Evaluación postural y análisis del equilibrio en principiantes de esquí nórdico / Postural evaluation and balance analysis in nordic skiing beginners

Se trata de un estudio de diseño transversal con evaluación del control postural antes-después de un curso de iniciación al esquí nórdico (5 jornadas, 6h/día), a un grupo experimental de 25 sujetos, participantes de la formación, y un grupo control de 8 sujetos. Se pretende evaluar el efecto de un entrenamiento inicial de esquí nórdico sobre el control postural en jóvenes deportistas. La estabilidad corporal se evaluó por medio de un test estabilométrico, calculando el área de desviación del centro de presiones por medio de una plataforma de fuerzas con sensores electrónicos. No se aprecian diferencias significativas en el grado de mejora del equilibrio entre el grupo experimental y el grupo control. Nuevos estudios con una mayor exigencia en la formación en esquí nórdico y un mayor número de sujetos en el grupo control son requeridos (AU)

Cross-sectional study with evaluation of postural control before and after a course of initiation to Nordic skiing (5 days, 6 hours per day). The sample consisted of an experimental group of 25 subjects participated in the training and a control group of 8 subjects. It is intended to evaluate the effect of initial training of Nordic skiing on postural control in young athletes. The physical stability was evaluated by a stabilometric test, calculating the deviation of the area of the center of pressures by the use of a force platform with electronic sensors. No significant differences were observed in the degree of improvement of the balance between the experimental group and the control group. New studies with a greater emphasis on training in Nordic skiing and a greater number of subjects in the control group are required (AU)

Tumor buding y el cáncer colorrectal / Budding tumour and colorectal cancer

Introduction: Tumour budding (TB) is defined as the presence of clusters of tumoural cells detaching from invasive margin of main tumor. It is an independent adverse prognostic factor in colorectal cancer. The aim of this study is to determinate if severity of tumor budding is associated with others prognostic factors in colorectal cancer. Materials and Methods: The study group is composed by 43 patients (27 males and 16 females; average age 73.4 years, (27-91) with colorectal cancer who underwent curative surgery. The histologic method of tumour budding used in this study was described by Nakamura. The applied statistical software was G-Stat 2.0. Statistical significance is accepted at p < 0.05. Results: High grade of TB was significantly associated with lymph node metastasis (p = 0.027), infiltrative tumour-border configuration (p = 0.016), lymphvessels invasion (p = 0.02), perineural invasion (p = 0.009) and tumor deposits (p = 0.018). There was a significant association with low grade of TB and peritumoural lymphocytic infiltration (p = 0.004). Conclusions: High grade of TB is significantly associated with other adverse prognostic factors as lymph node metastasis, infiltrative tumour-border configuration, lymphvessels invasion, venous invasion, perineural invasion and tumor deposits; and low grade of TB with favorable prognostic factor as peritumoural lymphocytic infiltration in colorectal cancer. Tumour budding can help to identify hig-risk patients with colorectal cancer.

Introducción: El tumor budding (TB) es la presencia de células tumorales aisladas o en pequeños grupos situadas en el frente infiltrante del tumor. Su hallazgo en alto grado es un factor de mal pronóstico independiente del cáncer colorrectal. El objetivo de este trabajo es determinar si el grado de TB está asociado con otros factores pronósticos del cáncer colorrectal. Materiales y Métodos: Se analizaron retrospectivamente 43 pacientes (27 varones y 16 mujeres) con una edad media de 73,4 años (27-91) intervenidos por cáncer colorrectal. El método histológico utilizado para determinar la presencia de TB fue el descrito por Nakamura en 2005. El análisis estadístico se realizó con el programa G-Stat2.0. Las diferencias se consideraron significativas si p < 0,05. Resultados: La presencia de TB de alto grado se asocia significativamente con la presencia de metástasis a ganglios linfáticos (p = 0,027), patrón de crecimiento infiltrativo (p = 0,016), invasión linfática (p = 0,02), perineural (p = 0,009) y depósitos tumorales discontinuos (p = 0,018). El TB de bajo grado se relaciona con la presencia de reacción linfocitaria peritumoral (p = 0,004). Conclusiones: El tumor budding alto grado se asocia con otros factores de mal pronóstico, como metástasis a ganglios linfáticos, crecimiento infiltrativo, invasión linfática, perineural y depósitos tu-morales discontinuos; y el tumor budding bajo grado con factores de buen pronóstico del cáncer colorrectal como reacción linfocitaria peritumoral. El análisis del grado de tumor budding podría ayudar a identificar a pacientes de peor pronóstico con cáncer colorrectal.

High sphingomyelin levels induce lysosomal damage and autophagy dysfunction in Niemann Pick disease type A.

