Cloning and molecular analysis of paraxis: A {\it trans\/}-acting factor required for somite maturation

During vertebrate embryogenesis, cells from the paraxial mesoderm coalesce in a rostral-to-caudal progression to form the somites. Subsequent compartmentalization of the somites yields the sclerotome, myotome and dermatome, which give rise to the axial skeleton, axial musculature, and dermis, respectively. Recently, we cloned a novel basic-Helix-Loop-Helix (bHLH) protein, called scleraxis, which is expressed in the sclerotome, in mesenchymal precursors of bone and cartilage, and in connective tissues. This dissertation focuses on the cloning, expression and functional analysis of a bHLH protein termed paraxis, which is nearly identical to scleraxis within the bHLH region but diverges in both its amino and carboxyl termini. During the process of mouse embryogenesis, paraxis transcripts are first detected at about day 7.5 post coitum within the primitive mesoderm lying posterior to the head and heart primordia. Subsequently, paraxis expression progresses caudally through the paraxial mesoderm, immediately preceding somite formation. Paraxis is expressed at high levels in newly formed somites before the first detectable expression of the myogenic bHLH genes, and as the somite becomes compartmentalized, paraxis becomes downregulated within the myotome.^ To determine the function of paraxis during mammalian embryogenesis, mice were generated with a null mutation in the paraxis locus. Paraxis null mice survived until birth, but exhibited severe foreshortening along the anteroposterior axis due to the absence of vertebrae caudal to the midthoracic region. The phenotype also included axial skeletal defects, particularly shortened bifurcated ribs which were detached from the vertebral column, fused vertebrae and extensive truncation and disorganization caudal to the hindlimbs. Mutant neonates also lacked normal levels of trunk muscle and exhibited defects in the dermis as well as the stratification of the epidermis. Analysis of paraxis -/- mutant embryos has revealed a failure of the somites to both properly epithelialize and compartmentalize, resulting in defects in somite-derived cell lineages. These results suggest that paraxis is an essential component of the genetic pathway regulating somitogenesis. ^

Learning -dependent plasticity of movement representations in the primate primary motor cortex

Primary motor cortex (M1) is involved in the production of voluntary movement and contains a complete functional representation, or map, of the skeletal musculature. This functional map can be altered by pathological experiences, such as peripheral nerve injury or stroke, by pharmacological manipulation, and by behavioral experience. The process by which experience-dependent alterations of cortical function occur is termed plasticity. In this thesis, plasticity of M1 functional organization as a consequence of behavioral experience was examined in adult primates (squirrel monkeys). Maps of movement representations were derived under anesthesia using intracortical microstimulation, whereby a microelectrode was inserted into the cortex to electrically stimulate corticospinal neurons at low current levels and evoke movements of the forelimb, principally of the hand. Movement representations were examined before and at several times after training on behavioral tasks that emphasized use of the fingers. Two behavioral tasks were utilized that dissociated the repetition of motor activity from the acquisition of motor skills. One task was easy to perform, and as such promoted repetitive motor activity without learning. The other task was more difficult, requiring the acquisition of motor skills for successful performance. Kinematic analysis indicated that monkeys used a consistent set of forelimb movements during pellet extractions. Functional mapping revealed that repetitive motor activity during the easier task did not produce plastic changes in movement representations. Instead, map plasticity, in the form of selective expansions of task-related movement representations, was only produced following skill acquisition on the difficult task. Additional studies revealed that, in general, map plasticity persisted without further training for up to three months, in parallel with the retention of task-related motor skills. Also, extensive additional training on the small well task produced further improvements in performance, and further changes in movement maps. In sum, these experiments support the following three conclusions regarding the role of M1 in motor learning. First, behaviorally-driven plasticity is learning-dependent, not activity-dependent. Second, plastic changes in M1 functional representations represent a neural correlate of acquired motor skills. Third, the persistence of map plasticity suggests that M1 is part of the neural substrate for the memory of motor skills. ^

Plataforma terapeútica basada en Kinect para niños con parálisis cerebral infantil

