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Summary: Low-density cell culture of the postnatal cerebellum, combined with live imaging and single-cell tracking, allows the behavior of postnatal cerebellar neural stem cells (NSCs) and their progeny to be monitored. Cultured cerebellar NSCs maintain their neurogenic nature giving rise, in the same relative proportions that exist in vivo, to the neuronal progeny generated by the three postnatal cerebellar neurogenic niches. This protocol describes the identification of the nature of the progeny through both post-imaging immunocytochemistry and patch-clamp recordings.For complete details on the use and execution of this protocol, please refer to Paniagua-Herranz et al. (2020b).

Background: Evidence suggests that earlier diagnosis and initiation of treatment immediately after birth is critical for improved neurodevelopmental outcomes following neonatal encephalopathy (NE). Current diagnostic tests are, however, mainly restricted to clinical diagnosis with no molecular tests available. Purines including adenosine are released during brain injury such as hypoxia and are also present in biofluids. Whether blood purine changes can be used to diagnose NE has not been investigated to date.Methods: Blood purines were measured in a mouse model of neonatal hypoxia and infants with NE using a novel point-of-care diagnostic technology (SMARTChip) based on the summated electrochemical detection of adenosine and adenosine metabolites in the blood.Results: Blood purine concentrations were ∼2–3-fold elevated following hypoxia in mice [2.77 ± 0.48 μM (Control) vs. 7.57 ± 1.41 μM (post-hypoxia), p = 0.029]. Data in infants with NE had a 2–3-fold elevation when compared to healthy controls [1.63 ± 0.47 μM (Control, N = 5) vs. 4.87 ± 0.92 μM (NE, N = 21), p = 0.0155]. ROC curve analysis demonstrates a high sensitivity (81%) and specificity (80%) for our approach to identify infants with NE. Moreover, blood purine concentrations were higher in infants with NE and seizures [8.13 ± 3.23 μM (with seizures, N = 5) vs. 3.86 ± 0.56 μM (without seizures, N = 16), p = 0.044].Conclusion: Our data provides the proof-of-concept that measurement of blood purine concentrations via SMARTChip technology may offer a low-volume bedside test to support a rapid diagnosis of NE.

Circulating molecules have potential as biomarkers to support the diagnosis of epilepsy and to assist with differential diagnosis, for example, in conditions resembling epilepsy, such as in psychogenic non-epileptic seizures (PNES). The P2X7 receptor (P2X7R) is an important regulator of inflammation and mounting evidence supports its activation in the brain during epilepsy. Whether the P2X7R or P2X7R-dependent signaling molecules can be used as biomarkers of epilepsy has not been reported. P2X7R levels were analyzed by quantitative ELISA using plasma samples from controls and patients with temporal lobe epilepsy (TLE) or PNES. Moreover, blood cell P2X7R expression and P2X7R-dependent cytokine signature was measured following status epilepticus in P2X7R-EGFP reporter, wildtype, and P2X7R-knockout mice. P2X7R plasma levels were higher in TLE patients when compared with controls and patients with PNES. Plasma levels of the broad inflammatory marker protein C-Reactive protein (CRP) were similar between the three groups. Using P2X7R-EGFP reporter mice, we identified monocytes as the main blood cell type expressing P2X7R after experimentally evoked seizures. Finally, cytokine array analysis in P2X7R-deficient mice identified KC/GRO as a potential P2X7R-dependent plasma biomarker following status epilepticus and during epilepsy. Our data suggest that P2X7R signaling components may be a promising subclass of circulating biomarkers to support the diagnosis of epilepsy.

