Afadin downregulation by helicobacter pylori Induces epithelial to mesenchymal transition in gastric cells

Afadin is a cytoplasmic protein of the adherens junctions, which regulates the formation and stabilization of both the adherens and the tight junctions. Aberrant expression of Afadin has been shown in cancer and its loss has been associated with epithelial-tomesenchymal transition (EMT). EMT is characterized by the change from an epithelial to a mesenchymal phenotype, with modifications on the expression of adhesion molecules and acquisition of a migratory and invasive cell behavior. While it is known that Helicobacter pylori disrupts the tight and the adherens junctions and induces EMT, the effect of the bacteria on Afadin is still unknown. The aim of this study was to disclose the effect of H. pylori on Afadin and its impact in the induction of an EMT phenotype in gastric cells. Using two different cell lines, we observed that H. pylori infection decreased Afadin protein levels, independently of CagA, T4SS, and VacA virulence factors. H. pylori infection of cell lines recapitulated several EMT features, displacing and downregulating multiple proteins from cell–cell junctions, and increasing the expression of ZEB1, Vimentin, Slug, N-cadherin, and Snail. Silencing of Afadin by RNAi promoted delocalization of junctional proteins from the cell–cell contacts, increased paracellular permeability, and decreased transepithelial electrical resistance, all compatible with impaired junctional integrity. Afadin silencing also led to increased expression of the EMT marker Snail, and to the formation of actin stress fibers, together with increased cell motility and invasion. Finally, and in line with our in vitro data, the gastric mucosa of individuals infected with H. pylori showed decrease/loss of Afadin membrane staining at cell–cell contacts significantly more frequently than uninfected individuals. In conclusion, Afadin is downregulated by H. pylori infection in vitro and in vivo, and its downregulation leads to the emergence of EMT and to the acquisition of an aggressive phenotype in gastric cells, which can contribute to gastric carcinogenesis.This article is a result of the project NORTE-01-0145-FEDER-000029, supported by Norte Portugal Regional Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). i3S was financed by ERDF funds through the COMPETE 2020 and Portugal 2020, and by Portuguese funds through FCT – Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologia e Inovação (POCI-01-0145-FEDER-007274). MM, JM, and ML have fellowships from FCT (SFRH/BD/95631/2013, SFRH/BD/116965/2016, and SFRH/BPD/110065/2015)

Análise de genes expressos durante estádios finais da maturação de frutos de café.

A maturação uniforme dos frutos do cafeeiro relaciona-se diretamente com a qualidade da bebida. Diferentes floradas em um mesmo cafeeiro propiciam frutos em estádios desiguais de maturação podendo resultar em uma maior dificuldade na colheita, maior gasto com mão de obra e queda na qualidade do produto. Em frutos climatéricos, o processo final da maturação é desencadeado por um grande acúmulo de etileno, seguido por mudança bioquímicas e fisiológicas que promovem principalmente a desestabilização da parede celular dos frutos, composta principalmente por compostos pécticos. As pectinas são degradadas devido à solubilização e despolimerização da parede celular vegetal em decorrência da ação de enzimas como: pectinametilesterase, poligalacturonase, xiloglucanases, xilanases, pectinaliases e bgalactosidases. Nosso objetivo principal é caracterizar e entender as mudanças na expressão gênica e enzimática que ocorrem durante a maturação dos frutos de café, visando melhorar a uniformidade da maturação. Neste trabalho foram feitos estudos de expressão de genes que codificam para ACC oxidase, expansina, pectinametilesterase e poligalacturonases, através da análise ?in silico? do banco de dados do Projeto Genoma Café, e experimentos de expressão dos genes e enzimas. Análise de transcritos demonstrou um aumento de ACC oxidase em frutos verdecana seguido da atividade de pectinametilesterases em estágios intermediários de maturação (frutos avermelhados). A atividade da PG foi detectada nos estádios finais de maturação, principalmente em frutos cereja

In vitro degradation and gas production of brachiaria grass with levels of biodiesel byproducts.

ABSTRACT This study aimed to evaluate the in vitro degradability and gas production in diets containing byproducts from the national biodiesel industry (castor bean, canola, forage radish and black sunflower) replacing Brachiaria grass in four levels (0, 30, 50 and 70%). The inoculum for in vitro incubation was obtained from three fistulated Holstein cows. The experimental design was 4 x 4 factorial completely randomized experimental design consisting of four byproducts and four levels. All byproducts studied had a significant effect (p < 0.05) on in vitro digestibility. The castor bean byproducts promoted the lowest cumulative gas production at the end of 48 hours incubation. Regarding digestibility, the byproducts of canola and radish at 70% level did not affect the degradability of dry matter

Genetic diversity of the grapevine (Vitis vinifera L.) cultivars most utilized for wine production in Portugal

