The corticotrophin-releasing factor/urocortin system regulates white fat browning in mice through paracrine mechanisms

Objectives: The corticotrophin-releasing factor (CRF)/urocortin system is expressed in the adipose tissue of mammals, but its functional role in this tissue remains unknown. Methods: Pharmacological manipulation of the activity of CRF receptors, CRF1 and CRF2, was performed in 3T3L1 white pre-adipocytes and T37i brown pre-adipocytes during in vitro differentiation. The expression of genes of the CRF/urocortin system and of markers of white and brown adipocytes was evaluated along with mitochondrial biogenesis and cellular oxygen consumption. Metabolic evaluation of corticosterone-deficient or supplemented Crhr1-null (Crhr1−/−) mice and their wild-type controls was performed along with gene expression analysis carried out in white (WAT) and brown (BAT) adipose tissues. Results: Peptides of the CRF/urocortin system and their cognate receptors were expressed in both pre-adipocyte cell lines. In vitro pharmacological studies showed an inhibition of the expression of the CRF2 pathway by the constitutive activity of the CRF1 pathway. Pharmacological activation of CRF2 and, to a lesser extent, inhibition of CRF1 signaling induced molecular and functional changes indicating transdifferentiation of white pre-adipocytes and differentiation of brown pre-adipocytes. Crhr1−/− mice showed increased expression of CRF2 and its agonist Urocortin 2 in adipocytes that was associated to brown conversion of WAT and activation of BAT. Crhr1−/− mice were resistant to diet-induced obesity and glucose intolerance. Restoring physiological circulating corticosterone levels abrogated molecular changes in adipocytes and the favorable phenotype of Crhr1−/− mice. Conclusions: Our findings suggest the importance of the CRF2 pathway in the control of adipocyte plasticity. Increased CRF2 activity in adipocytes induces browning of WAT, differentiation of BAT and is associated with a favorable metabolic phenotype in mice lacking CRF1. Circulating corticosterone represses CRF2 activity in adipocytes and may thus regulate adipocyte physiology through the modulation of the local CRF/urocortin system. Targeting CRF receptor signaling specifically in the adipose tissue may represent a novel approach to tackle obesity

Metabolic reprogramming: a new relevant pathway in adult adrenocortical tumors

Adrenocortical carcinomas (ACCs) are complex neoplasias that may present unexpected clinical behavior, being imperative to identify new biological markers that can predict patient prognosis and provide new therapeutic options. The main aim of the present study was to evaluate the prognostic value of metabolism-related key proteins in adrenocortical carcinoma. The immunohistochemical expression of MCT1, MCT2, MCT4, CD147, CD44, GLUT1 and CAIX was evaluated in a series of 154 adult patients with adrenocortical neoplasia and associated with patients’ clinicopathological parameters. A significant increase in was found for membranous expression of MCT4, GLUT1 and CAIX in carcinomas, when compared to adenomas. Importantly MCT1, GLUT1 and CAIX expressions were significantly associated with poor prognostic variables, including high nuclear grade, high mitotic index, advanced tumor staging, presence of metastasis, as well as shorter overall and disease free survival. In opposition, MCT2 membranous expression was associated with favorable prognostic parameters. Importantly, cytoplasmic expression of CD147 was identified as an independent predictor of longer overall survival and cytoplasmic expression of CAIX as an independent predictor of longer disease-free survival. We provide evidence for a metabolic reprogramming in adrenocortical malignant tumors towards the hyperglycolytic and acid-resistant phenotype, which was associated with poor prognosis.This work was supported by FAPESP to Maria Claudia Nogueira Zerbini (2013-26344-8). SG received the post-doctoral fellowship UMINHO/BPD/18/2014.info:eu-repo/semantics/publishedVersio

Combining machine learning and SMILEs to classify, better understand, and project changes in ENSO events

The El Niño Southern Oscillation (ENSO) occurs in three phases: neutral, warm (El Niño) and cool (La Niña). While classifying El Niño and La Niña is relatively straightforward, El Niño events can be broadly classified into two types: Central Pacific (CP) and Eastern Pacific (EP). Differentiating between CP and EP events is currently dependent on both the method and observational dataset used. In this study, we create a new classification scheme using supervised machine learning trained on 18 observational and reanalysis products. This builds on previous work by identifying classes of events using the temporal evolution of sea surface temperature in multiple regions across the tropical Pacific. By applying this new classifier to seven single model initial-condition large ensembles (SMILEs) we investigate both the internal variability and forced changes in each type of ENSO event, where events identified behave similar to those observed. It is currently debated whether the observed increase in the frequency of CP events after the late 1970s is due to climate change. We found it to be within the range of internal variability in the SMILEs. When considering future changes, we do not project a change in CP frequency or amplitude under a strong warming scenario (RCP8.5/SSP370) and we find model differences in EP El Niño and La Niña frequency and amplitude projections. Finally, we find that models show differences in projected precipitation and SST pattern changes for each event type that do not seem to be linked to the Pacific mean state SST change, although the SST and precipitation changes in individual SMILEs are linked. Our work demonstrates the value of combining machine learning with climate models, and highlights the need to use SMILEs when evaluating ENSO in climate models due to the large spread of results found within a single model due to internal variability alone

