Análise comparativa entre isolados de Fonsecaea sp na modulação da resposta imune do hospedeiro : papel das vesículas e componentes extracelulares

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Instituto de Ciências Biológicas, Departamento de Biologia Molecular, 2021.A cromoblastomicose (CBM) é uma micose cutânea e subcutânea crônica, causada por fungos negros, dimórficos e filamentosos da família Dematiaceae. Fonsecaea pedrosoi é considerado uma das principais espécies causadoras da doença no Brasil, apesar de que F. monophora, F. nubica e F. pugnacius terem sido descritas como outras espécies capazes de causar CBM em humanos. Todas estas espécies possuem a capacidade de transitar entre as formas saprofíticas, conídios e hifas, e a forma patogênica do fungo, as células muriformes (MCs). Fatores de virulência produzidos por fungos, como vesículas extracelulares (EVs) e moléculas extracelulares presentes no meio condicionado (CM) vem sendo mais estudados nos últimos anos, apesar de existirem poucas informações sobre o papel destas moléculas extracelulares durante a CBM. Este trabalho visa compreender os padrões imunoestimulatórios in vitro e in vivo causados por EVs e CMs produzidos por F. pedrosoi, F. monophora, F. nubica e F. erecta isolados a partir de conídios e hifas cultivados em meio rico (RM) e meio mínimo (MM), além de analisar o papel dessas moléculas quando produzidas na presença ou ausência de MCs. Em relação as diferentes condições nutricionais de cultivo, foi demonstrado que EVs produzidas em RM tem maior capacidade de estimular a produção de TNF, IL-1β e IL-10 por BMDMs em comparação a EVs produzidas em MM na presença ou na ausência de MCs. Tais resultados são diferentes dos observados para EVs de C. neoformans produzidas em RM e MM, em que EVs isoladas de MM demonstram estimular mais a produção de citocinas pró-inflamatórias do que em RM. Já os CMs demonstram ser mais pró-inflamatórios do que as EVs, principalmente os isolados a partir de MM na ausência de MCs, aumentando os níveis de secreção das citocinas TNF, IL-1β e IL-10 in vitro. Por fim, os CMs produzidos por F. pedrosoi e F. erecta foram utilizados como tratamento in vivo e demonstraram agravar a progressão da CBM em modelo murino, aumentando o diâmetro da área da lesão, o número de unidades formadoras de colônia e os níveis das citocinas pró-inflamatórias TNF e IL-1β em relação ao controle positivo tratado com PBS. Já foi observado que as moléculas presentes no CM produzidos pela forma capsular de C. neoformans conseguem inibir a ativação do inflamassoma NLRP3 e promover a proliferação celular in vitro, indicando que as moléculas presentes nos CMs de fungos patogênicos possuem potencial de imunomodular a resposta imune do hospedeiro. Desta forma, observa-se que as EVs e os CMs produzidos por diferentes espécies do gênero Fonsecaea possuem a capacidade tanto de estimular, quanto de inibir a secreção de citocinas importantes na resposta imune inata como TNF, IL-1β e IL-10, demonstrando serem moléculas com capacidade de virulência e possivelmente, de modular a progressão da CBM em humanos.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).Chromoblastomycosis (CBM) is a chronic cutaneous and subcutaneous mycosis caused by black, dimorphic and filamentous fungi of the Dematiaceae family. Fonsecaea pedrosoi is considered one of the main species causing the disease, although F. monophora, F. nubica and F. pugnacious are considered as other species capable of causing CBM in humans. All these species have an ability to transition between saprophytic forms, such as conidia and hyphae and the pathogenic form of the fungus, the murine cells (MCs). Virulence factors produced by fungi, such as extracellular vesicles (EVs) and conditioned media (CM) have been more studied in recent years, although there is little information about the roles of these extracellular molecules during CBM. This work aims to understand the in vitro and in vivo immunostimulatory patterns caused by EVs and CMs obtained by F. pedrosoi, F. monophora, F. nubica and F. erecta isolated from conidia and hyphae cultivated in rich medium (RM) and minimal medium (MM), in addition to analyzing the role of these molecules when produced in the presence or absence of MCs. Regarding the different nutritional conditions of cultivation, it was shown that EVs produced in RM have a greater capacity to stimulate the production of TNF, IL-1β and IL-10 by BMDMs when compared to EVs produced in MM in the presence or absence of MCs. These results are different from those observed for EVs of C. neoformans produced in RM and MM, in which EVs isolated from MM are shown to stimulate more the production of pro-inflammatory cytokines than in RM. The CMs, on the other hand, demonstrate to be more pro-inflammatory than EVs, especially those isolated from MM in the absence of MCs, which increased the levels of cytokine secretion of TNF, IL-1β and IL-10 in vitro. Finally, the CMs produced from F. pedrosoi and F. erecta were used as an in vivo treatment and where shown to aggravate the progression of CBM in a murine model, increasing the diameter of the lesion area, the number of colony forming units and the levels of pro-inflammatory cytokines such as TNF and IL-1β, compared to the positive control treated with PBS. The role of the molecules presented in the CM produced by capsular C. neoformans has already been observed to inhibit inflamassome NLRP3 activation and promote cell proliferation in vitro, indicating that theses extracellular molecules produced by pathogenic fungi have the potential to immunomodulate the host’s immune response. Thus, it is observed that the EVs produced by different species of the Fonsecaea genus have the ability to both stimulate and inhibit important cytokines in the innate immune response, proving to be molecules with high virulence capacity and possible mediators of the CBM progression in humans

