Full genome analysis of microglial activation; ramifications of TREM2

Neuroinflammation is a pathological hallmark of Alzheimer's disease (AD) and it is well established that microglia, the brain's resident phagocytes, are pivotal for the immune response observed in AD. In the healthy brain, microglia attack and remove pathogens and cell debris, but have been shown to become reactive in AD. An apparent link between microglia and AD is Amyloid β (Aβ), which accumulates in the plaques observed in the brains of AD patients and has been reported as a microglia activator. Genome Wide Association Studies (GWAS) have allowed the identification of more than 20 genetic risk associations to AD. Many of these associations highlight the importance of immune pathways (and others) in AD. More recently, the identification of mutations in TREM2 (Triggering Receptor Expressed on Myeloid Cells 2), a gene exclusively expressed by microglia in the brain, has brought microglial activation and dysfunction back to the attention of the AD community. The main focus of this study is to understand microglial activation elicited by different stimuli including Aβ1-42 monomers, oligomers and fibrils- with regards to their inflammatory activation status (M1, M2 or other) and whole-genome expression profile. To this end, the mouse-derived BV2 cell line was used to assess gene expression changes during microglial activation. Data shows that M1 and M2 activators alter gene expression of AD-associated genes in a manner that is potentially detrimental for AD progression. A second objective of this thesis was to use the CRISPR/Cas9 gene editing technology for the generation of Trem2-deficient BV2 cell lines. As a result, Trem2 +/- (haploinsufficient) and Trem2 -/- (knockout) BV2 cell lines were generated. Subsequently, these cell lines were characterised in terms of their phagocytic, proliferation, migration, cytokine release capacities and whole genome expression. In consequence, this study provides new and wellcharacterised in vitro models for the study of Trem2 function

Propuesta de un Plan de mejora para incrementar la eficiencia de la gestión de pedidos automatizando los procesos de una editorial de soluciones educativa

La presente investigación se ha llevado a cabo en una empresa perteneciente a la industria editorial, la cual se encarga de la edición, comercialización y distribución de textos escolares, está ubicada en Lima y tiene como objetivo ser la mejor empresa de soluciones educativas a nivel nacional, brindando las mejores herramientas y soporte a los estudiantes. En las últimas campañas escolares, el área de operaciones presentó problemas con el manejo de la gestión de pedidos, ya que no cuentan con un sistema el cual les permita reducir los errores de cara al cliente. Por esto, se vieron en la necesidad de implementar un software para agilizar y reducir los errores en el proceso de Gestión de pedidos brindando un servicio de calidad de cara al cliente. Por consiguiente, la presente investigación propuso la aplicación de un sistema BPMS, ya que con estas herramientas se podrá realizar la unificación de la tecnología de la información con el entorno operativo de la organización. De igual forma, con este sistema se pretendió automatizar las etapas de modelado, ejecución, control y seguimiento, con el fin de detallar de forma clara el proceso de la presente investigación. Con esta propuesta se obtuvieron mejores resultados de los indicadores (reducción de la tasa de devoluciones, incremento en la eficiencia de generación de contratos, incremento en la agilización de evaluaciones de clientes). Ya que mediante el uso del software implementado se optimizó la Gestión de pedidos en esta empresa. Finalmente, con la propuesta del plan de mejora se lograron obtener resultados mejorados con una automatización de procesos que incrementó la eficiencia de la gestión de pedidos, lo que fue demostrado con la reducción de 24.52% en los tiempos que demora la gestión de un pedido. De igual forma, el tiempo de evaluación de un cliente se redujo en 24.94 minutos. Así mismo, los pedidos aumentaron en un 35% con respecto a la campaña del 2021 - 2022, mientras que las notas de crédito se redujeron en un 7,69%. Por último, la eficiencia administrativa de contratos aumentó a 0,71 contratos/minuto. Todo ello automatizando los procesos de una editorial de soluciones educativa

Amyloid precursor protein processing in human neurons with an allelic series of the PSEN1 intron 4 deletion mutation and total presenilin-1 knockout

Mutations in presenilin-1 (PSEN1), encoding the catalytic subunit of the amyloid precursor protein-processing enzyme γ-secretase, cause familial Alzheimer’s disease. However, the mechanism of disease is yet to be fully understood and it remains contentious whether mutations exert their effects predominantly through gain or loss of function. To address this question, we generated an isogenic allelic series for the PSEN1 mutation intron 4 deletion; represented by control, heterozygous and homozygous mutant induced pluripotent stem cells in addition to a presenilin-1 knockout line. Induced pluripotent stem cell-derived cortical neurons reveal reduced, yet detectable amyloid-beta levels in the presenilin-1 knockout line, and a mutant gene dosage-dependent defect in amyloid precursor protein processing in PSEN1 intron 4 deletion lines, consistent with reduced processivity of γ-secretase. The different effects of presenilin-1 knockout and the PSEN1 intron 4 deletion mutation on amyloid precursor protein-C99 fragment accumulation, nicastrin maturation and amyloid-beta peptide generation support distinct consequences of familial Alzheimer’s disease-associated mutations and knockout of presenilin-1 on the function of γ-secretase

