The team's influence on physical and technical demands of elite goalkeepers in LaLiga: a longitudinal study in professional soccer.

This study examined physical and technical demands and the influence of the team's level on elite goalkeepers' performance during six consecutive seasons in Spanish Professional Soccer League. The goalkeepers' performance data were obtained by analyzing a total of 3,874 matches using a multiple-camera computerized tracking system. The physical and technical match variables registered were: distance traveled; distance Sprinted and the number of sprints; total number of passes; successful passes; pass percentage; recovered balls; lost balls; ratio lost balls: recovered balls, and number of saves. The results showed that the number of saves made has shown a significant reduction (p < 0.001). When comparing between the teams' level, the goalkeepers of the worst classified teams showed a greater distance traveled by sprint (+3.72 m, IC95%: 1.00-6.44, ES: 0.41, p = 0.008). In conclusion, the results the influence of the team's level on the technical and physical parameters of the goalkeepers during the last six seasons.

Fenestrations and preferential flow routes in the prelaminar optic nerve through wet scanning electron microscope and perfusion of tracers.

PURPOSE: We study the vitreous interface of the optic disc to delimit the passages for the flow of fluids through the prelaminar tissue of porcine eyes. METHODS: Wet scanning electron microscope (SEM), conventional SEM and transmission electron microscope (TEM) were used to explore the surface of the optic nerve of the pig. The vitreous cavity was perfused with a fluorescent marker and colloidal gold at controlled pressure. Samples of perfused optic nerve head were cryosectioned and observed with the confocal laser microscope (lectin) or resin embedded and observed under TEM (gold). RESULTS: Fenestrations were present under the SEM in all three regions of the vitreous interface. SEM results were confirmed at the TEM level and under the wet-SEM. Perfusion experiments traced the flow of a fluorescent molecule delineating routes of preferential flow with origin in the fenestrations. Colloidal gold marked the site of entrance in the prelaminar tissue identifying major fenestrations in the basal membrane. CONCLUSIONS: Interchange of fluid between the optic nerve and the vitreous cavity in the pig is facilitated by fenestrations of varied sizes in the basal membrane and preferential flow routes through the prelaminar tissue. Preferential flow routes exist in the extracellular spaces of Elschnig and Kuhn' astrocytes and give a sharply distinct image when compared with flow through zones in which astrocytes envelope axons. Escape routes may be instrumental in preventing oedemas of the optic nerve head, but they could also serve as entrance doors for fluids from the vitreous and aqueous and play a pathogenic role in ageing and glaucoma.

Mapping the surface astrocytes of the optic disc: a fluid-conducting role of the astrocytic covering of the central vessels.

BACKGROUND: The vitreous interface of the optic nerve has been studied to delimit the covering of Elschnig's astrocytes and interstitial pathways of flow through the prelaminar region. METHODS: Perfusion of the prelaminar tissue under controlled pressure with a fluorescent marker injected into the vitreous cavity in pig eyes. The prelaminar region of the optic nerve and adjacent retina was fixed and flatmounted or frozen and cryosectioned and examined with the confocal laser microscope. Samples were also prepared for conventional transmission electron microscopy. RESULTS: The surface of the vitreous surface of the optic nerve is covered by a cobblestone-like pavement made of astrocytic projections. Intensely stained passages of different thickness indicate the presence of wide interconnected intercellular spaces in the covering of Elschnig's astrocytes. Those passages are absent in the intervascular areas occupied by axons and axon-linked astrocytes. CONCLUSIONS: Delineation of the astrocytic pavement and the preferred flow routes formed by wide extracellular spaces are conspicuous features of the prelaminar region when examined with the confocal laser microscope and the help of sticky fluorescent tracer. This suggests that excess extracellular fluid can be interchanged with the vitreous by a network of interconnected extracellular spaces or preferred flow routes. Some pathogenic mechanisms can be related to fluid interchange in the optic nerve head.

Cell-cell adhesion in the prelaminar region of the optic nerve head: a possible target for ionic stress.