A less expensive NiMnGa based Heusler alloy for magnetic refrigeration

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)We present a study of the substitution of Mn by Cu on the compound Ni2Mn1-xCuxGa0.9Al0.1, showing that the substitution of a small amount of Al on the Ga site does not affect the magnetic and magnetocaloric potential compared to Ni-2(Mn,Cu)Ga alloy. The samples were prepared with 10% substitution of Al and with Cu concentrations of x = 0.0, 0.2, and 0.3. Magnetization measurements as a function of temperature performed from 10 to 400 K, with an applied field of 0.02 T showed a ferromagnetic state, with critical temperature T-c = 295 and 300 K for the samples with Cu, x = 0.2 and 0.3, respectively. For the sample without Cu, a complex behavior is observed at T-c = 370 K, with martensitic transition at 220 K and a premartensitic at 250 K. Analysis of x-rays diffractograms at room temperature show a L2(1) structure for x = 0.0, while for x = 0.2 a mixture of L2(1) and martensitic is present, and the sample with x = 0.3 it is in a fully martensitic phase. Heat capacity measurements were performed in order to calculate magnetocaloric effect in the samples. The results indicate that in Ni(Mn,Cu)Ga alloys, a partial substitution of Ga by Al still produce a high refrigerant capacity while reducing the costs of fabrication. (C) 2012 American Institute of Physics. [doi:10.1063/1.3675064]1117Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Anti-atherogenic properties associated with the antioxidant activity from the hydrophilic extracts of Halimeda incrassata (Chlorophyta, Bryopsidales)

