Whole body modelling of musculoskeletal interactions during whole body vibration to inform rehabilitation intervention design

Background: A major secondary complication which can arise in individuals with spinal cord injury (SCI) is disuse-related bone loss as a result of long-term paralysis and immobilisation. An emerging rehabilitation technique used to treat this musculoskeletal degeneration is whole body vibration (WBV). This can be applied to patients in different body positions on a WBV platform in order to stimulate different muscle groups and in turn apply muscle forces to target bones. To treat the disuse-related bone loss, the hypothesis is that WBV intervention can stimulate bone formation indirectly via targeted muscle action, and/or directly if vibration acts as a mechanostimulus on the bone. Aim & Objectives: The aim of this study was to develop whole body computational models of WBV intervention, to inform the design of intervention protocols in SCI patients. Effects of muscle loss were simulated, and activation and forces of different muscles analysed for a number of proposed configurations on the WBV platform. Methods: WBV intervention was simulated using the AnyBody Technology Modelling software by implementing and adapting the currently available standing model. Different body positions (standing, knee flexed standing, squatting) and parameters of WBV such as frequency and amplitude were modelled and analysed, and the muscle actions simulated. Results: Realistic muscle activities compared to the literature were found in all body position configurations without the WBV simulation. When modelling the WBV intervention, only the squatting body position and side-alternating WBV plate were found to give accurate results. The activities of several muscles were recorded in this configuration under a variety of frequencies and amplitudes. Finally muscle forces were analysed with changes in frequency and amplitude and found to cause a corresponding change in the loading of regions the bones of the ilium, tibia, femur, ischium, fibula, sacrum and coccyx. The aim is for the results of this model to be used in the future to inform WBV protocol development for musculoskeletal rehabilitation in SCI and other target patient groups.Background: A major secondary complication which can arise in individuals with spinal cord injury (SCI) is disuse-related bone loss as a result of long-term paralysis and immobilisation. An emerging rehabilitation technique used to treat this musculoskeletal degeneration is whole body vibration (WBV). This can be applied to patients in different body positions on a WBV platform in order to stimulate different muscle groups and in turn apply muscle forces to target bones. To treat the disuse-related bone loss, the hypothesis is that WBV intervention can stimulate bone formation indirectly via targeted muscle action, and/or directly if vibration acts as a mechanostimulus on the bone. Aim & Objectives: The aim of this study was to develop whole body computational models of WBV intervention, to inform the design of intervention protocols in SCI patients. Effects of muscle loss were simulated, and activation and forces of different muscles analysed for a number of proposed configurations on the WBV platform. Methods: WBV intervention was simulated using the AnyBody Technology Modelling software by implementing and adapting the currently available standing model. Different body positions (standing, knee flexed standing, squatting) and parameters of WBV such as frequency and amplitude were modelled and analysed, and the muscle actions simulated. Results: Realistic muscle activities compared to the literature were found in all body position configurations without the WBV simulation. When modelling the WBV intervention, only the squatting body position and side-alternating WBV plate were found to give accurate results. The activities of several muscles were recorded in this configuration under a variety of frequencies and amplitudes. Finally muscle forces were analysed with changes in frequency and amplitude and found to cause a corresponding change in the loading of regions the bones of the ilium, tibia, femur, ischium, fibula, sacrum and coccyx. The aim is for the results of this model to be used in the future to inform WBV protocol development for musculoskeletal rehabilitation in SCI and other target patient groups

Wearable fusion system for assessment of motor function in lesion-symptom mapping studies

Lesion-symptom mapping studies are a critical component of addressing the relationship between brain and behaviour. Recent developments have yielded significant improvements in the imaging and detection of lesion profiles, but the quantification of motor outcomes is still largely performed by subjective and low-resolution standard clinical rating scales. This mismatch means than lesion-symptom mapping studies are limited in scope by scores which lack the necessary accuracy to fully quantify the subcomponents of motor function. The first study conducted aimed to develop a new automated system of motor function which addressed the limitations inherent in the clinical rating scales. A wearable fusion system was designed that included the attachment of inertial sensors to record the kinematics of upper extremity. This was combined with the novel application of mechanomyographic sensors in this field, to enable the quantification of hand/wrist function. Novel outputs were developed for this system which aimed to combine the validity of the clinical rating scales with the high accuracy of measurements possible with a wearable sensor system. This was achieved by the development of a sophisticated classification model which was trained on series of kinematic and myographic measures to classify the clinical rating scale. These classified scores were combined with a series of fine-grained clinical features derived from higher-order sensor metrics. The developed automated system graded the upper-extremity tasks of the Fugl-Meyer Assessment with a mean accuracy of 75\% for gross motor tasks and 66\% for the wrist/hand tasks. This accuracy increased to 85\% and 74\% when distinguishing between healthy and impaired function for each of these tasks. Several clinical features were computed to describe the subcomponents of upper extremity motor function. This fine-grained clinical feature set offers a novel means to complement the low resolution but well-validated standardised clinical rating scales. A second study was performed to utilise the fine-grained clinical feature set calculated in the previous study in a large-scale region-of-interest lesion-symptom mapping study. Statistically significant regions of motor dysfunction were found in the corticospinal tract and the internal capsule, which are consistent with other motor-based lesion-symptom mapping studies. In addition, the cortico-ponto-cerebellar tract was found to be statistically significant when testing with a clinical feature of hand/wrist motor function. This is a novel finding, potentially due to prior studies being limited to quantifying this subcomponent of motor function using standard clinical rating scales. These results indicate the validity and potential of the clinical feature set to provide a more detailed picture of motor dysfunction in lesion-symptom mapping studies.Open Acces

