M1 Macrophages Activate Notch Signalling in Epithelial Cells: Relevance in Crohn's Disease

Background: The Notch signalling pathway plays an essential role in mucosal regeneration, which constitutes a key goal of Crohn's disease (CD) treatment. Macrophages coordinate tissue repair and several phenotypes have been reported which differ in the expression of surface proteins, cytokines and hypoxia-inducible factors (HIFs). We analysed the role of HIFs in the expression of Notch ligands in macrophages and the relevance of this pathway in mucosal regeneration. Methods: Human monocytes and U937-derived macrophages were polarized towards the M1 and M2 phenotypes and the expression levels of HIF-1α, HIF-2α, Jagged 1 (Jag1) and delta-like 4 (Dll4) were evaluated. The effects of macrophages on the expression of hairy and enhancer of split-1 (HES1, the main target of Notch signalling) and intestinal alkaline phosphatase (IAP, enterocyte marker) in epithelial cells in co-culture were also analysed. Phenotype macrophage markers and Notch signalling were evaluated in the mucosa of CD patients. Results: M1 macrophages were associated with HIF-1-dependent induction of Jag1 and Dll4, which increased HES1 protein levels and IAP activity in co-cultured epithelial cells. In the mucosa of CD patients a high percentage of M1 macrophages expressed both HIF-1α and Jag1 while M2 macrophages mainly expressed HIF-2α and we detected a good correlation between the ratio of M1/M2 macrophages and both HES1 and IAP protein levels. Conclusion: M1, but not M2, macrophages are associated with HIF-1-dependent induction of Notch ligands and activation of epithelial Notch signalling pathway. In the mucosa of chronic CD patients, the prevalence of M2 macrophages is associated with diminution of Notch signalling and impaired enterocyte differentiation. Key Words: MacrophagesCrohn's diseasemucosal healingNotch signallin

Effectiveness and safety of integrase strand transfer inhibitors in Spain: a prospective real-world study

IntroductionSecond-generation integrase strand transfer inhibitors (INSTIs) are preferred treatment options worldwide, and dolutegravir (DTG) is the treatment of choice in resource-limited settings. Nevertheless, in some resource-limited settings, these drugs are not always available. An analysis of the experience with the use of INSTIs in unselected adults living with HIV may be of help to make therapeutic decisions when second-generation INSTIs are not available. This study aimed to evaluate the real-life effectiveness and safety of dolutegravir (DTG), elvitegravir/cobicistat (EVG/c), and raltegravir (RAL) in a large Spanish cohort of HIV-1-infected patients. MethodsReal-world study of adults living with HIV who initiated integrase INSTIs DTG, EVG/c, and RAL-based regimens in three settings (ART-naive patients, ART-switching, and ART-salvage patients). The primary endpoint was the median time to treatment discontinuation after INSTI-based regimen initiation. Proportion of patients experiencing virological failure (VF) (defined as two consecutive viral loads (VL) & GE;200 copies/mL at 24 weeks or as a single determination of VL & GE;1,000 copies/mL while receiving DTG, EVG/c or RAL, and at least 3 months after INSTI initiation) and time to VF were also evaluated. ResultsVirological effectiveness of EVG/c- and RAL-based regimens was similar to that of DTG when given as first-line and salvage therapy. Treatment switching for reasons other than virological failure was more frequent in subjects receiving EVG/c and, in particular, RAL. Naive patients with CD4+ nadir <100 cells/& mu;L were more likely to develop VF, particularly if they initiated RAL or EVG/c. In the ART switching population, initiation of RAL and EVG/c was associated with both VF and INSTI discontinuation. There were no differences in the time to VF and INSTI discontinuation between DTG, EVG/c and RAL. Immunological parameters improved in the three groups and for the three drugs assessed. Safety and tolerability were consistent with expected safety profiles. DiscussionWhereas second-generation INSTIs are preferred treatment options worldwide, and DTG is one of the treatment of choices in resource-limited settings, first-generation INSTIs may still provide high virological and immunological effectiveness when DTG is not available

Darunavir/cobicistat/emtricitabine/tenofovir alafenamide versus dolutegravir /abacavir/lamivudine in antiretroviral-naïve adults (SYMTRI): a multicenter randomized open-label study (PReEC/RIS-57)