Niemann Pick disease type A (NPA), which is caused by loss of function mutations in the acid sphingomyelinase (ASM) gene, is a lysosomal storage disorder leading to neurodegeneration. Yet, lysosomal dysfunction and its consequences in the disease are poorly characterized. Here we show that undegraded molecules build up in neurons of acid sphingomyelinase knockout mice and in fibroblasts from NPA patients in which autophagolysosomes accumulate. The latter is not due to alterations in autophagy initiation or autophagosome-lysosome fusion but because of inefficient autophago-lysosomal clearance. This, in turn, can be explained by lysosomal membrane permeabilization leading to cytosolic release of Cathepsin B. High sphingomyelin (SM) levels account for these effects as they can be induced in control cells on addition of the lipid and reverted on SM-lowering strategies in ASM-deficient cells. These results unveil a relevant role for SM in autophagy modulation and characterize autophagy anomalies in NPA, opening new perspectives for therapeutic interventions.

Carbamazepine induces mitotic arrest in mammalian Vero cells.

We reported recently that the anticonvulsant drug carbamazepine, at supratherapeutic concentrations, exerts antiproliferative effects in mammalian Vero cells, but the underlying mechanism has not been elucidated. This motivates us to examine rigorously whether growth arrest was associated with structural changes in cellular organization during mitosis. In the present work, we found that exposure of the cells to carbamazepine led to an increase in mitotic index, mainly due to the sustained block at the metaphase/anaphase boundary, with the consequent inhibition of cell proliferation. Indirect immunofluorescence, using antibodies directed against spindle apparatus proteins, revealed that mitotic arrest was associated with formation of monopolar spindles, caused by impairment of centrosome separation. The final consequence of the spindle defects induced by carbamazepine, depended on the duration of cell cycle arrest. Following the time course of accumulation of metaphase and apoptotic cells during carbamazepine treatments, we observed a causative relationship between mitotic arrest and induction of cell death. Conversely, cells released from the block of metaphase by removal of the drug, continued to progress through mitosis and resume normal proliferation. Our results show that carbamazepine shares a common antiproliferative mechanism with spindle-targeted drugs and contribute to a better understanding of the cytostatic activity previously described in Vero cells. Additional studies are in progress to extend these initial findings that define a novel mode of action of carbamazepine in cultured mammalian cells.

Cytotoxicity of butylated hydroxyanisole in Vero cells.

Butylated hydroxyanisole (BHA) is perhaps the most extensively used synthetic antioxidant in the food and cosmetic industry, although considerable controversy exists in the literature regarding the safety of this compound. Most in vitro studies describing the effects of BHA have been performed in cancer cells, but it is unclear whether normal cells are equally susceptible to BHA exposure. The present study investigate the toxic potential of BHA in mammalian cells, using biochemical and morphological parameters, which reveal interference with structures essential for cell survival, proliferation and/or function. Cell growth inhibition was assessed by using colorimetric assays, whereas cellular alterations after BHA exposure, were evaluated using conventional light and fluorescence microscopy. Low doses of BHA exerted a significant cytotoxic effect, associated with loss of mitochondrial function. As the concentration of BHA was increased, morphological alterations in critical subcellular targets such as lysosomes, mitochondria and actin cytoskeleton, were observed. In parallel, BHA induced an irreversible loss of cell proliferative capacity, preceding apoptosis induction. Thus, the dose-dependent activity of BHA on Vero cells appears to be cytotoxic as well as cytostatic. Our observations, although simplified with respect to the in vivo situations, allowed the assessment of the specific damage at the cellular level, and provide some clue about the effects of BHA in non-tumoral mammalian cells.

Cytotoxic effects in mammalian Vero cells exposed to pentachlorophenol.

The effects of pentachlorophenol have been studied on diverse biological systems both in vivo and in vitro, however the cellular basis of the pronounced cytotoxicity of this organochlorine compound is poorly understood. In this work, morphological and biochemical analyses were carried out to identify the primary targets of pentachlorophenol toxicity in mammalian cells. Our results show that pentachlorophenol is a very potent cytotoxic drug that displays an unusual and interesting mode of action in Vero cells. Although this compound is a powerful uncoupler of oxidative phosphorylation, we present the novel finding that lysosome destabilization is an early cytotoxic response that precedes the mitochondrial dysfunction. In addition, soon after exposure to moderate doses of pentachlorophenol, a significant number of cells initiate an apoptotic death process identified by the condensed and fragmented state of their nuclei. These results demonstrate that there are multiple potential targets of PCP-induced toxicity in mammalian cells, and the need to develop further experimental studies for the risk assessment of this environmental pollutant.

From hemorheology to vascular mechanobiology: An overview.

Almost all of the cells of the human body are subjected to mechanical stresses. In endothelial cells, mechanical stresses can vary from some milli-Pascal (shear stress) to one ore more Pascal (hydrostatic pressure). Now it is know that mechanical stresses have a decisive part cellular physiology. However, if the main biological effects of mechanical stress are well related, the mechanisms allowed the relation between mechanical stress to physiological phenomenon remain nearly unknown (mechanotransduction phenomenon). In this work, through personal results and published works, the authors considers all the effects of mechanical stresses and the possible hypothesis.

Ultrastructural changes induced in HeLa cells after phototoxic treatment with harmine.

