Transcriptomic Profiles of Monocyte-Derived Macrophages in Response to Escherichia coli is Associated with the Host Genetics.

Reactive Nitrogen Species (RNS) are a group of bactericidal molecules produced by macrophages in response to pathogens in a process called oxidative burst. Nitric oxide (NO-) is a member of RNS produced from arginine by inducible Nitric Oxide Synthase (iNOS) enzyme. The activity of iNOS and production of NO- by macrophages following stimulation is one of the indicators of macrophage polarization towards M1/proinflammatory. Production of NO- by bovine monocyte-derived macrophage (MDM) and mouse peritoneal macrophages has been shown to be strongly associated with host genetic with the heritability of 0.776 in bovine MDM and 0.8 in mouse peritoneal macrophages. However, the mechanism of genetic regulation of macrophage response has remained less explored. In the current study, the transcriptome of bovine MDMs was compared between two extreme phenotypes that had been classified as high and low responder based on NO- production. The results showed that 179 and 392 genes were differentially expressed (DE) between high and low responder groups at 3 and 18 hours after exposure to Escherichia coli, respectively. A set of 11 Transcription Factors (TFs) (STAT1, IRF7, SPI1, STAT4, IRF1, HIF1A, FOXO3, REL, NFAT5, HIC1, and IRF4) at 3 hours and a set of 13 TFs (STAT1, IRF1, HIF1A, STAT4, ATF4, TP63, EGR1, CDKN2A, RBL1, E2F1, PRDM1, GATA3, and IRF4) at 18 hours after exposure to E. coli were identified to be differentially regulated between the high and low responder phenotypes. These TFs were found to be divided into two clusters of inflammatory- and hypoxia-related TFs. Functional analysis revealed that some key canonical pathways such as phagocytosis, chemotaxis, antigen presentation, and cell-to-cell signalling are enriched among the over-expressed genes by high responder phenotype. Based on the results of this study, it was inferred that the functional characteristics of bovine MDMs are associated with NO-based classification. Since NO- production is strongly associated with host genetics, this study for the first time shows the distinct proinflammatory profiles of macrophages are controlled by the natural genetic polymorphism in an outbred population. In addition, the results suggest that genetics can be considered as a new dimension in the current model of macrophage polarization which is currently described by the combination of stimulants, only

Perinatal Stroke: 11 Cases

Introdução: Os acidentes vasculares cerebrais (AVC) são causa de epilepsia e perturbações do neurodesenvolvimento. Têm tido uma incidencia crescente atribuível em parte à melhoria das tecnicas de imagens cerebrais. Alguns diagnósticos são efectuados retrospectivamente após o periodo neonatal. O objectivo da presente serie de casos clínicos foi identificar factores comuns que possam facilitar o diagnóstico no período neonatal. Material e métodos: Estudo retrospectivo efectuado com base na consulta dos processos clinicos dos recém-nascidos, com diagnóstico de AVC entre um de Janeiro de 2003 e 31 de Dezembro de 2013. Resultados: Foram identificados onze casos de AVC perinatal, num total 28382 nados-vivos. Dois casos foram diagnosticados no periodo fetal e nove no periodo neonatal. As convulsões foram a manifestação clinica mais frequente (8 em 11 casos). A mediana da idade de diagnóstico foi um dia e variou entre um e nove dias. A ecografia transfontanelar mostrou alterações em sete casos. A ressonancia magnetica nuclear cranio-encefalica mostrou alterações em todos os casos. Cinco AVC foram arteriais isquemicos, quatro hemorragicos e dois tromboses dos seios venosos. Em seis casos foram identificadas possiveis causas. Foram observadas complicações e sequelas em quatro dos casos. Discussão: As convulsões foram a manifestação clinica mais frequentemente encontrada.Em recém-nascidos de termo com convulsões sem historia de asfixia intraparto o AVC perinatal deverá ser diagnóstico de exclusão, mesmo na ausencia de alterações na ecografia transfontanelar

The impact of self-stigma in people with diagnosis of severe mental illness: a cross-sectional pilot study from a community psychiatry unit in Porto, Portugal

