Identification of aceNKPs, a committed common progenitor population of the ILC1 and NK cell continuum.

The development of innate lymphoid cell (ILC) transcription factor reporter mice has shown a previously unexpected complexity in ILC hematopoiesis. Using novel polychromic mice to achieve higher phenotypic resolution, we have characterized bone marrow progenitors that are committed to the group 1 ILC lineage. These common ILC1/NK cell progenitors (ILC1/NKP), which we call "aceNKPs", are defined as lineage-Id2+IL-7Rα+CD25-α4ß7-NKG2A/C/E+Bcl11b-. In vitro, aceNKPs differentiate into group 1 ILCs, including NK-like cells that express Eomes without the requirement for IL-15, and produce IFN-γ and perforin upon IL-15 stimulation. Following reconstitution of Rag2-/-Il2rg-/- hosts, aceNKPs give rise to a spectrum of mature ILC1/NK cells (regardless of their tissue location) that cannot be clearly segregated into the traditional ILC1 and NK subsets, suggesting that group 1 ILCs constitute a dynamic continuum of ILCs that can develop from a common progenitor. In addition, aceNKP-derived ILC1/NK cells effectively ameliorate tumor burden in a model of lung metastasis, where they acquired a cytotoxic NK cell phenotype. Our results identify the primary ILC1/NK progenitor that lacks ILC2 or ILC3 potential and is strictly committed to ILC1/NK cell production irrespective of tissue homing.

Tumor-Derived Lactic Acid Modulates Activation and Metabolic Status of Draining Lymph Node Stroma.

Communication between tumors and the stroma of tumor-draining lymph nodes (TDLN) exists before metastasis arises, altering the structure and function of the TDLN niche. Transcriptional profiling of fibroblastic reticular cells (FRC), the dominant stromal population of lymph nodes, has revealed that FRCs in TDLNs are reprogrammed. However, the tumor-derived factors driving the changes in FRCs remain to be identified. Taking an unbiased approach, we have shown herein that lactic acid (LA), a metabolite released by cancer cells, was not only secreted by B16.F10 and 4T1 tumors in high amounts, but also that it was enriched in TDLNs. LA supported an upregulation of Podoplanin (Pdpn) and Thy1 and downregulation of IL7 in FRCs of TDLNs, making them akin to activated fibroblasts found at the primary tumor site. Furthermore, we found that tumor-derived LA altered mitochondrial function of FRCs in TDLNs. Thus, our results demonstrate a mechanism by which a tumor-derived metabolite connected with a low pH environment modulates the function of fibroblasts in TDLNs. How lymph node function is perturbed to support cancer metastases remains unclear. The authors show that tumor-derived LA drains to lymph nodes where it modulates the function of lymph node stromal cells, prior to metastatic colonization.

Well-being and disease severity of multiple sclerosis patients following a physical activity program / Bem-estar e gravidade da doença de pacientes com esclerose múltipla seguindo um programa de atividade física

Abstract Introduction: Multiple sclerosis (MS) is a chronic disease of the central nervous system that mainly affects young adults, promoting a great impact on functionality. Fatigue is a very common symptom, associated with multiple impairments in sensitivity, muscle activity, neuromotor control, balance, cognition and problem-solving ability. MS leads to strong functional restrictions, particularly in the context of daily living activities, as well as in patient participation. Objective: To understand the implications of a self-regulation program in the perception of well-being and mental health in MS patients. Methods: A set of exercises was implemented for use in daily activities, supported by different studies with MS patients. Patients were asked to classify the severity of their disease and to use the Mental Health Inventory (MHI-38), at the beginning (time A) and at the end (time B) of the self-regulation program. We used the Statistical Package for the Social Sciences (SPSS) version 25. A non-parametric statistical hypothesis test (Wilcoxon test) was used to analyze the variables. Results: The mean age was 44 years old, with patients between the ages of 20 and 58. 58.3% were women, 37.5% were currently married, 67% were retired and the mean level of education was 12.5 years. The correlation between the perception of disease severity and psychological well-being before the self-regulation program (r = 0.26, p < 0.05) and after the intervention (r = 0.37, p < 0.01) suggests a low to moderate correlation. Conclusion: The implementation of the self-regulatory model, through the promotion of physical activity in patients with MS, had a positive impact of clinical rehabilitation, well-being, and perception of disease severity of these people.