Functional role of T-cell receptor nanoclusters in signal initiation and antigen discrimination

Antigen recognition by the T-cell receptor (TCR) is a hallmark of the adaptive immune system. When the TCR engages a peptide bound to the restricting major histocompatibility complex molecule (pMHC), it transmits a signal via the associated CD3 complex. How the extracellular antigen recognition event leads to intracellular phosphorylation remains unclear. Here, we used single-molecule localization microscopy to quantify the organization of TCR–CD3 complexes into nanoscale clusters and to distinguish between triggered and nontriggered TCR–CD3 complexes. We found that only TCR–CD3 complexes in dense clusters were phosphorylated and associated with downstream signaling proteins, demonstrating that the molecular density within clusters dictates signal initiation. Moreover, both pMHC dose and TCR–pMHC affinity determined the density of TCR–CD3 clusters, which scaled with overall phosphorylation levels. Thus, TCR–CD3 clustering translates antigen recognition by the TCR into signal initiation by the CD3 complex, and the formation of dense signaling-competent clusters is a process of antigen discrimination

GLUT1 expression in pediatric adrenocortical tumors: a promising candidate to predict clinical behavior

Background: Discrimination between benign and malignant tumors is a challenging process in pediatric adrenocortical tumors. New insights in the metabolic profile of pediatric adrenocortical tumors may contribute to this distinction, predict prognosis, as well as identify new molecular targets for therapy. The aim of this work is to characterize the expression of the metabolism-related proteins MCT1, MCT2, MCT4, CD147, CD44, GLUT1 and CAIX in a series of pediatric adrenocortical tumors. Methods: A total of 50 pediatric patients presenting adrenocortical tumors, including 41 clinically benign and 9 clinically malignant tumors, were included. Protein expression was evaluated using immunohistochemistry in samples arranged in tissue microarrays. Results: The immunohistochemical analysis showed a significant increase in plasma membrane expression of GLUT1 in malignant lesions, when compared to benign lesions (p=0.004), being the expression of this protein associated with shorter overall and disease-free survival (p=0.004 and p=0.001, respectively). Although significant differences were not observed for proteins other than GLUT1, MCT1, MCT4 and CD147 were highly expressed in pediatric adrenocortical neoplasias (around 90%). Conclusion: GLUT1 expression was differentially expressed in pediatric adrenocortical tumors, with higher expression in clinically malignant tumors, and associated with shorter survival, suggesting a metabolic remodeling towards a hyperglycolytic phenotype in this malignancy.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) to Maria Claudia Nogueira Zerbini (2013/26344-8) and under the scope of the project NORTE-01-0145-FEDER-000013, supported by the Northern Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER) and by FEDER, through the Competitiveness Factors Operational Programme (COMPETE), and by National funds, through the Foundation for Science and Technology (FCT), under the scope of the project POCI-01-0145-FEDER-007038info:eu-repo/semantics/publishedVersio

Meio Ambiente E Sociedade de Risco: Proteção Integral e Prevenção na Tutela Inibitória Além do Ilícito

TCC(graduação) - Universidade Federal de Santa Catarina. Centro de Ciências Jurídicas. Direito.Esta monografia tem como objetivo principal estudar a concepção de tutela inibitória e a possibilidade de sua aplicação na área do direito ambiental brasileiro, não somente diante da prática de atividades ilícitas, mas também de atividades lícitas, levando-se em conta a problemática do risco, sua tolerabilidade e intolerabilidade. Desse modo, no primeiro capítulo se apresentam noções sobre meio ambiente, teoria da sociedade de risco, de Ulrich Beck, e a preocupação da ordem jurídica com o risco. Já no segundo capítulo, destacam-se a prevenção e a proteção integral como valores essenciais e necessários em sede de direito ambiental, aliados às formas de tutela adequada dos direitos, estas incluídas entre os principais escopos do processo civil moderno. Por fim, no terceiro capítulo, abordam-se as linhas gerais da tutela inibitória, frisando-se seu feitio preventivo e específico, tal como propõe Luiz Guilherme Marinoni, inspirado na doutrina italiana. Procura-se verificar, utilizando-se o método indutivo, se é possível a utilização da teoria da tutela inibitória para se evitar o cometimento de ilícitos (e conseqüentemente a ocorrência eventual de danos), sua continuação ou repetição e, ainda, impedir a prática de lícitos, caso estes não encontrem mais amparo na tolerabilidade do risco a que expõem o meio ambiente

