Caracterización de biomarcadores en la recurrencia postquirúrgica de la enfermedad de Crohn : valor predictivo y/o implicación patogénica de los mismos

La recurrència postquirúrgica en els pacients amb malaltia de Crohn intervinguts és molt freqüent i, al seu torn, té implicacions terapèutiques importants. No s'han caracteritzat fins ara marcadors biològics que, pel seu valor predictiu, poguessin ser d'utilitat en el maneig postquirúrgic d'aquests pacients. En aquest estudi s'analitza el perfil evolutiu després de la cirurgia de diverses citocines (IL-2, IL-6, IL-10, IL1B, IFN-g i TNF-a), marcadors fecals (calprotectina) i paràmetres serològics (proteïna C reactiva, fibrinogen), per determinar la seva possible utilitat com a marcadors predictius de recurrència després de la cirurgia resectiva.La recurrencia postquirúrgica en los pacientes con enfermedad de Crohn intervenidos es muy frecuente y, a su vez, tiene implicaciones terapéuticas importantes. No se han caracterizado hasta la fecha marcadores biológicos que, por su valor predictivo, pudieran ser de utilidad en el manejo postquirúrgico de estos pacientes. En el presente estudio se analiza el perfil evolutivo tras la cirugía de diversas citoquinas (IL-2, IL-6, IL-10, IL1B, IFN-g y TNF-a), marcadores fecales (calprotectina) y parámetros serológicos (proteína C reactiva, fibrinógeno), para determinar su posible utilidad como marcadores predictivos de recurrencia tras la cirugía resectiva

Possible Biomarkers in Blood for Crohn’s Disease: Oxidative Stress and MicroRNAs—Current Evidences and Further Aspects to Unravel

Crohn’s disease (CD) is an inflammatory disorder characterised by a transmural inflammation of the intestinal wall. Although the physiopathology of the disease is not yet fully understood, it is clear that the immune response plays an important role in it. This hyperreactive immune system is accompanied by the presence of unregulated reactive oxygen species (ROS). These elements are modulated in normal conditions by different elements, including enzymes that function as antioxidant defences preventing the harmful effects of ROS. However, in CD there is an imbalance between ROS production and these antioxidant elements, resulting in oxidative stress (OxS) phenomena. In fact, now OxS is being considered more a potential etiological factor for Crohn’s disease rather than a concomitant effect in the disease. The persistence of the OxS can also be influencing the evolution of the disease. Furthermore, the epigenetic mechanisms, above all microRNAs, are being considered key elements in the pathogenesis of CD. These elements and the presence of OxS have also been linked to several diseases. We, therefore, describe in this review the most significant findings related to oxidative stress and microRNAs profiles in the peripheral blood of CD patients

Role of oxidative stress and antioxidant enzymes in Crohn’s disease,”

Abstract There is increasing interest in oxidative stress being a potential aetiological factor and/or a triggering factor in Crohn's disease, rather than a concomitant occurrence during the pathogenesis of the disease. Recent research has shown that the immune mononuclear cells of Crohn's disease patients are induced to produce hydrogen peroxide (H 2 O 2 ). Similarly, the regulation of antioxidant enzymes during disease in these cells has been unravelled, showing that SOD (superoxide dismutase) activity and GPx (glutathione peroxidase) activity is increased during active disease and returns to normal in remission phases. However, catalase remains constantly inhibited which supports the idea that catalase is not a redox-sensitive enzyme, but a regulator of cellular processes. ROS (reactive oxygen species) can be produced under the stimulus of different cytokines such as TNFα (tumour necrosis factor α). It has been shown in different experimental models that they are also able to regulate apoptosis and other cellular processes. The status of oxidative stress elements in Crohn's disease and their possible implications in regulating cellular processes are reviewed in the present paper

Influence of Neospora caninum intra-specific variability in the outcome of infection in a pregnant BALB/c mouse model

