Sphingosine-1-phosphate and ceramide-1-phosphate promote migration, pro-inflammatory and pro-fibrotic responses in retinal pigment epithelium cells.

Retinal pigment epithelium (RPE) cells, essential for preserving retina homeostasis, also contribute to the development of retina proliferative diseases, through their exacerbated migration, epithelial to mesenchymal transition (EMT) and inflammatory response. Uncovering the mechanisms inducing these changes is crucial for designing effective treatments for these pathologies. Sphingosine-1-phosphate (S1P) and ceramide-1-phosphate (C1P) are bioactive sphingolipids that promote migration and inflammation in several cell types; we recently established that they stimulate the migration of retina Müller glial cells (Simón et al., 2015; Vera et al., 2021). We here analyzed whether S1P and C1P regulate migration, inflammation and EMT in RPE cells. We cultured two human RPE cell lines, ARPE-19 and D407 cells, and supplemented them with either 5 µM S1P or 10 µM C1P, or their vehicles, for 24 h. Analysis of cell migration by the scratch wound assay showed that S1P addition significantly enhanced migration in both cell lines. Pre-treatment with W146 and BML-241, antagonists for S1P receptor 1 (S1P1) and 3 (S1P3), respectively, blocked exogenous S1P-induced migration. Inhibiting sphingosine kinase 1 (SphK1), the enzyme involved in S1P synthesis, significantly reduced cell migration and exogenous S1P only partially restored it. Addition of C1P markedly stimulated cell migration. Whereas inhibiting C1P synthesis did not affect C1P-induced migration, inhibiting S1P synthesis strikingly decreased it; noteworthy, addition of C1P promoted the transcription of SphK1. These results suggest that S1P and C1P stimulate RPE cell migration and their effect requires S1P endogenous synthesis. Both S1P and C1P increase the transcription of pro-inflammatory cytokines IL-6 and IL-8, and of EMT marker α-smooth muscle actin (α-SMA) in ARPE-19 cells. Collectively, our results suggest new roles for S1P and C1P in the regulation of RPE cell migration and inflammation; since the deregulation of sphingolipid metabolism is involved in several proliferative retinopathies, targeting their metabolism might provide new tools for treating these pathologies.

Ceramide-1-phosphate promotes the migration of retina Müller glial cells.

Müller glial cells, the major glial cell type in the retina, are activated by most retina injuries, leading to an increased proliferation and migration that contributes to visual dysfunction. The molecular cues involved in these processes are still ill defined. We demonstrated that sphingosine-1-phosphate (S1P), a bioactive sphingolipid, promotes glial migration. We now investigated whether ceramide-1-phosphate (C1P), also a bioactive sphingolipid, was involved in Müller glial cell migration. We evaluated cell migration in primary Müller glial cultures, prepared from newborn rat retinas, by the scratch wound assay. Addition of either 10 µM C8-ceramide-1-phosphate (C8-C1P) or 5 µM C16-C1P (a long chain, natural C1P) stimulated glial migration. Inhibiting PI3K almost completely blocked C8-C1P-elicited migration whereas inhibition of ERK1-2/MAPK pathway diminished it and p38MAPK inhibition did not affect it. Pre-treatment with a cytoplasmic phospholipase A2 (cPLA2) inhibitor markedly reduced C8-C1P-induced migration. Inhibiting ceramide kinase (CerK), the enzyme catalyzing C1P synthesis, partially decreased glial migration. Combined addition of S1P and C8-C1P promoted glial migration to the same extent as when they were added separately, suggesting they converge on their downstream signaling to stimulate Müller glia migration. These results suggest that C1P addition stimulated migration of glial Müller cells, promoting the activation of cPLA2, and the PI3K and ERK/MAPK pathways. They also suggest that CerK-dependent C1P synthesis was one of the factors contributing to glial migration, thus uncovering a novel role for C1P in controlling glial motility.

Sphingolipids as Emerging Mediators in Retina Degeneration.

