A selective defect in the glial wedge as part of the neuroepithelium disruption in hydrocephalus development in the mouse hyh model is associated with complete corpus callosum dysgenesis.

Introduction: Dysgenesis of the corpus callosum is present in neurodevelopmental disorders and coexists with hydrocephalus in several human congenital syndromes. The mechanisms that underlie the etiology of congenital hydrocephalus and agenesis of the corpus callosum when they coappear during neurodevelopment persist unclear. In this work, the mechanistic relationship between both disorders is investigated in the hyh mouse model for congenital hydrocephalus, which also develops agenesis of the corpus callosum. In this model, hydrocephalus is generated by a defective program in the development of neuroepithelium during its differentiation into radial glial cells. Methods: In this work, the populations implicated in the development of the corpus callosum (callosal neurons, pioneering axons, glial wedge cells, subcallosal sling and indusium griseum glial cells) were studied in wild-type and hyh mutant mice. Immunohistochemistry, mRNA in situ hybridization, axonal tracing experiments, and organotypic cultures from normal and hyh mouse embryos were used. Results: Our results show that the defective program in the neuroepithelium/radial glial cell development in the hyh mutant mouse selectively affects the glial wedge cells. The glial wedge cells are necessary to guide the pioneering axons as they approach the corticoseptal boundary. Our results show that the pioneering callosal axons arising from neurons in the cingulate cortex can extend projections to the interhemispheric midline in normal and hyh mice. However, pioneering axons in the hyh mutant mouse, when approaching the area corresponding to the damaged glial wedge cell population, turned toward the ipsilateral lateral ventricle. This defect occurred before the appearance of ventriculomegaly. Discussion: In conclusion, the abnormal development of the ventricular zone, which appears to be inherent to the etiology of several forms of congenital hydrocephalus, can explain, in some cases, the common association between hydrocephalus and corpus callosum dysgenesis. These results imply that further studies may be needed to understand the corpus callosum dysgenesis etiology when it concurs with hydrocephalus.

Therapeutic strategies to recover ependymal barrier after inflammatory damage: relevance for recovering neurogenesis during development.

The epithelium covering the surfaces of the cerebral ventricular system is known as the ependyma, and is essential for maintaining the physical and functional integrity of the central nervous system. Additionally, the ependyma plays an essential role in neurogenesis, neuroinflammatory modulation and neurodegenerative diseases. Ependyma barrier is severely affected by perinatal hemorrhages and infections that cross the blood brain barrier. The recovery and regeneration of ependyma after damage are key to stabilizing neuroinflammatory and neurodegenerative processes that are critical during early postnatal ages. Unfortunately, there are no effective therapies to regenerate this tissue in human patients. Here, the roles of the ependymal barrier in the context of neurogenesis and homeostasis are reviewed, and future research lines for development of actual therapeutic strategies are discussed.

Microbiome Alterations and Alzheimer's Disease: Modeling Strategies with Transgenic Mice.

In the last decade, the role of the microbiota-gut-brain axis has been gaining momentum in the context of many neurodegenerative and metabolic disorders, including Alzheimer's disease (AD) and diabetes, respectively. Notably, a balanced gut microbiota contributes to the epithelial intestinal barrier maintenance, modulates the host immune system, and releases neurotransmitters and/or neuroprotective short-chain fatty acids. However, dysbiosis may provoke immune dysregulation, impacting neuroinflammation through peripheral-central immune communication. Moreover, lipopolysaccharide or detrimental microbial end-products can cross the blood-brain barrier and induce or at least potentiate the neuropathological progression of AD. Thus, after repeated failure to find a cure for this dementia, a necessary paradigmatic shift towards considering AD as a systemic disorder has occurred. Here, we present an overview of the use of germ-free and/or transgenic animal models as valid tools to unravel the connection between dysbiosis, metabolic diseases, and AD, and to investigate novel therapeutical targets. Given the high impact of dietary habits, not only on the microbiota but also on other well-established AD risk factors such as diabetes or obesity, consistent changes of lifestyle along with microbiome-based therapies should be considered as complementary approaches.

Generation of Periventricular Reactive Astrocytes Overexpressing Aquaporin 4 Is Stimulated by Mesenchymal Stem Cell Therapy.