Vesicular storage of neurotransmitters, which allows their subsequent exocytotic release, is essential for chemical transmission in the central nervous system. Neurotransmitter uptake into secretory vesicles is carried out by vesicular transporters, which use the electrochemical proton gradient generated by a vacuolar H+-ATPase to drive neurotransmitter vesicular accumulation. ATP and other nucleotides are relevant extracellular signaling molecules that participate in a variety of biological processes. Although the active transport of nucleotides into secretory vesicles has been characterized from the pharmacological and biochemical point of view, the protein responsible for such vesicular accumulation remained unidentified for some time. In 2008, the human SLC17A9 gene, the last identified member of the SLC17 transporters, was found to encode the vesicular nucleotide transporter (VNUT). VNUT is expressed in various ATP-secreting cells and is able to transport a wide variety of nucleotides in a vesicular membrane potential-dependent manner. VNUT knockout mice lack vesicular storage and release of ATP, resulting in blockage of the purinergic transmission. This review summarizes the current studies on VNUT and analyzes the physiological relevance of the vesicular nucleotide transport in the central nervous system. The possible role of VNUT in the development of some pathological processes, such as chronic neuropathic pain or glaucoma is also discussed. The putative involvement of VNUT in these pathologies raises the possibility of the use of VNUT inhibitors for therapeutic purposes.

Calcium sulfate (CaSO4) is one of the most common evaporites found in the earth’s crust. It can be found as four main variations: gypsum (CaSO4∙2H2O), bassanite (CaSO4∙0.5H2O), soluble anhydrite, and insoluble anhydrite (CaSO4), being the key difference the hydration state of the sulfate mineral. Naica giant crystals’ growth starts from a supersaturated solution in a delicate thermodynamic balance close to equilibrium, where gypsum can form nanocrystals able to grow up to 11–12 m long. The growth rates are reported to be as slow as (1.4 ± 0.2) × 10−5 nm/s, taking thousands of years to form crystals with a unique smoothness and diaphaneity, which may or may not include solid or liquid inclusions. Conservation efforts can be traced back to other gypsum structures found prior to Naica’s. Furthermore, in the last two decades, several authors have explored the unique requirements in which these crystals grow, the characterization of their environment and microclimatic conditions, and the prediction of deterioration scenarios. We present a state-of-the-art review on the mentioned topics. Beyond the findings on the origin, in this work we present the current state and the foreseeable future of these astounding crystals.

Neonatal seizures are one of the most common comorbidities of neonatal encephalopathy, with seizures aggravating acute injury and clinical outcomes. Current treatment can control early life seizures; however, a high level of pharmacoresistance remains among infants, with increasing evidence suggesting current anti-seizure medication potentiating brain damage. This emphasises the need to develop safer therapeutic strategies with a different mechanism of action. The purinergic system, characterised by the use of adenosine triphosphate and its metabolites as signalling molecules, consists of the membrane-bound P1 and P2 purinoreceptors and proteins to modulate extracellular purine nucleotides and nucleoside levels. Targeting this system is proving successful at treating many disorders and diseases of the central nervous system, including epilepsy. Mounting evidence demonstrates that drugs targeting the purinergic system provide both convulsive and anticonvulsive effects. With components of the purinergic signalling system being widely expressed during brain development, emerging evidence suggests that purinergic signalling contributes to neonatal seizures. In this review, we first provide an overview on neonatal seizure pathology and purinergic signalling during brain development. We then describe in detail recent evidence demonstrating a role for purinergic signalling during neonatal seizures and discuss possible purine-based avenues for seizure suppression in neonates.

Adenosine triphosphate (ATP) is an important extracellular neurotransmitter that participates in several critical processes like cell differentiation, neuroprotection or axon guidance. Prior to its exocytosis, ATP must be stored in secretory vesicles, a process that is mediated by the Vesicular Nucleotide Transporter (VNUT). This transporter has been identified as the product of the SLC17A9 gene and it is prominently expressed in discrete brain areas, including the cerebellum. The main population of cerebellar neurons, the glutamatergic granule neurons, depends on purinergic signaling to trigger neuroprotective responses. However, while nucleotide receptors like P2X7 and P2Y13 are known to be involved in neuroprotection, the mechanisms that regulate ATP release in relation to such events are less clearly understood. In this work, we demonstrate that cerebellar granule cells express a functional VNUT that is involved in the regulation of ATP exocytosis. Numerous vesicles loaded with this nucleotide can be detected in these granule cells and are staining by the fluorescent ATP-marker, quinacrine. High potassium stimulation reduces quinacrine fluorescence in granule cells, indicating they release ATP via calcium dependent exocytosis. Specific subcellular markers were used to assess the localization of VNUT in granule cells, and the transporter was detected in both the axonal and somatodendritic compartments, most predominantly in the latter. However, co-localization with the specific lysosomal marker LAMP-1 indicated that VNUT can also be found in non-synaptic vesicles, such as lysosomes. Interestingly, the weak co-localization between VNUT and VGLUT1 suggests that the ATP and glutamate vesicle pools are segregated, as also observed in the cerebellar cortex. During post-natal cerebellar development, VNUT is found in granule cell precursors, co-localizing with markers of immature cells like doublecortin, suggesting that this transporter may be implicated in the initial stages of granule cell development.