Seaweeds are a source of natural antioxidants having potential application in oxidative stress and associated diseases. In this work, anti-atherogenic properties associated with the antioxidant activity from the hydrophilic extracts of Halimeda incrassata were studied. The phenolic content assessed inthe aqueous extract and fraction phenolic acids (FPA) was 0.13 ± 0.05 and 0.47 ± 0.09 mg of gallic acid equivalents (GAE)/g dry seaweed, respectively. In DPPH?, radical scavenging assay fractions exhibited a dependent concentration. The seaweeds extract inhibited the desoxirribose oxidation in the presenceor absence of EDTA (IC50 = 1.91± 0.09 mg/mL) (IC50 = 2.95 ± 0.01 mg/mL). In vivo antioxidant properties of FPA-H.incrassata were investigated in rats with a CCl4-induced liver injury. Pre-treatment with H.incrassata led to approximately 50% reductions in liver TBARS levels. The treatment with H. incrassataFPA also increased the activity of the CAT enzyme, which in turn resulted in an enhanced antioxidantdefense. The expression of Catalase by PCR-RT technique demonstrated a higher gene expression when compared with that which was observed in the CCl 4-treated group. Antiatherogenic properties were studied in the inhibition of lipoprotein oxidation mediated by Cu2+ or HRP/H2O2, free radicalscavenging, and metal ion chelation, and it was dose dependent with a higher concentration needed for the aqueous extract than for the FPA fraction. Antioxidant activity was also improved in macrophages as evaluated in the cell supernatant (by TBARS formation); and by luminol enhanced chemiluminescence after cell activation with zymosan; and a degree of cell lipoperoxidation wasdecreased by the Halimeda incrassata extract. The results of this work add to the antioxidant potential of the seaweed for its application in oxidative stress associated conditions.Fil: Vidal-Novoa, Alexis. Universidad de la Habana. Facultad de Biología; CubaFil: Costa-Mugica, Ariadna. Universidad de la Habana. Facultad de Biología; CubaFil: Zulueta Díaz, Yenisleidy de Las Mercedes. Universidad de la Habana. Facultad de Biología; Cuba. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; ArgentinaFil: Diaz-Gutierrez, Daylín. Universidad de la Habana. Facultad de Biología; CubaFil: de Oliveira e Silva, Ana, Mara. Universidade de Sao Paulo; BrasilFil: Vazquez, Ana María. Center For Molecular Inmunology; CubaFil: Claudina, Zaldívar-Munoz. Universidad de la Habana. Facultad de Biología; CubaFil: Dalva, Assuncao Portari de Mancini. Institute Butantan Sao Paulo; BrasilFil: Mancini-Filho, Jorge. Universidade de Sao Paulo; Brasi

The Atp-dependent Rna Helicase Hrpb Plays An Important Role In Motility And Biofilm Formation In Xanthomonas Citri Subsp Citri

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)RNA helicases are enzymes that catalyze the separation of double-stranded RNA (dsRNA) using the free energy of ATP binding and hydrolysis. DEAD/DEAH families participate in many different aspects of RNA metabolism, including RNA synthesis, RNA folding, RNA-RNA interactions, RNA localization and RNA degradation. Several important bacterial DEAD/DEAH-box RNA helicases have been extensively studied. In this study, we characterize the ATP-dependent RNA helicase encoded by the hrpB (XAC0293) gene using deletion and genetic complementation assays. We provide insights into the function of the hrpB gene in Xanthomonas citri subsp. citri by investigating the roles of hrpB in biofilm formation on abiotic surfaces and host leaves, cell motility, host virulence of the citrus canker bacterium and growth in planta. Results: The hrpB gene is highly conserved in the sequenced strains of Xanthomonas. Mutation of the hrpB gene (Delta hrpB) resulted in a significant reduction in biofilms on abiotic surfaces and host leaves. Delta hrpB also exhibited increased cell dispersion on solid medium plates. Delta hrpB showed reduced adhesion on biotic and abiotic surfaces and delayed development in disease symptoms when sprayed on susceptible citrus leaves. Quantitative reverse transcription-PCR assays indicated that deletion of hrpB reduced the expression of four type IV pili genes. The transcriptional start site of fimA (XAC3241) was determined using rapid amplification of 5'-cDNA Ends (5' RACE). Based on the results of fimA mRNA structure predictions, the fimA 5' UTR may contain three different loops. HrpB may be involved in alterations to the structure of fimA mRNA that promote the stability of fimA RNA. Conclusions: Our data show that hrpB is involved in adherence of Xanthomonas citri subsp. citri to different surfaces. In addition, to the best of our knowledge, this is the first time that a DEAH RNA helicase has been implicated in the regulation of type IV pili in Xanthomonas.1635INCT CitrusCAPES/PSDE fellowship [99999.002657/2014-07]CNPqCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Morphology and molecular evidence support the validity of Pogonias courbina (Lacepède, 1803) (Teleostei: Sciaenidae), with a redescription and neotype designation