Resumo Introdução: A esclerose múltipla (EM) é uma doença crônica do sistema nervoso central que afeta principalmente adultos jovens, promovendo um grande impacto na funcionalidade. A fadiga é um sintoma muito comum, associado a múltiplas deficiências na sensibilidade, atividade muscular, controle neuromotor, equilíbrio, cognição e capacidade de resolução de problemas. A EM leva a fortes restrições funcionais, principalmente no contexto das atividades de vida diária, bem como na participação do paciente. Objetivo: Compreender as implicações de um programa de autorregulação na percepção de bem-estar e saúde mental em pacientes com EM. Métodos: Um conjunto de exercícios foi implementado para uso nas atividades diárias, apoiado por diferentes estudos com pacientes com EM. Solicitou-se aos pacientes que classificassem a gravidade de sua doença e que utilizassem o Inventário de Saúde Mental (MHI-38), no início (tempo A) e no final (tempo B) do programa de autorregulação. Utilizou-se o Statistical Package for the Social Sciences (SPSS) versão 25. Um teste de hipótese estatística não paramétrica (teste de Wilcoxon) foi utilizado para analisar as variáveis. Resultados: A média de idade foi de 44 anos, com pacientes entre 20 e 58 anos. 58,3% eram mulheres, 37,5% eram casados, 67% eram aposentados e a escolaridade média era de 12,5 anos. A correlação entre a percepção da gravidade da doença e o bem-estar psicológico antes do programa de autorregulação (r = 0,26, p < 0,05) e após a intervenção (r = 0,37, p < 0,01) sugere uma correlação baixa a moderada. Conclusão: A implantação do modelo autorregulatório, por meio da promoção de atividade física em pacientes com EM, teve impacto positivo na reabilitação clínica, bem-estar e percepção da gravidade da doença dessas pessoas.

Impact of Locally Administered Carboxydextran-Coated Super-Paramagnetic Iron Nanoparticles on Cellular Immune Function.

Interstitially administered iron oxide particles are currently used for interoperative localization of sentinel lymph nodes (LNs) in cancer staging. Several studies have described concerns regarding the cellular accumulation of iron oxide nanoparticles relating them to phenotype and function deregulation of macrophages, impairing their ability to mount an appropriate immune response once an insult is present. This study aims to address what phenotypic and functional changes occur during lymphatic transit and accumulation of these particles. Data show that 60 nm carboxydextran-coated iron nanoparticles use a noncellular mechanism to reach the draining LNs and that their accumulation in macrophages induces transient phenotypic and functional changes. Nevertheless, macrophages recover their baseline levels of response within 7 days, and are still able to mount an appropriate response to bacterially induced inflammation.

Exploring the role of stromal osmoregulation in cancer and disease using executable modelling.

Osmotic regulation is a vital homoeostatic process in all cells and tissues. Cells initially respond to osmotic stresses by activating transmembrane transport proteins to move osmotically active ions. Disruption of ion and water transport is frequently observed in cellular transformations such as cancer. We report that genes involved in membrane transport are significantly deregulated in many cancers, and that their expression can distinguish cancer cells from normal cells with a high degree of accuracy. We present an executable model of osmotic regulation and membrane transport in mammalian cells, providing a mechanistic explanation for phenotype change in varied disease states, and accurately predicting behaviour from single cell expression data. We also predict key proteins involved in cellular transformation, SLC4A3 (AE3), and SLC9A1 (NHE1). Furthermore, we predict and verify a synergistic drug combination in vitro, of sodium and chloride channel inhibitors, which target the osmoregulatory network to reduce cancer-associated phenotypes in fibroblasts.

Macrophages are exploited from an innate wound healing response to facilitate cancer metastasis.

Tumour-associated macrophages (TAMs) play an important role in tumour progression, which is facilitated by their ability to respond to environmental cues. Here we report, using murine models of breast cancer, that TAMs expressing fibroblast activation protein alpha (FAP) and haem oxygenase-1 (HO-1), which are also found in human breast cancer, represent a macrophage phenotype similar to that observed during the wound healing response. Importantly, the expression of a wound-like cytokine response within the tumour is clinically associated with poor prognosis in a variety of cancers. We show that co-expression of FAP and HO-1 in macrophages results from an innate early regenerative response driven by IL-6, which both directly regulates HO-1 expression and licenses FAP expression in a skin-like collagen-rich environment. We show that tumours can exploit this response to facilitate transendothelial migration and metastatic spread of the disease, which can be pharmacologically targeted using a clinically relevant HO-1 inhibitor.

Registered report: Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through MET.