D/C/F/TAF is the reference for combination therapy based on protease inhibitors but has not been compared with regimens containing integrase inhibitors as initial ART. We could not demonstrate D/C/F/TAF noninferiority relative to DTG/ABC/3TC, although both regimens were similarly well tolerated. Background Darunavir/cobicistat/emtricitabine/tenofovir alafenamide (D/C/F/TAF) is the reference for combination therapy based on protease inhibitors due to its efficacy, tolerability, and convenience. Head-to-head randomized comparisons between D/C/F/TAF and combination therapy based on integrase inhibitors in antiretroviral-naive patients are lacking. Methods Adult (>18 years old) human immunodeficiency virus-infected antiretroviral-naive patients (HLA-B*5701 negative and hepatitis B virus negative), with viral load (VL) >= 500 c/mL, were centrally randomized to initiate D/C/F/TAF or dolutegravir/abacavir/lamivudine (DTG/3TC/ABC) after stratifying by VL and CD4 count. Clinical and analytical assessments were performed at weeks 0, 4, 12, 24, and 48. The primary endpoint was VL 100 000 copies/mL, and 13% had <200 CD4 cells/mu L. Median weight was 73 kg and median body mass index was 24 kg/m(2). At 48 weeks, 79% (D/C/F/TAF) versus 82% (DTG/3TC/ABC) had VL <50 c/mL (difference, -2.4%; 95% confidence interval [CI], -11.3 to 6.6). Eight percent versus four percent experienced virologic failure but no resistance-associated mutations emerged. Four percent versus six percent had drug discontinuation due to adverse events. In the per-protocol analysis, 94% versus 96% of patients had VL <50 c/mL (difference, -2%; 95% CI, -8.1 to 3.5). There were no differences in CD4 cell count or weight changes. Conclusions We could not demonstrate the noninferiority of D/C/F/TAF relative to DTG/ABC/3TC as initial antiretroviral therapy, although both regimens were similarly well tolerated

Progastrin Represses the Alternative Activation of Human Macrophages and Modulates Their Influence on Colon Cancer Epithelial Cells

Macrophage infiltration is a negative prognostic factor for most cancers but gastrointestinal tumors seem to be an exception. The effect of macrophages on cancer progression depends on their phenotype, which may vary between M1 (pro-inflammatory, defensive) to M2 (tolerogenic, pro-tumoral). Gastrointestinal cancers often become an ectopic source of gastrins and macrophages present receptors for these peptides. The aim of the present study is to analyze whether gastrins can affect the pattern of macrophage infiltration in colorectal tumors. We have evaluated the relationship between gastrin expression and the pattern of macrophage infiltration in samples from colorectal cancer and the influence of these peptides on the phenotype of macrophages differentiated from human peripheral monocytes in vitro. The total number of macrophages (CD68+ cells) was similar in tumoral and normal surrounding tissue, but the number of M2 macrophages (CD206+ cells) was significantly higher in the tumor. However, the number of these tumor-associated M2 macrophages correlated negatively with the immunoreactivity for gastrin peptides in tumor epithelial cells. Macrophages differentiated from human peripheral monocytes in the presence of progastrin showed lower levels of M2-markers (CD206, IL10) with normal amounts of M1-markers (CD86, IL12). Progastrin induced similar effects in mature macrophages treated with IL4 to obtain a M2-phenotype or with LPS plus IFNγ to generate M1-macrophages. Macrophages differentiated in the presence of progastrin presented a reduced expression of Wnt ligands and decreased the number and increased cell death of co-cultured colorectal cancer epithelial cells. Our results suggest that progastrin inhibits the acquisition of a M2-phenotype in human macrophages. This effect exerted on tumor associated macrophages may modulate cancer progression and should be taken into account when analyzing the therapeutic value of gastrin immunoneutralization

Effectiveness and safety of integrase strand transfer inhibitors in Spain: a prospective real-world study

IntroductionSecond-generation integrase strand transfer inhibitors (INSTIs) are preferred treatment options worldwide, and dolutegravir (DTG) is the treatment of choice in resource-limited settings. Nevertheless, in some resource-limited settings, these drugs are not always available. An analysis of the experience with the use of INSTIs in unselected adults living with HIV may be of help to make therapeutic decisions when second-generation INSTIs are not available. This study aimed to evaluate the real-life effectiveness and safety of dolutegravir (DTG), elvitegravir/cobicistat (EVG/c), and raltegravir (RAL) in a large Spanish cohort of HIV-1-infected patients.MethodsReal-world study of adults living with HIV who initiated integrase INSTIs DTG, EVG/c, and RAL-based regimens in three settings (ART-naïve patients, ART-switching, and ART-salvage patients). The primary endpoint was the median time to treatment discontinuation after INSTI-based regimen initiation. Proportion of patients experiencing virological failure (VF) (defined as two consecutive viral loads (VL) ≥200 copies/mL at 24 weeks or as a single determination of VL ≥1,000 copies/mL while receiving DTG, EVG/c or RAL, and at least 3 months after INSTI initiation) and time to VF were also evaluated.ResultsVirological effectiveness of EVG/c- and RAL-based regimens was similar to that of DTG when given as first-line and salvage therapy. Treatment switching for reasons other than virological failure was more frequent in subjects receiving EVG/c and, in particular, RAL. Naïve patients with CD4+ nadir &lt;100 cells/μL were more likely to develop VF, particularly if they initiated RAL or EVG/c. In the ART switching population, initiation of RAL and EVG/c was associated with both VF and INSTI discontinuation. There were no differences in the time to VF and INSTI discontinuation between DTG, EVG/c and RAL. Immunological parameters improved in the three groups and for the three drugs assessed. Safety and tolerability were consistent with expected safety profiles.DiscussionWhereas second-generation INSTIs are preferred treatment options worldwide, and DTG is one of the treatment of choices in resource-limited settings, first-generation INSTIs may still provide high virological and immunological effectiveness when DTG is not available

