Decellularized human colorectal cancer matrices polarize macrophages towards an anti-inflammatory phenotype promoting cancer cell invasion via CCL18.

Macrophages are frequently identified in solid tumors, playing important roles in cancer progression. Their remarkable plasticity makes them very sensitive to environmental factors, including the extracellular matrix (ECM). In the present work, we investigated the impact of human colorectal tumor matrices on macrophage polarization and on macrophage-mediated cancer cell invasion. Accordingly, we developed an innovative 3D-organotypic model, based on the decellularization of normal and tumor tissues derived from colorectal cancer patients' surgical resections. Extensive characterization of these scaffolds revealed that DNA and other cell constituents were efficiently removed, while native tissue characteristics, namely major ECM components, architecture and mechanical properties, were preserved. Notably, normal and tumor decellularized matrices distinctly promoted macrophage polarization, with macrophages in tumor matrices differentiating towards an anti-inflammatory M2-like phenotype (higher IL-10, TGF-ß and CCL18 and lower CCR7 and TNF expression). Matrigel invasion assays revealed that tumor ECM-educated macrophages efficiently stimulated cancer cell invasion through a mechanism involving CCL18. Notably, the high expression of this chemokine at the invasive front of human colorectal tumors correlated with advanced tumor staging. Our approach evidences that normal and tumor decellularized matrices constitute excellent scaffolds when trying to recreate complex microenvironments to understand basic mechanisms of disease or therapeutic resistance.

Evaluation of physico-chemical characteristics of fresh, refrigerated and frozen Lacaune ewes' milk / Avaliação das características físico-químicas de leite fresco, resfriado e congelado de ovelhas Lacaune

The production of ewe milk is seasonal and milk yield per animal is low, even in specialized animals. This study aimed to verify the possibility of preserving bulk tank milk for seven days under cooling (5°C) and freezing (-5°C), verify the influence of cooling treatments and of the months of the year on the physical and chemical characteristics of the product. The chemical composition of milk, including the fat, protein, lactose and total solids contents, was not altered by cooling and freezing. Protein and lactose contents varied according to the months of the year. The average percentage and standard deviation of fat, protein, lactose and total solids was 8.10±1.30, 5.22±0.37, 4.43±0.23 and 19.34±1.54, respectively. The density, pH, titratable acidity, as well as alcohol and heat stability tests were significantly influenced by the treatments used (P < 0.05), but no differences were found between fresh and frozen milk. Prolonged refrigeration caused an increase in acidity and decrease in pH, with a consequent reduction in the stability of milk. These results demonstrated that freezing does not affect the chemical composition and physical characteristics of milk in nature and it could be a solution for the producer and the sheep milk industry...

O leite ovino, mesmo no caso de raças especializadas, apresenta sazonalidade de produção e baixa produtividade por fêmea. Em função disso, o objetivo do presente estudo foi verificar a possibilidade de conservação do leite fresco, por um período de sete dias, sob refrigeração e congelamento, verificando a influência dos tratamentos aplicados, e dos meses do ano, sobre as características físico-químicas do produto. A composição química, incluindo gordura, proteína, lactose e sólidos totais, não sofreu alterações com o resfriamento e congelamento do leite, contudo os teores de proteína e lactose variaram durante os meses do ano. O percentual médio de gordura, proteína, lactose e sólidos totais foi de 8,10±1,30, 5,22±0,37, 4,43±0,23 e 19,34±1,54, respectivamente. A densidade, o pH, a acidez titulável e as provas de estabilidade ao álcool e ao calor sofreram influência significativa dos tratamentos aplicados (P<0,05), não havendo diferenças entre o leite fresco e o leite congelado. O resfriamento prolongado causou aumento da acidez e diminuição do pH, com a consequente diminuição da estabilidade do leite. Os resultados do presente estudo demonstram que o congelamento não afeta a composição química e as características físicas do leite in natura, podendo ser uma solução para o produtor e para a indústria de leite ovino...

Macrophages stimulate gastric and colorectal cancer invasion through EGFR Y(1086), c-Src, Erk1/2 and Akt phosphorylation and smallGTPase activity.

