. Utilizing a combination of protein pull-down assays, mass spectrometry, Western blotting, and immunofluorescence microscopy, this study identifies and describes the interaction between LRRK2 and microtubules. The Roc or GTPase-like domain of LRRK2 is sufficient for interaction with alpha/beta-tubulin heterodimers. This interaction occurs in a guanine nucleotide-independent manner, suggesting that tubulin might not be an effector of the LRRK2 GTPase domain. The R1441C pathogenic mutation, located within the Roc domain, retains interaction with alpha/beta-tubulin heterodimers, suggesting that disruption of this interaction likely is not the mechanism whereby the R1441C mutation leads to disease. At a subcellular level, endogenous LRRK2 protein was found to colocalize with alpha/beta-tubulin in primary hippocampal neurons. These findings are significant in that they link LRRK2 with microtubules, a structural component of the cell that is critically involved in the pathogenesis of several neurodegenerative diseases, including PD.

The Parkinson's disease-associated protein, leucine-rich repeat kinase 2 (LRRK2), is an authentic GTPase that stimulates kinase activity.

Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the leading cause of autosomal dominant Parkinson's disease (PD). LRRK2, a member of the ROCO protein family, contains both Ras GTPase-like (Roc) and kinase (MAPKKK) domains, as well as other functional motifs. Here, we have identified LRRK2 as the first mammalian ROCO protein that is an authentic and functional GTPase, defined by the ability to bind GTP and undergo intrinsic GTP hydrolysis. Furthermore, the Roc domain is sufficient for this native GTPase activity and binds and hydrolyzes GTP indistinguishably from the Ras-related small GTPase, Rac1. The PD-associated mutation, R1441C, located within the Roc domain, leads to an increase in LRRK2 kinase activity and a decrease in the rate of GTP hydrolysis, compared to the wild-type protein, in an in vitro assay. This finding suggests that the R1441C mutation may help stabilize an activated state of LRRK2. Additionally, LRRK2-mediated phosphorylation is stimulated upon binding of non-hydrolyzable GTP analogs, suggesting that LRRK2 is an MAPKKK-activated intramolecularly by its own GTPase. Since GTPases and MAPKKKs are upstream regulators of multiple signal transduction cascades, LRRK2 may play a central role in integrating pathways involved in neuronal cell signaling and the pathogenesis of PD.

An activating mutant of Cdc42 that fails to interact with Rho GDP-dissociation inhibitor localizes to the plasma membrane and mediates actin reorganization.

Cdc42 is a member of the Rho family of GTPases and plays an important role in the regulation of actin cytoskeletal organization. Activation of Cdc42 and associated signal transduction cascades are dependent upon proper localization of this GTPase. The studies described herein address the hypothesis that Rho GDP-dissociation inhibitor, RhoGDI, plays an essential role in the translocation of Cdc42 to signaling complexes at the plasma membrane and is essential for Cdc42-mediated actin cytoskeletal rearrangements. An activating mutant of Cdc42 that is RhoGDI-binding defective (Cdc42(G12V/R66E)) is characterized and used as a tool to study the functional importance of the Cdc42-RhoGDI interaction. Overexpression of mycCdc42(G12V/R66E) in COS-7 cells results in actin cytoskeletal rearrangements that are indistinguishable from those stimulated by overexpression of mycCdc42(G12V). In addition, the G12V activating mutant of Cdc42 was overexpressed in mesangial cells that are null for RhoGDI expression. MycCdc42(G12V) stimulation of filopodia formation in these cells was indistinguishable from that observed in wild-type mesangial cells. Taken together, the results presented herein indicate that although RhoGDI is a critical regulator of guanine nucleotide binding, cycling of Cdc42 between membranes and the cytosol and cellular transformation, it is not essential for Cdc42-mediated organization of the actin cytoskeleton.

An activating mutant of Rac1 that fails to interact with Rho GDP-dissociation inhibitor stimulates membrane ruffling in mammalian cells.

Rac1, a member of the Rho family of small GTP-binding proteins, is involved in the regulation of the actin cytoskeleton via activation of lamellipodia and membrane ruffle formation. RhoGDI (Rho-family-specific GDP-dissociation inhibitor) forms a complex with Rho proteins in the cytosol of mammalian cells. It not only regulates guanine nucleotide binding to Rho proteins, but may also function as a molecular shuttle to carry Rho proteins from an inactive cytosolic pool to the membrane for activation. These studies tested if RhoGDI is necessary for the translocation of Rac1 from the cytosol to the plasma membrane for the formation of membrane ruffles. We describe a novel mutant of Rac1, R66E (Arg66-->Glu), that fails to bind RhoGDI. This RhoGDI-binding-defective mutation is combined with a Rac1-activating mutation G12V, resulting in a double-mutant [Rac1(G12V/R66E)] that fails to interact with RhoGDI in COS-7 cells, but remains constitutively activated. This double mutant stimulates membrane ruffling to a similar extent as that observed after epidermal growth factor treatment of non-transfected cells. To confirm that Rac1 can signal ruffle formation in the absence of interaction with RhoGDI, Rac1(G12V) was overexpressed in cultured mesangial cells derived from a RhoGDI knockout mouse. Rac1-mediated membrane ruffling was indistinguishable between the RhoGDI(-/-) and RhoGDI(+/+) cell lines. In both the COS-7 and cultured mesangial cells, Rac1(G12V) and Rac1(G12V/R66E) co-localize with membrane ruffles. These findings suggest that interaction with RhoGDI is not essential in the mechanism by which Rac1 translocates to the plasma membrane to stimulate ruffle formation.