Soluble Fibrinogen Triggers Non-cell Autonomous ER Stress-Mediated Microglial-Induced Neurotoxicity

Aberrant or chronic microglial activation is strongly implicated in neurodegeneration, where prolonged induction of classical inflammatory pathways may lead to a compromised blood-brain barrier (BBB) or vasculature, features of many neurodegenerative disorders and implicated in the observed cognitive decline. BBB disruption or vascular disease may expose the brain parenchyma to “foreign” plasma proteins which subsequently impact on neuronal network integrity through neurotoxicity, synaptic loss and the potentiation of microglial inflammation. Here we show that the blood coagulation factor fibrinogen (FG), implicated in the pathogenesis of dementias such as Alzheimer’s disease (AD), induces an inflammatory microglial phenotype as identified through genetic microarray analysis of a microglial cell line, and proteome cytokine profiling of primary microglia. We also identify a FG-mediated induction of non-cell autonomous ER stress-associated neurotoxicity via a signaling pathway that can be blocked by pharmacological inhibition of microglial TNFα transcription or neuronal caspase-12 activity, supporting a disease relevant role for plasma components in neuronal dysfunction

Gastronomía típica como identidad cultural

Introduction. Currently the typical gastronomy is considered an important heritage for the development of tourism, the tasting of typical dishes generates a cultural and sensorial experience where diners have a connection with the gastronomy of the area. Objective. Analyze the typical gastronomy as a cultural identity of the inhabitants of Pelileo. Methodology. The research had a mixed design, that is, quantitative using a questionnaire as a tool and qualitative using the registration forms of the National Institute of Cultural Heritage (INPC) with a field study. The questionnaire was sent to 383 inhabitants of Pelileo, the questionnaire was prepared on the Google forms platform and a reliability analysis was carried out for its application. The collection of information on typical gastronomy in 179 establishments in Pelileo was carried out using registration cards (INPC), a field of study that is as descriptive as it is deductive. Thanks to the registration form (INPC), the most typical dishes of the canton were identified, such as: roast guinea pig, roast rabbit, fried, sweet empanadas, baked, tamales, chawarmisqui, chicken broth and wind empanadas. Results. Among the results of the survey, it should be noted that most of the inhabitants of the canton show an inclination on the effective dimension with an average of 3.80 corresponding to 19.84%. Conclusions. Emotions are superimposed on other more rational aspects; therefore, a greater degree of relevance is required to inform the typically representative food by the possible means.Introducción. Actualmente la gastronomía típica es considerada como un patrimonio importante para el desarrollo del turismo, la degustación de platos típicos genera una experiencia cultural y sensorial donde los comensales tienen una conexión con la gastronomía de la zona. Objetivo. Analizar la gastronomía típica como identidad cultural de los habitantes de Pelileo. Metodología. La investigación tuvo un diseño mixto, es decir, cuantitativo utilizando un cuestionario como herramienta y cualitativo utilizando los formularios de registro del Instituto Nacional de Patrimonio Cultural (INPC) con un estudio de campo. El cuestionario fue enviado a 383 pobladores de Pelileo, el cuestionario fue elaborado en la plataforma de formularios de Google y se realizó un análisis de confiabilidad para su aplicación. El levantamiento de información sobre la gastronomía típica en 179 establecimientos de Pelileo se realizó mediante fichas de registro (INPC), campo de estudio que es tan descriptivo como deductivo. Gracias a la ficha de registro (INPC), se identificaron los platos típicos más típicos del cantón como son: cuy asado, conejo asado, fritada, empanadas de dulce, hornado, tamales, chawarmisqui, caldo de gallina y empanadas de viento. Resultados. Entre los resultados de la encuesta, cabe señalar que la mayoría de los habitantes del cantón muestran una inclinación sobre la dimensión efectiva con una media de 3.80 correspondiente al 19.84 % Conclusiones. Las emociones se superponen a otros aspectos más racionales, se requiere por tanto un mayor grado de relevancia para informar por los medios posibles la comida típicamente representativos

Multifunctional receptor-targeted nanocomplexes for magnetic resonance imaging and transfection of tumours.

The efficient targeted delivery of nucleic acids in vivo provides some of the greatest challenges to the development of genetic therapies. We aim to develop nanocomplex formulations that achieve targeted transfection of neuroblastoma tumours that can be monitored simultaneously by MRI. Here, we have compared nanocomplexes comprising self-assembling mixtures of liposomes, plasmid DNA and one of three different peptide ligands derived from ApoE, neurotensin and tetanus toxin for targeted transfection in vitro and in vivo. Neurotensin-targeted nanocomplexes produced the highest levels of transfection and showed a 4.7-fold increase in transfected luciferase expression over non-targeted nanocomplexes in Neuro-2A cells. Transfection of subcutaneous Neuro-2A tumours in vivo with neurotensin-targeted nanocomplexes produced a 9.3-fold increase in gene expression over non-targeted controls. Confocal microscopy analysis elucidated the time course of DNA delivery with fluorescently labelled nanocomplex formulations in cells. It was confirmed that addition of a gadolinium lipid conjugate contrast agent allowed real time in vivo monitoring of nanocomplex localisation in tumours by MRI, which was maintained for at least 24 h. The peptide-targeted nanocomplexes developed here allow for the specific enhancement of targeted gene therapy both in vitro and in vivo, whilst allowing real time monitoring of delivery with MRI

Lipid peptide nanocomplexes for gene delivery and magnetic resonance imaging in the brain.