The novel mouse mutant, chuzhoi, has disruption of Ptk7 protein and exhibits defects in neural tube, heart and lung development and abnormal planar cell polarity in the ear

Background The planar cell polarity (PCP) signalling pathway is fundamental to a number of key developmental events, including initiation of neural tube closure. Disruption of the PCP pathway causes the severe neural tube defect of craniorachischisis, in which almost the entire brain and spinal cord fails to close. Identification of mouse mutants with craniorachischisis has proven a powerful way of identifying molecules that are components or regulators of the PCP pathway. In addition, identification of an allelic series of mutants, including hypomorphs and neomorphs in addition to complete nulls, can provide novel genetic tools to help elucidate the function of the PCP proteins. Results We report the identification of a new N-ethyl-N-nitrosourea (ENU)-induced mutant with craniorachischisis, which we have named chuzhoi (chz). We demonstrate that chuzhoi mutant embryos fail to undergo initiation of neural tube closure, and have characteristics consistent with defective convergent extension. These characteristics include a broadened midline and reduced rate of increase of their length-to-width ratio. In addition, we demonstrate disruption in the orientation of outer hair cells in the inner ear, and defects in heart and lung development in chuzhoi mutants. We demonstrate a genetic interaction between chuzhoi mutants and both Vangl2Lp and Celsr1Crsh mutants, strengthening the hypothesis that chuzhoi is involved in regulating the PCP pathway. We demonstrate that chuzhoi maps to Chromosome 17 and carries a splice site mutation in Ptk7. This mutation results in the insertion of three amino acids into the Ptk7 protein and causes disruption of Ptk7 protein expression in chuzhoi mutants. Conclusions The chuzhoi mutant provides an additional genetic resource to help investigate the developmental basis of several congenital abnormalities including neural tube, heart and lung defects and their relationship to disruption of PCP. The chuzhoi mutation differentially affects the expression levels of the two Ptk7 protein isoforms and, while some Ptk7 protein can still be detected at the membrane, chuzhoi mutants demonstrate a significant reduction in membrane localization of Ptk7 protein. This mutant provides a useful tool to allow future studies aimed at understanding the molecular function of Ptk7

Expression of avian prickle genes during early development and organogenesis

Chicken homologues of prickle-1 (pk-1) and prickle-2 (pk-2) were isolated to gain insight into the extent of planar cell polarity signaling during avian embryogenesis. Bioinformatics analyses demonstrated homology and showed that pk-1 and pk-2 exhibited conserved synteny with ADAMTS20 and ADAMTS9, GON-related zinc metalloproteases. Expression of pk-1 and pk-2 was established during embryogenesis and early organogenesis, using in situ hybridization and sections of chicken embryos. At early stages, pk-1 was expressed in Hensen's node, primitive streak, ventral neural tube, and foregut. In older embryos, pk-1 transcripts were detected in dorsolateral epithelial somites, dorsomedial lip of dermomyotomes, and differentiating myotomes. Furthermore, pk-1 expression was seen in lateral body folds, limb buds, and ventral metencephalon. pk-2 was expressed in Hensen's node and neural ectoderm at early stages. In older embryos, pk-2 expression was restricted to ventromedial epithelial somites, except in the most recently formed somite pair, and limb bud mesenchyme

The PCP genes Celsr1 and Vangl2 are required for normal lung branching morphogenesis