Previous assays in pregnant animals have demonstrated the effect of different host factors and timing of infection on the outcome of neosporosis during pregnancy. However, the influence of Neospora caninum isolate itself has been poorly investigated. Here, we compared the effects on clinical outcome and vertical transmission observed in a pregnant mouse model following infection with 10 different N. caninum isolates. The isolates in our study included the Nc-Liv isolate and nine N. caninum isolates obtained from calves. Female BALB/c mice were inoculated with 2 × 106 tachyzoites at day 7 of pregnancy. Morbidity and mortality, in both dams and offspring during the course of infection, and transmission to progeny at day 30 postpartum were evaluated. The serum IgG1 and IgG2a production in dams were also examined. All dams showed elevated IgG1 and IgG2a responses, confirming N. caninum infection, although signs of disease were only exhibited in dams infected with 4 of the 10 isolates (Nc-Spain 4H, Nc-Spain 5H, Nc-Spain 7 and Nc-Liv). In neonates, clinical signs were observed in all N. caninum-infected groups, and neonatal mortality rates varied from greater than 95% with the isolates mentioned above to less than 32.5% with the other isolates. Vertical transmission rates, as assessed by parasite PCR-detection in neonate brains, also varied from 50% to 100% according to the isolate implicated. These results confirm the wide pathogenic and transmission variability of N. caninum. The intra-specific variability observed herein could help us explain the differences in the outcome of the infection in the natural host

MICa/b-dependent activation of natural killer cells by CD64+ inflammatory type 2 dendritic cells contributes to autoimmunity

Artículo escrito por un elevado número de autores, solo se referencian el que aparece en primer lugar, el nombre del grupo de colaboración, si le hubiere, y los autores pertenecientes a la UAMPrimary Sjögren's syndrome (pSS) is an inflammatory autoimmune disorder largely mediated by type I and II interferon (IFN). The potential contribution of innate immune cells, such as natural killer (NK) cells and dendritic cells (DC), to the pSS pathology remains understudied. Here, we identified an enriched CD16+ CD56hi NK cell subset associated with higher cytotoxic function, as well as elevated proportions of inflammatory CD64+ conventional dendritic cell (cDC2) subtype that expresses increased levels of MICa/b, the ligand for the activating receptor NKG2D, in pSS individuals. Circulating cDC2 from pSS patients efficiently induced activation of cytotoxic NK cells ex vivo and were found in proximity to CD56+ NK cells in salivary glands (SG) from pSS patients. Interestingly, transcriptional activation of IFN signatures associated with the RIG-I/DDX60 pathway, IFN I receptor, and its target genes regulate the expression of NKG2D ligands on cDC2 from pSS patients. Finally, increased proportions of CD64hi RAE-1+ cDC2 and NKG2D+CD11b+CD27+ NK cells were present in vivo in the SG after poly I:C injection. Our study provides novel insight into the contribution and interplay of NK and cDC2 in pSS pathology and identifies new potential therapy targetsRTI2018-097485-A-I00, PID2021-127899OB-I0

MICa/b-dependent activation of natural killer cells by CD64+ inflammatory type 2 dendritic cells contributes to autoimmunity.

Primary Sjögren's syndrome (pSS) is an inflammatory autoimmune disorder largely mediated by type I and II interferon (IFN). The potential contribution of innate immune cells, such as natural killer (NK) cells and dendritic cells (DC), to the pSS pathology remains understudied. Here, we identified an enriched CD16+ CD56hi NK cell subset associated with higher cytotoxic function, as well as elevated proportions of inflammatory CD64+ conventional dendritic cell (cDC2) subtype that expresses increased levels of MICa/b, the ligand for the activating receptor NKG2D, in pSS individuals. Circulating cDC2 from pSS patients efficiently induced activation of cytotoxic NK cells ex vivo and were found in proximity to CD56+ NK cells in salivary glands (SG) from pSS patients. Interestingly, transcriptional activation of IFN signatures associated with the RIG-I/DDX60 pathway, IFN I receptor, and its target genes regulate the expression of NKG2D ligands on cDC2 from pSS patients. Finally, increased proportions of CD64hi RAE-1+ cDC2 and NKG2D+ CD11b+ CD27+ NK cells were present in vivo in the SG after poly I:C injection. Our study provides novel insight into the contribution and interplay of NK and cDC2 in pSS pathology and identifies new potential therapy targets.S

Endoscopic treatment (endoscopic balloon dilation/self-expandable metal stent) vs surgical resection for the treatment of de novo stenosis in Crohn's disease (ENDOCIR study): an open-label, multicentre, randomized trial.