El sistema SONRIE (Sistema de terapia, basadO en KiNect, paRa nIños con parálisis cErebral), realizado como Proyecto Fin de Grado por Dña. Estefanía Sampedro Sánchez, se desarrolló con el fin de permitir el proceso de rehabilitación de los músculos faciales en niños con Parálisis Cerebral Infantil (PCI). SONRIE se compone de una plataforma de juegos cuyo objetivo es lograr una mejora terapéutica en la musculatura orofacial de niños diagnosticados de PCI con edades comprendidas entre los 4 y los 12 años. El escenario de aplicación del sistema SONRIE son las escuelas de integración que tienen escolarizados alumnos diagnosticados con este trastorno. La posibilidad de rehabilitación de los músculos faciales mediante tratamientos que se apoyan en el uso de sistemas telemáticos, junto con el empleo de tecnologías actuales (Realidad Virtual, Realidad Aumentada y Serious Games) supone una gran innovación en el entorno de la neuro-rehabilitación, entendida como el proceso de terapia que permite optimizar la participación de una persona en la sociedad, alcanzando un grado de bienestar óptimo. El trabajo realizado en este Proyecto Fin de Grado pretende escalar el sistema SONRIE, mediante el análisis, diseño y desarrollo de un Framework encargado de facilitar, ampliar y validar el uso adecuado del sistema SONRIE en entornos escolares a través de la integración de nuevas tecnologías. La plataforma desarrollada en este proyecto, permite dotar de dinamismo y persistencia a la plataforma de juegos, ofreciendo a los usuarios de SONRIE (principalmente fisioterapeutas y rehabilitadores que trabajan en entornos escolares) un sistema de terapia para niños con PCI accesible vía web. En este Proyecto Fin de Grado se describe el conjunto de componentes software desarrollados con el fin de proporcionar un entorno web que escale el sistema SONRIE, convirtiéndolo en un sistema de terapia efectivo, completo y usable. ABSTRACT. The SONRIE system (Sistema de terapia, basadO en KiNect, paRa nIños con parálisis cErebral), performed as a final project by Miss Estefanía Sampedro, was developed in order to allow the rehabilitation process of the facial muscles of children with Cerebral Palsy (CP). SONRIE consists of a gaming platform which aims to achieve a therapeutic improvement in the orofacial musculature on children diagnosed with CP aged between 4 and 12 years. The application scenario of the SONRIE system are the integration schools that have students diagnosed with this disorder. The possibility of rehabilitation of facial muscles through treatments based on the use of telematics systems, together with the use of new technologies (Virtual Reality, Augmented Reality and Serious Games) is a great innovation in the neuro-rehabilitation environment, understood as the therapy process that optimizes the participation of a person in the society, reaching an optimum level of welfare. The work done in this final project aims to scale the SONRIE system, through the analysis, design and development of a framework in charge of facilitating, extending and validating the proper use of the SONRIE system in school environments, through the integration of new technologies. The platform developed in this project, can provide dynamism and persistence to the gaming platform, offering to the SONRIE users (mainly physiotherapists and rehabilitators who work in school settings) a therapy system for children with CP accessible via web. In this final project are described the software components developed in order to provide a web environment that scales the SONRIE system, making it an effective, complete and usable therapy system.

Pax3 modulates expression of the c-Met receptor during limb muscle development.

Pax3 is a transcription factor whose expression has been used as a marker of myogenic precursor cells arising in the lateral somite destined to migrate to and populate the limb musculature. Accruing evidence indicates that the embryologic origins of axial and appendicular muscles are distinct, and limb muscle abnormalities in both mice and humans harboring Pax3 mutations support this distinction. The mechanisms by which Pax3 affects limb muscle development are unknown. The tyrosine kinase receptor for hepatocyte growth factor/scatter factor encoded by the c-met protooncogene is also expressed in limb muscle progenitors and, like Pax-3, is required in the mouse for limb muscle development. Here, we show that c-met expression is markedly reduced in the lateral dermomyotome of Splotch embryos lacking Pax3. We show that Pax3 can stimulate c-met expression in cultured cells, and we identify a potential Pax3 binding site in the human c-MET promoter that may contribute to direct transcriptional regulation. In addition, we have found that several cell lines derived from patients with rhabdomyosarcomas caused by a t(2;13) chromosomal translocation activating PAX3 express c-MET, whereas those rhabdomyosarcoma cell lines examined without the translocation do not. These results are consistent with a model in which Pax3 modulates c-met expression in the lateral dermomyotome, a function that is required for the appropriate migration of these myogenic precursors to the limb where the ligand for c-met (hepatocyte growth factor/scatter factor) is expressed at high levels.

Control of somite patterning by signals from the lateral plate.