"Introduction Self-stigma refers to the process in which a person internalizes negative stereotypes, beliefs, and prejudices about their mental illness, adopting a stigmatized view of themselves. Severe mental illness is one of the most socially exclusive stigmata and is associated with poor clinical and functional outcomes and social withdrawal. Objectives In Portugal, investigation regarding self-stigma is scarce. In this study, we aim to evaluate the impact of self-stigma among people with diagnosis of severe mental illness (SMI). For this goal we assess the prevalence of self-stigma of psychiatric patients with diagnosis SMI; and investigate the correlates of elevated self-stigma levels. Methods Fifty-one outpatients with SMI, were recruited from a community psychiatry unit from Porto, Portugal. After informed consent, evaluations included sociodemographic data, illness characteristics, and self-reported standardized scales. Self-stigma (ISMI), self-esteem (RSES) and quality of life (WHO-QoL) were assessed. Data analyses were performed using the SPSS version 28.0 (IBM Corp., Armonk, NY). p-values<0.05 were considered significant. Results From the study sample, 66.7% were male, with mean age of 44.8±11.0 and 56.9% were single. 33.3% reported living with their parents while 31.4% were living with a partner/spouse. The majority of participants had a diagnosis of schizophrenia (60.8%). Concerning the level of education, 58.8% completed basic education, but most patients were retired due to illness (62.7%). In this study, moderate to high self-stigma levels was found in 31.4% participants. Proportion of elevated self-stigma was significantly higher in unemployed/retired patients vs. those who were active (39.0% vs. 0%; P=0.021). No significant correlations were found with age, level of education, age at diagnosis, duration of illness, and number of hospitalizations. In the correlations analysis, a negative correlation between self-stigma and self-esteem (rho=-0.745; P<0.001), as well as self-stigma and quality of life (rho=-0.585; P<0.001) was found. A positive relationship between self-esteem and quality of life (rho=0.551; P<0.001) was found. Conclusions This study investigates, for the first time, the prevalence of self-stigma among outpatients with SMI in a community psychiatric unit from Porto. Our findings suggest a high prevalence of elevated levels of self-stigma among these patients. A significant association with being unemployed/retired was also found. Our results support previous evidence that internalized stigma is strongly associated with diminished self-esteem and impaired quality of life, in particular those aspects related to physical and psychological complaints. Targeting internalized stigma and self-esteem among patient with SMI will likely improve their quality of life, besides improving their clinical and functional outcomes.

Identification of clinical predictors of flare in systemic lupus erythematosus patients: a 24-month prospective cohort study

Objective. SLE has a relapsing-remitting course with disease activity flares over time. This study aims to identify clinical predictors of SLE flares.Methods. This prospective cohort study over 24 months included all SLE patients on follow-up at one academic lupus clinic. Flare was defined as an increase in SLEDAI-2K score ≥4 points. Baseline clinical and demographic parameters were compared using survival analysis for time-to-flare outcome with univariate log-rank tests. Variables with significant differences were further evaluated as predictors with multivariate Cox regression models adjusting for potential confounding or contributing factors and hazard ratio (HR) calculation.Results. A total of 202 SLE patients were included. Over the follow-up period, 1083 visits were documented and 16.8% of patients presented with flares. In multivariate analysis, the following parameters emerged as flare predictors: SLE diagnosis up to 25 years of age (HR = 2.14, P = 0.03), lupus nephritis previous to baseline visit (HR = 4.78, P < 0.0001) and immunosuppressor treatment for severe SLE (HR = 3.22, P < 0.001). Baseline disease activity, disease duration and treatment with prednisone or HCQ were not predictive factors.Conclusion. Patients with an SLE diagnosis before age 25 years, lupus nephritis or immunosuppressor treatment for severe SLE present greater HRs for flares, suggesting the need for tighter clinical monitoring. Current immunosuppressive strategies seem to be inefficient in providing flare prevention

Schizotypy: The Way Ahead

Background: Empirical evidence suggests that schizotypy is a useful construct for analyzing and understanding psychotic disorders. However, several issues remain to be resolved. Method: This selective, critical review, addresses some questions and limitations, and discusses future directions of work. Results: First, we present a conceptual outline and discuss the evidence from translational and interdisciplinary studies on schizotypy. Next, we examine and discuss newer analytical and methodological approaches, including network and machine learning approaches. We also discuss newer psychometric identification approaches, such as those using biobehavioral and ambulatory assessment. Next, we review recent cross-cultural studies in schizotypy research. Finally, we identify new challenges and directions and draw conclusions. Conclusions: This selective, critical review suggests that new methods can contribute to the construction of a solid scientific model of schizotypy as a risk construct. // Esquizotipia: el Camino a Seguir. Antecedentes: la evidencia empírica ha demostrado que la esquizotipia es un constructo útil para analizar y comprender los trastornos psicóticos. Sin embargo, todavía quedan por resolver varias cuestiones. Método: en esta revisión selectiva y crítica se abordan algunas limitaciones, se discuten interrogantes y se comentan direcciones futuras de trabajo. Resultados: en primer lugar, se presenta una delimitación conceptual y se comenta la evidencia acumulada en diferentes estudios y niveles de análisis en el campo de la esquizotipia. A continuación, se examinan nuevos modelos psicopatológicos, como el modelo de red, y se presentan las diferentes herramientas desarrolladas y validadas para su evaluación. Seguidamente, se abordan algunas inquietudes metodológicas de fondo y se presentan nuevas técnicas y procedimientos psicométricos, como la evaluación ambulatoria y bioconductual. También se analizan algunos de los problemas inherentes en la investigación entre países y culturas. Finalmente, se establecen las conclusiones y se abordan nuevos desafíos y direcciones futuras de investigación. Conclusiones: esta revisión selectiva y crítica plantea que es necesario continuar trabajando en la construcción de un modelo científico sólido y refutable e incorporar nuevas pruebas científicas en el campo de la esquizotipia