The sphingolipids ceramide (Cer), sphingosine-1-phosphate (S1P), sphingosine (Sph), and ceramide-1-phosphate (C1P) are key signaling molecules that regulate major cellular functions. Their roles in the retina have gained increasing attention during the last decade since they emerge as mediators of proliferation, survival, migration, neovascularization, inflammation and death in retina cells. As exacerbation of these processes is central to retina degenerative diseases, they appear as crucial players in their progression. This review analyzes the functions of these sphingolipids in retina cell types and their possible pathological roles. Cer appears as a key arbitrator in diverse retinal pathologies; it promotes inflammation in endothelial and retina pigment epithelium (RPE) cells and its increase is a common feature in photoreceptor death in vitro and in animal models of retina degeneration; noteworthy, inhibiting Cer synthesis preserves photoreceptor viability and functionality. In turn, S1P acts as a double edge sword in the retina. It is essential for retina development, promoting the survival of photoreceptors and ganglion cells and regulating proliferation and differentiation of photoreceptor progenitors. However, S1P has also deleterious effects, stimulating migration of Müller glial cells, angiogenesis and fibrosis, contributing to the inflammatory scenario of proliferative retinopathies and age related macular degeneration (AMD). C1P, as S1P, promotes photoreceptor survival and differentiation. Collectively, the expanding role for these sphingolipids in the regulation of critical processes in retina cell types and in their dysregulation in retina degenerations makes them attractive targets for treating these diseases.

Ceramide Induces the Death of Retina Photoreceptors Through Activation of Parthanatos.

Ceramide (Cer) has a key role inducing cell death and has been proposed as a messenger in photoreceptor cell death in the retina. Here, we explored the pathways induced by C2-acetylsphingosine (C2-Cer), a cell-permeable Cer, to elicit photoreceptor death. Treating pure retina neuronal cultures with 10 µM C2-Cer for 6 h selectively induced photoreceptor death, decreasing mitochondrial membrane potential and increasing the formation of reactive oxygen species (ROS). In contrast, amacrine neurons preserved their viability. Noteworthy, the amount of TUNEL-labeled cells and photoreceptors expressing cleaved caspase-3 remained constant and pretreatment with a pan-caspase inhibitor did not prevent C2-Cer-induced death. C2-Cer provoked polyADP ribosyl polymerase-1 (PARP-1) overactivation. Inhibiting PARP-1 decreased C2-Cer-induced photoreceptor death; C2-Cer increased polyADP ribose polymer (PAR) levels and induced the translocation of apoptosis inducing factor (AIF) from mitochondria to photoreceptor nuclei, which was prevented by PARP-1 inhibition. Pretreatment with a calpain and cathepsin inhibitor and with a calpain inhibitor reduced photoreceptor death, whereas selective cathepsin inhibitors granted no protection. Combined pretreatment with a PARP-1 and a calpain inhibitor evidenced the same protection as each inhibitor by itself. Neither autophagy nor necroptosis was involved in C2-Cer-elicited death; no increase in LDH release was observed upon C2-Cer treatment and pretreatment with inhibitors of necroptosis and autophagy did not rescue photoreceptors. These results suggest that C2-Cer induced photoreceptor death by a novel, caspase-independent mechanism, involving activation of PARP-1, decline of mitochondrial membrane potential, calpain activation, and AIF translocation, all of which are biochemical features of parthanatos.

Primary health care: the experience of nurses.

OBJECTIVE: to understand the meaning of nursing care in primary health care from the perspective of Chilean nurses. METHOD: this was a qualitative study based on the social phenomenology of Alfred Schutz. Data was collected between January and April 2013, through interviews with 13 primary health care nurses in Chile. RESULTS: the nurses perceived primary care as a gratifying experience, considering it an encounter of subjectivities. However, they felt burdened with multiple functions and by the hierarchical pressure to achieve targets. They strived to implement innovative care, expressed by the desire to go beyond traditional care practices, and improve the efficiency of management at the various levels of health care. CONCLUSION: it is important to discuss the results of the present study in the context of health care and especially nursing education, with the goal of better preparing nurses who will deliver care at the primary health care level.