The Reproducibility Project: Cancer Biology seeks to address growing concerns about reproducibility in scientific research by conducting replications of selected experiments from a number of high-profile papers in the field of cancer biology. The papers, which were published between 2010 and 2012, were selected on the basis of citations and Altmetric scores (Errington et al., 2014). This Registered Report describes the proposed replication plan of key experiments from "Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through MET" by Peinado and colleagues, published in Nature Medicine in 2012 (Peinado et al., 2012). The key experiments being replicated are from Figures 4E, as well as Supplementary Figures 1C and 5A. In these experiments, Peinado and colleagues show tumor exosomes enhance metastasis to bones and lungs, which is diminished by reducing Met expression in exosomes (Peinado et al., 2012). The Reproducibility Project: Cancer Biology is a collaboration between the Center for Open Science and Science Exchange and the results of the replications will be published in eLife.

Preditores da adesão à medicação na Epilepsia: Estudo Longitudinal / Medication adherence predictors in Epilepsy: Longitudinal Study

Introdução: A não adesão à medicação na epilepsia é prevalente, pelo que a compreensão dos fatores associados deve ser promovida. Objetivos: Analisar a capacidade preditiva das estratégias de coping e da espiritualidade em relação à adesão à medicação ao longo do tempo. Metodologia: Foram avaliados 60 indivíduos através de um Questionário Sociodemográfico e Clínico, a Medida de Adesão aos Tratamentos, o COPER e a Escala de Avaliação de Espiritualidade em Contextos de Saúde, em dois momentos. Resultados A espiritualidade Momento 1 não se relacionou com adesão à medicação no Momento 3, e as estratégias de coping: Desinvestimento Comportamental e Aceitação no Momento 1 predizem negativamente a adesão à medicação no Momento 2. Conclusão: A relação entre a adesão à medicação, estratégias de coping e espiritualidade varia emfunção do tempo, o que deve ser considerado ao nível da intervenção...

Introduction: The nonmedication adherence in epilepsy is prevalent by which the understanding of associated factors shouldbe promoted. Objectives: To analyze the predictive capacity of the coping strategies and spirituality in relationto medication adherence over time. Methodology: A sociodemographic and clinical questionnaire, medical adherence to treatment, COPER and spirituality evaluation scale in health contexts was undertaken to evaluate 60 individuals with epilepsy, in three moments. Results: Spirituality at Moment 1 is not related to medication adherence at Moment 3, and the coping strategies: behavioral disinvestment and acceptance at Moment 1 predict medication adherence at moment 2. Conclusion: The relation between medication adherence, coping strategies and spirituality change according to time, which should be considered on the level intervention...

Optimism on quality of life in Portuguese chronic patients: moderator/mediator?

OBJECTIVE: optimism is an important variable that has consistently been shown to affect adjustment to quality of life in chronic diseases. This study aims to clarify if dispositional optimism exerts a moderating or a mediating influence on the personality traits-quality of life association, in Portuguese chronic patients. METHODS: multiple regression models were used to test the moderation and mediation effects of dispositional optimism in quality of life. A sample of 729 patients was recruited in Portugal's main hospitals and completed self-reported questionnaires assessing socio-demographic and clinical variables, personality, dispositional optimism, quality of life (QoL) and subjective well-being (SWB). RESULTS: the results of the regression models showed that dispositional optimism did not moderate the relationships between personality traits and quality of life. After controlling for gender, age, education level and severity of disease perception, the effects of personality traits on QoL and in SWB were mediated by dispositional optimism (partially and completely), except for the links between neuroticism/openness to experience and physical health. CONCLUSION: dispositional optimism is more likely to play a mediating, rather than a moderating role in personality traits-quality of life pathway in Portuguese chronic patients, suggesting that "the expectation that good things will happen" contributes to a better quality of life and subjective well-being.

Optimism on quality of life in Portuguese chronic patients: moderator/mediator? / O papel do otimismo na qualidade de vida de pessoas portuguesas com doenças crônicas: moderador/mediador?

Objective: optimism is an important variable that has consistently been shown to affect adjustment to quality of life in chronic diseases. This study aims to clarify if dispositional optimism exerts a moderating or a mediating influence on the personality traits-quality of life association, in Portuguese chronic patients. Methods: multiple regression models were used to test the moderation and mediation effects of dispositional optimism in quality of life. A sample of 729 patients was recruited in Portugal's main hospitals and completed self-reported questionnaires assessing socio-demographic and clinical variables, personality, dispositional optimism, quality of life (QoL) and subjective well-being (SWB). Results: the results of the regression models showed that dispositional optimism did not moderate the relationships between personality traits and quality of life. After controlling for gender, age, education level and severity of disease perception, the effects of personality traits on QoL and in SWB were mediated by dispositional optimism (partially and completely), except for the links between neuroticism/openness to experience and physical health. Conclusion: dispositional optimism is more likely to play a mediating, rather than a moderating role in personality traits-quality of life pathway in Portuguese chronic patients, suggesting that "the expectation that good things will happen" contributes to a better quality of life and subjective well-being. .