Aquaporin-4 (AQP4) plays a crucial role in brain water circulation and is considered a therapeutic target in hydrocephalus. Congenital hydrocephalus is associated with a reaction of astrocytes in the periventricular white matter both in experimental models and human cases. A previous report showed that bone marrow-derived mesenchymal stem cells (BM-MSCs) transplanted into the lateral ventricles of hyh mice exhibiting severe congenital hydrocephalus are attracted by the periventricular astrocyte reaction, and the cerebral tissue displays recovery. The present investigation aimed to test the effect of BM-MSC treatment on astrocyte reaction formation. BM-MSCs were injected into the lateral ventricles of four-day-old hyh mice, and the periventricular reaction was detected two weeks later. A protein expression analysis of the cerebral tissue differentiated the BM-MSC-treated mice from the controls and revealed effects on neural development. In in vivo and in vitro experiments, BM-MSCs stimulated the generation of periventricular reactive astrocytes overexpressing AQP4 and its regulatory protein kinase D-interacting substrate of 220 kDa (Kidins220). In the cerebral tissue, mRNA overexpression of nerve growth factor (NGF), vascular endothelial growth factor (VEGF), hypoxia-inducible factor-1 (HIF1α), and transforming growth factor beta 1 (TGFß1) could be related to the regulation of the astrocyte reaction and AQP4 expression. In conclusion, BM-MSC treatment in hydrocephalus can stimulate a key developmental process such as the periventricular astrocyte reaction, where AQP4 overexpression could be implicated in tissue recovery.

Does Small-Scale Livestock Production Use a High Technological Level to Survive? Evidence from Dairy Production in Northeast-ern Michoacán, Mexico.

The objective of this study was to identify the technological level used by dairy farmers in the northeastern region of Michoacán, Mexico, through a characterisation of small-scale dairy production units, as well as to learn about the socioeconomic conditions that have enabled them to survive in the current context. A semi-structured interview was applied to 114 production units, chosen by stratified random sampling. The interview included technological, production and socioeconomic aspects. Twenty-eight variables were initially explored and 12 were used for multivariate analysis, which included Principal Component Analysis, Hierarchical Cluster Analysis and K-means Cluster. The characterisation carried out showed that the production units that predominate in northeastern Michoacán have survived with a low technological level, having as strengths the diversification of their activities and the use of family labour. On the contrary, production units with a high technological level and high productivity are few and less diversified. This shows the need to generate differentiated public policies for each cluster, aimed at strengthening the aspects that have allowed them to survive and guaranteeing a market for their production, before promoting the use of technologies.

Increased levels of tumour necrosis factor alpha (TNFα) but not transforming growth factor-beta 1 (TGFß1) are associated with the severity of congenital hydrocephalus in the hyh mouse.

AIMS: Here, we tested the hypothesis that glial responses via the production of cytokines such as transforming growth factor-beta 1 (TGFß1) and tumour necrosis factor alpha (TNFα), which play important roles in neurodegenerative diseases, are correlated with the severity of congenital hydrocephalus in the hyh mouse model. We also searched for evidence of this association in human cases of primary hydrocephalus. METHODS: Hyh mice, which exhibit either severe or compensated long-lasting forms of hydrocephalus, were examined and compared with wild-type mice. TGFß1, TNFα and TNFαR1 mRNA levels were quantified using real-time PCR. TNFα and TNFαR1 were immunolocalized in the brain tissues of hyh mice and four hydrocephalic human foetuses relative to astroglial and microglial reactions. RESULTS: The TGFß1 mRNA levels were not significantly different between hyh mice exhibiting severe or compensated hydrocephalus and normal mice. In contrast, severely hydrocephalic mice exhibited four- and two-fold increases in the mean levels of TNFα and TNFαR1, respectively, compared with normal mice. In the hyh mouse, TNFα and TNFαR1 immunoreactivity was preferentially detected in astrocytes that form a particular periventricular reaction characteristic of hydrocephalus. However, these proteins were rarely detected in microglia, which did not appear to be activated. TNFα immunoreactivity was also detected in the glial reaction in the small group of human foetuses exhibiting hydrocephalus that were examined. CONCLUSIONS: In the hyh mouse model of congenital hydrocephalus, TNFα and TNFαR1 appear to be associated with the severity of the disease, probably mediating the astrocyte reaction, neurodegenerative processes and ischaemia.

Alignment of the measurement scale mark during immersion hydrometer calibration using an image processing system.

The present work presents an improved method to align the measurement scale mark in an immersion hydrometer calibration system of CENAM, the National Metrology Institute (NMI) of Mexico, The proposed method uses a vision system to align the scale mark of the hydrometer to the surface of the liquid where it is immersed by implementing image processing algorithms. This approach reduces the variability in the apparent mass determination during the hydrostatic weighing in the calibration process, therefore decreasing the relative uncertainty of calibration.