Spatiotemporal Characteristics of the Largest HIV-1 CRF02_AG Outbreak in Spain: Evidence for Onward Transmissions

Background and Aim: The circulating recombinant form 02_AG (CRF02_AG) is the predominant clade among the human immunodeficiency virus type-1 (HIV-1) non-Bs with a prevalence of 5.97% (95% Confidence Interval-CI: 5.41–6.57%) across Spain. Our aim was to estimate the levels of regional clustering for CRF02_AG and the spatiotemporal characteristics of the largest CRF02_AG subepidemic in Spain.Methods: We studied 396 CRF02_AG sequences obtained from HIV-1 diagnosed patients during 2000–2014 from 10 autonomous communities of Spain. Phylogenetic analysis was performed on the 391 CRF02_AG sequences along with all globally sampled CRF02_AG sequences (N = 3,302) as references. Phylodynamic and phylogeographic analysis was performed to the largest CRF02_AG monophyletic cluster by a Bayesian method in BEAST v1.8.0 and by reconstructing ancestral states using the criterion of parsimony in Mesquite v3.4, respectively.Results: The HIV-1 CRF02_AG prevalence differed across Spanish autonomous communities we sampled from (p &lt; 0.001). Phylogenetic analysis revealed that 52.7% of the CRF02_AG sequences formed 56 monophyletic clusters, with a range of 2–79 sequences. The CRF02_AG regional dispersal differed across Spain (p = 0.003), as suggested by monophyletic clustering. For the largest monophyletic cluster (subepidemic) (N = 79), 49.4% of the clustered sequences originated from Madrid, while most sequences (51.9%) had been obtained from men having sex with men (MSM). Molecular clock analysis suggested that the origin (tMRCA) of the CRF02_AG subepidemic was in 2002 (median estimate; 95% Highest Posterior Density-HPD interval: 1999–2004). Additionally, we found significant clustering within the CRF02_AG subepidemic according to the ethnic origin.Conclusion: CRF02_AG has been introduced as a result of multiple introductions in Spain, following regional dispersal in several cases. We showed that CRF02_AG transmissions were mostly due to regional dispersal in Spain. The hot-spot for the largest CRF02_AG regional subepidemic in Spain was in Madrid associated with MSM transmission risk group. The existence of subepidemics suggest that several spillovers occurred from Madrid to other areas. CRF02_AG sequences from Hispanics were clustered in a separate subclade suggesting no linkage between the local and Hispanic subepidemics

Aplicação e interpretação uniforme dos instrumentos do direito internacional dos negócios

O presente artigo tem como objetivo verificar a possibilidade de aplicação de uma interpretação uniforme do direito internacional dos negócios tanto no âmbito da União Europeia como a nível mundial. Devido às limitações normativas dos ordenamentos jurídicos nacionais, as variadas interpretações jurisdicionais divergentes suscitam sérios problemas quanto à aplicação uniforme do direito, ameaçando a segurança jurídica e a previsibilidade das decisões judiciais. Por meio da análise das disposições do Regulamento Roma I que designa a lei aplicável às obrigações contratuais de natureza civil ou comercial no espaço da União Europeia e da Convenção de Viena sobre os contratos de compra e venda internacional de mercadorias, será inquirido como estes textos oferecem instrumentos aos operadores jurídicos para alcançar uma maior uniformização da interpretação e atividade jurídica desta seara do direito internacional

Impacto de la actitud frente a los síntomas en la mortalidad temprana por infarto de miocardio

