Identification of a Potential Entry-Fusion Complex Based on Sequence Homology of African Swine Fever and Vaccinia Virus.

African swine fever virus (ASFV) belongs to the family of Asfarviridae, part of the group of nucleocytoplasmic large DNA viruses (NCLDV). Little is known about the internalization of ASFV in the host cell and the fusion membrane events that take place at early stages of the infection. Poxviruses, also members of the NCLDV and represented by vaccinia virus (VACV), are large, enveloped, double-stranded DNA viruses. Poxviruses were considered unique in having an elaborate entry-fusion complex (EFC) composed of 11 highly conserved proteins integrated into the membrane of mature virions. Recent advances in methodological techniques have again revealed several connections between VACV EFC proteins. In this study, we explored the possibility of an analogous ASFV EFC by identifying ten candidate proteins exhibiting structural similarities with VACV EFC proteins. This could reveal key functions of these ASFV proteins, drawing attention to shared features between the two virus families, suggesting the potential existence of an ASFV entry-fusion complex.

Kidins220 sets the threshold for survival of neural stem cells and progenitors to sustain adult neurogenesis.

In the adult mammalian brain, neural stem cells (NSCs) located in highly restricted niches sustain the generation of new neurons that integrate into existing circuits. A reduction in adult neurogenesis is linked to ageing and neurodegeneration, whereas dysregulation of proliferation and survival of NSCs have been hypothesized to be at the origin of glioma. Thus, unravelling the molecular underpinnings of the regulated activation that NSCs must undergo to proliferate and generate new progeny is of considerable relevance. Current research has identified cues promoting or restraining NSCs activation. Yet, whether NSCs depend on external signals to survive or if intrinsic factors establish a threshold for sustaining their viability remains elusive, even if this knowledge could involve potential for devising novel therapeutic strategies. Kidins220 (Kinase D-interacting substrate of 220 kDa) is an essential effector of crucial pathways for neuronal survival and differentiation. It is dramatically altered in cancer and in neurological and neurodegenerative disorders, emerging as a regulatory molecule with important functions in human disease. Herein, we discover severe neurogenic deficits and hippocampal-based spatial memory defects accompanied by increased neuroblast death and high loss of newly formed neurons in Kidins220 deficient mice. Mechanistically, we demonstrate that Kidins220-dependent activation of AKT in response to EGF restraints GSK3 activity preventing NSCs apoptosis. We also show that NSCs with Kidins220 can survive with lower concentrations of EGF than the ones lacking this molecule. Hence, Kidins220 levels set a molecular threshold for survival in response to mitogens, allowing adult NSCs growth and expansion. Our study identifies Kidins220 as a key player for sensing the availability of growth factors to sustain adult neurogenesis, uncovering a molecular link that may help paving the way towards neurorepair.

Elucidation of the Cellular Interactome of African Swine Fever Virus Fusion Proteins and Identification of Potential Therapeutic Targets.

African swine fever virus (ASFV) encodes more than 150 proteins, most of them of unknown function. We used a high-throughput proteomic analysis to elucidate the interactome of four ASFV proteins, which potentially mediate a critical step of the infection cycle, the fusion and endosomal exit of the virions. Using affinity purification and mass spectrometry, we were able to identify potential interacting partners for those ASFV proteins P34, E199L, MGF360-15R and E248R. Representative molecular pathways for these proteins were intracellular and Golgi vesicle transport, endoplasmic reticulum organization, lipid biosynthesis, and cholesterol metabolism. Rab geranyl geranylation emerged as a significant hit, and also Rab proteins, which are crucial regulators of the endocytic pathway and interactors of both p34 and E199L. Rab proteins co-ordinate a tight regulation of the endocytic pathway that is necessary for ASFV infection. Moreover, several interactors were proteins involved in the molecular exchange at ER membrane contacts. These ASFV fusion proteins shared interacting partners, suggesting potential common functions. Membrane trafficking and lipid metabolism were important categories, as we found significant interactions with several enzymes of the lipid metabolism. These targets were confirmed using specific inhibitors with antiviral effect in cell lines and macrophages.

New insights into the role of endosomal proteins for African swine fever virus infection.

