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International audience; PURPOSE:Dogs have been proposed as spontaneous animal models of human breast cancer, based on clinicopathologic similarities between canine and human mammary carcinomas. We hypothesized that a better knowledge of the natural history and prognostic factors of canine invasive mammary carcinomas would favor the design of preclinical trials using dogs as models of breast cancer.METHODS:The 2-year outcome of 350 female dogs with spontaneous invasive mammary carcinoma was studied. The investigated prognostic factors included age at diagnosis, pathologic tumor size, pathologic nodal stage, lymphovascular invasion, histological grade, and expression of Estrogen Receptor alpha (ERα), Progesterone Receptor, Ki-67, Human Epidermal Growth Factor Receptor 2, basal cytokeratins 5/6, and Epidermal Growth Factor Receptor. Multivariate survival analyses were performed using the Cox proportional hazards model.RESULTS:The overall survival after mastectomy was 11 months. Within 1 year post mastectomy, 41.5% of dogs (145/350) died from their mammary carcinoma. By multivariate analysis, the significant prognostic factors for overall survival included a pathologic tumor size larger than 20 mm [HR 1.47 (95% confidence interval 1.15-1.89)], a positive nodal stage [pN+, HR 1.89 (1.43-2.48)], a histological grade III [HR 1.32 (1.02-1.69)], ERα negativity [HR 1.39 (1.01-1.89)], a high Ki-67 proliferation index [HR 1.32 (1.04-1.67)], and EGFR absence [HR 1.33 (1.04-1.69)].CONCLUSION:The short natural history of spontaneous canine invasive mammary carcinomas and high rate of cancer-related death allow for rapid termination of preclinical investigations. The prognostic factors of invasive mammary carcinomas are remarkably similar in dogs and humans, highlighting the similarities in cancer biology between both species.

International audience; PURPOSE:Relevant animal models of human breast cancer are currently needed, especially for the aggressive triple-negative breast cancer subtype. Recent studies and our results (Part 1) indicate that spontaneous canine invasive mammary carcinomas (CMCs) resemble human breast cancer by clinics and pathology as well as behavior and prognostic indicators. We hypothesized that the current molecular classifications of human breast cancer, used for therapeutic decision, could be relevant to dogs.METHODS:Three hundred and fifty female dogs with spontaneous CMC and a 2-year follow-up were retrospectively included. By immunohistochemistry, CMCs were classified according to Nielsen (Clin Cancer Res 10:5367-5374, 2004) and Blows (PlosOne doi: 10.1371/journal.pmed.1000279, 2010) into the subtypes of human breast cancer.RESULTS:Four immunophenotypes were defined either according to Nielsen classification (luminal A 14.3%, luminal B 9.4%, triple-negative basal-like 58.6%, and triple-negative nonbasal-like 17.7% CMCs); or to Blows classification (luminal 1-: 11.4%, luminal 1+: 12.3%, Core basal phenotype: 58.6%, and five-negative phenotype: 17.7%). No HER2-overexpressing CMC as defined by a 3 + immunohistochemical score was observed in our cohort. By univariate and multivariate analyses, both immunophenotypical classifications applied to CMCs showed strong prognostic significance: luminal A or luminal 1+ CMCs showed a significantly longer disease-free interval (HR = 0.46), Overall (HR = 0.47), and Specific Survival (HR = 0.56) compared to triple-negative carcinomas, after adjustment for stage.CONCLUSIONS:In our cohort, triple-negative CMCs largely predominated (76%), were much more prevalent than in human beings, and showed an aggressive natural behavior after mastectomy. Dogs are thus potent valuable spontaneous models to test new therapeutic strategies for this particular subtype of breast cancer.

