Morphologie tridimensionnelle du bassin scoliotique

La scoliose idiopathique de l’adolescence (SIA) est une déformation tridimensionnelle de la colonne vertébrale et de la cage thoracique dont la cause est inconnue. Il semble que la ceinture pelvienne soit impliquée dans la pathogénie de la SIA, car des différences géométriques des os coxaux ont été observées. Notamment, une rotation du bassin ou une inclinaison latérale dans le sens de la courbe scoliotique ont été mises en évidence en plus des distorsions osseuses. Il est difficile de dissocier la rotation du bassin de son asymétrie, car la majorité des études porte sur des données radiologiques bidimensionnelles. Une analyse tridimensionnelle de la morphologie du bassin de patientes ayant une SIA, mais n’ayant pas reçu de traitement par corset ou chirurgie permettrait d’identifier le rôle du bassin dans la progression de la scoliose. Dix-sept jeunes filles atteintes de la SIA ont participé à cette étude pour lesquelles des radiographies bi-planaires en station debout étaient disponibles au moment du diagnostic par un chirurgien orthopédiste pédiatrique et à l’instant de la prescription d'un corset. Des radiographies postéro-antérieures et latérales avaient été prises au moyen du système EOS®. Douze repères anatomiques du bassin ont été identifiés sur les paires de radiographies, alors que quatre repères ont été identifiés sur la radiographie postéro-antérieure uniquement. Ces quatre derniers n’étaient pas identifiables sur la radiographie latérale à cause de la superposition des repères droits et gauches. La reconstruction tridimensionnelle du bassin a été réalisée à partir de deux clichés radiographiques des 12 premiers repères osseux. Au total, neuf paramètres tridimensionnels ont été calculés afin de quantifier l’asymétrie et la distorsion du bassin entre les deux temps donnés. Des paramètres bidimensionnels ont également été mesurés sur les quatre derniers repères osseux afin de documenter des déformations du bassin pertinentes à la pratique clinique, telle que la rotation axiale de celui-ci. Dans le but d'évaluer une possible asymétrie entre les os coxaux du bassin, les paramètres tridimensionnels du bassin gauche ont été comparés à ceux du côté droit à chaque temps, au moyen d'un test-t pour échantillon apparié. La morphologie pelvienne a été également évaluée par l'analyse multivariée (MANOVA) à mesures répétées à deux conditions (côté*temps). En conséquence, nous avons constaté une croissance osseuse statistiquement significative du bassin dans l’intervalle de temps entre le diagnostic de la scoliose et le port du corset (p=0,033). Une asymétrie significative entre les côtés gauche et droit du bassin (p=0,013) a également été constatée. En ce qui concerne les paramètres bidimensionnels, nous avons constaté une augmentation de la version pelvienne (p=0,024) au cours de la croissance des jeunes filles. Finalement, le bassin n'a pas présenté de distorsion, puisqu'une valeur de p de 0,763 a été observée. En conclusion, la croissance des jeunes filles atteintes de la scoliose idiopathique de l'adolescence est accompagnée d'une asymétrie morphologique entre les deux os coxaux du bassin. Cette asymétrie constatée au moment du diagnostic de la scoliose des filles a évolué jusqu'à l’instant où le port du corset a été prescrit. Quant aux paramètres bidimensionnels, nous pouvons conclure que la rotation du bassin vers l'arrière a augmenté au cours de la croissance des jeunes filles, produisant ainsi une rétroversion pelvienne dans le plan sagittal. La distorsion tridimensionnelle du bassin n'a toutefois pas été observée au cours de la croissance des jeunes filles.Adolescent idiopathic scoliosis (AIS) is a three-dimensional deformation of the spine and rib cage of unknown cause. It seems that the pelvis is involved in the pathogenesis of the AIS because geometric differences in the hip bones were observed. A pelvic rotation or a lateral tilt in the direction of the scoliotic curve was observed in addition to bone distortions. It is difficult to separate the pelvic rotation from its asymmetry since most studies are based on bi-dimensional radiologic data. A three-dimensional analysis of the pelvis morphology in patients with AIS, but no brace treatment nor surgery could better define the role of pelvis in the progression of the scoliosis. Seventeen young girls with AIS have participated in this study. Standing bi-planer radiographs were available at the time of diagnosis by a pediatric orthopedic surgeon and at the moment of the brace prescription. Postero-anterior and lateral X-rays were taken using the EOS® system. Twelve anatomical landmarks of the pelvis were identified on the radiograph pairs and four landmarks were identified on the postero-anterior radiography only. These were not identified on the lateral radiograph due to the superposition of the right and left landmarks. The three-dimensional reconstruction of the pelvis was performed using two radiographs of the first 12 bony landmarks. A total of nine three-dimensional parameters were calculated to quantify the asymmetry of the pelvis distortion between the two given times. Bi-dimensional parameters were also measured on the last four bony landmarks in order to document the pelvic deformations relevant to the clinical practice such as the rotation. With the objective of evaluating a possible asymmetry between the hip bones of the pelvis, the three-dimension parameters of the left pelvis were compared to those of the right side in each time using a paired sample t-test. The pelvic morphology was also evaluated using a multivariate analysis (MANOVA) with repeated measures on two conditions (side*time). A statistically significant bone growth of the pelvis in the time interval between the diagnosis of the scoliosis and the brace prescription (p=0.033) was observed in consequence. A significant asymmetry between the left and right sides of the pelvis (p=0.013) without distortion was also observed. Regarding the bi-dimensional parameters, we found an increase in pelvic release (p=0.024) during the growth of the girls. Finally, the pelvis did not show a distortion because a p value of 0.763 was observed. In conclusion, the growth of young girls with adolescent idiopathic scoliosis is related to a morphological asymmetry between the two hipbones of the pelvis. This asymmetry observed at the moment of the scolioisis diagnosis has evolved until de moment of the brace prescription. As for the bi-dimensional parameters, we can conclude that the rotation of the pelvis to the back increased during the growth of the girls, producing a pelvic retroversion in the sagittal plan. The three-dimensional distortion of the pelvis, however, has not been observed during the young girls growth