African swine fever virus (ASFV) infectious cycle starts with the viral adsorption and entry into the host cell. Then, the virus is internalized via clathrin/dynamin mediated endocytosis and macropinocytosis. Similar to other viruses, ASF virion is then internalized and incorporated into the endocytic pathway. While the endosomal maturation entails luminal acidification, the decrease in pH acts on the multilayer structure of the virion dissolving the outer capsid. Upon decapsidation, the inner viral membrane is exposed to interact with the limiting membrane of the late endosome for fusion. Viral fusion is then necessary for the egress of incoming virions from endosomes into the cytoplasm, however this remains an intriguing and yet essential process for infection, specifically for the egress of viral nucleic acid into the cytoplasm for replication. ASFV proteins E248R and E199L, located at the exposed inner viral membrane, might be implicated in the fusion step. An interaction between these viral proteins and cellular endosomal proteins such as the Niemann-Pick C type 1 (NPC1) and lysosomal membrane proteins (Lamp-1 and -2) was shown. Furthermore, the silencing of these proteins impaired ASFV infection. It was also observed that NPC1 knock-out cells using CRISPR jeopardized ASFV infection and that the progression and endosomal exit of viral cores was arrested within endosomes at viral entry. These results suggest that the interactions of ASFV proteins with some endosomal proteins might be important for the membrane fusion step. In addition to this, reductions on ASFV infectivity and replication in NPC1 KO cells were accompanied by fewer and smaller viral factories. Our findings pave the way to understanding the role of proteins of the endosomal membrane in ASFV infection.

Identification of Niemann-Pick C1 protein as a potential novel SARS-CoV-2 intracellular target.

Niemann-Pick type C1 (NPC1) receptor is an endosomal membrane protein that regulates intracellular cholesterol traffic. This protein has been shown to play an important role for several viruses. It has been reported that SARS-CoV-2 enters the cell through plasma membrane fusion and/or endosomal entry upon availability of proteases. However, the whole process is not fully understood yet and additional viral/host factors might be required for viral fusion and subsequent viral replication. Here, we report a novel interaction between the SARS-CoV-2 nucleoprotein (N) and the cholesterol transporter NPC1. Furthermore, we have found that some compounds reported to interact with NPC1, carbazole SC816 and sulfides SC198 and SC073, were able to reduce SARS-CoV-2 viral infection with a good selectivity index in human cell infection models. These findings suggest the importance of NPC1 for SARS-CoV-2 viral infection and a new possible potential therapeutic target to fight against COVID-19.

African Swine Fever Virus Ubiquitin-Conjugating Enzyme Is an Immunomodulator Targeting NF-κB Activation.

African swine fever virus (ASFV) is an acute and persistent swine virus with a high economic burden that encodes multiple genes to evade host immune response. In this work, we have revealed that early viral protein UBCv1, the only known conjugating enzyme encoded by a virus, modulates innate immune and inflammatory signaling. Transient overexpression of UBCv1 impaired activation of NF-κB and AP-1 transcription factors induced by several agonists of these pathways. In contrast, activation of IRF3 and ISRE signaling upon stimulation with TRIFΔRIP, cGAS/STING or RIG-I-CARD remained unaltered. Experiments aimed at mapping UBCv1 inhibitory activity indicated that this viral protein acts upstream or at the level step of IKKß. In agreement with this, UBCv1 was able to block p65 nuclear translocation upon cytokine stimulation, a key event in NF-ĸB signaling. Additionally, A549 stably transduced for UBCv1 showed a significant decrease in the levels of NF-ĸB dependent genes. Interestingly, despite the well-defined capacity of UBCv1 to conjugate ubiquitin chains, a mutant disabled for ubiquitylation activity retained similar immunomodulatory activity as the wild-type enzyme, suggesting that the two functions are segregated. Altogether these data suggest that ASFV UBCv1 manipulates the innate immune response targeting the NF-κB and AP-1 pathways and opens new questions about the multifunctionality of this enzyme.

Kidins220 deficiency causes ventriculomegaly via SNX27-retromer-dependent AQP4 degradation.