The family Sciaenidae comprises about 300 species. The black drum Pogonias cromis was the only valid species of the genus. Herein, Pogonias courbina Lacepède 1803 is rede-scribed based on morphological and molecular evidence and a neotype is designated. Pogonias courbina is distinguished by the following characters: the occurrence of characteristic thickening of the dorsal spines VII to XI in all specimens larger than 250 mm SL; all pterygiophores in the dorsal-fin laminar, thin; anal-fin pterygiophores slender excluded those of spines; lateral projections of gas bladder with few finger-like projections; genetic distance between both species 1%; exclusive occurrence of characters in six informative sites of COI (58 G; 214 G; 328 A; 331 A; 553C; 580 G). The method Automatic Barcode gap Discovery detected gaps in nucleotid distance congruent with the NJ, MP, and ML tree analysis. Also, advertisement calls are three times shorter in duration in P. courbina than in P. cromis. In addition, two monophyletic groups for P. cromis and P. courbina appear in trees obtained with different methodologies, emphasizing the absence of shared haplotypes. A gap of about 8000 km occurs in the distribution of both species along coastal areas of the Atlantic Ocean.Fil: Azpelicueta, María de las Mercedes. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Área Zoología; ArgentinaFil: Delpiani, Sergio Matías. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; ArgentinaFil: Cione, Alberto Luis. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Departamento Paleontología; ArgentinaFil: Oliveira, Claudio. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Marceniuk, Alexandre Pires. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Díaz de Astarloa, Juan Martín. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentin

Investigation of the effect of nozzle design on rheological bioprinting properties using computational fluid dynamics

Bioprinting is the utilization of techniques derived from three-dimensional printing to generate complex biologicalstructures which may replace natural tissues or organs. It employs high spatial resolution depositionof different cell types, growth factors and biomaterials. Those together form bioinks, which are the bioprintinginputs, analogously to conventional inks with regard to inkjet printing. In extrusion bioprinting, continuousbioink filaments are deposited layer by layer on a surface by means of an extruder nozzle, employing thedisplacement of a piston or pneumatic pressure. If mechanical stresses applied on a cell membrane exceed acritical value, which depends on the cell type, the cell membrane may disrupt. Computational fluid dynamics(CFD) simulations of the bioink extrusion were done to evaluate shear stresses caused by the internal pressureof extruder nozzles during bioprinting. Different three-dimensional conical nozzle designs were testedby varying angles of convergence, lengths, input diameters and output diameters of the nozzles. The powerlawmodel, with constants k = 109.73 Pa·s0,154 and n = 0.154, was used to describe the expected non-Newtonian behavior of the bioink. Shear stresses and shear rates were evaluated for each nozzle design consideringdifferent pressures or velocities as boundary conditions at the nozzle entrance. The maximum wallshear stress value on each different nozzle varied between 1,038 Pa and 4,915 Pa. The results indicated whichdetails of nozzle geometry are most relevant in order to optimize bioprinting. The best conditions for bioinkrheology were also evaluated to ensure good printability and high cell viability.Keywords: bioink, bioprinting, biofabrication, 3D printing, CFD

NOVOS DANOS DA RESPONSABILIDADE CIVIL NO DIREITO BRASILEIRO

O presente artigo apresenta as novas modalidades de danos que surgem de alguns julgados brasileiros, sua necessidade e possibilidade jurídica. Para tanto, faz o estudo de danos no contexto da dignidade da pessoa humana, como vêm sendo examinadas algumas dessas modalidades e como se caracterizam. Com isto, analisamos a real existência de novos danos ou se estes são apenas designações do mesmo dano moral

