Understanding how the Effectors HopD1 and HopG1 from the Bacterial Pathogen Pseudomonas syringae pv. tomato DC3000 target the Arabidopsis thaliana Protein AtNHR2B to cause Disease in Plants

The pathogenicity of Pseudomonas syringae is associated with the type III secretion system (T3SS), a complex of proteins assembled in the inner and outer bacterial membranes that traverses the plant cell wall to deliver bacterial proteins into the cytoplasm of plant cells. The effector proteins translocated into the plant cells are called Hops (Hypersensitive response and pathogenicity outer proteins). Bacterial effectors target plant immune proteins to suppress defense responses and enhance bacterial parasitism. The Arabidopsis thaliana nonhost resistance 2B (AtNHR2B), a recently identified immune protein, is degraded after inoculation with the adapted pathogen of Arabidopsis, P. syringae pv tomato DC3000 (Pst DC3000), but not by the non-adapted pathogen P. syringae pv. tabaci (Pstab). Several Pst DC3000 effectors, including HopG1 and HopD1 interact with AtNHR2B in planta. Characterization of the effectors presence in plants upon inoculation with Pstab showed that transgenic expression of HopG1-FLAG triggered cell death, high electrolyte leakage levels and increased production of mitochondrial ROS. In contrast, HopG1-FLAG expression in combination with AtNHR2B-GFP caused susceptibility to Pstab as shown by the development of disease symptoms and the significant increase in bacterial growth. Together, these results suggest that HopG1 targets AtNHR2B to interfere with plant immune response upon bacterial infection. In contrast, transgenic plants expressing HopD1-HA alone or in combination with AtNHR2B-GFP, showed disease symptoms after inoculation with Pstab, that normally does not cause disease in wild-type Col-0 plants. Moreover, Pstab grew significantly more in transgenic plants overexpressing HopD1-HA than in wild-type Col-0. Interestingly, Arabidopsis plants expressing the bacterial effector HopD1-HA alone or in combination with AtNHR2B-GFP were deficient in callose deposition and showed a downregulation of the callose synthase gene PMR4. Altogether, these results suggest that HopD1 interferes with callose deposition and by doing so hinders defense responses

Modelo de dados territorial e ambiental: Metodologia, análise e resultados

En el marco de los proyectos llevados a cabo sobre el área transfronteriza Alentejo- Extremadura, y posteriormente, con la ampliación de la región Centro, se han tratado y armonizado una serie de indicadores con el fin de comparar estas regiones de una forma homogénea y estandarizada. Los vectores territorial y ambiental forman parte del Sistema de Indicadores (SI-OTALEX) estructurado y diseñado para ser un sistema abierto y flexible que forme parte de la IDE-OTALEX (www.ideotalex.eu), e integra información homogeneizada y actualizada de este área transfronteriza

Stress and Pain: Predictive (Neuro)Pattern Identification for Chronic Back Pain; A Longitudinal Observational Study

Introduction: Low back pain (LBP) leads to considerable impairment of quality of life worldwide and is often accompanied by psychosomatic symptoms. Objectives: First, to assess the association between stress and chronic low back pain (CLBP) and its simultaneous appearance with fatigue and depression as a symptom triad. Second, to identify the most predictive stress-related pattern set for CLBP for a 1-year diagnosis. Methods: In a 1-year observational study with four measurement points, a total of 140 volunteers (aged 18-45 years with intermittent pain) were recruited. The primary outcomes were pain [characteristic pain intensity (CPI), subjective pain disability (DISS)], fatigue, and depressive mood. Stress was assessed as chronic stress, perceived stress, effort reward imbalance, life events, and physiological markers [allostatic load index (ALI), hair cortisol concentration (HCC)]. Multiple linear regression models and selection procedures for model shrinkage and variable selection (least absolute shrinkage and selection operator) were applied. Prediction accuracy was calculated by root mean squared error (RMSE) and receiver-operating characteristic curves. Results: There were 110 participants completed the baseline assessments (28.2 ± 7.5 years, 38.1% female), including HCC, and a further of 46 participants agreed to ALI laboratory measurements. Different stress types were associated with LBP, CLBP, fatigue, and depressive mood and its joint occurrence as a symptom triad at baseline; mainly social-related stress types were of relevance. Work-related stress, such as "excessive demands at work" [b = 0.51 (95%CI -0.23, 1.25), p = 0.18] played a role for upcoming chronic pain disability. "Social overload" [b = 0.45 (95%CI -0.06, 0.96), p = 0.080] and "over-commitment at work" [b = 0.28 (95%CI -0.39, 0.95), p = 0.42] were associated with an upcoming depressive mood within 1-year. Finally, seven psychometric (CPI: RMSE = 12.63; DISS: RMSE = 9.81) and five biomarkers (CPI: RMSE = 12.21; DISS: RMSE = 8.94) could be derived as the most predictive pattern set for a 1-year prediction of CLBP. The biomarker set showed an apparent area under the curve of 0.88 for CPI and 0.99 for DISS. Conclusion: Stress disrupts allostasis and favors the development of chronic pain, fatigue, and depression and the emergence of a "hypocortisolemic symptom triad," whereby the social-related stressors play a significant role. For translational medicine, a predictive pattern set could be derived which enables to diagnose the individuals at higher risk for the upcoming pain disorders and can be used in practice