Endocrinology in the time of COVID-19: Management of Cushing’s syndrome

Clinical evaluation should guide those needing immediate investigation. Strict adherence to COVID-19 protection measures is necessary. Alternative ways of consultations (telephone, video) should be used. Early discussion with regional/national experts about investigation and management of potential and existing patients is strongly encouraged. Patients with moderate or severe clinical features need urgent investigation and management. Patients with active Cushing’s syndrome, especially when severe, are immunocompromised and vigorous adherence to the principles of social isolation is recommended. In patients with mild features or in whom a diagnosis is less likely, clinical re-evaluation should be repeated at 3 and 6 months or deferred until the prevalence of SARS-CoV-2 has significantly decreased; however, those individuals should be encouraged to maintain social distancing. Diagnostic pathways may need to be very different from usual recommendations in order to reduce possible exposure to SARS-CoV-2. When extensive differential diagnostic testing and/or surgery is not feasible, it should be deferred and medical treatment should be initiated. Transsphenoidal pituitary surgery should be delayed during high SARS-CoV-2 viral prevalence. Medical management rather than surgery will be the used for most patients, since the short- to mid-term prognosis depends in most cases on hypercortisolism rather than its cause; it should be initiated promptly to minimize the risk of infection in these immunosuppressed patients. The risk/benefit ratio of these recommendations will need re-evaluation every 2–3 months from April 2020 in each country (and possibly local areas) and will depend on the local health care structure and phase of pandemic

Geopolítica contemporânea e temas socioespaciais: a atuação do Brasil nas Conferências socioespaciais das Nações Unidas

O mundo multipolar pós-Guerra Fria, moldado pela Globalização, trouxe novas facetas para a Geopolítica e, modificou sob o ponto de vista geoestratégico, as táticas dos Estados para obtenção de poder no cenário internacional. A cooperação internacional procura substituir o foco conflituoso do mundo bipolar e os Estados veem, nesse novo paradigma, a possibilidade de alcançar seus objetivos e resultados desejados de novas maneiras, com o uso do soft power ou poder brando, que designa a habilidade de influenciar os outros a fazer o que você deseja pela atração em vez de coerção. Fomentar o poder brando pode ser uma das justificativas para que inúmeros países atuem e participem de cúpulas e acordos de cooperação incentivando os direitos humanos, o meio ambiente ou o desenvolvimento

Can single molecule localization microscopy be used to map closely spaced RGD nanodomains?

Cells sense and respond to nanoscale variations in the distribution of ligands to adhesion receptors. This makes single molecule localization microscopy (SMLM) an attractive tool to map the distribution of ligands on nanopatterned surfaces. We explore the use of SMLM spatial cluster analysis to detect nanodomains of the cell adhesion-stimulating tripeptide arginine-glycine-aspartic acid (RGD). These domains were formed by the phase separation of block copolymers with controllable spacing on the scale of tens of nanometers. We first determined the topology of the block copolymer with atomic force microscopy (AFM) and then imaged the localization of individual RGD peptides with direct stochastic optical reconstruction microscopy (dSTORM). To compare the data, we analyzed the dSTORM data with DBSCAN (density-based spatial clustering application with noise). The ligand distribution and polymer topology are not necessary identical since peptides may attach to the polymer outside the nanodomains and/or coupling and detection of peptides within the nanodomains is incomplete. We therefore performed simulations to explore the extent to which nanodomains could be mapped with dSTORM. We found that successful detection of nanodomains by dSTORM was influenced by the inter-domain spacing and the localization precision of individual fluorophores, and less by non-specific absorption of ligands to the substratum. For example, under our imaging conditions, DBSCAN identification of nanodomains spaced further than 50 nm apart was largely independent of background localisations, while nanodomains spaced closer than 50 nm required a localization precision of ~11 nm to correctly estimate the modal nearest neighbor distance (NDD) between nanodomains. We therefore conclude that SMLM is a promising technique to directly map the distribution and nanoscale organization of ligands and would benefit from an improved localization precision