The body musculature of higher vertebrates is composed of the epaxial muscles, associated with the vertebral column, and of the hypaxial muscles of the limbs and ventro-lateral body wall. Both sets of muscles arise from different cell populations within the dermomyotomal component of the somite. Myogenesis first occurs in the medial somitic cells that will form the epaxial muscles and starts with a significant delay in cells derived from the lateral somitic moiety that migrate to yield the hypaxial muscles. The newly formed somite is mostly composed of unspecified cells, and the determination of somitic compartments toward specific lineages is controlled by environmental cues. In this report, we show that determinant signals for lateral somite specification are provided by the lateral plate. They result in a blockade of the myogenic program, which maintains the lateral somitic cells as undifferentiated muscle progenitors expressing the Pax-3 gene, and represses the activation of the MyoD family genes. In vivo, this mechanism could account for the delay observed in the onset of myogenesis between muscles of the epaxial and hypaxial domains.

Morris's Human anatomy; a complete systematic treatise by English and American authors,

pt. 1. General morphogeny. Osteology. Articulations.--pt. 2. The musculature. The organs of circulation. The lymphatics.--pt. 3. The nervous system. Organs of special sense.--pt. 4. The organs of digestion. The respitory organs. The urinary and reproductive organs. The ductless glands. The skin and mammary gland.--pt.5. Surgical and topographical anatomy.

Simultaneous fixation using glutaraldehyde and osmium tetroxide or potassium ferricyanide-reduced osmium for the preservation of monogenean flatworms: An assessment for Merizocotyle icopae

Simultaneous fixation was investigated for a marine organism: the monogenean flatworm ectoparasite Merizocotyle icopae. Four protocols for primary fixation were compared: 3% glutaraldehyde alone in OAM cacodylate buffer for a minimum of 2 hours; 1% glutaraldehyde in combination with 1% osmium tetroxide, both in 0.1M cacodylate buffer, until tissues darkened (5-20 minutes); 1% glutaraldehyde in OAM cacodylate buffer in combination with 0.5% potassium ferricyanide-reduced osmium until tissues darkened (5-20 minutes); 1% glutaraldehyde in combination with 1% osmium tetroxide, both in 0.1M cacodylate buffer, for 30 minutes. The study confirms that the standard method for transmission electron microscopic fixation (first listed protocol) routinely applied to platyhelminths is optimal for ultrastructural preservation, but some simultaneous fixation methods (second and third listed protocols) are acceptable when rapid immobilization is needed. Scanning electron microscopic preparations may be improved using simultaneous primary fixation. (C) 2004 Wilcy-Liss, Inc.

Nervous and muscle system development in Phascolion strombus (Sipuncula)

Recent interpretations of developmental gene expression patterns propose that the last common metazoan ancestor was segmented, although most animal phyla show no obvious signs of segmentation. Developmental studies of non-model system trochozoan taxa may shed light on this hypothesis by assessing possible cryptic segmentation patterns. In this paper, we present the first immunocytochemical data on the ontogeny of the nervous system and the musculature in the sipunculan Phascolion strombus. Myogenesis of the first anlagen of the body wall ring muscles occurs synchronously and not subsequently from anterior to posterior as in segmented spiralian taxa (i.e. annelids). The number of ring muscles remains constant during the initial stages of body axis elongation. In the anterior-posteriorly elongated larva, newly formed ring muscles originate along the entire body axis between existing myocytes, indicating that repeated muscle bands do not form from a posterior growth zone. During neurogenesis, the Phascolion larva expresses a non-metameric, paired, ventral nerve cord that fuses in the mid-body region in the late-stage elongated larva. Contrary to other trochozoans, Phascolion lacks any larval serotonergic structures. However, two to three FMRFamide-positive cells are found in the apical organ. In addition, late larvae show commissure-like neurones interconnecting the two ventral nerve cords, while early juveniles exhibit a third, medially placed FMRFamidergic ventral nerve. Although we did not find any indications for cryptic segmentation, certain neuro-developmental traits in Phascolion resemble the conditions found in polychaetes (including echiurans) and myzostomids and support a close relationship of Sipuncula and Annelida.