Objetivo: o otimismo tem sido demonstrado como uma variável importante no ajustamento da qualidade de vida de pessoas com doenças crônicas. O estudo tem como objetivo verificar se o otimismo exerce um efeito moderador ou mediador entre os traços de personalidade e a qualidade de vida, em portugueses com doenças crônicas. Métodos: os modelos de regressão linear múltipla foram usados para avaliar o efeito de moderação e mediação do otimismo na qualidade de vida. A amostra, constituída por 729 doentes, recrutados nos principais hospitais de Portugal responderam a questionários de autorresposta avaliando questões sócio-demográficas e clínicas, personalidade, otimismo disposicional, qualidade de vida e bem-estar subjetivo. Resultados: os resultados encontrados mostraram que o otimismo disposicional não exerce um papel moderador entre os traços de personalidade e a qualidade de vida. Controlando por idade, sexo, nível de escolaridade e percepção da severidade da doença, o efeito dos traços de personalidade na qualidade de vida e no bem-estar subjetivo foi mediado pelo otimismo (parcial e total), expecto para as associações, neuroticismo/abertura à experiência e à saúde física. Conclusão: o otimismo disposicional exerce apenas um papel mediador entre os traços de personalidade e qualidade de vida, em pessoas com doenças crônicas, sugerindo que 'a expectativa de que coisas boas vão acontecer' contribui para uma melhor qualidade de vida e melhor bem-estar subjetivo. .

Psychosocial factors as predictors of quality of life in chronic Portuguese patients.

BACKGROUND: Chronic illnesses are diseases of long duration and generally of slow progression. They cause significant quality of life impairment. The aim of this study was to analyse psychosocial predictors of quality of life and of subjective well-being in chronic Portuguese patients. METHODS: Chronic disease patients (n = 774) were recruited from central Portuguese Hospitals. Participants completed self-reported questionnaires assessing socio-demographic, clinical, psychosocial and outcome variables: quality of life (HRQL) and subjective well-being (SWB). MANCOVA analyses were used to test psychosocial factors as determinants of HRQL and SWB. RESULTS: After controlling for socio-demographic and clinical variables, results showed that dispositional optimism, positive affect, spirituality, social support and treatment adherence are significant predictors of HRQL and SWB. Similar predictors of quality of life, such as positive affect, treatment adherence and spirituality, were found for subgroups of disease classified by medical condition. CONCLUSIONS: The work identifies psychosocial factors associated with quality of life. The predictors for the entire group of different chronic diseases are similar to the ones found in different chronic disease subgroups: positive affect, social support, treatment adherence and spirituality. Patients with more positive affect, additional social support, an adequate treatment adherence and a feel-good spirituality, felt better with the disease conditions and consequently had a better quality of life. This study contributes to understanding and improving the processes associated with quality of life, which is relevant for health care providers and chronic diseases support.

Dispositional optimism is unidimensional or bidimensional? the Portuguese revised life orientation test

The aim of the study is to adapt and then discuss the appropriateness of the Life Orientation Test as a one or two dimension scale. The research includes two studies; one is composed of a sequential sample of 280 people with multiple sclerosis, 71 % female, and another includes a convenience sample of 615 individuals from the community, 51.1% female. Because the construct is built upon a theoretical assumption that has one dimension, we examine the hypothesis of one or two factor solutions through confirmatory factor analysis, and the two-dimension solution premise demonstrates better adjustment for both samples. The other psychometric properties explored show appropriate results for the Portuguese sample, and similar to the original ones; the Test therefore seems appropriate for use in cross cultural studies. Based on our results, we discuss whether the questionnaire is a one or two dimension instrument, concluding that it appears appropriate to accept the recommendations of the original authors to use it as a one-dimensional tool and, when necessary, to use both dimensions (AU)