Nanoparticle-Delivered Multimeric Soluble CD40L DNA Combined with Toll-Like Receptor Agonists as a Treatment for Melanoma

Stimulation of CD40 or Toll-Like Receptors (TLR) has potential for tumor immunotherapy. Combinations of CD40 and TLR stimulation can be synergistic, resulting in even stronger dendritic cell (DC) and CD8+ T cell responses. To evaluate such combinations, established B16F10 melanoma tumors were injected every other day X 5 with plasmid DNA encoding a multimeric, soluble form of CD40L (pSP-D-CD40L) either alone or combined with an agonist for TLR1/2 (Pam3CSK4 ), TLR2/6 (FSL-1 and MALP2), TLR3 (polyinosinic-polycytidylic acid, poly(I:C)), TLR4 ( monophosphoryl lipid A, MPL), TLR7 (imiquimod), or TLR9 (Class B CpG phosphorothioate oligodeoxynucleotide, CpG). When used by itself, pSP-D-CD40L slowed tumor growth and prolonged survival, but did not lead to cure. Of the TLR agonists, CpG and poly(I:C) also slowed tumor growth, and the combination of these two TLR agonists was more effective than either agent alone. The triple combination of intratumoral pSP-D-CD40L + CpG + poly(I:C) markedly slowed tumor growth and prolonged survival. This treatment was associated with a reduction in intratumoral CD11c+ dendritic cells and an influx of CD8+ T cells. Since intratumoral injection of plasmid DNA does not lead to efficient transgene expression, pSP-D-CD40L was also tested with cationic polymers that form DNA-containing nanoparticles which lead to enhanced intratumoral gene expression. Intratumoral injections of pSP-D-CD40L-containing nanoparticles formed from polyethylenimine (PEI) or C32 (a novel biodegradable poly(B-amino esters) polymer) in combination with CpG + poly(I:C) had dramatic antitumor effects and frequently cured mice of B16F10 tumors. These data confirm and extend previous reports that CD40 and TLR agonists are synergistic and demonstrate that this combination of immunostimulants can significantly suppress tumor growth in mice. In addition, the enhanced effectiveness of nanoparticle formulations of DNA encoding immunostimulatory molecules such as multimeric, soluble CD40L supports the further study of this technology for tumor immunotherapy

Short-Term High-Fat Diet Consumption Reduces Hypothalamic Expression of the Nicotinic Acetylcholine Receptor α7 Subunit (α7nAChR) and Affects the Anti-inflammatory Response in a Mouse Model of Sepsis