Primary health care: the experience of nurses / O cuidado na Atenção Primária à Saúde: a vivência de enfermeiras / El cuidado en la Atención Primaria de Salud: vivencia de las enfermeras

ABSTRACT Objective: to understand the meaning of nursing care in primary health care from the perspective of Chilean nurses. Method: this was a qualitative study based on the social phenomenology of Alfred Schutz. Data was collected between January and April 2013, through interviews with 13 primary health care nurses in Chile. Results: the nurses perceived primary care as a gratifying experience, considering it an encounter of subjectivities. However, they felt burdened with multiple functions and by the hierarchical pressure to achieve targets. They strived to implement innovative care, expressed by the desire to go beyond traditional care practices, and improve the efficiency of management at the various levels of health care. Conclusion: it is important to discuss the results of the present study in the context of health care and especially nursing education, with the goal of better preparing nurses who will deliver care at the primary health care level.

RESUMO Objetivo: compreender o significado atribuído aos cuidados de enfermagem na Atenção Básica de Saúde por enfermeiras chilenas. Método: pesquisa qualitativa baseada na fenomenologia social de Alfred Schutz. Os dados foram coletados entre janeiro e abril de 2013, por meio de entrevistas com 13 enfermeiras de unidades de Atenção Básica em uma cidade do Chile. Resultados: as enfermeiras percebem o cuidado prestado como uma experiência gratificante, considerando-o um encontro de subjetividades. No entanto, elas se sentem sobrecarregadas pelas múltiplas funções que realizam e pelas pressões hierárquicas para cumprir metas. Elas se esforçam para implementar cuidados inovadores que possam superar o cuidado tradicional e aumentar a eficiência da gestão nos vários níveis de atenção à saúde. Conclusão: é importante discutir estes resultados no contexto da assistência e especialmente na formação, com o objetivo de melhorar a preparação de enfermeiros trabalhando nesse nível de atenção.

RESUMEN Objetivo: comprender el significado de realizar cuidados de enfermería en la Atención Primaria de Salud desde la perspectiva de las enfermeras chilenas. Método: investigación cualitativa con enfoque desde la fenomenología social de Alfred Schütz. La recolección de datos fue realizada entre enero y abril del 2013, a través de entrevista con 13 enfermeras de Atención Primaria en Chile. Resultados: las enfermeras perciben el cuidado como una experiencia gratificante considerando un encuentro de subjetividades. Sin embargo, se sienten sobrecargadas por múltiples funciones a realizar y por presiones jerárquicas en el logro de las metas. Aspiran implementar cuidados innovadores manifiestados por el deseo de superar el cuidado tradicional y la eficiencia de la gestión de los diversos niveles de atención de la salud. Conclusión: es importante discutir estos resultados en el contexto de la asistencia y especialmente en la formación, con el fin de preparar mejor a las enfermeras que brindarán cuidados en este nivel de atención.

Desempeño de las enfermeras en Atención Primaria de Salud: un Enfoque Fenomenológico / Performance of primary health care nurses: a phenomenological approach

Se buscó comprender el desempeño de las enfermeras chilenas en la Atención Primaria de Salud. Investigación cualitativa con enfoque desde la fenomenología social. La recolección de datos fue realizada entre enero y abril del 2013, a través de entrevista, con 13 enfermeras. El desempeño de las enfermeras muestra que ellas se ven con más autonomia profesional, mayor cercania a los usuarios, sin embargo, se sienten desgastadas en el trabajo. Sus expectativas incluyen el deseo de reconocimiento de su labor y una mejor gestión del servicio. Este estudio aporta como contribuciones la posibilidad de pensar nuevos diseños para las prácticas profesionales y de gestión en la Atención Primaria de Salud, desde la perspectiva de las enfermeras chilenas. Estos diseños se deben basar en la reorganización del proceso de trabajo y alcanzar los espacios de la macropolítica, influyendo el desempeño profesional de la enfermera en este nivel de atención de salud.