Facteurs de risque associés au statut de troupeau positif à Mycobacterium avium ssp. paratuberculosis

La paratuberculose (PTB) est une maladie entérique chronique, contagieuse et incurable qui affecte les ruminants et est causée par Mycobacterium avium ssp paratuberculosis (MAP). Les pertes économiques et l’association entre le MAP et la maladie de Crohn maintiennent un intérêt pour la paratuberculose. Les animaux s’infectent à un jeune âge, principalement par la voie féco-orale. Après une longue période d’incubation (jusqu'à plusieurs années), les vaches débutent l’excrétion fécale de MAP en absence de signes cliniques, perpétuant ainsi l’infection dans le troupeau. Les pratiques de gestion qui limitent l’exposition aux matières fécales contenant le MAP des animaux susceptibles sont plus efficaces pour réduire la prévalence que la simple élimination des animaux positifs. Les objectifs de ce mémoire sont : 1) Examiner et résumer de façon critique la littérature scientifique disponible sur les pratiques de gestion (mesurées à l'aide d'un questionnaire d’analyse de risque) associées au statut du troupeau pour MAP, et 2) identifier l'association entre les pratiques de gestion utilisées et le statut du troupeau pour MAP (déterminé à l'aide de culture bactériologique d'échantillons environnementaux) dans les troupeaux laitiers du Québec. Pour le premier objectif, une revue globale de la littérature a été réalisée. Nous avons inclus des études qui ont évalué les facteurs de risque de PTB en utilisant un questionnaire d'analyse de risque (QAR) et mesuré l'association entre les facteurs de risque et le statut du troupeau pour MAP. Pour le deuxième objectif, une étude cas-témoins a été conçue. Un total de 26 troupeaux où MAP a été isolé d'au moins 1 échantillon environnemental et 91 troupeaux témoins (aucun cas clinique de paratuberculose et négatifs lors de 2 prélèvements environnementaux annuels consécutifs) ont été sélectionnés. Une régression logistique multivariée a été utilisée pour évaluer l'association entre les facteurs de risque sélectionnés et le statut du troupeau pour MAP. En tout, 21 études transversales, 5 études cas-témoins et 3 études longitudinales répondaient aux critères d'inclusion. La taille du troupeau était significativement associée à un statut de troupeau positif à MAP dans 12 (dont 4 avec faible risque de biais (RB)) sur 18, l'introduction de nouveaux animaux était significativement associée à un statut de troupeau positif à MAP dans 10 (dont 4 avec faible RB) sur 24 études, l'histoire de PTB était significativement associée à un statut de troupeau positif à MAP dans 6 (dont 4 à faible RB) sur 13, et la gestion du colostrum et du lait était significativement associée à un statut de troupeau positif à MAP dans 5 (aucun à faible RB) sur 18. Dans les troupeaux laitiers du Québec la taille du troupeau (OR = 1,17; IC à 95%: 1,02-1,33) et la proportion de vaches achetées par année au cours des 5 dernières années (OR = 5,44 IC à 95%: 1,23-23,98) étaient significativement associées à un statut de troupeau MAP positif. Les résultats de ce mémoire fournissent une grande compilation des informations disponibles sur les facteurs de risque associés au statut du troupeau pour MAP et évalués à l'aide d'un QAR. Certains facteurs de risque sont apparemment plus consistants d'une étude à l'autre. Cependant, les résultats doivent être interprétés à la lumière de la qualité et du risque de biais de chaque étude. Les pratiques de gestion visant à empêcher l'introduction de nouveaux animaux dans le troupeau et à réduire le contact des veaux nouveau-nés avec les animaux adultes ou leurs excréments sont des éléments clés pour minimiser l'introduction et la transmission du MAP dans un troupeau. Ces éléments devraient être priorisés dans les programmes de contrôle.Paratuberculosis is a chronic and contagious enteric disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Control of paratuberculosis is justified given the associated economic losses and the potential role of MAP in Crohn’s disease in humans. Cattle usually become infected at a young age, primarily by the fecal-oral route. After a long incubation period (up to several years), infected cows may start shedding MAP without showing clinical signs, thus perpetuating MAP infections on the farm. Management procedures that limit exposure of susceptible animals to MAP are more effective at reducing disease prevalence than simply testing and culling MAP infected cows. Any management practices that expose (directly or indirectly) susceptible animals to fecal material from MAP shedders can be considered a risk factor for infection. The objectives of this master’s thesis are: 1) critically review the available scientific literature that evaluates the association between management practices (measured by a risk assessment questionnaire (RAQ)) and MAP herd status, et 2) identify the association between management practices and MAP herd status (determined using bacteriological culture of environmental samples) of dairy herds in Québec, Canada. A systematic review was performed to answer the first objective. We included studies that assessed PTB risk factors using a RAQ and measured the association between risks factors and MAP herd status. For the second objective, a case-control study was designed. A total of 26 case herds in which MAP had been isolated from at least 1 environmental sample in each herd and 91 control herds (no clinical cases of paratuberculosis and negative on 2 consecutive yearly environmental samplings) were selected. Multivariable logistic regression was used to evaluate the association between selected risk factors and MAP herd status. Twenty-one cross-sectional, 5 case control and 3 longitudinal studies met the inclusion criteria. Herd size was significantly associated with MAP herd status in 12 (4 with low RoB) out of 18 studies, introduction of new animals was significantly associated with MAP herd status in 10 (4 with low RoB) out of 24, history of PTB was significantly associated with MAP herd status in 6 (4 with low RoB) out of 13, and management of colostrum and milk were significantly associated with MAP herd status in 5 (none with low RoB) out of 18. For Québec dairy herds, herd size (OR=1.17; 95% CI: 1.02-1.33) and proportion of cows purchased per year in the last 5 years (OR=5.44; 95% CI: 1.23-23.98) were significantly associated with a positive MAP herd status. The results of this master’s thesis provide a large compilation of available information about risk factors associated with MAP herd status evaluated using a RAQ. Some risk factors are apparently more consistent across studies. However, results should be interpreted in the light of the quality and risk of bias of each study. Management practices aiming to prevent the introduction of new animals into the herd and to reduce the contact of newborn calves with adult animals or their feces are key elements to minimize MAP introduction and transmission into a herd. These elements should be prioritized in control programs