Sepsis is one of the leading causes of death in hospitalized patients and the chronic and low-grade inflammation observed in obesity seems to worsen susceptibility and morbidity of infections. However, little is known with respect to a short-term high-fat diet (HFD) and its role in the development of sepsis. Here, we show for the first time, that short-term HFD consumption impairs early nicotinic acetylcholine receptor α7 subunit (α7nAChR)- mediated signaling, one of the major components of the cholinergic anti-inflammatory pathway, with a focus on hypothalamic inflammation and innate immune response. Mice were randomized to a HFD or standard chow (SC) for 3 days, and sepsis was subsequently induced by a lethal intraperitoneal (i.p.) injection of lipopolysaccharide (LPS) or by cecal ligation and puncture (CLP) surgery. In a separate experiment, both groups received LPS (i.p.) or LPS (i.p.) in conjunction with the selective α7nAChR agonist, PNU-282987 (i.p. or intracerebroventricular; i.c.v.), and were sacrificed 2 h after the challenge. Short-term HFD consumption significantly reduced the α7nAChR mRNA and protein levels in the hypothalamus and liver (p < 0.05). Immunofluorescence microscopy demonstrated lower cholinergic receptor nicotinic α7 subunit (α7nAChR)+ cells in the arcuate nucleus (ARC) (α7nAChR+ cells in SC = 216 and HFD = 84) and increased F4/80+ cells in the ARC (2.6-fold) and median eminence (ME) (1.6-fold), which can contribute to neuronal damage. Glial fibrillary acidic protein (GFAP)+ cells and neuronal nuclear antigen (NeuN)+ cells were also increased following consumption of HFD. The HFD-fed mice died quickly after a lethal dose of LPS or following CLP surgery (2-fold compared with SC). The LPS challenge raised most cytokine levels in both groups; however, higher levels of TNF-α (Spleen and liver), IL-1β and IL-6 (in all tissues evaluated) were observed in HFD-fed mice. Moreover, PNU-282987 administration (i.p. or i.c.v.) reduced the levels of inflammatory markers in the hypothalamus following LPS injection. Nevertheless, when the i.c.v. injection of PNU-282987 was performed the anti-inflammatory effect was much smaller in HFD-fed mice than SC-fed mice. Here, we provide evidence that a short-term HFD impairs early α7nAChR expression in central and peripheral tissues, contributing to a higher probability of death in sepsis

In Vivo Gene Knockdown in Rat Dorsal Root Ganglia Mediated by Self-Complementary Adeno-Associated Virus Serotype 5 Following Intrathecal Delivery

We report here in adult rat viral vector mediate-gene knockdown in the primary sensory neurons and the associated cellular and behavior consequences. Self-complementary adeno-associated virus serotype 5 (AAV5) was constructed to express green fluorescent protein (GFP) and a small interfering RNA (siRNA) targeting mammalian target of rapamycin (mTOR). The AAV vectors were injected via an intrathecal catheter. We observed profound GFP expression in lumbar DRG neurons beginning at 2-week post-injection. Of those neurons, over 85% were large to medium-diameter and co-labeled with NF200, a marker for myelinated fibers. Western blotting of mTOR revealed an 80% reduction in the lumbar DRGs (L4–L6) of rats treated with the active siRNA vectors compared to the control siRNA vector. Gene knockdown became apparent as early as 7-day post-injection and lasted for at least 5 weeks. Importantly, mTOR knockdown occurred in large (NF200) and small-diameter neurons (nociceptors). The viral administration induced an increase of Iba1 immunoreactivity in the DRGs, which was likely attributed to the expression of GFP but not siRNA. Rats with mTOR knockdown in DRG neurons showed normal general behavior and unaltered responses to noxious stimuli. In conclusion, intrathecal AAV5 is a highly efficient vehicle to deliver siRNA and generate gene knockdown in DRG neurons. This will be valuable for both basic research and clinic intervention of diseases involving primary sensory neurons

Current concepts on hemodynamic support and therapy in septic shock

ABSTRACTSevere sepsis and septic shock represent a major healthcare challenge. Much of the improvement in mortality associated with septic shock is related to early recognition combined with timely fluid resuscitation and adequate antibiotics administration. The main goals of septic shock resuscitation include intravascular replenishment, maintenance of adequate perfusion pressure and oxygen delivery to tissues. To achieve those goals, fluid responsiveness evaluation and complementary interventions - i.e. vasopressors, inotropes and blood transfusion - may be necessary. This article is a literature review of the available evidence on the initial hemodynamic support of the septic shock patients presenting to the emergency room or to the intensive care unit and the main interventions available to reach those targets, focusing on fluid and vasopressor therapy, blood transfusion and inotrope administration

Short-term high-fat diet consumption reduces hypothalamic expression of the nicotinic acetylcholine receptor alpha 7 subunit (alpha 7nAChR) and affects the anti-inflammatory response in a mouse model of sepsis