In the present work, we have continued our studies on harmine phototoxicity in human tumour cells. The toxicity of harmine in the dark was analysed by a quantitative neutral red uptake assay, and subcellular sensitive targets following harmine photosensitization were de fi ned by electron microscopic analysis of HeLa cells. The results obtained indicated that this compound shows a clear dose-dependent cytotoxic effect in the dark. The combined treatment with suitable doses of harmine and UV radiation was very effective at an early stage, although maximal cell killing appeared 48 h after photodynamic activation. Ultrastructural examination of HeLa cells immediately after the photodynamic treatment revealed lysosomal destabilization and profound cytoplasmic vacuolization that evolved to cytolysis, which is typical of necrotic cell death. It is concluded that harmine could be a valuable photosensitizer whose biological applications merit further evaluation.

The strength of integrin binding between neutrophils and endothelial cells.

The firm adhesion of activated polymorphonuclear neutrophils to endothelial cells in blood vessels is achieved through binding of the integrin intercellular adhesion molecule. To contribute to the better understanding of this adhesion step, our investigation is aimed at the relationship between integrin expression and the strength of neutrophil binding to endothelial cells. Flow cytometry and 3D scanning microscopy are used to study integrin expression and distribution, respectively. It is found that CD11b/CD18 integrin expression is localized in clusters distributed irregularly over the neutrophil surface. After cell activation, the cluster distribution polarizes, increasing the local CD11b/CD18 density concurrently with nearly doubled integrin expression. The neutrophil adhesion efficiency is measured in a flow chamber coated successively by various substrates, including endothelial cells in an activated state. Analysis of the flow dependence of the number of attached cells reveals the prevailing number of neutrophils with stronger binding to the endothelium when both cells are in the activated state in comparison with non-activated cells.

[Leukocyte adhesion on a fibrinogen-coated surface under static conditions: experimentation and creation of a model]. / Adhésion des leucocytes sur une surface recouverte de fibrinogène en conditions statiques: expíence et modélisation]

The adhesion of polymorphonuclear leukocytes (PMNs) on the vascular endothelium is a complex process that occurs during different biological and pathological events and involves numerous molecules. The adhesion cascade is induced after PMN stimulation by various molecular or cellular signals. Fibrinogen is one of the substrates for CD11b/CD18 B2-integrins expressed at the PMN surface; fibrinogen-neutrophil binding is induced by inflammatory reactions. In order to understand this process, we have carried out studies on the basis of preliminary experiments on red blood cells and synthetic particles. The modelization of quiescent PMNs adhesion on a fibrinogen substrate was investigated with a sedimentation cell chamber. Two different physiological conditions were tested: the activated state of PMN by a synthetic pro-inflammatory activator (FMLP). The activated state of PMNs was both quantified by flow cytometry and controlled by fluorescence microscopy. The results suggest that quiescent neutrophils deposit in accordance with the ballistic deposition model. This random adsorption model differs from random sequential adsorption (RSA) in that the cells arriving at the surface are able to roll along cells previously adsorbed introducing the notion of gravitational attraction of cells. The preliminary results obtained with stimulated PMN do not allow to choose between one of this two deposition models. Nevertheless, the qualitative and quantitative effects of FMLP on neutrophils were demonstrated by modifications of adhesion molecules expression.

Hemorheology and vascular endothelial cells.

The vascular endothelium is a biologically active monolayer of cells provided an interface between blood and tissues. Vascular endothelial cells (ECs) have two functional states, which are allowed by their different properties: (i) vaso regulating properties: ECs releases vasomotor components, as endothelin (vaso constriction), prostacyclin and nitrite oxide (vaso dilatation); (ii) antithrombotic and hemostatic properties; and (iii) anti-adhesion properties. The endothelium is normally antithrombotic and anti-adhesive to ensure blood fluidity. During many cardio-vascular diseases, these properties may be reversed. Thus, the ECs have a determinant role in hemodynamic control through these various metabolic activities. Otherwise, many studies have demonstrated that local blood flow conditions have a crucial role on the EC properties (mechanotransduction concept). The knowledge of the properties of ECs and the control of the phenomena which define their functions is a key element in the cardiovascular diseases understanding.

Modelisation of leukocyte adhesion on a fibrinogen coated surface in static conditions.

The adhesion of polymorphonuclear leukocytes (PMNs) on the vascular endothelium is a complex process that occurs during biological and pathological events and involves a large family of molecules. This phenomenom could be approached by a modelisation study of the adhesion of PMNs on a biological substrate, fibrinogen. Two different physiological conditions were tested such as the activated state of PMNs with a synthetic pro-inflammatory activator (N-Formyl-Methionyl-Leucyl-Phenylalanine, FMLP). The activated state of PMNs was both quantified by flow cytometry and controlled by fluorescence microscopy. The results suggest that quiescent PMNs deposit in accordance with the ballistic deposition model. The preliminary results obtained with FMLP-stimulated PMNs show a different deposit process compared to quiescent PMNs but do not allow to determine exactly a deposition model.