Branchial vascular pathways in two species of Tetraodontiformes and the concept of secondary vessels and nutrient arteries

The vascular organisation of the branchial basket was examined in two Tetraodontiform fishes; the three-barred porcupinefish, Dicotylichthys punctulatus and the banded toadfish, Marylina pleurosticta by scanning electron microscopy of vascular casts and standard histological approaches. In D. punctulatus, interarterial anastomoses (iaas) originated at high densities from the efferent filamental and branchial arteries, subsequently re-anastomosing to form progressively larger secondary vessels. Small branches of this system entered the filament body, where it was interspersed between the intrafilamental vessels. Large-bore secondary vessels ran parallel with the efferent branchial arteries, and were found to constitute an additional arterio-arterial pathway, in that these vessels exited the branchial basket in company with the mandibular, the carotid and the afferent and efferent branchial arteries, from where they gave rise to capillary beds after exit. Secondary vessels were not found to supply filament muscle; rather these tissues were supplied by single specialised vessels running in parallel between the efferent and afferent branchial arteries in both species examined. Although the branchial vascular anatomy was generally fairly similar for the two species examined, iaas were not found to originate from any branchial component in the banded toadfish, M. pleurosticta, which instead showed a moderate frequency of iaas on other vessels in the cephalic region. It is proposed that four independent vascular pathways may be present within the teleostean gill filament, the conventional arterio-arterial pathway across the respiratory lamellae; an arterio-arterial system of secondary vessels supplying the filament and non-branchial tissues; a system of vessels supplying the filament musculature; and the intrafilamental vessels (central venous sinus). The present study demonstrates that phylogenetic differences in the arrangement of the branchial vascular system occur between species of the same taxon.

Differential activity of regions of transversus abdominis during trunk rotation

The role of the abdominal muscles in trunk rotation is not comprehensively understood. This study investigated the electromyographic (EMG) activity of anatomically distinct regions of the abdominal muscles during trunk rotation in six subjects with no history of spinal pain. Fine-wire electrodes were inserted into the right abdominal wall; upper region of transversus abdominis (TrA), middle region of TrA, obliquus internus abdominis (OI) and obliquus externus abdominis (OE), and lower region of TrA and OI. Surface electrodes were placed over right rectus abdominis (RA). Subjects performed trunk rotation to the left and right in sitting by rotating their pelvis relative to a fixed thorax. EMG activity was recorded in relaxed supine and sitting, and during an isometric hold at end range. TrA was consistently active during trunk rotation, with the recruitment patterns of the upper fascicles opposite to that of the middle and lower fascicles. During left rotation, there was greater activity of the lower and middle regions of contralateral TrA and the lower region of contralateral OI. The upper region of ipsilateral TrA and OE were predominately active during right rotation. In contrast, there was no difference in activity of RA and middle OI between directions (although middle OI was different between directions for all but one subject). This study indicates that TrA is active during trunk rotation, but this activity varies between muscle regions. These normative data will assist in understanding the role of TrA in lumbopelvic control and movement, and the effect of spinal pain on abdominal muscle recruitment.

Effect of cyclic pneumatic soft tissue compression on simulated distal radius fractures

We investigated the effect of pneumatic pressure applied to the proximal musculature of the sheep foreleg on load at the site of a transverse osteotomy of the distal radius. The distal radii of 10 fresh sheep foreleg specimens were osteotomized and a pressure sensor was inserted between the two bone fragments. An inflatable cuff, connected to a second pressure sensor, was positioned around the proximal forelimb musculature and the leg then was immobilized in a plaster cast. The inflatable cuff was inflated and deflated repeatedly to various pressures. Measurements of the cuff pressure and corresponding change in pressure at the osteotomy site were recorded. The results indicated that application of pneumatic pressure to the proximal foreleg musculature produced a corresponding increase in load at the osteotomy site. For the cuff pressures tested (109.8-238.4 mm Hg), there was a linear correlation with the load at the osteotomy site with a gradient of 12 mm Hg/N. It is conceivable, based on the results of this study, that a technique could be developed to provide dynamic loading to accelerate fracture healing in the upper limb of humans.

Regional morphology of the transversus abdominis and obliquus internus and externus abdominis muscles

Background. The mechanisms by which the abdominal muscles move and control the lumbosacral spine are not clearly understood. Descriptions of abdominal morphology are also conflicting and the regional anatomy of these muscles has not been comprehensively examined. The aim of this study was to investigate the morphology of regions of transversus abdominis and obliquus internus and externus abdominis. Methods. Anterior and posterolateral abdominal walls were dissected bilaterally in 26 embalmed human cadavers. The orientation, thickness and length of the upper, middle and lower fascicles of transversus abdominis and obliquus internus abdominis, and the upper and middle fascicles of obliquus externus abdominis were measured. Findings. Differences in fascicle orientation, thickness and length were documented between the abdominal muscles and between regions of each muscle. The fascicles of transversus abdominis were horizontal in the upper region, with increasing inferomedial orientation in the middle and lower regions. The upper and middle fascicles of obliquus internus abdominis were oriented superomedially and the lower fascicles inferomedially. The mean vertical dimension of transversus abdominis that attaches to the lumbar spine via the thoracolumbar fascia was 5.2 (SD 2.1) cm. Intramuscular septa were observed between regions of transversus abdominis, and obliquus internus abdominis could be separated into two distinct layers in the lower and middle regions. Interpretation. This study provides quantitative data of morphological differences between regions of the abdominal muscles, which suggest variation in function between muscle regions. Precise understanding of abdominal muscle anatomy is required for incorporation of these muscles into biomechanical models. Furthermore, regional variation in their morphology may reflect differences in function. (C) 2004 Elsevier Ltd. All rights reserved.