Several psychiatric, neurologic and neurodegenerative disorders present increased brain ventricles volume, being hydrocephalus the disease with the major manifestation of ventriculomegaly caused by the accumulation of high amounts of cerebrospinal fluid (CSF). The molecules and pathomechanisms underlying cerebral ventricular enlargement are widely unknown. Kinase D interacting substrate of 220 kDa (KIDINS220) gene has been recently associated with schizophrenia and with a novel syndrome characterized by spastic paraplegia, intellectual disability, nystagmus and obesity (SINO syndrome), diseases frequently occurring with ventriculomegaly. Here we show that Kidins220, a transmembrane protein effector of various key neuronal signalling pathways, is a critical regulator of CSF homeostasis. We observe that both KIDINS220 and the water channel aquaporin-4 (AQP4) are markedly downregulated at the ventricular ependymal lining of idiopathic normal pressure hydrocephalus (iNPH) patients. We also find that Kidins220 deficient mice develop ventriculomegaly accompanied by water dyshomeostasis and loss of AQP4 in the brain ventricular ependymal layer and astrocytes. Kidins220 is a known cargo of the SNX27-retromer, a complex that redirects endocytosed plasma membrane proteins (cargos) back to the cell surface, thus avoiding their targeting to lysosomes for degradation. Mechanistically, we show that AQP4 is a novel cargo of the SNX27-retromer and that Kidins220 deficiency promotes a striking and unexpected downregulation of the SNX27-retromer that results in AQP4 lysosomal degradation. Accordingly, SNX27 silencing decreases AQP4 levels in wild-type astrocytes whereas SNX27 overexpression restores AQP4 content in Kidins220 deficient astrocytes. Together our data suggest that the KIDINS220-SNX27-retromer-AQP4 pathway is involved in human ventriculomegaly and open novel therapeutic perspectives.

African Swine Fever Virus Ubiquitin-Conjugating Enzyme Interacts With Host Translation Machinery to Regulate the Host Protein Synthesis.

African Swine Fever virus (ASFV) causes one of the most relevant emerging diseases affecting swine, now extended through three continents. The virus has a large coding capacity to deploy an arsenal of molecules antagonizing the host functions. In the present work, we have studied the only known E2 viral-conjugating enzyme, UBCv1 that is encoded by the I215L gene of ASFV. UBCv1 was expressed as an early expression protein that accumulates throughout the course of infection. This versatile protein, bound several types of polyubiquitin chains and its catalytic domain was required for enzymatic activity. High throughput mass spectrometry analysis in combination with a screening of an alveolar macrophage library was used to identify and characterize novel UBCv1-host interactors. The analysis revealed interaction with the 40S ribosomal protein RPS23, the cap-dependent translation machinery initiation factor eIF4E, and the E3 ubiquitin ligase Cullin 4B. Our data show that during ASFV infection, UBCv1 was able to bind to eIF4E, independent from the cap-dependent complex. Our results provide novel insights into the function of the viral UBCv1 in hijacking cellular components that impact the mTORC signaling pathway, the regulation of the host translation machinery, and the cellular protein expression during the ASFV lifecycle.

Differential regulation of Kidins220 isoforms in Huntington's disease.

Huntington's disease (HD) is an inherited progressive neurodegenerative disease characterized by brain atrophy particularly in the striatum that produces motor impairment, and cognitive and psychiatric disturbances. Multiple pathogenic mechanisms have been proposed including dysfunctions in neurotrophic support and calpain-overactivation, among others. Kinase D-interacting substrate of 220 kDa (Kidins220), also known as ankyrin repeat-rich membrane spanning (ARMS), is an essential mediator of neurotrophin signaling. In adult brain, Kidins220 presents two main isoforms that differ in their carboxy-terminal length and critical protein-protein interaction domains. These variants are generated through alternative terminal exon splicing of the conventional exon 32 (Kidins220-C32) and the recently identified exon 33 (Kidins220-C33). The lack of domains encoded by exon 32 involved in key neuronal functions, including those controlling neurotrophin pathways, pointed to Kidins220-C33 as a form detrimental for neurons. However, the functional role of Kidins220-C33 in neurodegeneration or other pathologies, including HD, has not been explored. In the present work, we discover an unexpected selective downregulation of Kidins220-C33, in the striatum of HD patients, as well as in the R6/1 HD mouse model starting at early symptomatic stages. These changes are C33-specific as Kidins220-C32 variant remains unchanged. We also find the early decrease in Kidins220-C33 levels takes place in neurons, suggesting an unanticipated neuroprotective role for this isoform. Finally, using ex vivo assays and primary neurons, we demonstrate that Kidins220-C33 is downregulated by mechanisms that depend on the activation of the protease calpain. Altogether, these results strongly suggest that calpain-mediated Kidins220-C33 proteolysis modulates onset and/or progression of HD.