Comparación de las pruebas de permutación y asintóticas aplicadas en tablas de contingencia de dimensión rxc no ordenadas utilizadas en estudios biomédicos / Comparison between permutation and asintotics tests applied to unordered rxc dimension contingency tables used in biomedical surveys

Se realizó un estudio comparativo entre pruebas de permutación y asintóticas, aplicadas a tablas de contingencia de dimensión R×C no ordenadas, utilizando como medida de comparación la diferencia entre el p-valor exacto y asintótico. Se analizaron cinco (05) ejemplos que presentan tablas de contingencia no ordenadas, publicados en la literatura científica internacional relacionados con estudios biomédicos, con el objeto de mostrar bajo cuales condiciones ambos enfoques difieren o convergen para las pruebas de independencia de Pearson, Razón de Verosimilitud y Freeman-Halton. Los resultados mostraron que el comportamiento de las metodologías exacta y asintótica depende del tamaño de muestra, dimensión, balanceo y dispersión de la tabla de contingencia y prueba aplicada. Para los casos estudiados se encontró que los p-valores exactos y asintóticos presentaron diferencias notables para tamaños de muestras pequeños; sobre todo en tablas de contingencia desbalanceadas y dispersas; y mostraron convergencia de los p-valores asintóticos a los exactos en la medida que el tamaño de muestra y dimensión de la tabla era mayor.

A comparative study between permutation and asymptotic tests applied to unordered R×C dimension contingency tables was carried out, using the difference between the exact p-value and the asymptotic one as comparison measurement. Five (05) biomedical research-paper results based on unordered contingency tables were examined from international scientific literature, analyzing how different or equivalent they appear using Pearson, Likelihood ratio and Freeman-Halton independency tests. Results revealed that both methodologies, exact and asymptotic, behave depending on sample size, dimensions, balance and dispersion of the contingency table, as well as on the test applied. The exact and asymptotic p-values showed striking differences for small sample sizes mainly in unbalanced and sparse contingency tables, but they converged as the sample size and table dimensions increased.

Astrocytes acquire morphological and functional characteristics of ependymal cells following disruption of ependyma in hydrocephalus.

Hydrocephalic hyh mutant mice undergo a programmed loss of the neuroepithelium/ependyma followed by a reaction of periventricular astrocytes, which form a new cell layer covering the denuded ventricular surface. We present a comparative morphological and functional study of the newly formed layer of astrocytes and the multiciliated ependyma of hyh mice. Transmission electron microscopy, immunocytochemistry for junction proteins (N-cadherin, connexin 43) and proteins involved in permeability (aquaporin 4) and endocytosis (caveolin-1, EEA1) were used. Horseradish peroxidase (HRP) and lanthanum nitrate were used to trace the intracellular and paracellular transport routes. The astrocyte layer shares several cytological features with the normal multiciliated ependyma, such as numerous microvilli projected into the ventricle, extensive cell-cell interdigitations and connexin 43-based gap junctions, suggesting that these astrocytes are coupled to play an unknown function as a cell layer. The ependyma and the astrocyte layers also share transport properties: (1) high expression of aquaporin 4, caveolin-1 and the endosome marker EEA1; (2) internalization into endocytic vesicles and early endosomes of HRP injected into the ventricle; (3) and a similar paracellular route of molecules moving between CSF, the subependymal neuropile and the pericapillary space, as shown by lanthanum nitrate and HRP. A parallel analysis performed in human hydrocephalic foetuses indicated that a similar phenomenon would occur in humans. We suggest that in foetal-onset hydrocephalus, the astrocyte assembly at the denuded ventricular walls functions as a CSF-brain barrier involved in water and solute transport, thus contributing to re-establish lost functions at the brain parenchyma-CSF interphase.

New ependymal cells are born postnatally in two discrete regions of the mouse brain and support ventricular enlargement in hydrocephalus.

A heterogeneous population of ependymal cells lines the brain ventricles. The evidence about the origin and birth dates of these cell populations is scarce. Furthermore, the possibility that mature ependymal cells are born (ependymogenesis) or self-renewed (ependymal proliferation) postnatally is controversial. The present study was designed to investigate both phenomena in wild-type (wt) and hydrocephalic α-SNAP mutant (hyh) mice at different postnatal stages. In wt mice, proliferating cells in the ventricular zone (VZ) were only found in two distinct regions: the dorsal walls of the third ventricle and Sylvian aqueduct (SA). Most proliferating cells were monociliated and nestin+, likely corresponding to radial glial cells. Postnatal cumulative BrdU-labeling showed that most daughter cells remained in the VZ of both regions and they lost nestin-immunoreactivity. Furthermore, some labeled cells became multiciliated and GLUT-1+, indicating they were ependymal cells born postnatally. Postnatal pulse BrdU-labeling and Ki-67 immunostaining further demonstrated the presence of cycling multiciliated ependymal cells. In hydrocephalic mutants, the dorsal walls of the third ventricle and SA expanded enormously and showed neither ependymal disruption nor ventriculostomies. This phenomenon was sustained by an increased ependymogenesis. Consequently, in addition to the physical and geometrical mechanisms traditionally explaining ventricular enlargement in fetal-onset hydrocephalus, we propose that postnatal ependymogenesis could also play a role. Furthermore, as generation of new ependymal cells during postnatal stages was observed in distinct regions of the ventricular walls, such as the roof of the third ventricle, it may be a key mechanism involved in the development of human type 1 interhemispheric cysts.