Differential motor speech outcomes in children treated for mid-line cerebellar tumour

Primary objective: To investigate the nature of the motor speech impairments and dysarthria that can arise subsequent to treatment for childhood mid-line cerebellar tumours (CMCT). Research design: The motor speech ability of six cases of children with CMCT was analysed using perceptual and physiological measures and compared with that of a group of non-neurologically impaired children matched for age and sex. Main outcome and results: Three of the children with CMCT were perceived to exhibit dysarthric speech, while the remaining three were judged to have normal speech. The speech disorder in three of the children with CMCT was marked by deviances in prosody, articulation and phonation. The underlying pathophysiology was linked to cerebellar damage and expressed as difficulty in co-ordinating the motor speech musculature as required for speech production. These deficits were not identified in the three non-dysarthric children with CMCT. Conclusion: Differential motor speech outcomes occur for children treated for CMCT and these are discussed within the realm of possible mechanisms responsible for these differences. The need for further investigation of the risk factors for development of motor speech impairment in children treated for CMCT is also highlighted.

Postural activity of the abdominal muscles varies between regions of these muscles and between body positions

The abdominal muscles have an important role in control and movement of the lumbar spine and pelvis. Given there is new evidence of morphological and functional differences between distinct anatomical regions of the abdominal muscles, this study investigated whether there are regional differences in postural activity of these muscles and whether recruitment varies between different body positions. Eleven subjects with no history of low back pain that affected function or for which they sought treatment participated in the study. Electromyographic (EMG) activity of the upper, middle and lower regions of transversus abdominis (TrA), the middle and lower regions of obliquus internus abdominis (OI) and the middle region of obliquus externus abdominis (OE) was recorded using intramuscular electrodes. All subjects performed rapid, unilateral shoulder flexion in standing and six subjects also moved their upper limb in sitting. There were regional differences in the postural responses of TrA with limb movement. Notably, the onset of EMG of the upper region was later than that of the lower and middle regions. There were no differences in the EMG onsets of lower and middle TrA or OI. The postural responses of the abdominal muscles were also found to differ between body positions, with recruitment delayed in sitting compared to standing. This study showed that there is regional differentiation in TrA activity with challenges to postural control and that body position influences the postural responses of the abdominal muscles. These results may reflect variation in the contribution of abdominal muscle regions to stability of the trunk. (c) 2004 Elsevier B.V. All rights reserved.

The relationship between knee strength and functional stability before and after anterior cruciate ligament reconstruction

Functional stability of the knee is dependent on an intact ligamentous system and the timely and efficient contraction of supporting musculature. The aim of this study was to assess the relationship between muscle strength and functional stability in 31 patients pre- and post-operatively, following a unilateral anterior cruciate ligament rupture. All subjects underwent reconstructive surgery using semitendonosis and gracilis tendons. Isokinetic strength assessment of quadriceps and hamstring muscles was performed at a rate of movement of 60% and 120degrees/s. Functional stability was determined by performance during five functional stability tests that included the shuttle run, side step, carioca, single and triple hop tests. Pearson's correlation coefficient statistics were applied to pre-operative and post-operative data respectively. These analyses demonstrated a significant positive correlation between quadriceps strength indices at both testing speeds and the two hop tests pre-operatively (p's < 0.007) and between quadriceps strength indices at both speeds and all five functional tests post-operatively (p's < 0.01). Assessed using Steiger's formula, there was a significant increase in the correlation between quadriceps strength indices and three functional tests post-operatively compared to pre-operatively (p < 0.05). No significant correlation between hamstring strength indices and functional scores existed pre- or post-operatively. This study has shown a significant correlation exists between quadriceps strength indices and functional stability both before and after surgery, this relationship does not reach significance between hamstring strength indices and functional stability. (C) 2002 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved.