Involvement of Glucosamine 6 Phosphate Isomerase 2 (GNPDA2) Overproduction in ß-Amyloid- and Tau P301L-Driven Pathomechanisms.

Alzheimer's disease (AD) is a neurodegenerative olfactory disorder affecting millions of people worldwide. Alterations in the hexosamine- or glucose-related pathways have been described through AD progression. Specifically, an alteration in glucosamine 6 phosphate isomerase 2 (GNPDA2) protein levels has been observed in olfactory areas of AD subjects. However, the biological role of GNPDA2 in neurodegeneration remains unknown. Using mass spectrometry, multiple GNPDA2 interactors were identified in human nasal epithelial cells (NECs) mainly involved in intraciliary transport. Moreover, GNPDA2 overexpression induced an increment in NEC proliferation rates, accompanied by transcriptomic alterations in Type II interferon signaling or cellular stress responses. In contrast, the presence of beta-amyloid or mutated Tau-P301L in GNPDA2-overexpressing NECs induced a slowdown in the proliferative capacity in parallel with a disruption in protein processing. The proteomic characterization of Tau-P301L transgenic zebrafish embryos demonstrated that GNPDA2 overexpression interfered with collagen biosynthesis and RNA/protein processing, without inducing additional changes in axonal outgrowth defects or neuronal cell death. In humans, a significant increase in serum GNPDA2 levels was observed across multiple neurological proteinopathies (AD, Lewy body dementia, progressive supranuclear palsy, mixed dementia and amyotrophic lateral sclerosis) (n = 215). These data shed new light on GNPDA2-dependent mechanisms associated with the neurodegenerative process beyond the hexosamine route.

Enhancer-AAVs allow genetic access to oligodendrocytes and diverse populations of astrocytes across species.

Proper brain function requires the assembly and function of diverse populations of neurons and glia. Single cell gene expression studies have mostly focused on characterization of neuronal cell diversity; however, recent studies have revealed substantial diversity of glial cells, particularly astrocytes. To better understand glial cell types and their roles in neurobiology, we built a new suite of adeno-associated viral (AAV)-based genetic tools to enable genetic access to astrocytes and oligodendrocytes. These oligodendrocyte and astrocyte enhancer-AAVs are highly specific (usually > 95% cell type specificity) with variable expression levels, and our astrocyte enhancer-AAVs show multiple distinct expression patterns reflecting the spatial distribution of astrocyte cell types. To provide the best glial-specific functional tools, several enhancer-AAVs were: optimized for higher expression levels, shown to be functional and specific in rat and macaque, shown to maintain specific activity in epilepsy where traditional promoters changed activity, and used to drive functional transgenes in astrocytes including Cre recombinase and acetylcholine-responsive sensor iAChSnFR. The astrocyte-specific iAChSnFR revealed a clear reward-dependent acetylcholine response in astrocytes of the nucleus accumbens during reinforcement learning. Together, this collection of glial enhancer-AAVs will enable characterization of astrocyte and oligodendrocyte populations and their roles across species, disease states, and behavioral epochs.

Endothelial NOX5 overexpression induces changes in the cardiac gene profile: potential impact in myocardial infarction?

Cardiovascular diseases and the ischemic heart disease specifically constitute the main cause of death worldwide. The ischemic heart disease may lead to myocardial infarction, which in turn triggers numerous mechanisms and pathways involved in cardiac repair and remodeling. Our goal in the present study was to characterize the effect of the NADPH oxidase 5 (NOX5) endothelial expression in healthy and infarcted knock-in mice on diverse signaling pathways. The mechanisms studied in the heart of mice were the redox pathway, metalloproteinases and collagen pathway, signaling factors such as NFκB, AKT or Bcl-2, and adhesion molecules among others. Recent studies support that NOX5 expression in animal models can modify the environment and predisposes organ response to harmful stimuli prior to pathological processes. We found many alterations in the mRNA expression of components involved in cardiac fibrosis as collagen type I or TGF-ß and in key players of cardiac apoptosis such as AKT, Bcl-2, or p53. In the heart of NOX5-expressing mice after chronic myocardial infarction, gene alterations were predominant in the redox pathway (NOX2, NOX4, p22phox, or SOD1), but we also found alterations in VCAM-1 and ß-MHC expression. Our results suggest that NOX5 endothelial expression in mice preconditions the heart, and we propose that NOX5 has a cardioprotective role. The correlation studies performed between echocardiographic parameters and cardiac mRNA expression supported NOX5 protective action.

NADPH Oxidase 5 (NOX5) Overexpression Promotes Endothelial Dysfunction via Cell Apoptosis, Migration, and Metabolic Alterations in Human Brain Microvascular Endothelial Cells (hCMEC/D3).

NADPH oxidases (NOX) constitute the main reactive oxygen species (ROS) source in blood vessels. An oxidative stress situation due to ROS overproduction can lead into endothelial dysfunction, a molecular mechanism that precedes cardiovascular diseases (CVDs) such as atherosclerosis, myocardial infarction, and stroke. NOX5 is the last discovered member of the NOX family, studied in a lesser extent due to its absence in the rodent genome. Our objective was to describe the phenotypic alterations produced by an oxidative stress situation derived from NOX5 overexpression in an endothelial in vitro model. The in vitro model consists of the hCMEC/D3 cell line, derived from brain microvascular endothelium, infected with a recombinant NOX5-ß adenovirus. After an initial proteomic analysis, three phenotypic alterations detected in silico were studied: cell proliferation and apoptosis, general and mitochondrial metabolism, and migration capacity. NOX5 infection of hCMEC/D3 generates a functional protein and an increase in ROS production. This model produced changes in the whole cell proteome. The in silico analysis together with in vitro validations demonstrated that NOX5 overexpression inhibits proliferation and promotes apoptosis, metabolic alterations and cell migration in hCMEC/D3 cells. NOX5 overexpression in endothelial cells leads to phenotypic changes that can lead to endothelial dysfunction, the onset of atherosclerosis, myocardial infarction, and stroke.

Riesgo de enfermedad cardiovascular y cerebrovascular a largo plazo en mujeres con antecedente personal de abrupción placentaria

La salud materna y perinatal es una de las prioridades actuales de la salud global. La enfermedad cardiovascular y el accidente cerebrovascular son las principales causas de mortalidad materna. La abrupción placentaria sigue siendo una preocupación crítica para la morbilidad materna debido a que se ha asociado a enfermedad vascular a largo plazo. Sin embargo, no existe mucha literatura disponible en español ni evidencia reciente que haya dilucidado algunas interrogantes sobre este tópico. Entonces, el objetivo de esta revisión consiste en sintetizar evidencia reciente sobre el riesgo de enfermedad cardiovascular y cerebrovascular a largo plazo en mujeres con antecedente personal de abrupción placentaria. Se encontró que, a través de mecanismos fisiopatológicos complejos, que involucran la estructura y funcionalidad de la red vascular placentaria con posterior extensión de lesión vascular y producción de factores proinflamatorios y procoagulantes que permanecen después del parto, se precipita la aparición de eventos cardiovasculares mayores a mediano y largo plazo. Evidencia de alta calidad ha revelado que el riesgo de sufrir de complicaciones maternas en aquellas mujeres con abrupción placentaria es de 2,14, que se eleva aún más para aquellas con desprendimiento severo. A mediano y largo plazo, el riesgo de mortalidad por cardiopatía coronaria es de 2,64, y de 1,70 para desorden cerebrovascular, con igual riesgo tanto para el tipo isquémico como hemorrágico. Entonces, se puede concluir que el riesgo cardiovascular y cerebrovascular es inminente en mujeres con antecedente de abrupción placentaria, dado por numerosos mecanismos fisiopatológicos vasculares. No obstante, este riesgo se eleva considerablemente al asociarse con factores modificables tradicionales y no tradicionales.

Maternal and perinatal health is one of today's global health priorities. Cardiovascular disease and stroke are the leading causes of maternal mortality. Placental abruption remains a critical concern for maternal morbidity because it has been associated with long-term vascular disease. However, there is neither much literature available in Spanish nor recent evidence elucidating some questions on this topic. Thus, this review aims to synthesize recent evidence on the long-term risk of cardiovascular and cerebrovascular disease in women with a personal history of placental abruption. It was found that, through complex pathophysiological mechanisms involving the structure and functionality of the placental vascular network with subsequent extension of vascular injury and production of proinflammatory and procoagulant factors which remain after delivery, major cardiovascular events are precipitated in the medium and long term. High-quality evidence has shown that the risk of maternal complications in women with placental abruption accounts for 2.14, rising even higher for those with severe placental abruption. In the medium and long term, the mortality risk caused by coronary heart diseases is 2.64 and by cerebrovascular disorders is 1.70, with equal risk for both ischemic and hemorrhagic strokes. It can therefore be concluded that cardiovascular and cerebrovascular risk is imminent in women with a history of placental abruption due to a number of vascular pathophysiological mechanisms. However, this risk is considerably increased when associated with traditional and non-traditional modifiable factors.

Enhancer viruses for combinatorial cell-subclass-specific labeling.

Rapid cell type identification by new genomic single-cell analysis methods has not been met with efficient experimental access to these cell types. To facilitate access to specific neural populations in mouse cortex, we collected chromatin accessibility data from individual cells and identified enhancers specific for cell subclasses and types. When cloned into recombinant adeno-associated viruses (AAVs) and delivered to the brain, these enhancers drive transgene expression in specific cortical cell subclasses. We extensively characterized several enhancer AAVs to show that they label different projection neuron subclasses as well as a homologous neuron subclass in human cortical slices. We also show how coupling enhancer viruses expressing recombinases to a newly generated transgenic mouse, Ai213, enables strong labeling of three different neuronal classes/subclasses in the brain of a single transgenic animal. This approach combines unprecedented flexibility with specificity for investigation of cell types in the mouse brain and beyond.

NADPH Oxidase 5 Induces Changes in the Unfolded Protein Response in Human Aortic Endothelial Cells and in Endothelial-Specific Knock-in Mice.

Oxidative stress constitutes a key molecular mechanism in the development of cardiovascular diseases. A potential relationship between reactive oxygen species (ROS) driven by the NADPH oxidase family (NOX) and the unfolded protein response (UPR) has been postulated. Nevertheless, there is a lack of information about the crosstalk between NOX5 homologue and the UPR in a cardiovascular context. The main aim was to analyze NOX5-mediated ROS effects in the UPR and its importance in cardiovascular diseases. To this effect, we used an adenoviral NOX5-ß overexpression model in human aortic endothelial cells (HAEC) and a conditional endothelial NOX5 knock-in mouse. Using expression arrays, we investigated NOX5-induced genomic changes in HAEC. Compared with the control HAEC, 298 genes were differentially expressed. Gene ontology analysis revealed the activation of numerous cellular routes, the most relevant being the UPR pathway. Using real-time PCR and Western Blot experiments, we confirmed that NOX5 overexpression induced changes in the expression of the UPR components, which were associated with increased apoptosis. Moreover, in endothelial-specific NOX5 knock-in mice, we found changes in the expression of the UPR components genes. In these mice, myocardial infarction was performed by permanent coronary artery ligation; however, NOX5 expression was not associated with differences in the UPR components mRNA levels. In these animals, we found significant associations between the UPR components gene expression and echocardiographic parameters. Our data support the idea that NOX5-derived ROS may modulate the UPR pathway in endothelial cells, which might play a relevant role in cardiac physiology.

Induction of Cyclooxygenase-2 by Overexpression of the Human NADPH Oxidase 5 (NOX5) Gene in Aortic Endothelial Cells.

Oxidative stress is a main molecular mechanism that underlies cardiovascular diseases. A close relationship between reactive oxygen species (ROS) derived from NADPH oxidase (NOX) activity and the prostaglandin (PG) biosynthesis pathway has been described. However, little information is available about the interaction between NOX5 homolog-derived ROS and the PG pathway in the cardiovascular context. Our main goal was to characterize NOX5-derived ROS effects in PG homeostasis and their potential relevance in cardiovascular pathologies. For that purpose, two experimental systems were employed: an adenoviral NOX5-ß overexpression model in immortalized human aortic endothelial cells (TeloHAEC) and a chronic infarction in vivo model developed from a conditional endothelial NOX5 knock-in mouse. NOX5 increased cyclooxygenase-2 isoform (COX-2) expression and prostaglandin E2 (PGE2) production through nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) in TeloHAEC. Protein kinase C (PKC) activation and intracellular calcium level (Ca++) mobilization increased ROS production and NOX5 overexpression, which promoted a COX-2/PGE2 response in vitro. In the chronic infarction model, mice encoding endothelial NOX5 enhanced the cardiac mRNA expression of COX-2 and PGES, suggesting a COX-2/PGE2 response to NOX5 presence in an ischemic situation. Our data support that NOX5-derived ROS may modulate the COX-2/PGE2 axis in endothelial cells, which might play a relevant role in the pathophysiology of heart infarction.