The interactions between cancer cells and their microenvironment are crucial for malignant progression, as they modulate invasion-related activities. Tumor-associated macrophages are generally considered allies in the process of tumor progression in several types of cancer, although their role on gastric and colorectal carcinomas is still poorly understood. In this report, we studied the influence of primary human macrophages on gastric and colorectal cancer cells, considering invasion, motility/migration, proteolysis and activated intracellular signaling pathways. We demonstrated that macrophages stimulate cancer cell invasion, motility and migration, and that these effects depend on matrix metalloproteinase (MMP) activity and on the activation of epidermal growth factor receptor (EGFR) (at the residue Y(1086)), PLC-γ (phospholipase C-gamma) and Gab1 (GRB2-associated binding protein-1), as evidenced by siRNA (small interference RNA) experiments. Epidermal growth factor (EGF)-immunodepletion impaired macrophage-mediated cancer cell invasion and motility, suggesting that EGF is the pro-invasive and pro-motile factor produced by macrophages. Macrophages also induced gastric and colorectal cancer cell phosphorylation of Akt, c-Src and ERK1/2, and led to an increase of RhoA and Cdc42 activity. Interestingly, whereas macrophage-mediated cancer cell c-Src and ERK1/2 phosphorylation occurred downstream EGFR activation, Akt phosphorylation seems to be a parallel event, taking place in an EGFR-independent manner. The involvement of EGF, EGFR-downstream signaling partners and MMPs in macrophage-mediated invasion provides novel insights into the molecular crosstalk established between cancer cells and macrophages, opening new perspectives for the design of new and more efficient therapeutic strategies to counteract cancer cell invasion.

Ensaios de penetração de Salmonella Enteritidis em ovos de galinha com diferentes qualidades de casca, submetidos ou não a lavagem industrial e a duas temperaturas de armazenagem / Penetration of Salmonella Enteritidis in chicken eggs, comparing different shell quality, industrial washing and temperature of storage

Estudou-se o comportamento da Salmonella Enteritidis quando em contato com a superfície de ovos de consumo, classificados de acordo com a qualidade da casca ou com tratamentos (lavados ou não), usando fezes de aves como veículo de contaminação, e armazenados a 8ºC e 70 por cento de umidade relativa ou a 30ºC e 90 por cento de umidade relativa. A 8ºC, o microrganismo permaneceu viável na superfície dos ovos, ao longo do tempo, enquanto a 30ºC foi possível identificar a sua inibição e até sua eliminação da superfície. A contaminação do conteúdo de ovos com defeito de casca foi mais frequente quando os ovos foram submetidos a temperaturas inadequadas de armazenagem (30ºC).

Salmonella Enteriditis behavior was studied when present on the surface of consume eggs, classified according to the shell quality or treatments (washed or not). Stools of fowls were used as a vehicle for contamination. The eggs were stored at 8ºC and 70 percent relative humidity or at 30ºC and 90 percent of relative humidity. At 8ºC, the microorganism stayed viable on the surface of the eggs throughout the study, while at 30ºC it was inhibited and eliminated from the egg shell. The contamination of content of the eggs with shell defect was more frequent when they were submitted to inappropriate temperatures of storage (30ºC).

Isolated or associated experimental contamination of albumen and egg yolk for Salmonella Enteritidis and Escherichia coli: influence of temperature and storage time / Contaminação experimental de albúmen e gema de ovos por Salmonella Enteritidis e Escherichia coli isoladas ou associadas: influência da temperatura e tempo de armazenagem

The behavior of Salmonella enteritidis (SE) and Escherichia coli in albumen and yolk of chicken eggs submitted to two different temperatures, 8ºC and 30ºC, when separately inoculated or in the form of mixed cultures was studied. There was no difference in the behaviour of the tested agents even when inoculated together. Even under high temperature, E. coli did not multiply in the albumen, demonstrating the inhibition power for that microorganism, while SE increased its population up to three logarithmic cycles. In egg yolk, SE demonstrated psychrotrophic characteristics.

Avaliou-se o comportamento de Salmonella enteritidis (SE) e Escherichia coli em albúmen e gema de ovos de galinha, submetidos a duas diferentes temperaturas, 8 e 30ºC, quando inoculadas isoladamente ou na forma de culturas mistas. Não houve diferença no comportamento dos agentes testados mesmo quando inoculadas em conjunto. Mesmo sob temperatura alta, E. coli não se multiplicou no albúmen, demonstrando o poder inibidor de seus constituintes para esse microrganismo, enquanto a SE aumentou sua população em até três ciclos logarítmicos. Em gema de ovo, SE demonstrou características psicrotróficas.