International audience; Herein, we describe the synthesis of an aptadendrimer by covalent bioconjugation of a gallic acid–triethylene glycol (GATG) dendrimer with the G-quadruplex (G4) AT11 aptamer (a modified version of AS1411) at the surface. We evaluated the loading and interaction of an acridine orange ligand, termed C8, that acts as an anticancer drug and binder/stabilizer of the G4 structure of AT11. Dynamic light scattering experiments demonstrated that the aptadendrimer was approximately 3.1 nm in diameter. Both steady-state and time-resolved fluorescence anisotropy evidenced the interaction between the aptadendrimer and C8. Additionally, we demonstrated that the iodine atom of the C8 ligand acts as an effective intramolecular quencher in solution, while upon complexation with the aptadendrimer, it adopts a more extended conformation. Docking studies support this conclusion. Release experiments show a delivery of C8 after 4 h. The aptadendrimers tend to localize in the cytoplasm of various cell lines studied as demonstrated by confocal microscopy. The internalization of the aptadendrimers is not nucleolin-mediated or by passive diffusion, but via endocytosis. MTT studies with prostate cancer cells and non-malignant cells evidenced high cytotoxicity mainly due to the C8 ligand. The rapid internalization of the aptadendrimers and the fluorescence properties make them attractive for the development of potential nanocarriers.

Beeswax adulteration in the apiculture sector represents a growing problem worldwide due to the lack of clearly defined purity criteria, the absence of official quality (authenticity) controls, and the inconsistency of the analytical methods used for adulteration detection. Although beeswax authentication is implemented in other regulatory sectors (pharmaceutical and food industry), the classical physico-chemical analytical methods used for determination of beeswax purity exhibit inconsistencies for the detection of adulterants. In this study, an inventory was made on a comprehensive set of analytical methods and the corresponding purity criteria used for the detection of the most common beeswax adulterants (paraffin, stearin and/or stearic acid) from existing legislations and scientific literature. The selected analytical methods (classical physico-chemical, and advanced instrumental, i.e. chromatographic and spectroscopic analytical techniques) were weighted by three independent experts against two criteria: feasibility and analytical performance in detecting targeted adulterants. Classical methods for which measurement data were available (melting point and acid/saponification/ester values for paraffin-adulterated vs. non-adulterated beeswax samples) were retained and further validated by a receiver operating characteristic (ROC) analysis. These methods were also validated by generating the corresponding calibration curves for paraffin detection using paraffin-beeswax mixtures containing different proportions of paraffin (ranging from 5 to 95%, w/w). The results of the ROC analysis revealed that a tentative detection of paraffin in beeswax can be achieved by a combination of at least two physico-chemical methods. However, for a reliable detection of the most common adulterants in beeswax, physico-chemical methods should be complemented with advanced analytical tools. i.e. GC-MS, HTGC-FID (MS) and/or FTIR-ATR spectroscopy, depending on the expected adulterant.

International audience; Objectives. Epidemiological data concerning third-generation cephalosporin (3GC) resistance in wild fauna are scarce. The aim of this study was to characterize the resistance genes, their genetic context, and clonal relatedness in 17 Escherichia coli resistant to 3GC isolated from wild animals. Methods. The isolates were characterized by short-read whole genome sequencing, and long-read sequencing was used for the hybrid assembly of plasmid sequences. Results. The 3GC resistance gene most identified in the isolates was the extended-spectrum β-lactamases (ESBL)-encoding gene blaCTX-M-1 (82.3%), followed by blaCTX-M-32 (5.9%), blaCTX-M-14 (5.9%), and blaSHV-12 (5.9%). E. coli isolates mainly belonged to the sequence types (STs) rarely reported from humans. The single nucleotide polymorphism (SNP)-based typing showed that most E. coli genomes from wild animals (wild boars, birds of prey, and buzzards) formed clonal clusters (