Gadolinium-labelled nanocomplexes offer prospects for the development of real-time, non-invasive imaging strategies to visualise the location of gene delivery by MRI. In this study, targeted nanoparticle formulations were prepared comprising a cationic liposome (L) containing a Gd-chelated lipid at 10, 15 and 20% by weight of total lipid, a receptor-targeted, DNA-binding peptide (P) and plasmid DNA (D), which electrostatically self-assembled into LPD nanocomplexes. The LPD formulation containing the liposome with 15% Gd-chelated lipid displayed optimal peptide-targeted, transfection efficiency. MRI conspicuity peaked at 4h after incubation of the nanocomplexes with cells, suggesting enhancement by cellular uptake and trafficking. This was supported by time course confocal microscopy analysis of transfections with fluorescently-labelled LPD nanocomplexes. Gd-LPD nanocomplexes delivered to rat brains by convection-enhanced delivery were visible by MRI at 6 h, 24 h and 48 h after administration. Histological brain sections analysed by laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) confirmed that the MRI signal was associated with the distribution of Gd(3+) moieties and differentiated MRI signals due to haemorrhage. The transfected brain cells near the injection site appeared to be mostly microglial. This study shows the potential of Gd-LPD nanocomplexes for simultaneous delivery of contrast agents and genes for real-time monitoring of gene therapy in the brain

Human induced pluripotent stem cell-derived microglia-like cells harboring TREM2 missense mutations show specific deficits in phagocytosis

Dysfunction of microglia, the brain’s immune cells, is linked to neurodegeneration. Homozygous missense mutations in TREM2 cause Nasu-Hakola disease (NHD), an early-onset dementia. To study the consequences of these TREM2 variants, we generated induced pluripotent stem cell-derived microglia-like cells (iPSC-MGLCs) from patients with NHD caused by homozygous T66M or W50C missense mutations. iPSC-MGLCs expressed microglial markers and secreted higher levels of TREM2 than primary macrophages. TREM2 expression and secretion were reduced in variant lines. LPS-mediated cytokine secretion was comparable between control and TREM2 variant iPSC-MGLCs, whereas survival was markedly reduced in cells harboring missense mutations when compared with controls. Furthermore, TREM2 missense mutations caused a marked impairment in the phagocytosis of apoptotic bodies, but not in Escherichia coli or zymosan substrates. Coupled with changes in apoptotic cell-induced cytokine release and migration, these data identify specific deficits in the ability of iPSC-MGLCs harboring TREM2 missense mutations to respond to specific pathogenic signals

Efecto de los inhibidores selectivos de la recaptación de serotonina en los linfocitos de pacientes con depresión

Depression is the most frequent mental disorder worldwide whose pathology is complex due to the psychological and pathophysiological factors that it involves. Pharmacological treatment is essential, nevertheless, it presents high percentages of resistance to which several investigations have identified that alterations in the immune system play a decisive role. For this reason, the objective of this article is to review the current bibliography on the effects of selective serotonin reuptake inhibitors (SSRIs), the first line of pharmacological management, on changes in the immune system in patients with depression. The alteration of lymphocytes and cytokines in the pathophysiology of depression and against SSRIs is evident. However, extensive research is required to establish normal immune patterns and their affectation against these factors. This will allow the generation of new pharmacological targets that reduce the resistance, chronicity and recurrence of depression.La depresión es el trastorno mental más frecuente a nivel mundial cuya patología es compleja por los factores psicológicos y fisiopatológicos que conlleva. El tratamiento farmacológico es fundamental, no obstante, presenta altos porcentajes de resistencia ante lo cual varias investigaciones han identificado que las alteraciones del sistema inmune juegan un papel decisivo. Es por ello que el objetivo de este artículo es revisar la bibliografía actual sobre los efectos de los inhibidores selectivos de la recaptación de serotonina (ISRS), primera línea de manejo farmacológico, sobre las alteraciones del sistema inmune en pacientes con depresión. Es evidente la alteración de los linfocitos y citoquinas en la fisiopatología de la depresión y frente a los ISRS. Sin embargo, se requieren amplias investigaciones para establecer los patrones inmunes normales y su afectación frente a estos factores. Esto permitirá generar nuevas metas farmacológicas que disminuyan la resistencia, cronicidad y recurrencia de la depresión