Lipid Exchange Factors at Membrane Contact Sites in African Swine Fever Virus Infection.

African swine fever (ASF) is a hemorrhagic fever of wild and domestic pigs with a high rate of mortality. Originally endemic in Africa, this disease is currently disseminating in Europe and China, causing a large socioeconomic impact. ASF is caused by a DNA virus, African swine fever virus (ASFV). There is no vaccine available against ASFV, limiting the options for disease control. ASFV reorganizes intracellular membranes to generate viral factories (VFs) in order to amplify its genome. However, little is known about the process involved in the formation of these viral replication organelles. Membrane contact sites (MCSs) allow nonvesicular lipids and ion exchange between organelles. Lipid exchange to form VFs apparently requires a number of proteins at MCSs, such as the oxysterol-binding protein (OSBP), the acyl-coenzyme A binding domain containing 3 (ACBD3) and the phosphatidylinositol-phosphate-4-kinase III beta (PI4Kß). Itraconazole (ITZ) is an antifungal agent that targets sterol-transport molecules such as OSBP and OSBP-related protein 4 (ORP4). 25-Hydroxycholesterol (25-HC) inhibits lipid transport by high affinity binding OSBP. In this work, we analyzed the antiviral function of ITZ and 25-HC against ASFV in Vero cell cultures using the cell-adapted Ba71V isolate. ITZ and 25-HC decreased significantly ASFV replication. Our study revealed OSBP distribution in cytoplasmic membranes in uninfected Vero cells and to the periphery of VFs in infected cells. In addition, we showed that OSBP and OSBP-related proteins, PI4Kß and ACBD3 were recruited to VFs in the context ASFV infection.

Author Correction: Excitotoxic inactivation of constitutive oxidative stress detoxification pathway in neurons can be rescued by PKD1.

The original version of this Article contained an error in the spelling of the author Álvaro Sebastián-Serrano, which was incorrectly given as Álvaro Sebastián Serrano. This has now been corrected in both the PDF and HTML versions of the Article.

Excitotoxic inactivation of constitutive oxidative stress detoxification pathway in neurons can be rescued by PKD1.

Excitotoxicity, a critical process in neurodegeneration, induces oxidative stress and neuronal death through mechanisms largely unknown. Since oxidative stress activates protein kinase D1 (PKD1) in tumor cells, we investigated the effect of excitotoxicity on neuronal PKD1 activity. Unexpectedly, we find that excitotoxicity provokes an early inactivation of PKD1 through a dephosphorylation-dependent mechanism mediated by protein phosphatase-1 (PP1) and dual specificity phosphatase-1 (DUSP1). This step turns off the IKK/NF-κB/SOD2 antioxidant pathway. Neuronal PKD1 inactivation by pharmacological inhibition or lentiviral silencing in vitro, or by genetic inactivation in neurons in vivo, strongly enhances excitotoxic neuronal death. In contrast, expression of an active dephosphorylation-resistant PKD1 mutant potentiates the IKK/NF-κB/SOD2 oxidative stress detoxification pathway and confers neuroprotection from in vitro and in vivo excitotoxicity. Our results indicate that PKD1 inactivation underlies excitotoxicity-induced neuronal death and suggest that PKD1 inactivation may be critical for the accumulation of oxidation-induced neuronal damage during aging and in neurodegenerative disorders.

Endocarditis infecciosa derecha en portadores de dispositivos cardiacos: perfil clínico y pronóstico / Right-sided infective endocarditis in cardiac device carriers: Clinical profile and prognosis

