Prévalence et signification fonctionnelle des mouvements périodiques des jambes

Les mouvements périodiques des jambes sont de courts mouvements involontaires qui surviennent de façon périodique au cours du sommeil ou de l’éveil. Ils sont présents dans certains troubles du sommeil, mais également chez des sujets sans plainte reliée au sommeil. Le premier objectif de cette thèse visait une meilleure description de la prévalence de ces mouvements. Nous avons montré que chez les sujets sans plainte de sommeil, la prévalence des mouvements périodiques des jambes en sommeil augmentait de façon importante à partir d’environ 40 ans, tandis que l’index des mouvements périodiques des jambes à l’éveil évoluait avec l’âge selon une courbe en U. Chez les sujets atteints de narcolepsie, on retrouvait davantage de mouvements périodiques des jambes que chez les sujets témoins, mais leur patron d’évolution avec l’âge était similaire. Le deuxième objectif de cette thèse visait l’étude des mouvements périodiques des jambes en relation avec le système nerveux autonome cardiovasculaire. Nous avons non seulement confirmé la présence d’une tachycardie suivie d’une bradycardie lors des mouvements périodiques des jambes durant le sommeil chez les patients atteints du syndrome d’impatiences musculaires à l’éveil et chez les sujets sans plainte de sommeil, mais nous avons également décrit ces mêmes changements de la fréquence cardiaque, quoiqu’avec une plus faible amplitude, chez les sujets atteints de narcolepsie. Finalement, nous avons montré pour la première fois que les mouvements périodiques des jambes en sommeil des sujets atteints du syndrome d’impatiences musculaires à l’éveil et des sujets sans plainte de sommeil étaient aussi associés à des augmentations importantes et significatives de la pression artérielle.Periodic leg movements are short involuntary movements occurring periodically during sleep or wakefulness. They occur in some sleep disorders, but also in healthy subjects not complaining of sleep problems. The first objective of this thesis was to provide a better description of the prevalence of these movements. In healthy non-complaining subjects, the prevalence of periodic leg movements during sleep increased dramatically from about age 40, whereas the age-related evolution of periodic leg movements during wakefulness followed a U curve. In narcoleptic patients there were more periodic leg movements than in control subjects, but their evolution with age showed the same pattern. The second objective of this thesis was to study periodic leg movements in relationship with cardiovascular autonomic nervous system. We not only confirmed that periodic leg movements during sleep were associated with a tachycardia followed by a bradycardia in restless legs syndrome patients and in healthy non-complaining subjects, but that these heart rate changes were also present in narcoleptic patients, albeit of a lower amplitude. Finally, we showed for the first time that periodic leg movements during sleep in restless legs syndrome patients and in healthy non-complaining subjects were also associated with significant and important rises of blood pressure

Determination of intracellular protein-ligand binding affinity by competition binding in-cell NMR

Structure-based drug development suffers from high attrition rates due to the poor activity of lead compounds in cellular and animal models caused by low cell penetrance, off-target binding or changes in the conformation of the target protein in the cellular environment. The latter two effects cause a change in the apparent binding affinity of the compound, which is indirectly assessed by cellular activity assays. To date, direct measurement of the intracellular binding affinity remains a challenging task. In this work, in-cell NMR spectroscopy was applied to measure intracellular dissociation constants in the nanomolar range by means of protein-observed competition binding experiments. Competition binding curves relative to a reference compound could be retrieved either from a series of independent cell samples or from a single real-time NMR bioreactor run. The method was validated using a set of sulfonamide-based inhibitors of human carbonic anhydrase II with known activity in the subnanomolar to submicromolar range. The intracellular affinities were similar to those obtained in vitro, indicating that these compounds selectively bind to the intracellular target. In principle, the approach can be applied to any soluble intracellular target that gives rise to measurable chemical shift changes upon ligand binding

Blood management in fast-track orthopedic surgery: An evidence-based narrative review