The lungs are generated by branching morphogenesis as a result of reciprocal signalling interactions between the epithelium and mesenchyme during development. Mutations that disrupt formation of either the correct number or shape of epithelial branches affect lung function. This, in turn, can lead to congenital abnormalities such as cystadenomatoid malformations, pulmonary hypertension or lung hypoplasia. Defects in lung architecture are also associated with adult lung disease, particularly in cases of idiopathic lung fibrosis. Identifying the signalling pathways which drive epithelial tube formation will likely shed light on both congenital and adult lung disease. Here we show that mutations in the planar cell polarity (PCP) genes Celsr1 and Vangl2 lead to disrupted lung development and defects in lung architecture. Lungs from Celsr1Crsh and Vangl2Lp mouse mutants are small and misshapen with fewer branches, and by late gestation exhibit thickened interstitial mesenchyme and defective saccular formation. We observe a recapitulation of these branching defects following inhibition of Rho kinase, an important downstream effector of the PCP signalling pathway. Moreover, epithelial integrity is disrupted, cytoskeletal remodelling perturbed and mutant endoderm does not branch normally in response to the chemoattractant FGF10. We further show that Celsr1 and Vangl2 proteins are present in restricted spatial domains within lung epithelium. Our data show that the PCP genes Celsr1 and Vangl2 are required for foetal lung development thereby revealing a novel signalling pathway critical for this process that will enhance our understanding of congenital and adult lung diseases and may in future lead to novel therapeutic strategies

Nuclear mRNA Degradation Pathway(s) Are Implicated in Xist Regulation and X Chromosome Inactivation

A critical step in X-chromosome inactivation (XCI), which results in the dosage compensation of X-linked gene expression in mammals, is the coating of the presumptive inactive X chromosome by the large noncoding Xist RNA, which then leads to the recruitment of other factors essential for the heterochromatinisation of the inactive X and its transcriptional silencing. In an approach aimed at identifying genes implicated in the X-inactivation process by comparative transcriptional profiling of female and male mouse gastrula, we identified the Eif1 gene involved in translation initiation and RNA degradation. We show here that female embryonic stem cell lines, silenced by RNA interference for the Eif1 gene, are unable to form Xist RNA domains upon differentiation and fail to undergo X-inactivation. To probe further an effect involving RNA degradation pathways, the inhibition by RNA interference of Rent1, a factor essential for nonsense-mediated decay and Exosc10, a specific nuclear component of the exosome, was analysed and shown to similarly impair Xist upregulation and XCI. In Eif1-, Rent1-, and Exosc10-interfered clones, Xist spliced form(s) are strongly downregulated, while the levels of unspliced form(s) of Xist and the stability of Xist RNA remain comparable to that of the control cell lines. Our data suggests a role for mRNA nuclear degradation pathways in the critical regulation of spliced Xist mRNA levels and the onset of the X-inactivation process

A role for core planar polarity proteins in cell contact-mediated orientation of planar cell division across the mammalian embryonic skin

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. © The Author(s) 2017. Supplementary information accompanies this paper at doi:10.1038/s41598-017-01971-2.The question of how cell division orientation is determined is fundamentally important for understanding tissue and organ shape in both healthy or disease conditions. Here we provide evidence for cell contact-dependent orientation of planar cell division in the mammalian embryonic skin. We propose a model where the core planar polarity proteins Celsr1 and Frizzled-6 (Fz6) communicate the long axis orientation of interphase basal cells to neighbouring basal mitoses so that they align their horizontal division plane along the same axis. The underlying mechanism requires a direct, cell surface, planar polarised cue, which we posit depends upon variant post-translational forms of Celsr1 protein coupled to Fz6. Our hypothesis has parallels with contact-mediated division orientation in early C. elegans embryos suggesting functional conservation between the adhesion-GPCRs Celsr1 and Latrophilin-1. We propose that linking planar cell division plane with interphase neighbour long axis geometry reinforces axial bias in skin spreading around the mouse embryo body.Peer reviewe

Seven-Pass Transmembrane Cadherins: Roles and Emerging Mechanisms in Axonal and Dendritic Patterning

The Flamingo/Celsr seven-transmembrane cadherins represent a conserved subgroup of the cadherin superfamily involved in multiple aspects of development. In the developing nervous system, Fmi/Celsr control axonal blueprint and dendritic morphogenesis from invertebrates to mammals. As expected from their molecular structure, seven-transmembrane cadherins can induce cell–cell homophilic interactions but also intracellular signaling. Fmi/Celsr is known to regulate planar cell polarity (PCP) through interactions with PCP proteins. In the nervous system, Fmi/Celsr can function in collaboration with or independently of other PCP genes. Here, we focus on recent studies which show that seven-transmembrane cadherins use distinct molecular mechanisms to achieve diverse functions in the development of the nervous system

A widespread family of bacterial cell wall assembly proteins

Teichoic acids and acidic capsular polysaccharides are major anionic cell wall polymers (APs) in many bacteria, with various critical cell functions, including maintenance of cell shape and structural integrity, charge and cation homeostasis, and multiple aspects of pathogenesis. We have identified the widespread LytR–Cps2A–Psr (LCP) protein family, of previously unknown function, as novel enzymes required for AP synthesis. Structural and biochemical analysis of several LCP proteins suggest that they carry out the final step of transferring APs from their lipid-linked precursor to cell wall peptidoglycan (PG). In Bacillus subtilis, LCP proteins are found in association with the MreB cytoskeleton, suggesting that MreB proteins coordinate the insertion of the major polymers, PG and AP, into the cell wall