Antecedentes y objetivos: Debido al aumento en las indicaciones de implante de dispositivos cardiacos y el envejecimiento de la población, la epidemiología de la endocarditis infecciosa derecha (EID) ha cambiado drásticamente, siendo hoy en día los portadores de dispositivos cardiacos el principal grupo afectado. El objetivo de este trabajo es describir la epidemiología, el perfil clínico y los resultados de la EID en portadores de dispositivos cardiacos. Pacientes y métodos: Se incluyeron episodios de endocarditis infecciosa definitiva diagnosticados consecutivamente en 3 centros terciarios entre marzo de 1995 y septiembre de 2014. Se realizó un análisis retrospectivo de 85 variables, seguimiento de un año y análisis univariante de la mortalidad hospitalaria. Resultados: Entre 1.182 episodios, 100 portadores de dispositivos cardiacos presentaron EID (8,5%). La edad media±DE fue de 67±14 años. Los estafilococos fueron los principales microorganismos responsables (coagulasa negativos 44%, aureus 31%), y de ellos, el 37% eran resistentes a meticilina. Los dispositivos cardiacos fueron retirados en el 95% de los pacientes. La mortalidad hospitalaria y al año fue del 8 y el 4%, respectivamente. El análisis univariante demostró que la insuficiencia renal al ingreso (OR 6,2; IC 95% 1,3-30,3), el shockséptico (OR 8,9; IC 95% 1,7-47,9) y la infección persistente durante la evolución (OR 19,4; IC 95% 3-125,7) aumentan la mortalidad hospitalaria, mientras que la retirada del dispositivo es un factor protector (OR 0,08; IC 95% 0,02-0,39). Conclusiones: La EID sobre dispositivos cardiacos tiene una baja mortalidad intrahospitalaria y al año. Los estafilococos coagulasa negativos son responsables de casi la mitad de los episodios y la incidencia de resistencia a meticilina es elevada. La retirada de los dispositivos disminuye la mortalidad hospitalaria (AU)

Background and objectives: Due to the widespread indications for device implants and the population aging, right-sided infective endocarditis (RSIE) epidemiology has dramatically changed, being nowadays, cardiac device carriers the main affected group. The aim of this work is to describe the epidemiology, clinical profile and outcomes of RSIE in cardiac device carriers. Patients and methods: We included definitive infective endocarditis episodes consecutively diagnosed in 3 tertiary centers from March 1995 to September 2014. A retrospective analysis of 85 variables, one-year follow up and univariate analysis of in-hospital mortality was conducted. Results: Among 1,182 episodes, 100 cardiac device carriers presented with RSIE (8.5%). Mean age±SD was 67±14 years. Staphylococcus spp. were the main causative microorganisms (coagulase-negative 44%, aureus 31%) and 37% were methicillin-resistant. Cardiac devices were removed in 95% of patients. In-hospital mortality was 8% and one-year mortality was 4%. Univariate analysis demonstrated that renal failure at admission (OR 6.2; 95% CI 1.3-30.3), septic shock (OR 8.9; 95% CI 1.7-47.9) and persistent infection during clinical course (OR 19.4; 95% CI 3-125.7) increase in-hospital mortality while device removal is a protective factor (OR 0.08; 95% CI 0.02-0.39). Conclusions: RSIE have low in-hospital and one-year mortality. Coagulase-negative Staphylococci is responsible of almost half of the episodes and methicillin-resistant incidence is high. Device removal is mandatory since it decreases in-hospital mortality (AU)

Right-sided infective endocarditis in cardiac device carriers: Clinical profile and prognosis. / Endocarditis infecciosa derecha en portadores de dispositivos cardiacos: perfil clínico y pronóstico.

BACKGROUND AND OBJECTIVES: Due to the widespread indications for device implants and the population aging, right-sided infective endocarditis (RSIE) epidemiology has dramatically changed, being nowadays, cardiac device carriers the main affected group. The aim of this work is to describe the epidemiology, clinical profile and outcomes of RSIE in cardiac device carriers. PATIENTS AND METHODS: We included definitive infective endocarditis episodes consecutively diagnosed in 3 tertiary centers from March 1995 to September 2014. A retrospective analysis of 85 variables, one-year follow up and univariate analysis of in-hospital mortality was conducted. RESULTS: Among 1,182 episodes, 100 cardiac device carriers presented with RSIE (8.5%). Mean age±SD was 67±14 years. Staphylococcus spp. were the main causative microorganisms (coagulase-negative 44%, aureus 31%) and 37% were methicillin-resistant. Cardiac devices were removed in 95% of patients. In-hospital mortality was 8% and one-year mortality was 4%. Univariate analysis demonstrated that renal failure at admission (OR 6.2; 95% CI 1.3-30.3), septic shock (OR 8.9; 95% CI 1.7-47.9) and persistent infection during clinical course (OR 19.4; 95% CI 3-125.7) increase in-hospital mortality while device removal is a protective factor (OR 0.08; 95% CI 0.02-0.39). CONCLUSIONS: RSIE have low in-hospital and one-year mortality. Coagulase-negative Staphylococci is responsible of almost half of the episodes and methicillin-resistant incidence is high. Device removal is mandatory since it decreases in-hospital mortality.