Pinus nigra J.F. Arnold has an ecological and economic interest in Europe, but many of the planted populations have an unknown origin and infraspecific taxonomy. Six Portuguese P. nigra populations characterised with microsatellites revealed high intra-population diversity structured into two clusters with low differentiation that might suggest two provenances or infraspecific taxa. Despite compared with foreign samples from different subspecies, we were not able to infer about the origin or infraspecific taxonomy of the Portuguese populations based on the pooled microsatellite data . Many of the European Pinus nigra J.F. Arnold forests were planted with the material of unknown origin. Efforts have been made to determine their provenances and infraspecific taxonomy regarding their relevance for defining strategies of genetic resources conservation, germplasm use, forest management and genetic improvement. The Portuguese P. nigra populations are allochthonous, and their provenance and infraspecific taxonomy are unknown. With this work, we intended to characterise the Portuguese P. nigra populations regarding its genetic diversity, structure and relationships using simple sequence repeat (SSR) markers and/or to infer about its origin and infraspecific taxonomy by comparing the SSR patterns of foreign P. nigra samples of known taxonomic classification. A total of 224 Portuguese P. nigra individuals from six populations were characterised using 14 SSR markers developed specifically to different Pinus sp., including P. nigra, by other authors. Thirteen SSR loci were selected and showed 100% of polymorphism among individuals. The genetic diversity was higher within (95%) rather than among (5%) populations. The Portuguese individuals were structured into two main clusters (K = 2) with low genetic differentiation (FST = 0.04), and the foreign samples were genetically distant from the Portuguese individuals. The six Portuguese P. nigra populations revealed high genetic diversity and seemed to be structured into two main clusters with low differentiation suggesting two provenances or infraspecific taxa. Nonetheless, the comparative analyses with foreign samples did not allow a clear inference about its origin and/or infraspecific taxonomy. Additional foreign samples and/or molecular markers will be tested to pursue these goals.

Wildfire is the main disturbance in forested ecosystems of southern Europe and is due to complex interactions between climate-weather, fuels and people. Warmer and drier conditions projected in this region are expected to profoundly affect wildfires, which will impact ecosystems and humans. We review the scientific literature addressing the assessment of climate change impacts on wildfires in southern Europe, with a twofold objective: (i) report the trends in wildfire danger and activity projected under warming climate in southern Europe and (ii) discuss the limitations of wildfire projections under the specific biogeographical context of southern Europe.We identified 22 projection studies that examined future wildfire danger or wildfire activity at local, regional or continental scale. Under the scenario with the highest greenhouse gas emissions, we found that projections studies estimate an increase in future fire danger and burnt areas varying, on average, from 2 to 4 % and from 15 to 25 % per decade, respectively. Fire-prone area expansion to the north and to Mediterranean mountains is a concern, while climate-induced burnt area increase might be limited by fuel availability in the most arid areas. While all studies agreed on the direction of changes, further comparisons on the magnitude of increase remained challenging because of heterogeneous methodological choices between projections studies (climate models, projection period, spatial scale and fire metrics). We then described three main sources of uncertainty that may affect the reliability of wildfire projections: climate projections, climate-fire models, and the influences of fuel load/structure and human related factors on the climate-fire relationships. We finally suggest research directions to address some of these issues for the purpose of refining fire danger and fire activity projections in southern Europe.

During the last decades, climate and land use changes led to an increased prevalence of megafires in Mediterranean-type climate regions (MCRs). Here, we argue that current wildfire management policies in MCRs are destined to fail. Focused on fire suppression, these policies largely ignore ongoing climate warming and landscape-scale buildup of fuels. The result is a 'firefighting trap' that contributes to ongoing fuel accumulation precluding suppression under extreme fire weather, and resulting in more severe and larger fires. We believe that a 'business as usual' approach to wildfire in MCRs will not solve the fire problem, and recommend that policy and expenditures be rebalanced between suppression and mitigation of the negative impacts of fire. This requires a paradigm shift: policy effectiveness should not be primarily measured as a function of area burned (as it usually is), but rather as a function of avoided socio-ecological damage and loss., This work was financed by national funds through FCT—Foundation for Science and Technology, within the scope of project PCIF/AGT/0136/2017 (People&Fire: reducing risk, living with risk) and PTDC/AGR-FOR/2586/2014 (RurIntFIre: Fire in the Rural-Urban Interface: characterisation, risk mapping, and fuel break design). FM was funded through contract IF/01053/2015 (FCT). JMCP was supported by the Forest Research Centre, a research unit funded by Foundation for Science and Technology I.P. (FCT), Portugal (UID/AGR/00239/2019). PF work was carried under project UID/AGR/04033/2019 supported by FCT. JMM acknowledges funding from Ministerio de Ciencia, Innovación y Universidades (project CGL2016-78357-R)