Background and purpose: Innovations able to maintain patient safety while reducing the amount of transfusion add value to orthopedic procedures. Opportunities for improvement arise especially in elective procedures, as long as room for planning is available. Although many strategies have been proposed, there is no consensus about the most successful combination. The purpose of this investigation is to identify information to support blood management strategies in fast-track total joint arthroplasty (TJA) pathway, to (i) support clinical decision making according to current evidence and best practices, and (ii) identify critical issues which need further research. Methods and materials: We identified conventional blood management strategies in elective orthopedic procedures. We performed an electronic search about blood management strategies in fast-track TJA. We designed tables to match every step of the former with the latter. We submitted the findings to clinicians who operate using fast-track surgery protocols in TJA at our research hospital. Results: Preoperative anemia detection and treatment, blood anticoagulants/aggregants consumption, transfusion trigger, anesthetic technique, local infiltration analgesia, drainage clamping and removals, and postoperative multimodal thromboprophylaxis are the factors which can add best value to a fast-track pathway, since they provide significant room for planning and prediction. Conclusion: The difference between conventional and fast-track pathways does not lie in the contents of blood management, which are related to surgeons/surgeries, materials used and patients, but in the way these contents are integrated into each other, since elective orthopedic procedures offer significant room for planning. Further studies are needed to identify optimal regimens

Modeling the human tibio-femoral joint using ex vivo determined compliance matrices.

Several approaches have been used to devise a model of the human tibio-femoral joint for embedment in lower limb musculoskeletal models. However, no study has considered the use of cadaveric 6x6 compliance (or stiffness) matrices to model the tibio-femoral joint under normal or pathological conditions. The aim of this paper is to present a method to determine the compliance matrix of an ex vivo tibio-femoral joint for any given equilibrium pose. Experiments were carried out on a single ex vivo knee, first intact and, then, with the anterior cruciate ligament (ACL) transected. Controlled linear and angular displacements were imposed in single degree-of-freedom (DoF) tests to the specimen and resulting forces and moments measured using an instrumented robotic arm. This was done starting from seven equilibrium poses characterized by the following flexion angles: 0°, 15°, 30°, 45°, 60°, 75°and 90°. A compliance matrix for each of the selected equilibrium poses and for both the intact and ACL deficient specimen was calculated. The matrix, embedding the experimental load-displacement relationship of the examined DoFs, was calculated using a linear least squares inversion based on a QR decomposition, assuming symmetric and positive-defined matrices. Single compliance matrix terms were in agreement with the literature. Results showed an overall increase of the compliance matrix terms due to the ACL transection (2.6 ratio for rotational terms at full extension) confirming its role in the joint stabilization. Validation experiments were carried out by performing a Lachman test (the tibia is pulled forward) under load control on both the intact and ACL-deficient knee and assessing the difference (error) between measured linear and angular displacements and those estimated using the appropriate compliance matrix. This error increased non-linearly with respect to the values of the load. In particular, when an incremental posterior-anterior force up to 6 N was applied to the tibia of the intact specimen, the errors on the estimated linear and angular displacements were up to 0.6 mm and 1.5°, while for a force up to 18 N the errors were 1.5 mm and 10.5°, respectively. In conclusion, the method used in this study may be a viable alternative to characterize the tibio-femoral load-dependent behavior in several applications

Structural Basis for the Interaction of the Myosin Light Chain Mlc1p with the Myosin V Myo2p IQ Motifs

Calmodulin, regulatory, and essential myosin light chain are evolutionary conserved proteins that, by binding to IQ motifs of target proteins, regulate essential intracellular processes among which are efficiency of secretory vesicles release at synapsis, intracellular signaling, and regulation of cell division. The yeast Saccharomyces cerevisiae calmodulin Cmd1 and the essential myosin light chain Mlc1p share the ability to interact with the class V myosin Myo2p and Myo4 and the class II myosin Myo1p. These myosins are required for vesicle, organelle, and mRNA transport, spindle orientation, and cytokinesis. We have used the budding yeast model system to study how calmodulin and essential myosin light chain selectively regulate class V myosin function. NMR structural analysis of uncomplexed Mlc1p and interaction studies with the first three IQ motifs of Myo2p show that the structural similarities between Mlc1p and the other members of the EF-hand superfamily of calmodulin-like proteins are mainly restricted to the C-lobe of these proteins. The N-lobe of Mlc1p presents a significantly compact and stable structure that is maintained both in the free and complexed states. The Mlc1p N-lobe interacts with the IQ motif in a manner that is regulated both by the IQ motifs sequence as well as by light chain structural features. These characteristic allows a distinctive interaction of Mlc1p with the first IQ motif of Myo2p when compared with calmodulin. This finding gives us a novel view of how calmodulin and essential light chain, through a differential binding to IQ1 of class V myosin motor, regulate this activity during vegetative growth and cytokinesis