Eventos adversos en un hospital pediátrico de tercer nivel de Bogotá / Adverse Events in a third level pediatric hospital in Bogotá / Eventos adversos num hospital pediátrico de terceiro nível de Bogotá

Resumen En Colombia la seguridad del paciente es una prioridad en la atención en salud; los eventos adversos e incidentes son una muestra de atención insegura en las instituciones. El compromiso interdisciplinario es fundamental para trabajo proactivo en promoción de prácticas seguras, prevención y detección de fallas latentes y activas en un hospital. Objetivo: determinar la frecuencia de eventos adversos mediante revisión de historias clínicas de un hospital pediátrico de tercer nivel de Bogotá, con el fin de fomentar prácticas seguras. Metodología: Investigación cuantitativa, transversal y descriptiva, dirigida a pacientes hospitalizados el 16 de mayo de 2013, sobre factores de riesgo, antecedentes hospitalarios, eventos adversos y complicaciones relacionados al cuidado de la salud. Resultados: Hubo 169 casos clasificados para algún suceso, de los cuales 59 Eventos Adversos, 57 incidentes, y 53 descartados por falsos positivos. Discusión: Se evidencia responsabilidad de prevención y control de infecciones por los profesionales de la salud, encargados de minimizar el riesgo del paciente vulnerable y, ante todo, proporcionar un manejo y mantenimiento adecuado de dispositivos médicos invasivos. Conclusiones: El 34,4% de historias clínicas presentaban algún tipo de evento adverso o incidente, revelando que algunos de estos fueron causados por la asistencia hospitalaria.

Abstract In Colombia, patients' safety is a priority in healthcare. Adverse events and incidents are an example of unsafe healthcare in institutions. An interdisciplinary commitment is fundamental in proactive work to promote safe practices, prevention, and detection of latent and active failures in hospital. Objective: To determine the frequency of Adverse Events by reviewing clinical histories in a third level pediatric hospital in Bogotá to foster safe practices. Methodology: A descriptive cross-sectional quantitative research was conducted focusing on hospitalized patients on May 16, 2013, working risk factors, hospital backgrounds, adverse events, and healthcare related complications. Results: There were 169 cases classified for a given event, of which 59 were Adverse Events, 57 were incidents and 53 were discarded as false positives. Discussion: Researchers evidenced the responsibility in infection prevention and control by healthcare professionals in charge of minimizing a vulnerable patient's risk, and above all, they provided a suitable use and maintenance of invasive medical devices. Conclusions: 34.4% of the clinical histories show some type of adverse event or incident, revealing evidence that in some cases they were caused by hospital care.

Resumo Na Colômbia a seguridade do paciente é uma prioridade na atenção em saúde; os eventos adversos e incidentes são uma mostra de atenção insegura nas instituições. O compromisso interdisciplinar e fundamental para trabalho proativo em promoção de práticas seguras, prevenção e detecção de falhas latentes e ativas num hospital. Objetivo: Determinar a frequência de eventos adversos mediante revisão de histórias clínicas dum hospital pediátrico de terceiro nível de Bogotá, com o fim de fomentar práticas seguras. Metodologia: Investigação quantitativa, transversal e descritiva, dirigida a pacientes hospitalizados o 16 de maio de 2013, sobre fatores de risco, antecedentes hospitalários, eventos adversos e complicações relacionados ao cuidado da saúde. Resultados: Houve 169 casos classificados para algum acontecimento, dos quais 59 Eventos Adversos, 57 incidentes, e 53 descartados por serem falsos positivos. Discussão: Se evidencia responsabilidade de prevenção e controle de infecções pelos professionais da saúde, encarregados de minimizar o risco do paciente vulnerável e, por acima de todo, proporcionar um manejo e mantimento adequado de dispositivos médicos invasivos. Conclusões: O 34.4% das histórias clínicas apresentavam algum jeito de Evento Adverso ou incidente, revelando evidência de que alguns destes foram causados pela assistência hospitalar.