El objetivo del estudio es adaptar y discutir la adecuación de la prueba de Orientación de la Vida en una o dos escalas de dimensión. La investigación engloba dos estudios, uno constituido por una muestra secuencial de 280 personas con esclerosis múltiple, 71% mujeres y otro con una muestra de conveniencia de la comunidad de 615 individuos, 51,1% del sexo femenino. Como el constructo se asienta sobre la presunción teórica de que tiene una dimensión, inspeccionamos la hipótesis de una o dos soluciones de factor a través del análisis factorial confirmatorio y la hipótesis de dos dimensiones manifiesta un mejor ajuste para ambas muestras. Las otras propiedades psicométricas exploradas muestran los resultados apropiados para la muestra portuguesa, y semejantes a los originales. Parece apropiado para los estudios culturales transversales. Basándonos en nuestros resultados, discutimos si el cuestionario es un instrumento de una o dos dimensiones, concluyéndose que parece conveniente seguir las recomendaciones de los autores originales, para utilizarlo como un instrumento unidimensional y, si fuera necesario necesario, utilizar cada una de las dimensiones (AU)

Piezoelectric biosensors for biorecognition analysis: application to the kinetic study of HIV-1 Vif protein binding to recombinant antibodies.

In this work three piezoelectric sensors modified with anti-HIV-1 Vif (virion infectivity factor) single fragment antibodies (4BL scFV), single domains (VH) and camelized single domains (VHD) were constructed and used to detect HIV1 Vif in liquid samples. Dithio-bis-succinimidyl-undecanoate (DSU) and 11-hydroxy-1-undecanethiol (HUT) mixed self assembled monolayers (SAM) were generated at the sensors surface onto which the antibodies were immobilized. All sensors detected specifically the target HIV1-Vif antigen in solution and no unspecific binding was monitored. Impedance analysis was performed to quantify electroacoustic and viscoelastic interferences during antibody immobilization and antigen recognition. The elimination of such interferences enabled the quantitative use of the piezoelectric immunosensors to estimate the antibody surface density as well as antigen binding and equilibrium constants. In spite of the possible limitation regarding mass transport and other related molecular phenomena, which were not considered in the binding model used, this work demonstrates the usefulness of piezoelectric biosensors in biorecognition analysis and evidences the advantages on using simultaneous impedance analysis to bring analytical significance to measured data, and thus to improve piezoelectric sensors sensitivity and applicability.

Recombinant single-chain variable fragment and single domain antibody piezoimmunosensors for detection of HIV1 virion infectivity factor.

In this paper recombinant single-chain fragments (scFv-4BL), and single domain antibodies (4BL-V(H)) and (4BL-V(H)D) generated against HIV1 virion infectivity factor (Vif) are used to develop piezoimmunosensors for HIV1 recognition. Mixed self assembled monolayers were generated at the surface of gold coated crystal sensors to which scFv-4BL, 4BL-V(H), or 4BL-V(H)D were immobilized. Impedance analysis was used to discriminate interfering signals from frequency variation data and to increase the sensor sensitivity. The elimination of interfering signals enabled the quantification of the amount of immobilized protein and gave some indication on the viscoelasticity of immobilized biofilms. All the modified sensors were able to specifically recognize HIV1 Vif in liquid samples. The results indicate that lower sensitivities are obtained with 4BL-V(H) single domain antibodies, possibly due to its higher hydrophobic character. The sensitivity obtained when using scFv-4BL was reestablished when using the more hydrophilic 4BL-V(H)D single domain. 4BL-V(H)D piezoimunosensors were effective in recognizing HIV1 Vif from protein mixtures and from cell extracts of human embryonic kidney cells expressing HIV1 Vif. The results presented in this paper demonstrate the potential applicability of the developed piezoimmunosensors to monitor HIV1 infection evolution.

Molecular construction of bionanoparticles: chimaeric SIV p17-HIV I p6 nanoparticles with minimal viral protein content.

VLPs (virus-like particles) are promising delivery vectors for molecular therapy, since they combine the major advantages of viral vectors with significantly fewer viral vector disadvantages. The present paper describes the molecular construction of chimaeric VLPs based on minimal SIV (simian immunodeficiency virus) and HIV1 components. A chimaeric protein was constructed by fusion of SIV matrix protein (p17) and HIV1 p6 protein, and we demonstrated that the chimaeric proteins assemble as 80 nm nanoparticles containing approximately 7700 chimaeric protein units. Chimaeric VLPs are released from HEK-293T cells (human embryonic kidney cells expressing the large T-antigen of simian virus 40) and are fully encapsulated with lipid membrane. Chimaeric VLPs are produced at 3.7-fold higher levels when compared with SIV p17 VLPs owing to duplication of a PTAP (Pro-Thr-Ala-Pro) domain previously shown as essential for virus particle release. The chimaeric VLPs constructed in the present paper were efficiently pseudotyped with vesicular-stomatitis-virus glycoprotein, as shown by immunoprecipitation assays.