Background: Stenosis is one of the most common complications in patients with Crohn's disease (CD). Endoscopic balloon dilation (EBD) is the treatment of choice for a short stenosis adjacent to the anastomosis from previous surgery. Self-expandable metal stents (SEMS) may be a suitable treatment option for longer stenoses. To date, however, there is no scientific evidence as to whether endoscopic (EBD/SEMS) or surgical treatment is the best approach for de novo or primary stenoses that are less than 10 cm in length. Methods/design: Exploratory study as "proof-of-concept", multicentre, open-label, randomized trial of the treatment of de novo stenosis in the CD; endoscopic treatment (EBD/SEMS) vs surgical resection (SR). The type of endoscopic treatment will initially be with EDB; if a therapeutic failure occurs, then a SEMS will be placed. We estimate 2 years of recruitment and 1 year of follow-up for the assessment of quality of life, costs, complications, and clinical recurrence. After the end of the study, patients will be followed up for 3 years to re-evaluate the variables over the long term. Forty patients with de novo stenosis in CD will be recruited from 15 hospitals in Spain and will be randomly assigned to the endoscopic or surgical treatment groups. The primary aim will be the evaluation of the patient quality of life at 1 year follow-up (% of patients with an increase of 30 points in the 32-item Inflammatory Bowel Disease Questionnaire (IBDQ-32). The secondary aim will be evaluation of the clinical recurrence rate, complications, and costs of both treatments at 1-year follow-up. Discussion: The ENDOCIR trial has been designed to determine whether an endoscopic or surgical approach is therapeutically superior in the treatment of de novo stenosis in CD

Programa nacional de mejora y conservación de los recursos genéticos de la encina y el alcornoque frente a la seca

Como respuesta a la gravedad de los procesos de decaimiento y mortalidad de la encina y el alcornoque, que amenazan la sostenibilidad de sus masas en la Península Ibérica, en el año 2019 se constituyó el subgrupo de trabajo “Mejora genética y fisiológica”, que forma parte del grupo de trabajo “Seca”, coordinado por el MITECO. Este subgrupo incluye a científicos y técnicos de varias administraciones y numerosos centros de investigación y Universidades y una empresa pública, expertos en mejora genética, fitopatología, ecología, propagación vegetativa, bioquímica y biología molecular. El programa tiene como objetivo la selección de genotipos de Quercus ilex y Q. suber tolerantes al estrés hídrico y a la podredumbre radical provocada por Phytophthora cinnamomi. A partir del estudio de 18 poblaciones y de 194 árboles “escape” seleccionados en focos de seca, se espera seleccionar un material apto para restaurar zonas afectadas por problemas de decaimiento. Se presenta una síntesis de las primeras actividades de caracterización e identificación de árboles escape en focos de seca, ensayos de invernadero para el estudio de la variabilidad poblacional en cuanto a vigor y tolerancia al estrés, recogida de muestras vegetales y edáficas para el análisis genético, molecular y de microbioma, identificación de marcadores moleculares asociados a resiliencia y micropropagación del material de mayor valor potencial

Caracterización de biomarcadores en la recurrencia post-quirúrgica de la enfermedad de Crohn: valor predictivo e implicación patogénica