Cannabinoid Receptors Modulate Neuronal Morphology and AnkyrinG Density at the Axon Initial Segment.

Neuronal polarization underlies the ability of neurons to integrate and transmit information. This process begins early in development with axon outgrowth, followed by dendritic growth and subsequent maturation. In between these two steps, the axon initial segment (AIS), a subcellular domain crucial for generating action potentials (APs) and maintaining the morphological and functional polarization, starts to develop. However, the cellular/molecular mechanisms and receptors involved in AIS initial development and maturation are mostly unknown. In this study, we have focused on the role of the type-1 cannabinoid receptor (CB1R), a highly abundant G-protein coupled receptor (GPCR) in the nervous system largely involved in different phases of neuronal development and differentiation. Although CB1R activity modulation has been related to changes in axons or dendrites, its possible role as a modulator of AIS development has not been yet explored. Here we analyzed the potential role of CB1R on neuronal morphology and AIS development using pharmacological and RNA interference approaches in cultured hippocampal neurons. CB1R inhibition, at a very early developmental stage, has no effect on axonal growth, yet CB1R activation can promote it. By contrast, subsequent dendritic growth is impaired by CB1R inhibition, which also reduces ankyrinG density at the AIS. Moreover, our data show a significant correlation between early dendritic growth and ankyrinG density. However, CB1R inhibition in later developmental stages after dendrites are formed only reduces ankyrinG accumulation at the AIS. In conclusion, our data suggest that neuronal CB1R basal activity plays a role in initial development of dendrites and indirectly in AIS proteins accumulation. Based on the lack of CB1R expression at the AIS, we hypothesize that CB1R mediated modulation of dendritic arbor size during early development indirectly determines the accumulation of ankyrinG and AIS development. Further studies will be necessary to determine which CB1R-dependent mechanisms can coordinate these two domains, and what may be the impact of these early developmental changes once neurons mature and are embedded in a functional brain network.

Mitral Regurgitation After Transcatheter Aortic Valve Replacement: Prognosis, Imaging Predictors, and Potential Management.

OBJECTIVES: This study sought to analyze the clinical impact of the degree and improvement of mitral regurgitation in TAVR recipients, validate the main imaging determinants of this improvement, and assess the potential candidates for double valve repair with percutaneous techniques. BACKGROUND: Many patients with severe aortic stenosis present with concomitant mitral regurgitation (MR). Cardiac imaging plays a key role in identifying prognostic factors of MR persistence after transcatheter aortic valve replacement (TAVR) and for planning its treatment. METHODS: A total of 1,110 patients with severe aortic stenosis from 6 centers who underwent TAVR were included. In-hospital to 6-month follow-up clinical outcomes according to the degree of baseline MR were evaluated. Off-line analysis of echocardiographic and multidetector computed tomography images was performed to determine predictors of improvement, clinical outcomes, and potential percutaneous alternatives to treat persistent MR. RESULTS: Compared with patients without significant pre-TAVR MR, 177 patients (16%) presented with significant pre-TAVR MR, experiencing a 3-fold increase in 6-month mortality (35.0% vs. 10.2%; p < 0.001). After TAVR, the degree of MR improved in 60% of them. A mitral annular diameter of >35.5 mm (odds ratio: 9.0; 95% confidence interval: 3.2 to 25.3; p < 0.001) and calcification of the mitral apparatus by multidetector computed tomography (odds ratio: 11.2; 95% confidence interval: 4.03 to 31.3; p < 0.001) were independent predictors of persistent MR. At least 14 patients (1.3% of the entire cohort, 13.1% of patients with persistent MR) met criteria for percutaneous mitral repair with either MitraClip (9.3%) or a balloon-expandable valve (3.8%). CONCLUSIONS: Significant MR is not uncommon in TAVR recipients and associates with greater mortality. In more than one-half of patients, the degree of MR improves after TAVR, which can be predicted by characterizing the mitral apparatus with multidetector computed tomography. According to standardized imaging criteria, at least 1 in 10 patients whose MR persists after TAVR could benefit from percutaneous mitral procedures, and even more could be treated with MitraClip after dedicated pre-imaging evaluation.