El estudio consiste en relacionar el agente causal de las deformaciones cefálicas con el área cultural, el cierre sutural y las formas craneanas en el contexto del crecimiento craneofacial del poblador peruano precolombino, comprendidos en la clasificación tipológica de Weiss. Muestra: dieciocho cráneos representativos de grupos étnicos precolombinos del antiguo Perú comprendido dentro de la clasificación tipológica de Weiss, seleccionados de la curaduría de restos humanos del Museo Nacional de Arqueología, Antropología e Historia del Perú. Metodología: Se procedió con el reconocimiento de los cráneos y el registro de los códigos respectivos adoptada por la curaduría según las asociaciones culturales, regionales determinadas. Luego los cráneos fueron fotografiados en todas sus normas anatómicas. La información obtenida fue a través de la observación de imágenes de alta resolución. Resultados y conclusiones: Las deformaciones cefálicas son congruentes con la ley de Virchow. En el Perú precolombino se resume dos filums, el Llautu asociado a culturas andinas y la Cuna a pueblos costeños. El Llautu es concordante con el cierre precoz de la sutura sagital y en consecuencia el cerebro como materia orgánica influye en el bregma y la sutura coronal estimulando la osificación de un cráneo dolicocéfalo de aspecto anular. La cuna es concordante con el cierre precoz de la sutura coronal y en consecuencia el cerebro en crecimiento estimula las membranas suturales: sagital, parietoesfenoidal y parietotemporal orientando la osificación de un cráneo braquicéfalo de aspecto tabular.

El objetivo del presente estudio fue comparar mediante radiografías cefalométricas, una muestra de adolescentes mestizas con las medidas estandarizadas de caucásicos de Ricketts, utilizado como patrón y determinar si hay diferencias significativas entre los valores encontrados. La población se basó en mujeres adolescentes del nivel secundario del colegio Rosa de Santa María del distrito de Breña, entre 11 a 16 años de edad. De la cual se escogió una muestra por conveniencia de 32 alumnas, con características del mestizaje peruano según patrones antropológicos. Se consideraron características extraorales e intraorales para la selección de la muestra, se utilizó el equipo de radiografía cefalométrico y una cámara fotográfica SRL para obtener un registro fotográfico de las características faciales. Los resultados fueron procesados usando el paquete estadístico SPSS v.17, expresando los resultados bajo medidas descriptivas y utilizando la prueba estadística Z para encontrar diferencias al 95% de confianza. Se concluyó que estas diferencias se hacen más evidentes en los siguientes valores: línea estética “E” 0,5 mm, ángulo ínter incisivo 124°, punto “A” a la línea N-Pg hasta en 4 mm, longitud anterior de la base craneal 56 mm, lo cual confirma la hipótesis que sí hay diferencias significativas entre la muestra de mestizas peruanas con las del grupo caucásico de Ricketts.

En el presente articulo se desarrolla el procedimiento de diagnóstico y tratamiento de una maloclusion clase II división I de Angle, que se desarrollo en la clínica del post grado de ortodoncia de la facultad de odontología de la UNMSM, mediante la técnica edgewise,la misma que fue creada por el Dr. Edward Angle. El caso clínico se describen mediante las fotografías extra e intraorales, los análisis cefalométricos y los resultados obtenidos se pueden evaluar mediante las superposiciones cefalométricas, lo cual indica tanto el crecimiento craneofacial como los efectos del tratamiento.