A pesar de un conocimiento cada vez mayor sobre la patogenia de la enfermedad de Crohn (EC) y el desarrollo de esquemas de tratamiento más potentes que incluyen los fármacos biológicos, las tasas de cirugía en estos pacientes siguen siendo elevadas. Así, un 50-60% de los pacientes con EC requieren de una resección intestinal en los primeros 10 años tras el diagnóstico, y un 25% de éstos acabarán requiriendo de una segunda cirugía dentro de los 5 años siguientes. Dado que la cirugía no supone un tratamiento curativo, la reaparición de lesiones tras la resección de todo el tramo intestinal afecto (fenómeno conocido como recurrencia post-quirúrgica) es muy frecuente en estos pacientes, a pesar incluso de la intervención terapéutica. Entre los tratamientos empleados para prevenir la recurrencia de la EC se incluyen la mesalazina, los inmunomoduladores tiopurínicos y en la actualidad los fármacos biológicos o anti-TNF. En los últimos años, diferentes estudios han confirmado la eficacia del tratamiento biológico en la prevención de la recurrencia, especialmente en pacientes de alto riesgo, con una tendencia actual a su introducción precoz tras la cirugía. Sin embargo, estos tratamientos implican una tasa nada desdeñable de efectos adversos, además de un elevado coste por paciente. Por otro lado, no está claramente establecido qué pacientes se benefician de un tratamiento más intensivo tras la cirugía ni la duración de mantenimiento del mismo. Por todo ello, surge la necesidad de desarrollar nuevas herramientas y estrategias que permitan identificar el riesgo de una recurrencia precoz tras la cirugía y optimizar el manejo clínico de estos pacientes. En el momento actual, la ileocolonoscopia continúa siendo el gold estándar para el seguimiento y diagnóstico de la recurrencia, a pesar de suponer una técnica invasiva, costosa, no exenta de riesgos y en general mal tolerada por los pacientes. Esto es así porque hasta el momento no disponemos de buenos marcadores predictores de recurrencia tras la cirugía, y porque la utilidad de la valoración clínica y analítica para el diagnóstico de la recurrencia es limitada. Por este motivo, existe especial interés en el empleo de biomarcadores no invasivos y de fácil obtención, que sirvan para monitorizar a estos pacientes tras la cirugía y permitan detectar precozmente el desarrollo de recurrencia. En este sentido, entre los marcadores fecales, la calprotectina ha mostrado previamente gran utilidad clínica en diferentes escenarios de la enfermedad y, aunque su papel en el contexto post-quirúrgico no ha sido evaluado en profundidad en series amplias de carácter prospectivo, la evidencia disponible sugiere también su utilidad como marcador de recurrencia, aunque quedan aspectos por definir con claridad para su aplicación en la práctica. La existencia de otros posibles biomarcadores en sangre periférica tampoco ha sido explorada en este escenario de la EC. Por otro lado, el escenario clínico de la recurrencia supone una condición patogénica de novo, por lo que los pacientes con EC intervenidos constituyen el mejor modelo natural in vivo para poder estudiar e identificar los posibles mecanismos patogénicos implicados en la reaparición de la enfermedad. Con todo ello, el objetivo de la presente tesis doctoral se centra en la caracterización prospectiva de marcadores no invasivos, a partir de muestras biológicas de fácil obtención (sangre periférica y heces), que dispongan de capacidad predictiva de recurrencia en el contexto post-quirúrgico de la EC, y permitan identificar y monitorizar los cambios que acontecen en el desarrollo de la inflamación de novo. Igualmente, se pretende conocer el perfil de la respuesta inmune que dirige este proceso y los posibles elementos epigenéticos reguladores, así como su implicación patogénica en la génesis de la recurrencia. Finalmente, se plantea el desarrollo de una herramienta de predicción (índice predictivo combinado) empleando los biomarcadores identificados, con potencial aplicación en la práctica clínica para un mejor manejo de estos pacientes.Despite increasing awareness of the pathogenesis of Crohn's disease (CD) and the development of more potent therapeutic regimens including biological drugs, surgery rates in these patients remain high. Thus, 50-60% of patients with CD require bowel resection in the first 10 years after diagnosis, and 25% of these patients will require a second surgery within the next 5 years. Since surgery is not curative, the appearance of new intestinal lesions after resection is very frequent in these patients despite the therapeutic intervention. This phenomenon is known as post-operative recurrence. Treatments used to prevent recurrence of CD after surgery include mesalazine, thiopurines, and nowadays, biological or anti-TNF drugs. In recent years, several studies have confirmed the efficacy of biological treatment in the prevention of recurrence, especially in high-risk patients, with a current trend towards their early introduction after surgery. However, these drugs imply a not insignificant rate of adverse effects, in addition to a high cost per patient. On the other hand, it is not clearly established which patients benefit from a more intensive treatment after surgery or the duration of maintenance of the same. Therefore, the need arises to develop new tools and strategies to identify the risk of an early recurrence and to optimize the clinical management of these patients. Currently, ileocolonoscopy continues to be the gold standard for the follow-up and diagnosis of recurrence, despite being an invasive technique, expensive, non-risk-free and generally poorly tolerated by patients. This is because we do not have good predictive markers of recurrence after surgery until now, and because the usefulness of clinical and analytical assessment for the diagnosis of recurrence is limited. For this reason, there is a special interest in the use of non-invasive and easily obtained biomarkers, which serve to monitor these patients after surgery and allow early detection of post-operative recurrence. In this sense, fecal markers such as calprotectin have previously shown great clinical utility in different scenarios of the disease. In the post-surgical context, available evidence also suggests its potential utility as a marker of recurrence, although some aspects for its application in clinical practice remain to be clearly defined. Other possible biomarkers in peripheral blood have also not been explored in this EC scenario. On the other hand, post-operative recurrence supposes a new pathogenic condition, so that the CD patients underwent surgery are the best natural model in vivo to study and identify the pathogenic mechanisms involved in the reappearance of the disease. The aim of the present study is the prospective characterization of non-invasive markers in peripheral blood and feces, which have a predictive capacity for post-operative recurrence in CD, to identify and monitor the changes that occur in the development of the novo inflammation. Likewise, we intend to know the profile of the immune response that leads this process and possible regulatory epigenetic elements, as well as its pathogenic implication in the genesis of recurrence. Finally, we propose the development of a predictive tool (predictive combined index) using the identified biomarkers, with potential application in clinical practice for a better management of these patients