Increased phagocytic NADPH oxidase activity associates with coronary artery calcification in asymptomatic men.

Vascular calcification is a common feature in atherosclerosis and associates with cardiovascular events. Oxidative stress may be involved in the pathogenesis of vascular calcification. Previous studies have shown that the phagocytic NADPH oxidase is associated with atherosclerosis. The objective of the present study was to investigate the association between phagocytic NADPH oxidase-mediated superoxide production and coronary artery calcium (CAC). NADPH oxidase-mediated superoxide production was determined by chemiluminescence and CAC by computed tomography in 159 asymptomatic men free of overt clinical atherosclerosis. Multivariate linear regression analyses were used to assess the relationship between CAC and NADPH oxidase-mediated superoxide production. Compared with individuals in the lowest score of CAC (= 0 Agatston units), those in the upper score (>400 Agatston units) showed higher superoxide production (p < 0.05). In correlation analysis, superoxide production positively (p < 0.01) correlated with CAC, which in multivariate analysis remained significant after adjusting for age, HDL-cholesterol, triglycerides, body mass index, smoking, arterial hypertension and diabetes mellitus. In conclusion, in a population of men without clinically overt atherosclerotic disease, increased NADPH oxidase-mediated superoxide production associated with enhanced CAC. Albeit descriptive, these findings suggest a potential involvement of phagocytic NADPH oxidase-mediated oxidative stress in CAC.

Esquemas del tratamiento para pacientes con labio y/o paladar fisurado y análisis de casos de pacientes del hospital Lorencita Villegas de Santos / Schemes of treatment for patient sustained fissured lip or palate and cases analysis of patients of the Hospital Lorencita Villegas de Santos

El labio y /o paladar fisurado, es la más común malformación congénita de cabeza y cuello. Su etiología es multifactorial; está determinada por predisposición genética, más factores ambientales. El labio fisurado se presenta en uno de cada 900 nacimientos vivos, afectando varones en mayor proporción; el paladar fisurado afecta uno de cada 2000 nacimientos y las niñas se ven mas afectadas. En la mayoria de los casos revisados durante el desarrollo de ésta investigación, se presentó la combinación de labio y paladar fisurado. Los factores exógenos, incluídos el uso de insecticidas, Fenitoína o Fármacos para el tratamiento del cáncer durante el embarazo, han sido incluídos en la etiología de fisuras faciales. Algunos casos de labio y paladar fisurado, han sido asociados con sindromes congénitos que afectan otras partes del cuerpo. La fisura del labio y/o paladar, esta establecida dentro de los 5 a 8 primeras semanas de vida intrauterina. En general estos defectos parecen tener causas y efectos ambientales mixtos. Las hendiduras del labio y proceso alveolar que se continuan a través del paladar, suelen transmitirse a través de la primera generación. Pueden producirse por falta de unión de procesos maxilares con el proceso nasal, o por penetración incompleta del mesodermo dentro de las membranas epiteliales del proceso nasal medio, del proceso nasal lateral y de los procesos maxilares. Los pacientes con labio y/o paladar fisurado requeriran supervisión médica desde el nacimiento y el manejo por parte de un equipo multidisciplinario que incluye cirujanos plásticos, ortodoncista, cirujano oral, foniatras y psicólogos entre otros. El manejo por parte de un equipo conduce no solo al mejoramiento de cada paciente individualmente sino que brinda una perspectiva de manejo adecuado de todos los pacientes que presentan esta amlformación. Las técnicas modernas para la reparación de labio fisurado estiman que la deformidad afecta no solo la piel sino el músculo orbicular y la maxila subyacente la cual tiende a ser hipoplásica. Como es sabido que todas la técnicas que existen para el tratamiento de estos pacientes son numerosas; podriamos deciacion medica