Hollow porous implants filled with mesoporous silica particles as a two-stage antibiotic-eluting device.

A new type of implantable drug eluting device is presented, consisting of a bed of mesoporous microparticles packed inside a reservoir with a porous wall. This provides two sets of variables for drug release control that can be tailored independently. The first is related to the microparticles (packing density, size and pore structure) and the second to the reservoir (pore diameter and thickness of the wall, permeation area). In this work the concept is developed into a working model, used to fight bacterial (Staphylococcus aureus) growth by releasing linezolid that had previously been adsorbed on silica microparticles. These particles were placed inside the hollow interior of a porous medical grade stainless steel pin mimicking those used in traumatology and in orthopedic surgery. The mechanical behavior of the porous drug-eluting pin was tested and found satisfactory.

Disruption of the neurogenic niche in the subventricular zone of postnatal hydrocephalic hyh mice.

Neural stem cells persist after embryonic development in the subventricular zone (SVZ) niche and produce new neural cells during postnatal life; ependymal cells are a key component associated with this neurogenic niche. In the animal model of human hydrocephalus, the hyh mouse, the ependyma of the lateral ventricles is progressively lost during late embryonic and early postnatal life and disappears from most of the ventricular surface throughout its life span. To determine the potential consequences of this loss on the SVZ, we characterized the abnormalities in this neurogenic niche in hyh mice. There was overall disorganization and a marked reduction of proliferative cells in the SVZ of both newborn and adult hyh hydrocephalic mice in vivo; neuroblasts were displaced to the ventricular surface, and their migration through the rostral migratory stream was reduced. The numbers of resident neural progenitor cells in hyh mice were also markedly reduced, but they were capable of proliferating, forming neurospheres, and differentiating into neurons and glia in vitro in a manner indistinguishable from that of wild-type progenitor cells. These findings suggest that the reduction of proliferative activity observed in vivo is not caused by a cell autonomous defect of SVZ progenitors but is a consequence of a reduced number of these cells. Furthermore, the overall tissue disorganization of the SVZ and displacement of neuroblasts imply alterations in the neurogenic niche of postnatal hyh mice.

Effect of Nitinol surface treatments on its physico-chemical properties.

The main purpose of this work is the study of different physicochemical treatments on Nitinol slabs and wires, with the aim of inducing the formation of a TiO(2) surface film capable of increasing the corrosion resistance of the material and of reducing the release of Ni when the Nitinol samples were immersed in simulated body fluid (SBF). To this end, a battery of measurements (surface roughness, contact angle, electrochemical corrosion, chemical analysis as a function of depth, and Ni release to SBF) has been used to characterize Nitinol commercial samples, as received, and also after the different treatments performed. The results clearly indicate the effectiveness of the passivation TiO(2) layer as a barrier against Ni leaching, and the detrimental effects of any processes (such as polishing or cutting) that result in exposure of areas not coated by the TiO(2) film. Chemical methods such as oxidation in nitric acid or hydrothermal treatment of the samples (by prolonged immersion in boiling water) seem to provide simple and efficient ways of forming TiO(2) films of adequate thickness on the Nitinol surface.

Patterned neuropathologic events occurring in hyh congenital hydrocephalic mutant mice.

Hyh mutant mice develop long-lasting hydrocephalus and represent a good model for investigating neuropathologic events associated with hydrocephalus. The study of their brains by use of lectin binding, bromodeoxyuridine labeling, immunochemistry, and scanning electron microscopy revealed that certain events related to hydrocephalus followed a well-defined pattern. A program of neuroepithelium/ependyma denudation was initiated at embryonic day 12 and terminated at the end of the second postnatal week. After the third postnatal week the denuded areas remained permanently devoid of ependyma. In contrast, a selective group of ependymal areas resisted denudation throughout the lifespan. Ependymal denudation triggered neighboring astrocytes to proliferate. These astrocytes expressed particular glial markers and formed a superficial cell layer replacing the lost ependyma. The loss of the neuroepithelium/ependyma layer at specific regions of the ventricular walls and at specific stages of brain development would explain the fact that only certain brain structures had abnormal development. Therefore, commissural axons forming the corpus callosum and the hippocampal commissure displayed abnormalities, whereas those forming the anterior and posterior commissures did not; and the brain cortex was not homogenously affected, with the cingular and frontal cortices being the most altered regions. All of these telencephalic alterations developed at stages when hydrocephalus was not yet patent at the lateral ventricles, indicating that abnormal neural development and hydrocephalus are linked at the etiologic level, rather than the former being a consequence of the latter. All evidence collected on hydrocephalic hyh mutant mice indicates that a primary alteration in the neuroepithelium/ependyma cell lineage triggers both hydrocephalus and abnormalities in telencephalic development.