When centers can fail: a close second opportunity

This paper presents the p-next center problem, which aims to locate p out of n centers so as to minimize the maximum cost of allocating customers to backup centers. In this problem it is assumed that centers can fail and customers only realize that their closest (reference) center has failed upon arrival. When this happens, they move to their backup center, i.e., to the center that is closest to the reference center. Hence, minimizing the maximum travel distance from a customer to its backup center can be seen as an alternative approach to handle humanitarian logistics, that hedges customers against severe scenario deteriorations when a center fails. For this extension of the p-center problem we have developed several different integer programming formulations with their corresponding strengthenings based on valid inequalities and variable fixing. The suitability of these formulations for solving the p-next center problem using standard software is analyzed in a series of computational experiments. These experiments were carried out using instances taken from the previous discrete location literature.Peer ReviewedPostprint (author’s final draft

Variabilidad del ADN cloroplástico y su contribución al conocimiento del estado de conservación de Ulmus glabra en el Sistema Central

En el presente trabajo se ha analizado la diversidad existente en el ADN cloroplástico (ADNcp) de 16 poblaciones distribuidas a lo largo del Sistema Central. Para ello, se ha empleado la técnica de PCR-RFLP (amplificación de fragmentos específicos y posterior digestión con enzimas de restricción) sobre 38 regiones del ADNcp (Grivet et al., 2001)

Genetic Diversity in Relict and Fragmented Populations of Ulmus glabra Hudson in the Central System of the Iberian Peninsula

