Impact of metabolic disorders on the structural, functional, and immunological integrity of the blood-brain barrier: Therapeutic avenues

: Mounting evidence has linked the metabolic disease to neurovascular disorders and cognitive decline. Using a murine model of a high-fat high-sugar diet mimicking obesity-induced type 2 diabetes mellitus (T2DM) in humans, we show that pro-inflammatory mediators and altered immune responses damage the blood-brain barrier (BBB) structure, triggering a proinflammatory metabolic phenotype. We find that disruption to tight junctions and basal lamina due to loss of control in the production of matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) causes BBB impairment. Together the disruption to the structural and functional integrity of the BBB results in enhanced transmigration of leukocytes across the BBB that could contribute to an initiation of a neuroinflammatory response through activation of microglia. Using a humanized in vitro model of the BBB and T2DM patient post-mortem brains, we show the translatable applicability of our results. We find a leaky BBB phenotype in T2DM patients can be attributed to a loss of junctional proteins through changes in inflammatory mediators and MMP/TIMP levels, resulting in increased leukocyte extravasation into the brain parenchyma. We further investigated therapeutic avenues to reduce and restore the BBB damage caused by HFHS-feeding. Pharmacological treatment with recombinant annexin A1 (hrANXA1) or reversion from a high-fat high-sugar diet to a control chow diet (dietary intervention), attenuated T2DM development, reduced inflammation, and restored BBB integrity in the animals. Given the rising incidence of diabetes worldwide, understanding metabolic-disease-associated brain microvessel damage is vital and the proposed therapeutic avenues could help alleviate the burden of these diseases

Amphiphilic polylactide-poly(ethylene oxide)-poly(propylene oxide) block copolymers : self-assembly behavior and cell affinity

Polylactide (PLA) is a biodegradable polyester recognized for its potential use as a biomedical material. Poly(ethylene oxide) (PEO) and copolymers based on PEO and poly(propylene oxide) (PPO) are biocompatible polyethers widely applied in the biomedical field, particularly as macromolecular nonionic surfactants. In this work, PLA blocks were attached to the PEO and to the PEO and PPO‐based triblock copolymer PEO–PPO–PEO, through ring‐opening polymerization of racemic lactide (rac ‐LA) to obtain the amphiphilic triblock PLA–PEO–PLA and pentablock PLA–PEO–PPO–PEO–PLA copolymers containing hydrophilic/hydrophobic blocks with variable block mass ratios. The copolymers were evaluated for chemical composition, molar mass, and thermal properties, and they were used to prepare self‐assemble aggregates in water from tetrahydrofuran polymer solutions. The combination of scattering light experiments and microscopy techniques revealed the spherical morphology of the aggregates with diameters around 180–200 nm, which comprises a hydrophobic PLA core and a hydrophilic polyether shell. The aggregates are nontoxic to human cervical cancer cell line — HeLa cells, as determined by MTS assay, and the aggregates are potential candidates to be applied in the encapsulation of hydrophobic compounds561922032213CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP444392/2014-92010/17804-7; 2012/24821–0; 2015/25406–5The authors would like to acknowledge the financial support of the São Paulo Research Foundation – FAPESP (processes 2010/17804–7, 2012/24821–0, and 2015/25406–5) and of the Brazilian National Council for Scientific and Technological Development (CNPq, process 444392/2014–9). The authors thank the Brazilian Nanotechnology National Laboratory (LNNano) at the Brazilian Center for Research in Energy and Materials (CNPEM) for the use of electron microscopy and atomic force microcopy facilities. They also thank Prof. Dr Cátia C. C. O. Megiatto (Institute of Chemistry, UNICAMP) for providing HeLa cells for biological assays and Dr Annelize Zambon (Institute of Chemistry, UNICAMP) for training and technical suppor

Generation and characterization of a spontaneously immortalized endothelial cell line from mice microcirculation

Endothelial cells from microvasculature are directly involved in a large number of vascular diseases; however, culture of these cells is problematic, since most methodologies employ proteolytic enzymes or mechanical techniques, leading to cell damage and contamination of endothelial cultures with other cellular types. Besides, primary cultured cells have a short life span in vitro and undergo replicative senescence after 3-4 passages, limiting long-term studies. in the present work we report the generation of a spontaneously immortalized endothelial culture obtained from mice pulmonary capillaries. Firstly, primary (third passage) and immortalized (100th) cultures were established. Further, monoclonal populations were obtained by serial dilutions from immortalized cultures. Cells were analyzed according to: (1) morphological appearance, (2) expression of specific endothelial markers by fluorescent staining [von Willebrand Factor (vWF), endothelial nitric oxide synthase (eNOS), angiotensin converting enzyme (ACE) and Ulex europaeus (UEA-1)] and by flow cytometry (endoglin, VE-cadherin and VCAM-1), and (3) release of nitric oxide (NO), assessed by the specific fluorescent dye DAF-2 DA, and prostacyclin (PGI(2)), quantified by enzyme immune assay. in both cultures cells grew in monolayers and presented cobblestone appearance at confluence. Positive staining for vWF, eNOS, ACE and UEA-1 was detected in cloned as well as in early-passage cultured cells. Similarly, cultures presented equal expressions of endoglin, VE-cadherin and VCAM-1. Values of NO and PGI(2) levels did not differ between cultures. From these results we confirm that the described spontaneously immortalized endothelial cell line is capable of unlimited growth and retains typical morphological and functional properties exhibited by primary cultured cells. Therefore, the endothelial cell line described in the present study can become a suitable tool in the field of endothelium research and can be useful for the investigation of production of endothelial mediators, angiogenesis and inflammation. (c) 2013 Elsevier Inc. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade Federal de São Paulo, Inst Ciencias Ambientais Quim & Farmaceut, BR-09913030 São Paulo, BrazilUniv São Paulo, Inst Ciencias Biomed, Dept Imunol, BR-05508900 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biofis, BR-04039032 São Paulo, BrazilUniversidade Federal de São Paulo, Inst Ciencias Ambientais Quim & Farmaceut, BR-09913030 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biofis, BR-04039032 São Paulo, BrazilFAPESP: FAPESP 2007/59039-2FAPESP: 2008/06676-8FAPESP: 2010/01404-0Web of Scienc