The objective of this study was to identify the performance of Chilean primary health care nurses. This was a qualitative study based on the approach of social phenomenology. Data collection was conducted between January and April 2013 through interviews with 13 nurses. Nurse performance showed that they experience greater professional autonomy and are closer to users; however, they suffered from professional burnout. Their expectations included the desire to be recognized for their work and better service management. Contributions of this study include the possibility of considering new designs for professional management practices in primary health care from the perspective of Chilean nurses. These designs should be based on the reorganization of work processes and by occupying spaces in macropolitics, influencing the professional performance of nurses at this level of health care.

Treatment in vitro with PPARα and PPARγ ligands drives M1-to-M2 polarization of macrophages from T. cruzi-infected mice.

Trypanosoma cruzi, the etiological agent of Chagas' disease, induces a persistent inflammatory response. Macrophages are a first line cell phenotype involved in the clearance of infection. Upon parasite uptake, these cells increase inflammatory mediators like NO, TNF-α, IL-1ß and IL-6, leading to parasite killing. Although desired, inflammatory response perpetuation and exacerbation may lead to tissue damage. Peroxisome proliferator-activated receptors (PPARs) are ligand-dependent nuclear transcription factors that, besides regulating lipid and carbohydrate metabolism, have a significant anti-inflammatory effect. This is mediated through the interaction of the receptors with their ligands. PPARγ, one of the PPAR isoforms, has been implicated in macrophage polarization from M1, the classically activated phenotype, to M2, the alternatively activated phenotype, in different models of metabolic disorders and infection. In this study, we show for the first time that, besides PPARγ, PPARα is also involved in the in vitro polarization of macrophages isolated from T. cruzi-infected mice. Polarization was evidenced by a decrease in the expression of NOS2 and proinflammatory cytokines and the increase in M2 markers like Arginase I, Ym1, mannose receptor and TGF-ß. Besides, macrophage phagocytic activity was significantly enhanced, leading to increased parasite load. We suggest that modulation of the inflammatory response by both PPARs might be due, at least in part, to a change in the profile of inflammatory macrophages. The potential use of PPAR agonists as modulators of overt inflammatory response during the course of Chagas' disease deserves further investigation.

Percepción de la calidad del cuidado de enfermería en pacientes oncológicos hospitalizados / Perception of quality of nursing care in patients hospitalized in oncology services

Introducción: Se presenta la evaluación de la calidad del cuidado de enfermería desde la percepción de los pacientes oncológicos hospitalizados en una IPS de último nivel. Objetivo: Determinar la percepción de la calidad del cuidado según la dimensión de satisfacción y experiencia con los cuidados en los pacientes que reciben tratamiento en la unidad de oncología de una institución de salud. Materiales y Métodos: Estudio descriptivo de corte transversal, con pacientes hospitalizados en la unidad de oncología de una institución de salud, a través de la aplicación del Cuestionario de calidad de cuidados de enfermería (Cucace), versión en castellano de Rosario Alonso y colaboradores, 2005, versión en inglés: NSNS (Newcastle Satisfacción with Nursing Scales) de Thomas LH, 1996; basado en las dimensiones de experiencias y satisfacción con el cuidado de enfermería. Resultados: Se obtuvo un alfa de Cronbach de 0,96. Los pacientes del estudio están conformes con el cuidado y la atención brindada por las enfermeras durante su estancia en la terapia. La valoración que dieron los pacientes sobre el cuidado recibido de parte de las enfermeras en general fue excelente con un porcentaje de 58.7%, seguido de muy bueno con 29.3%, bueno con 10.7% y regular con 1.3%. Se obtuvo una puntuación media de 80.4 sobre 100 para la dimensión de experiencia con el cuidado y de 82.7 para la dimensión de la satisfacción con los cuidados. Discusión y Conclusiones: El proceso y desarrollo de esta investigación demostró que el Cuestionario de percepción de la calidad del cuidado de enfermería, CUCACE en sus dos dimensiones de experiencia y satisfacción de los cuidados fue asimilado fácilmente por los pacientes hospitalizados en el área oncológica de la FOSCAL, siendo entendible y comprensible, con un buen nivel de confiabilidad lo cual lo hace adecuado para la evaluación de la satisfacción y la experiencia del paciente hospitalizado y así medir la calidad del cuidado brindado por enfermería.