The wood of Pinus nigra populations planted in Portugal, comparatively to Pinus pinaster , has higher total extractive content, lower Klason lignin and H/G ratio, and similar mechanical properties, presenting advantages for industrial purposes. P. nigra was used in the reforestation of mountainous areas in Portugal, but its wood chemical and mechanical properties were never studied. This work intends to evaluate the chemical and mechanical wood properties of the P. nigra populations planted in Portugal, to relate these properties with previously characterised physical features and to compare these data with other European P. nigra stands and species, namely, P. pinaster. Wood chemical and mechanical properties were analysed in 90 trees from six Portuguese sites, using near-infrared (NIR) spectrometry and the three-point bending test. The wood of the P. nigra populations planted in Portugal presented average values of total extractive content = 9.4%, Klason lignin = 26.69%, MORRad = 14.93 MPa and MOERad = 1200.98 MPa. Ring density showed no significant correlation with ring width. The P. nigra populations planted in Portugal presented qualitative and quantitative properties similar to P. pinaster wood, the main resinous species in Portugal. Facing the lack of raw material for wood industry due to frequent forest fires in the Mediterranean region, P. nigra could be used to reforest mountainous areas of those regions.

The technological quality of wheat flour is defined by a range of dough characteristics relevant to the breadmaking processes and practices of individual countries and for particular products. The influence of storage protein diversity on wheat quality has been widely documented in the last three decades. The present chapter focuses on several aspects of wheat quality that merit more attention. The huge genetic diversity of wheat storage proteins means that all the possible allelic combinations and their interactions are too numerous to be tested in terms of their influence on the major quality parameters. However it is still relevant to describe the variation in rheological and viscoelastic properties of gluten in relation to its component proteins, glutenin and gliadin. Although gluten plays a major role in determining the properties of dough, the abundance of the two major storage protein fractions does not solely explain the observed variation in those properties. We therefore examine the influence of some genetic factors, including those affecting the protein composition, on the variation in the glutenin polymer sizes. Some examples will be given to illustrate how end-use quality can be improved by taking advantage of the available genetic resources in parallel with molecular genome analyses with the dual aim of widening the scope of characteristics that can be harnessed in breeding and ensuring consistent wheat quality in changing agro-climatic situations. The known alleles of the major genes are highlighted in the context of the challenges that the research community is facing regarding wheat allele nomenclature, exchange of gene bank material and the numerous quality attributes of interest. Finally, important research objectives are proposed for breeding future wheats with grain protein quality and technological properties tailored for different food products.

Phlebotomine sand fly-borne pathogens such as Leishmania spp. and phleboviruses are emerging threats to humans and animals worldwide. The aim of this work was to evaluate the exposure of cats from Portugal to Toscana virus (TOSV) and Sandfly Fever Sicilian virus (SFSV) and assess the associated risk factors. The possible association between exposure to Phlebotomus perniciosus saliva with TOSV and SFSV was also investigated. Out of 369 cats tested, 18 (4.9%, n = 365) were seropositive for TOSV, and eight (2.2%, n = 367) were seropositive for SFSV. Multivariate logistic regression analysis showed that cats presenting clinical signs that were compatible with leishmaniosis and antibodies to TOSV had a significantly higher risk of being SFSV seropositive. The presence of antibodies to sand fly-borne viruses in cats indicate that these animals are frequently exposed to sand flies and transmitted pathogens. Data suggest that cats can be used to qualitatively monitor human exposure to TOSV and SFSV in endemic areas. The clinical impact of SFSV in cats’ health should be investigated. The identification of the sand fly species responsible for the circulation of TOSV and SFSV in nature and the evaluation of the vectorial competence of P. perniciosus to SFSV should also be addressed.