Phenotyping clonal populations of glioma stem cell reveals a high degree of plasticity in response to changes of microenvironment

The phenotype of glioma-initiating cells (GIC) is modulated by cell-intrinsic and cell-extrinsic factors. Phenotypic heterogeneity and plasticity of GIC is an important limitation to therapeutic approaches targeting cancer stem cells. Plasticity also presents a challenge to the identification, isolation, and propagation of purified cancer stem cells. Here we use a barcode labelling approach of GIC to generate clonal populations over a number of passages, in combination with phenotyping using the established stem cell markers CD133, CD15, CD44, and A2B5. Using two cell lines derived from isocitrate dehydrogenase (IDH)-wildtype glioblastoma, we identify a remarkable heterogeneity of the phenotypes between the cell lines. During passaging, clonal expansion manifests as the emergence of a limited number of barcoded clones and a decrease in the overall number of clones. Dual-labelled GIC are capable of forming traceable clonal populations which emerge after as few as two passages from mixed cultures and through analyses of similarity of relative proportions of 16 surface markers we were able to pinpoint the fate of such populations. By generating tumour organoids we observed a remarkable persistence of dominant clones but also a significant plasticity of stemness marker expression. Our study presents an experimental approach to simultaneously barcode and phenotype glioma-initiating cells to assess their functional properties, for example to screen newly established GIC for tumour-specific therapeutic vulnerabilities

SRC inhibition prevents P-cadherin mediated signaling and function in basal-like breast cancer cells

BACKGROUND: Basal-like breast cancer (BLBC) is a poor prognosis subgroup of triple-negative carcinomas that still lack specific target therapies and accurate biomarkers for treatment selection. P-cadherin is frequently overexpressed in these tumors, promoting cell invasion, stem cell activity and tumorigenesis by the activation of Src-Family kinase (SRC) signaling. Therefore, our aim was to evaluate if the treatment of BLBC cells with dasatinib, the FDA approved SRC inhibitor, would impact on P-cadherin induced tumor aggressive behavior. METHODS: P-cadherin and SRC expression was evaluated in a series of invasive Breast Cancer and contingency tables and chi-square tests were performed. Cell-cell adhesion measurements were performed by Atomic Force Microscopy, where frequency histograms and Gaussian curves were applied. 2D and 3D cell migration and invasion, proteases secretion and self-renew potential were evaluated in vitro. Student's t-tests were used to determine statistically significant differences. The cadherin/catenin complex interactions were evaluated by in situ proximity-ligation assay, and statistically significant results were determined by using Mann-Whitney test with a Bonferroni correction. In vivo xenograft mouse models were used to evaluate the impact of dasatinib on tumor growth and survival. ANOVA test was used to evaluate the differences in tumor size, considering a confidence interval of 95%. Survival curves were estimated by the Kaplan-Meier's method, using the log-rank test to assess significant differences for mice overall survival. RESULTS: Our data demonstrated that P-cadherin overexpression is significantly associated with SRC activation in breast cancer cells, which was also validated in a large series of primary tumor samples. SRC activity suppression with dasatinib significantly prevented the in vitro functional effects of P-cadherin overexpressing cells, as well as their in vivo tumorigenic and metastatic ability, by increasing mice overall survival. Mechanistically, SRC inhibition affects P-cadherin downstream signaling, rescues the E-cadherin/p120-catenin complex to the cell membrane, recovering cell-cell adhesion function. CONCLUSIONS: In conclusion our findings show that targeting P-cadherin/SRC signaling and functional activity may open novel therapeutic opportunities for highly aggressive and poor prognostic basal-like breast cancer.This work was funded by Laço Grant 2014, by FEDER - Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020 - Operacional Programme for Competitiveness and Internationalisation (POCI), Portugal 2020, and by FCT - Fundação para a Ciência e a Tecnologia/ Ministério da Ciência, Tecnologia e Ensino Superior under the projects PTDC/SAU-GMG/120049/ 2010-FCOMP-01-0124-FEDER-021209, PEst-C/SAU/LA0003/2013, NORTE-01- 0145-FEDER-000029 and POCI-01-0145-FEDER-016390. FCT funded the research grants of ASR (SFRH/BPD/75705/2011), ARN (SFRH/BD/100380/2014), BS (SFRH/ BPD/104208/2014), AFV (SFRH/BPD/90303/2012), as well as JP with Programa IFCT 2013 (FCT Investigator). IPATIMUP integrates the i3S Research Unit, which is partially supported by FCT in the framework of the project “Institute for Research and Innovation in Health Sciences” (POCI-01-0145-FEDER-007274)

Incidence of WHO stage 3 and 4 conditions following initiation of Anti-Retroviral Therapy in resource limited settings

To determine the incidence of WHO clinical stage 3 and 4 conditions during early anti-retroviral therapy (ART) in resource limited settings (RLS)