Por un futuro mejor: programa de promoción de la salud para Afrodescendientes / Por um futuro melhor: programa de promoção da saúde para Afro-descendentes / For a better future: program of health promotion for Afro-descend

Introducción: Las familias afrocolombianas se caracterizan por vivir en contextos de alta vulnerabilidad, que requieren su fortalecimiento con programas en promoción de la salud. Objetivo: Construir un programa de promoción de la salud, con familias afrocolombianas en situación de pobreza en Tumaco, año 2012. Materiales y Métodos: Investigación - Acción - Participativa, conformando un grupo y comisión de seguimiento, esto permitió la adaptación de la propuesta de Martí a través de cuatro etapas en el proceso. Resultados: El programa de Promoción de la Salud fue una construcción colectiva entre el grupo Investigación Acción Participación, la comisión de seguimiento y las familias, realizada en cuatro fases: Etapa 1: Inicio y preparación del campo. Etapa 2. Diagnóstico: Aproximación al universo de las familias afrocolombianas. Etapa 3. Construcción y aplicación de programa de promoción de la salud: “Del dicho al hecho superamos los trechos”. Etapa 4. Evaluación y proyección del programa: “Evaluando vamos terminando”. Discusión y Conclusiones: La construcción social del programa de promoción de la salud, así como el abordaje pedagógico y didáctico para el desarrollo del mismo, se desarrolló en función del contexto cultural, desde la perspectiva de la Investigación Acción Participación. El programa “Por un futuro mejor” permitió crear nuevos conocimientos a familias, líderes y profesionales de la salud a partir de la realidad e intercambio de saberes, con acciones dirigidas a fortalecer el amor propio, la comunicación, el entorno y la participación, con el fin de mejorar las condiciones de salud.

Introdução: As famílias afro-colombianas caracterizam-se por viver em contextos de alta vulnerabilidade, que requerem seu fortalecimento com programas em promoção da saúde. Objetivo: Construir um programa da promoção da saúde, com famílias afro-colombianas em situação de pobreza em Tumaco, ano 2012. Materiais e Métodos: Investigação-Ação-Participativa, formando um grupo e a comissão de seguimento, este permitiu a adaptação da proposta do Martí através de quatro etapas no processo. Resultados: O programa de Promoção da Saúde foi uma construção coletiva entre o Grupo Investigação Ação Participação, a comissão de seguimento e as famílias, realizado em quatro fases: Etapa 1: Inicio e preparação do campo. Etapa 2. Diagnóstico: Aproximação ao universo das famílias afro-colombianas. Etapa 3. Construção e aplicação do programa de promoção da saúde: “Do dizer ao fazer, superamos as distâncias”. Passo 4. Avaliação e projeção do programa: "Avaliando vamos terminando" Discussão e Conclusões: A construção social do programa de promoção da saúde, bem como a abordagem pedagógica e didática para o desenvolvimento do mesmo, desenvolveu-se em função do contexto cultural, desde a perspectiva da Investigação Ação Participação. O programa “Por um futuro melhor” permitiu criar novos conhecimentos às famílias, líderes e profissionais da saúde a partir da realidade e intercâmbio de saberes, com ações dirigidas a fortalecer o amor próprio, a comunicação, o entorno e a participação, com o fim de melhorar as condições de saúde.

Introduction: Afro-Colombian families live in contexts characterized by high vulnerability that require strengthening programs in health promotion. Objective: To build a program of health promotion, with Afro-Colombian families in poverty in Tumaco, 2012. Materials and Methods: Research - Action - Participatory, forming a group and monitoring committee, this allowed the adaptation of the proposal Martí through four stages in the process. Results: The program of Health Promotion was a collective effort between the Research Action Group Participation, the monitoring committee and families, carried out in four stages: Stage 1: Initiation and preparation of the field. Step 2. Diagnosis: An approach to the universe of Afro-Colombian families. Stage 3. Construction and application of program health promotion: "Easier said than done overcome the stretches". Step 4. Evaluation and projection of the program: "Evaluating we finished." Discussion and Conclusions: The social construction of program health promotion as well as the educational and training to develop the same approach, developed depending on the cultural context, from the perspective of Action Research Participation. The "For a better future" allowed families to create new knowledge, leaders and health professionals from reality and knowledge exchange, with actions aimed at strengthening self-esteem, communication, the environment and participation, to improve health conditions.