Portugal is very rich in grapevine biodiversity but nowadays only a small number of cultivars (13 black and 3 white) with defined enological characteristics are being used. However, there are still 51 cultivars with significant expression in the country which have a great potential to be used in the establishment of new vineyards. Considering the importance of identifying and understanding the genetic relatedness of the main cultivars used for wine making in Portugal, those cultivars were genotyped with the 6 SSRs loci VVMD5, VVMD7, VVMD27, VrZag62, VrZag79 and VVS2 that are recommended by the Office de la Vigne et du Vin for the elaboration of the worldwide database containing the alleles of all the known grapevine cultivars. A total of 49 alleles were detected with an average allele number of 8.17 per locus. The SSR profile of the 51 cultivars, at the 6 evaluated loci, indicates that all can be distinguished from each other. In addition, this study produces information on the relationship of these 51 grapevine cultivars with cultivars of other important Mediterranean wine producing regions

Perfil de ácidos graxos no Longissimus dorsi de ovinos Santa Inês em crescimento alimentados com diferentes níveis de energia metabolizável.

O presente estudo objetivou avaliar a influência de rações com diferentes níveis de energia metabolizável sobre o perfil de ácidos graxos no músculo longissimus dorsi de cordeiros Santa Inês. Foram utilizados 20 cordeiros, com idade e peso de 50 dias e 13,0 ± 0,56 kg, respectivamente, alimentados com rações contendo diferentes níveis energéticos: 2,08; 2,28; 2,47 e 2,69 Mcal/kgMS. A duração do experimento foi definida pelo tempo necessário para que a média do peso de todos os animais de cada tratamento alcançasse 28 kg, ocasião em que os mesmos foram abatidos. Após o abate, para a análise do perfil de ácidos graxos foi retirada da meia carcaça esquerda, uma amostra do músculo longissimus dorsi compreendida entre a 12ª e 13ª costelas. Para a análise do perfil de ácidos graxos foi utilizado cromatógrafo a gás Shimadzu GC 2010. As rações influenciaram os ácidos graxos cis,10-heptadecanóico e eicosanotrienóico (P<0,05), não influenciando os demais ácidos graxos encontrados. Os ácidos palmítico (C16:0) e esteárico (C18:0) contribuíram mais intensamente nos valores totais de ácidos graxos saturados. Os ácidos graxos C16:0; C16:1; C18:0; C18:1 e C18:2, constituíram mais 90% das áreas totais do cromatograma. Os níveis de energia dietéticos influenciam o teor do ácidos graxo saturado cis 10-heptadecanóico, bem como do ácido graxo poliinsaturado eicosatrienóico no músculo longissimus dorsi de cordeiros da raça Santa Inês. [Profile of fatty acids in Longissimus dorsi of growing Santa Inês sheep fed with different energy levels]. Abstract: his study evaluated the influence of diets with different metabolizable energy levels on the fatty acid profile of longissimus dorsi muscle of Santa Ines lambs. We used 20 lambs, age and weight at 50 days and 13.0 ± 0.56 kg, respectively, fed diets with different energy levels: 2.08, 2.28, 2.47 and 2.69 Mcal/kgDM. The duration of the experiment was defined by the time it takes for the average weight of all animals in each treatment reached 28 kg, at which time they were slaughtered. After slaughter, to analyze the fatty acid profile was removed from the left half carcass, a sample of the longissimus dorsi between the 12th and 13th ribs. To analyze the fatty acid profile was used a gas chromatograph Shimadzu GC 2010. The diets influenced fatty acids and cis,10-heptadecanoic eicosanotrienóico (P <0.05), not to influence the other fatty acids found. The palmitic acid (C16:0) and stearic (C18:0) contributed more strongly in the values of total saturated fatty acids. The fatty acid C16:0, C16:1, C18: 0, C18:1 and C18:2, constituted over 90% of the total area of chromatogram. The levels of dietary energy influence the content of saturated fatty acids cis 10-heptadecanoic and polyunsaturated fatty acid eicosatrienóico in the longissimus dorsi of Santa Ines lambs

Design and Development of a Magnus Hydrokinetic Rotor

The energy matrix diversification has become noticed in the latest years. Energy conversion of the free flow in rivers and canals into electrical energy has been a good complementation for the conventional generation. The first application of hydrokinetic turbines, commercially, were in Mississippi's river (Minnesota, USA), in 2008. The USAge of Magnus effect in hydrokinetic turbines occurred in an innovative manner. In this project, rotational cylinders actuate as blades of an axial hydrokinetic rotor, converting kinetic energy of the flow into potency in the rotor axle. This effect was initially observed in 1853 by Henrich Magnus and, since then, few researches were carried out to its application in hydraulic generation of energy. Therefore, tests in reduced-scale prototype and numerical simulations were made for the development and executive design of a hydrokinetic rotor. At the end of this study, a hydro generator with 62% hydraulic efficiency, considering the Betz Limit, was constructed