Ulmus glabra Hudson, or Wych elm, occurs as fragmented and relict natural populations in the Central System, which acts as a refugium in the Iberian Peninsula. Considering the importance of the Central System populations of U. glabra, the main objective was to assess their genetic diversity using nuclear microsatellite markers. A total of 360 different genotypes were detected in the 427 U. glabra individuals analyzed. Wych elm populations showed a highly significant genetic differentiation (24%; p = 0.0001). Of the 22 populations studied, population of Rozas de Puerto Real (ROZ) showed the highest values of effective number of alleles (2.803), mean Shannon?s diversity (1.047) and expected heterozygosity (0.590). Populations of ROZ and Mombeltrán (MOM) showed the highest values of observed heterozygosity (0.838 and 0.709, respectively), and highly negative values for inbreeding coefficient (?0.412 and ?0.575, respectively). Also, most of putative hybrids (50 of 55) were observed in these two populations. Demographic analysis revealed signals for recent (four populations) and ancestral (fifteen populations) bottlenecks. Fragmented populations with diminishing number of individuals, along with anthropogenic intervention and Dutch elm disease (DED), are the main threats to U. glabra populations. From a future perspective, the information generated can be considered in the formulation of conservation strategies for U. glabra populations in the Central System

Involvement of extracellular vesicles in the macrophage-tumor cell communication in head and neck squamous cell carcinoma

Exosomes; Larynx cancer cells; MacrophagesExosomes; Cèl·lules cancerígenes de la laringe; MacròfagsExosomas; Células cancerosas de laringe; MacrófagosBackground: Exosomes are cell-derived vesicles that mediate cellular communication in health and multiple diseases, including cancer. However, its role in head and neck cancer has been poorly defined. Here, we investigated the relevance of exosomes in the signaling between larynx cancer cells and macrophages. Methods: Exosomes from THP1 macrophages and BICR18 cells (a larynx squamous cell carcinoma cell line) were purified and their role in the cancer cell migration, macrophage phenotype and immunosuppressive activity was evaluated. The activation of STAT3 signal transduction in macrophages in response to exosomes obtained from cancer cells was also evaluated. Results: Macrophages foster the cancer cell migration and this effect is mediated by exosome signaling. On the other hand, exosomes also induce the expression of IL-10 in macrophages and PD-L1 in cancer cells, thus resulting in the promotion of an immunosuppressive environment. Moreover, we observed that the effects induced in cancer cells are mediated by the exosome-depending activation of STAT-3 signal transduction pathway. Conclusions: Our study indicates that exosomes released by both macrophages and cancer cells plays a critical role in tumor progression in larynx cancer and might be a potential target for therapeutic intervention in head and neck cancer.This work was supported by a research grant from the Carlos III Institute of Health with reference FIS PI16/00060 (D.C.), co-funded with European Union ERDF funds (European Regional Development Fund)

Síndrome hemolítico urémico atípico posterior a trasplante renal: presentación de un caso y revisión de la literatura

Haemolytic uremic syndrome (HUS) is a clinical entity characterized by the appearance of non immune hemolytic anemia, thrombocytopenia and acute renal failure. It is a disease belonging to the group of thrombotic microangiopathy (MAT) which are part of thrombotic thrombocytopenic purpura also (PTT) and some other MAT associated with other medical conditions formerly known as secondary MAT.Moreover, the variety known as atypical HUS (aHUS) is an ultra-orphan disease that frequently progresses to chronic renal failure (CRF) and is associated with high morbidity and mortality if not properly treated. If a patient presents its first clinical manifestation of aHUS later receive a cadaveric renal transplant which not only makes it an even more exotic case but involves more complexity in their management is presented.El síndrome hemolítico urémico (SHU) es una entidad clínica caracterizada por la aparición de anemia hemolítica no inmune, trombocitopenia e insuficiencia renal aguda. Se trata de una enfermedad perteneciente al grupo de las microangiopatías trombóticas (MAT) de la que hacen parte también la purpura trombocitopénica trombótica (PTT) y algunas otras MAT asociadas a otras condiciones médicas antes conocidas como MAT secundarias. Por otra parte, la variedad conocida como SHU atípico (SHUa) es una patología ultra-huérfana que frecuentemente evoluciona a insuficiencia renal crónica (IRC) y se asocia con elevada morbimortalidad si no recibe el tratamiento adecuado. Se examina el caso de un paciente que presenta su primera manifestación clínica de síndrome hemolítico urémico atípico después de trasplante renal cadavérico lo cual no solo lo hace un caso aún más exótico, sino que implica mayor complejidad en su manejo. &nbsp