Sustainable multifunctional phenolic lipids as potential therapeutics in Dentistry

Phenolic lipids components of the cashew nutshell liquid (CNSL) have molecular structures capable of chemical signalling that regulate gene expression, metabolism and inflammation. This study sets out to assess how CNSL derivatives impact oral bacteria, from an antibacterial and anti-collagenolytic perspective, as well as its biocompatibility with dental pulp stem cells. Two hemi-synthetic saturated CNSL derivative compounds were selected (LDT11-Anacardic Acids-derivative and LDT409-cardanol-derivative). Bacteriostatic activity was tested against Streptococcus mutans and Veillonella parvula. Antimicrobial capacity against preformed S. mutans biofilms was investigated using a collagen-coated Calgary Biofilm Device and confocal microscopy. Clostridium histolyticum, P. gingivalis and S. mutans biofilms were used to assess anti-collagenolytic activity. Biocompatibility with human dental pulp stromal cells (HDPSCs) was investigated (MTT for viability proportion, LDH assays for cell death rate). LDTs inhibited the bacterial growth, as well as partially inhibited bacterial collagenases in concentrations higher than 5 μg/mL. Dose–response rates of biofilm cell death was observed (LDT11 at 20, 50, 100 μg/mL = 1.0 ± 0.4, 0.7 ± 0.3, 0.6 ± 0.03, respectively). Maximum cytotoxicity was 30%. After 1 week, LDT409 had no HDPSCs death. HDPSCs viability was decreased after 24 h of treatment with LDT11 and LDT409, but recovered at 72 h and showed a massive increase in viability and proliferation after 1 week. LDTs treatment was associated with odontoblast-like morphology. In conclusion, LDT11 multifunctionality and biocompatibility, stimulating dental pulp stem cells proliferation and differentiation, indicates a potential as a bio-based dental material for regenerative Dentistry. Its potential as a bacterial collagenases inhibitor to reduce collagen degradation in root/dentinal caries can be further explored

EU GPP criteria for Food procurement, Catering services and vending machines

This report summarized the discussions held during the revision process of the Green Public Procurement criteria for Food procurement and catering services. The report includes a summary of the preliminary report that includes related information such as market analysis and technical analysis and those changes in the criteria structure and wording that have been proposed and discussed during the whole process. The main part of the report consists of the wording and rationale of each of the revised GPP criteria. This section is organized in three main blocks including the proposed criteria on Food, catering services and vending machines. Each criterion is classified as contractual performance clause, technical specification, selection criteria or award criteria and includes the wording and values proposed (in the explanatory notes) for the core and the comprehensive levels. The report closes with the life cycle considerations for this product group.JRC.B.5-Circular Economy and Industrial Leadershi

Gardens and Tourism for and beyond economic profit

Gardens and Tourism for and beyond economic profit é o segundo volume da Collection of Gardens and Landscape Studies, coordenada por Ana Duarte Rodrigues. Conta com textos de Alexandra Gago da Câmara, Ana Duarte Rodrigues, António Lamas, Antonio Perla de las Parras, Celso Mangucci, Desidério Batista, Filipe Benjamim, Ignacio Rodriguez Somovilla, Jean-Paul Brigand, Maria Isabel Donas Botto, Nuno Oliveira, Paulo Carvalho, Susana Silva e Victoria Soto Caba. Assumindo uma perspectiva multidisciplinar das áreas da história da arte, arquitectura paisagista, literatura e geografia, e reunindo académicos e profissionais que operam na área do Garden Tourism, este livro constitui um trabalho inaugural nos estudos de jardins e paisagem. Certamente que constitui um balanço baseado em casos de estudo das potencialidades do turismo de jardins em Portugal, mas também aponta caminhos e hipóteses de trabalho num futuro próximo. Constitui um convite a que todos os interessados na preservação e promoção deste património se unam para criar as ferramentas necessárias à sua dinamização e salvaguarda para o futuro