Costurando sonhos: a presença boliviana em São Paulo

Sinopse: Estima-se que a comunidade de bolivianos na cidade São Paulo seja de 200 mil pessoas. Imigrados da Bolívia em busca de trabalho, principalmente no ramo da costura, concentram-se nos bairros do Pari e Bom Retiro, onde há um grande número de confecções. Em sua maioria, os bolivianos consideram que as condições de vida no Brasil são melhores do que em seu país de origem. Muitos, no entanto, por não terem documentos e não falarem português, são explorados por donos de oficinas de costura, sendo até submetidos ao trabalho em condições análogas à escravidão. É comum vê-los procurando serviço nas ruas do Bom Retiro, de porta em porta nas oficinas de costura. Mas é comum também encontrá-los reunidos, aos sábados e domingos, em feiras e eventos da comunidade boliviana, com música, comidas e danças típicas. Costurando Sonhos mostra um pouco dessa pequena Bolívia em São Paulo e revela, a partir de depoimentos e entrevistas, as dificuldades de adaptação desses imigrantes em um novo contexto sociocultural. SÉRIE LUGARES DA INTOLERÂNCI

Chloroquine, an Endocytosis Blocking Agent, Inhibits Zika Virus Infection in Different Cell Models

Zika virus (ZIKV) infection in utero might lead to microcephaly and other congenital defects. Since no specific therapy is available thus far, there is an urgent need for the discovery of agents capable of inhibiting its viral replication and deleterious effects. Chloroquine is widely used as an antimalarial drug, anti-inflammatory agent, and it also shows antiviral activity against several viruses. Here we show that chloroquine exhibits antiviral activity against ZIKV in Vero cells, human brain microvascular endothelial cells, human neural stem cells, and mouse neurospheres. We demonstrate that chloroquine reduces the number of ZIKV-infected cells in vitro, and inhibits virus production and cell death promoted by ZIKV infection without cytotoxic effects. In addition, chloroquine treatment partially reveres morphological changes induced by ZIKV infection in mouse neurospheres

Annexin A1 attenuates microvascular complications through restoration of Akt signalling in a murine model of type 1 diabetes

British Heart Foundation (Award number: FS/13/58/30648) to GP; the Ministry of Education, Brazil (Grant number: 7326/2014-09) to RAL; University of Turin (Ricerca Locale Linea B 2015 and Linea A 2016) to MC; the William Harvey Research Foundation to CT; Bart’s and The London Charity Centre of Diabetic Kidney Disease (programme grant:577/2348) to MY and CT; and FISM Fondazione Italiana Sclerosi Multipla–Cod. 2014/R/21 to ES

The western south atlantic ocean in a high-CO2 world: current measurement capabilities and perspectives

An international multi-disciplinary group of 24 researchers met to discuss ocean acidification (OA) during the Brazilian OA Network/Surface Ocean-Lower Atmosphere Study (BrOA/SOLAS) Workshop. Fifteen members of the BrOA Network (www. broa. furg. br) authored this review. The group concluded that identifying and evaluating the regional effects of OA is impossible without understanding the natural variability of seawater carbonate systems in marine ecosystems through a series of long-term observations. Here, we show that the western South Atlantic Ocean (WSAO) lacks appropriate observations for determining regional OA effects, including the effects of OA on key sensitive Brazilian ecosystems in this area. The impacts of OA likely affect marine life in coastal and oceanic ecosystems, with further social and economic consequences for Brazil and neighboring countries. Thus, we present (i) the diversity of coastal and open ocean ecosystems in the WSAO and emphasize their roles in the marine carbon cycle and biodiversity and their vulnerabilities to OA effects; (ii) ongoing observational, experimental, and modeling efforts that investigate OA in the WSAO; and (iii) highlights of the knowledge gaps, infrastructure deficiencies, and OA-related issues in the WSAO. Finally, this review outlines long-term actions that should be taken to manage marine ecosystems in this vast and unexplored ocean region