PURPOSE: This paper aims to study the anterior surface of the optic nerve in relation to its ability to support a source of stress acting from the vitreous cavity. The intercellular junctions of the lining astrocytes mediated by cellular adhesion molecules (CAMs) may be the main targets for ionic stress. METHODS: The optic nerve of the domestic pig was prepared for light, confocal laser and transmission electron microscopy. Immunostaining was performed for antibodies against glial fibrillary acidic protein, neural cadherin (N-cadherin) and neural CAM (N-CAM). RESULTS: Only 1 type of intercellular junction was found among the bordering astrocytes, which was characterized as a zonula adherens. Unions between lining cells showed a positive immunogold effect and immunofluorescence against N-cadherin in the zonula adherens and membrane apposition. N-CAM was also present in areas of nonjunctional cellular adhesion. CONCLUSION: The stability of intercellular junctions of the nerve-vitreous boundary is sensitive to altered concentrations of Ca(2+). Since aqueous humor has half the Ca(2+) concentration of plasma, any contact of aqueous humor with the optic nerve head can interfere with the ionic concentration of calcium in the extracellular spaces. This mechanism may contribute to age-related changes and some types of glaucoma.

Electron microscopy reveals the presence of viruses in mouse embryonic fibroblasts but neither in human embryonic fibroblasts nor in human mesenchymal cells used for hESC maintenance: toward an implementation of microbiological quality assurance program in stem cell banks.

Human embryonic stem cells (hESCs) are expected to open up new avenues in regenerative medicine by allowing the generation of transplantable cells to be used in future cell replacement therapies. Maintenance of hESCs in the presence of xenogenic compounds is likely to prevent their use in future therapeutic applications in humans. Recently, it has been claimed that human foreskin-derived human embryonic fibroblast (HEFs) and human adult marrow cells have the ability to support prolonged expansion of hESCs in culture similar to murine feeders. Here, to minimize the use of xenogenic components for hESC maintenance, we performed transmission electron microscopy-based microbiological studies in an attempt to implement a microbiological Quality Assurance Program in Stem Cell Banks by determining the potential presence of viral particles in MEFs compared with human HEFs and bone marrow-derived mesenchymal cells. We observed in three out of nine MEF samples (33.3%) viruses belonging to the Retroviridae family. Within the Retroviridae family, these viruses have a C morphology, which indicates they belong to the subfamily Orthoretroviridae. In contrast, no viral particles could be observed in either the HEF samples (n = 5) or the human BM-derived mesenchymal cells (n = 9) analyzed. Based on these experimental microbiological data, we recommend the implementation of microbiological Quality Assurance Programs by means of transmission electron microscopy as a routine technique to assess the potential presence of viral particles in any feeder cell used in stem cell banks and support the use of human cells rather than murine cells as feeders to maintain hESC cultures in an undifferentiated state.

Expression of glucose transporters in the prelaminar region of the optic-nerve head of the pig as determined by immunolabeling and tissue culture.

BACKGROUND: To develop the use of cultured tissue of the prelaminar optic nerve of the pig to explore possible alterations of the astrocyte-axon metabolic pathways in glaucoma, we map the distribution of the glucose transporters GLUT1 and GLUT3 in fresh and cultured tissue. METHODS: We monitor cell survival in cultures of the prelaminar optic-nerve tissue, measuring necrosis and apoptosis markers biochemically as well as morphologically, and establish the presence of the glucose transporters GLUT1 and GLUT3. We map the distribution of these transporters with immunolabeling in histological sections of the optic nerve using confocal and electronic transmission microscopy. RESULTS: We find that the main death type in prelaminar culture is apoptosis. Caspase 7 staining reveals an increment in apoptosis from day 1 to day 4 and a reduction from day 4 to day 8. Western blotting for GLUT1 shows stability with increased culture time. CLSM micrographs locate GLUT1 in the columnar astrocytes and in the area of axonal bundles. Anti-GLUT3 predominantly labels axonal bundles. TEM immunolabeling with colloidal gold displays a very specific distribution of GLUT-1 in the membranes of vascular endothelial cells and in periaxonal astrocyte expansions. The GLUT-3 isoform is observed with TEM only in axons in the axonal bundles. CONCLUSIONS: Tissue culture is suitable for apoptosis-induction experiments. The results suggest that glucose is transported to the axonal cleft intracytoplasmically and delivered to the cleft by GLUT1 transporters. As monocarboxylate transporters have been reported in the prelaminar region of the optic-nerve head, this area is likely to use both lactate and glucose as energy sources.