Objective: The P2X7 receptor (P2X7R) is an important contributor to neuroinflammation, responding to extracellularly released adenosine triphosphate. Expression of the P2X7R is increased in the brain in experimental and human epilepsy, and genetic or pharmacologic targeting of the receptor can reduce seizure frequency and severity in preclinical models. Experimentally induced seizures also increase levels of the P2X7R in blood. Here, we tested 18F‐JNJ‐64413739, a positron emission tomography (PET) P2X7R antagonist, as a potential noninvasive biomarker of seizure‐damage and epileptogenesis. Methods: Status epilepticus was induced via an intra‐amygdala microinjection of kainic acid. Static PET studies (30 min duration, initiated 30 min after tracer administration) were conducted 48 h after status epilepticus via an intravenous injection of 18F‐JNJ‐64413739. PET images were coregistered with a brain magnetic resonance imaging atlas, tracer uptake was determined in the different brain regions and peripheral organs, and values were correlated to seizure severity during status epilepticus. 18F‐JNJ‐64413739 was also applied to ex vivo human brain slices obtained following surgical resection for intractable temporal lobe epilepsy. Results: P2X7R radiotracer uptake correlated strongly with seizure severity during status epilepticus in brain structures including the cerebellum and ipsi‐ and contralateral cortex, hippocampus, striatum, and thalamus. In addition, a correlation between radiotracer uptake and seizure severity was also evident in peripheral organs such as the heart and the liver. Finally, P2X7R radiotracer uptake was found elevated in brain sections from patients with temporal lobe epilepsy when compared to control. Significance: Taken together, our data suggest that P2X7R‐based PET imaging may help to identify seizure‐induced neuropathology and temporal lobe epilepsy patients with increased P2X7R levels possibly benefitting from P2X7R‐based treatments. [ABSTRACT FROM AUTHOR]

El desarrollo legislativo y reglamentario europea se ha ido desarrollando de distinta manera en los diferentes Estados miembros (EM). Por su parte, la entrada en vigor de determinadas Directivas Europeas llevan asociado el desarrollo específico de algunos aspectos concretos, o la unificación de criterios, que pueden ser diferentes en varios EM.Los productos de construcción en contacto con agua de consumo humano están afectados por las circunstancias descritas anteriormente. Por una parte, distintos Estados miembros tienen establecidos diferentes criterios de aceptación para la instalación de productos en contacto con el agua de consumo, y por otra parte, tanto la Directiva sobre Productos de Construcción como la Directiva sobre agua de consumo humano afectan a distintos aspectos reglamentarios que han de desarrollarse para poder cumplir con lo establecido en estas directivas.Con el fin de adecuar la reglamentación de los distintos EM, que forman parte de la Unión Europea (UE), a los requisitos y especificaciones recogidos en las directivas mencionadas anteriormente, se vio la necesidad de desarrollar el Esquema de Aceptación Europeo para Productos de Construcción en Contacto con Agua de Consumo Humano (EAS).The quality of the water intended for human consumption is regulated through the Directive 98/83/EC (Drinking Water Directive). Products used in the construction of water supply systems can cause significant deterioration of the water quality and therefore many Member States operate national acceptance schemes for these products. These products are covered by the Directive 89/106/EEC (Construction Products Directive) which calls for the preparation of European specifications for construction products and the approximation of the regulatory provisions of the Member States.In order to minimise the differences in the national regulatory provisions and to enable the elaboration of European specifications for construction products in contact with water for human consumption, the regulatory authorities of the Member States have agreed to establish a European Acceptance Scheme (EAS) for construction products.

El artículo en referencia desarrolla la revisión de cuatro estudios de análisis cefalometricos con el método de análisis de Ricketts, en una población de 30 sujetos peruanos, un grupo de indios navajos, un grupo de negros americanos, 40 varones mejicanos, cuyos informes fueron enviados a la “Foundation for Orthodontic Research” (FOR), material que enviado al autor vía correo por el Dr Robert Ricketts. Estos cefalogramas muestran que los diferentes valores del análisis cefalometricos realizados en diferentes poblaciones raciales, sino difieren entre si en el 100% de los valores, si difieren en algunos factores, como el ángulo interincisivo, la ubicación del incisivo inferior a la línea A Pog, la línea estética “E”, la ubicación del punto “A” que se refiere a convexidad, así como algunos valores diferentes en la profundidad facial; estos valores tienen un significado muy importante para tomarlos en cuenta al formular los objetivos de tratamiento, que van a diferir en cada población racial, diferentes a las poblaciones caucásicas.