In this study, we present the first tree-ring chronology for the tropical tree species Copaifera lucens and its climatic signal in southeastern Brazil.Tree-ring width series were compared with local climate indices using a drought index (Standardized Precipitation Evapotranspiration Index -SPEI), in monthly, bi-monthly and four-monthly scales. We also calculated negative pointer years over the time-span of the tree-ring width. The radial growth of C. lucens showed a positive correlation with the SPEI of the current summer and autumn in all the three analyzed time scales, while the negative pointer years matched with drier years. The species was highly sensitive to very low summer precipitation, which may lead to a 49% reduction in growth.We conclude that the long-living C. lucens has a great potential for dendrochronological studies as it shows a marked climatic signal. Our study also reinforces the importance of rainfall in regulating radial growth in tropical forests and sheds light on the local climate influence on tree growth in recent decades.

Em todo o mundo, o mercado do vinho está a sofrer alterações fundamentais, cada vez mais competitivo, com a entrada de novas empresas produtoras de vinho. O vinho do Porto é um marco do vinho português. É um vinho fortificado produzido na Região Demarcada do Douro (uma das primeiras regiões vitivinícolas demarcadas e regulamentadas do mundo) em condições físicas e humanas muito específicas. O presente trabalho pretende verificar como a cultura (aferida pela nacionalidade) influencia o nível de importância dos atributos do vinho, no momento em que o consumidor decide comprar uma garrafa de vinho. Os dados primários foram obtidos através da aplicação de um questionário on-line. A amostra é constituída por 974 consumidores de vinho do Porto de seis nacionalidades diferentes: França, Espanha, Portugal, Roménia, Inglaterra e Brasil. O efeito da nacionalidade no comportamento dos consumidores, ao nível da importância dada aos atributos, é analisado recorrendo ao modelo logístico binário. Os resultados indicam que a região de origem, o prestígio da marca e o design são atributos mais importantes para os consumidores Portugueses (relativamente aos consumidores internacionais). Os consumidores internacionais estão dispostos a pagar mais por garrafa de vinho do Porto, não sendo o preço um fator influenciador determinante na escolha do vinho. os resultados suportam a necessidade de uma maior divulgação das marcas de vinho do Porto junto dos consumidores internacionais, já que este tipo de consumidores valorizam o produto através de uma maior disposição a pagar pelo mesmo. Worldwide the wine market is experiencing fundamental changes, with the entry of new wine producing companies, which generates greater competitiveness in domestic and international markets. The Port wine is a landmark of the Portuguese wine. It is a fortified wine produced in the Douro Demarcated Region (one of the first demarcated and regulated wine regions in the world) under very specific physical and human conditions. The present paper aims to ascertain how the culture (proxied by the nationality) influence the level of importance of wine attributes, when deciding to buy a bottle of wine.The primary data were gathered through an on-line questionnaire. A sample of 974 buyers/ consumers of the Port wine was obtained from six different nationalities: France, Spain, Portugal, Romania, England and Brazil. The effect of nationality on consumer behavior, in terms of importance given to attributes, was analyzed using the binary logistic model and bivariate statistics. The results indicate than origin region, the brand prestige and the design are not important attributes for the international consumers. However, international consumers are willing to pay more per bottle of Port win and price is not an influencing factor. Thus, it is necessary to increase the dissemination of Port wine brands to international consumers, since these consumers value the product through a greater willingness to pay for it. info:eu-repo/semantics/publishedVersion