Sepsis is one of the leading causes of death in hospitalized patients and the chronic and low-grade inflammation observed in obesity seems to worsen susceptibility and morbidity of infections. However, little is known with respect to a short-term high-fat diet (HFD) and its role in the development of sepsis. Here, we show for the first time, that short-term HFD consumption impairs early nicotinic acetylcholine receptor alpha 7 subunit (alpha 7nAChR)-mediated signaling, one of the major components of the cholinergic anti-inflammatory pathway, with a focus on hypothalamic inflammation and innate immune response. Mice were randomized to a HFD or standard chow (SC) for 3 days, and sepsis was subsequently induced by a lethal intraperitoneal (i.p.) injection of lipopolysaccharide (LPS) or by cecal ligation and puncture (CLP) surgery. In a separate experiment, both groups received LPS (i.p.) or LPS (i.p.) in conjunction with the selective a alpha nAChR agonist, PNU-282987 (i.p. or intracerebroventricular; i.c.v.), and were sacrificed 2 h after the challenge. Short-term HFD consumption significantly reduced the alpha 7nAChR mRNA and protein levels in the hypothalamus and liver (p < 0.05). Immunofluorescence microscopy demonstrated lower cholinergic receptor nicotinic alpha 7 subunit (alpha 7nAChR)+ cells in the arcuate nucleus (ARC) (alpha 7nAChR+ cells in SC = 216 and HFD = 84) and increased F4/80+ cells in the ARC (2.6-fold) and median eminence (ME) (1.6-fold), which can contribute to neuronal damage. Glial fibrillary acidic protein (GFAP)+ cells and neuronal nuclear antigen (NeuN)+ cells were also increased following consumption of HFD. The HFD-fed mice died quickly after a lethal dose of LPS or following CLP surgery (2-fold compared with SC). The LPS challenge raised most cytokine levels in both groups; however, higher levels of TNF-alpha (Spleen and liver), IL-1 beta and IL-6 (in all tissues evaluated) were observed in HFD-fed mice. Moreover, PNU-282987 administration (i.p. or i.c.v.) reduced the levels of inflammatory markers in the hypothalamus following LPS injection. Nevertheless, when the i.c.v. injection of PNU-282987 was performed the anti-inflammatory effect was much smaller in HFD-fed mice than SC-fed mice. Here, we provide evidence that a short-term HFD impairs early alpha 7nAChR expression in central and peripheral tissues, contributing to a higher probability of death in sepsis10CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informação16/23484-1; 13/07607-

Thoracic 9 Spinal Transection-Induced Model of Muscle Spasticity in the Rat: A Systematic Electrophysiological and Histopathological Characterization.

The development of spinal hyper-reflexia as part of the spasticity syndrome represents one of the major complications associated with chronic spinal traumatic injury (SCI). The primary mechanism leading to progressive appearance of muscle spasticity is multimodal and may include loss of descending inhibitory tone, alteration of segmental interneuron-mediated inhibition and/or increased reflex activity to sensory input. Here, we characterized a chronic thoracic (Th 9) complete transection model of muscle spasticity in Sprague-Dawley (SD) rats. Isoflurane-anesthetized rats received a Th9 laminectomy and the spinal cord was transected using a scalpel blade. After the transection the presence of muscle spasticity quantified as stretch and cutaneous hyper-reflexia was identified and quantified as time-dependent changes in: i) ankle-rotation-evoked peripheral muscle resistance (PMR) and corresponding electromyography (EMG) activity, ii) Hoffmann reflex, and iii) EMG responses in gastrocnemius muscle after paw tactile stimulation for up to 8 months after injury. To validate the clinical relevance of this model, the treatment potency after systemic treatment with the clinically established anti-spastic agents baclofen (GABAB receptor agonist), tizanidine (α2-adrenergic agonist) and NGX424 (AMPA receptor antagonist) was also tested. During the first 3 months post spinal transection, a progressive increase in ankle rotation-evoked muscle resistance, Hoffmann reflex amplitude and increased EMG responses to peripherally applied tactile stimuli were consistently measured. These changes, indicative of the spasticity syndrome, then remained relatively stable for up to 8 months post injury. Systemic treatment with baclofen, tizanidine and NGX424 led to a significant but transient suppression of spinal hyper-reflexia. These data demonstrate that a chronic Th9 spinal transection model in adult SD rat represents a reliable experimental platform to be used in studying the pathophysiology of chronic spinal injury-induced spasticity. In addition a consistent anti-spastic effect measured after treatment with clinically effective anti-spastic agents indicate that this model can effectively be used in screening new anti-spasticity compounds or procedures aimed at modulating chronic spinal trauma-associated muscle spasticity