Introducción y objetivos. El entorno y las circunstancias del paciente en el inicio de los síntomas del infarto agudo de miocardio (IAM) pueden condicionar su supervivencia. El objetivo es estudiar los aspectos relativos a la convivencia, las características y el retraso en las primeras acciones tomadas por los pacientes con IAM que sobrevivieron más de 1 h y analizar su relación con la mortalidad en distintos períodos. Pacientes y método. Se ha realizado un estudio de cohortes de base poblacional entre 1997 y 1998. La principal fuente de información ha sido el Registre Gironí del Cor (REGICOR) y, para los fallecidos antes de acceder a monitorización, los boletines estadísticos de defunción. Se estudiaron las características demográficas, los hábitos y los antecedentes, los síntomas, las alteraciones electrocardiográficas, el valor de las enzimas miocárdicas y los procedimientos terapéuticos y diagnósticos. Se analizó la mortalidad prehospitalaria, intrahospitalaria y global a los 28 días. Resultados. Se analizaron 1.097 casos: 652 (59,4%) supervivientes a 28 días, 171 muertes en hospitales (15,58%) y 274 muertes prehospitalarias (24,97%). Los pacientes que fueron directamente al hospital presentaron menor mortalidad (odds ratio [OR] = 0,32; intervalo de confianza [IC] del 95%, 0,17-0,59). Hubo mayor mortalidad a los 28 días entre los que tenían síntomas atípicos (OR = 5,52; IC del 95%, 2,90-10,50) y/o vivían institucionalizados (OR = 9,47; IC del 95%,1,05-84,9). Conclusiones. En ausencia de un servicio de ambulancias medicalizadas, los pacientes con un IAM que se dirigen directamente a un hospital y/o presentan síntomas típicos sobreviven en mayor proporción a los 28 días y en la fase prehospitalaria de la enfermedad, y los institucionalizados presentan una mayor mortalidad a los 28 días

Comparison between telephone and self-administration of Short Form Health Survey Questionnaire (SF-36) Estudio comparativo entre la encuesta telefónica y la autoaplicada del cuestionario de salud SF-36

Objective: The characteristics of the 36 item Medical Outcome Short Form Health Study Survey (SF-36) questionnaire, designed as a generic indicator of health status for the general population, allow it to be self-administered or used in personal or telephone interviews. The main objective of the study was to compare the telephone and self-administered modes of SF-36 for a population from Girona (Spain). Methods: A randomized crossover administration of the questionnaire design was used in a cardiovascular risk factor survey. Of 385 people invited to participate in the survey, 351 agreed to do so and were randomly assigned to two orders of administration (i.e., telephone-self and self-telephone); 261 completed both questionnaires. Scores were compared between administration modes using a paired t test. Internal consistency and agreement between modalities were analyzed by respectively applying Chronbach's alpha and intraclass correlation coefficients. The effect of the order of administration on the test-retest difference was analyzed by one-way ANOVA for repeated measurements. Results: Physical function, physical role and social functioning received significantly lower scores when the self-administered questionnaire was used prior to the telephone survey. When the initial survey was conducted by telephone, all Chronbach's alpha coefficients (except social functioning) scored over 0.70 in the self-administered modality. The intraclass correlation coefficient ranged from 0.41 to 0.83 for the telephone-self order and from 0.32 to 0.73 for the self-telephone order. No clinically significant effect was observed for the order of application. Conclusions: The results of the present study suggest that the telephone-administration mode of SF-36 is equivalent to and as valid as the self-administered mode.Objetivo: El cuestionario de salud SF-36 puede ser autoaplicado o utilizado en entrevistas personales o telefónicas. El objetivo principal de este trabajo fue comparar la aplicación telefónica del cuestionario y la versión autoaplicada en una población de Girona (España). Métodos: Diseño cruzado y aleatorizado para la aplicación de las dos formas del cuestionario. Se asignaron dos órdenes de aplicación de las encuestas (telefónica-autoaplicada y autoaplicada-telefónica). Un total de 261 personas completaron los cuestionarios. Las comparaciones entre modos de aplicación se realizaron mediante la prueba de la t de Student para datos apareados. La consistencia interna y la concordancia entre modos de aplicación se analizaron mediante los coeficientes * de Chronbach y de correlación intraclase, respectivamente. Su utilizó un modelo lineal general para medidas repetidas para evaluar el efecto del orden de la aplicación de los cuestionarios. Resultados: Cuando se utilizó primero el cuestionario autoaplicado, las escalas de función física, rol físico y función social resultaron en una menor puntuación. Todos los coeficientes * de Chronbach fueron superiores a 0,70, excepto para la escala de función social en la modalidad autoaplicada cuando se aplicó primero la encuesta telefónica. El rango de los coeficientes de correlación intraclase fue de 0,41 a 0,83 en la modalidad telefónica-autoaplicada y de 0,32 a 0,73 en la modalidad autoaplicada-telefónica. No se observó un efecto relevante del orden de aplicación