Introduction: It presents the evaluation of nursing care quality from the perception of cancer patients hospitalized in an end-level IPS. Objective: To determine the perceived quality of care depending on the size and experience satisfaction with care in patients receiving treatment in the oncology unit of a health institution. Materials and Methods: Cross sectional study with patients hospitalized in the oncology unit of a health institution, through the application of the questionnaire for nursing care quality (Cucace), Castilian version of Rosario Alonso et al, 2005, english version: NSNS (Newcastle Satisfaction with Nursing Scales) LH Thomas, 1996, based on the dimensions of experience and satisfaction with nursing care. Results: Cronbach alpha of 0.96. Study patients are satisfied with the care and attention given by nurses during their stay in therapy. The assessment given by the patients about the care received from nurses in general was excellent with a percentage of 58.7%, followed with 29.3% very good, good with 10.7% and 1.3% regular. We obtained a mean score of 80.4 out of 100 for the dimension of experience with the care and 82.7 for the dimension of satisfaction with care. Discussion and Conclusions: The process research and development of this questionnaire showed that the perceived quality of nursing care, CUCACE in its two dimensions of experience and satisfaction of care was easily assimilated by patients hospitalized in the oncology FOSCAL of being understood and understandable, with a good level of reliability which makes it suitable for the evaluation of satisfaction and inpatient experience and thus measure the quality of care provided by nurses.

Prevalencia del alcoholismo, comportamiento familiar y laboral, del personal policial que presta servicio en la unidad de vigilancia norte de la ciudad de Quito: 2008 / The prevalence of alcoholism, family and professional conduct of police personnel serving in the monitoring unit north of the city of Quito: 2008

El alcoholismo es una enfermedad crónica, progresiva y a menudo mortal. El alcohol es una droga que por su fácil acceso y poderosa propaganda que recibe, se ha convertido en un verdadero problema social en casi todos los países y en todas las edades a partir de la adolescencia. Las consecuencias por el consumo excesivo pueden ser graves e irreversibles. La presente investigación estuvo orientada a comprobar la prevalencia del alcoholismo como una enfermedad crónica marcada por la dependencia en el consumo del alcohol, dentro del ambiente laboral, sus consecuencias en las relaciones intrafamiliares, del personal de Oficiales, Clases, y Policías, que prestan sus servicios en la Unidad de Vigilancia Norte de Quito de la Policía Nacional. Se realizó un estudio epidemiológico analítico transversal en 176 policías que prestan sus servicios en la Unidad de Vigilancia Norte de la ciudad de Quito, con el propósito de identificar las dimensiones de la realidad del alcoholismo y su repercusión a nivel interpersonal y familiar en el segundo semestre del 2008. Los principales hallazgos encontrados fueron: la edad promedio de los policías es de ±...33 años, prevalece el género masculino con 90,3%, el estado civil de los policías fue el 44,9% casados, residen en Quito el 72,2%, la mayoría proceden de otras ciudades el 62%, con una frecuencia del consumo de alcohol más de 10 copas una vez al mes el 46%, los policías califican el ambiente laboral bueno y las relaciones familiares como muy buenas, disfrutan con su familia a diario el 43,8%, el 63,1% no tienen conflictos con su pareja por participar en reuniones sociales, el 81,2% no presentan conflictos con sus superiores por la ingesta de alcohol.