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[ENG]We evaluated the ecological significance of the boundary form between two patches with contrasting vegetation (mine grassland and adjacent forest) on woody colonization and forest expansion in open-cast coal mines in Northern Spain. Woody colonization and browsing traces were measured on three mine sites, along 24 transects that were laid out perpendicular to the forest-mine boundary and classified according to their shape (concave, convex, straight). Mine sites were colonized from the close forest by woody species, whose colonization intensity depends on the boundary form. The overall colonization intensity decreased with increasing distance to the forest and differed depending on the boundary form. The more intense colonization was found in concave boundaries and the strongest decrease in convex boundaries close to the forest, whereas straight boundaries showed an intermediate colonization pattern. Concave boundaries reached higher woody cover in the basal strata of the mines than convex (up to 2 m) or straight boundaries (up to 1 m) from 11 m to the forest edge, mainly by the presence of dense patches of Cytisus scoparius (L.) Link, with a scattered overstory of Genista florida L. These shrubs might reduce the browsing intensity and act as nurse plants facilitating the establishment of Quercus petraea (Matt.) Liebl. in mine areas at greater distances from the forest edge. The forest-mine boundary form does not affect the forest vertical structure that is homogenous and does not help explain the woody colonization pattern in the mines. We conclude that edge characteristics have a strong potential to be used in the restoration of native forests based on natural processes. The implications of our results for sessile oak (Quercus petraea (Matt.) Liebl.) forest expansion along edges in fragmented Mediterranean forest landscapes were discussed.

We evaluated the ecological significance of the boundary form between two patches with contrasting vegetation (mine grassland and adjacent forest) on woody colonization and forest expansion in open-cast coal mines in Northern Spain. Woody colonization and browsing traces were measured on three mine sites, along 24 transects that were laid out perpendicular to the forest-mine boundary and classified according to their shape (concave, convex, straight). Mine sites were colonized from the close forest by woody species, whose colonization intensity depends on the boundary form. The overall colonization intensity decreased with increasing distance to the forest and differed depending on the boundary form. The more intense colonization was found in concave boundaries and the strongest decrease in convex boundaries close to the forest, whereas straight boundaries showed an intermediate colonization pattern. Concave boundaries reached higher woody cover in the basal strata of the mines than convex (up to 2 m) or straight boundaries (up to 1 m) from 11 m to the forest edge, mainly by the presence of dense patches of Cytisus scoparius (L.) Link, with a scattered overstory of Genista florida L. These shrubs might reduce the browsing intensity and act as nurse plants facilitating the establishment of Quercus petraea (Matt.) Liebl. in mine areas at greater distances from the forest edge. The forest-mine boundary form does not affect the forest vertical structure that is homogenous and does not help explain the woody colonization pattern in the mines. We conclude that edge characteristics have a strong potential to be used in the restoration of native forests based on natural processes. The implications of our results for sessile oak (Quercus petraea (Matt.) Liebl.) forest expansion along edges in fragmented Mediterranean forest landscapes were discussed.

Seasonal influenza vaccines must be updated annually and suboptimally protect against strains mismatched to the selected vaccine strains. We previously developed a subunit vaccine antigen consisting of a stabilized trimeric influenza A group 1 hemagglutinin (H1) stem protein that elicits broadly neutralizing antibodies. Here, we further optimized the stability and manufacturability of the H1 stem antigen (H1 stem v2, also known as INFLUENZA G1 mHA) and characterized its formulation and potency with different adjuvants in vitro and in animal models. The recombinant H1 stem antigen (50 µg) was administered to influenza-naïve non-human primates either with aluminum hydroxide [Al(OH)3] + NaCl, AS01B, or SLA-LSQ formulations at week 0, 8 and 34. These SLA-LSQ formulations comprised of varying ratios of the synthetic TLR4 agonist ‘second generation synthetic lipid adjuvant’ (SLA) with liposomal QS-21 (LSQ). A vaccine formulation with aluminum hydroxide or SLA-LSQ (starting at a 10:25 µg ratio) induced HA-specific antibodies and breadth of neutralization against a panel of influenza A group 1 pseudoviruses, comparable with vaccine formulated with AS01B, four weeks after the second immunization. A formulation with SLA-LSQ in a 5:2 μg ratio contained larger fused or aggregated liposomes and induced significantly lower humoral responses. Broadly HA stem-binding antibodies were detectable for the entire period after the second vaccine dose up to week 34, after which they were boosted by a third vaccine dose. These findings inform about potential adjuvant formulations in clinical trials with an H1 stem-based vaccine candidate.

IntroductionIn human breast cancer, the invasion of tumor-draining lymph nodes (TDLNs) is an important step in disease progression and has predictive value [1]. TDLN dendritic cells (DCs), which are comprised of lymphoid-organ-resident and skin-derived migratory DCs, present tumor antigens to the naïve T cells and induce their activation and polarization into different functional subsets (Th1, Th2, Th17, Th22, Tfh, regulatory T cells) that will lead to antitumor T cell responses or to tolerance [2].ObjectiveSystematic comparison of the immune profile of Invaded (INV) versus Non-invaded (NI) TDLNs would help to identify those immunomodulatory mechanisms associated to the presence of the tumor that could condition the response to immunotherapy.Material and methodsTDLNs from 70 untreated breast cancer patients undergoing surgery at Institut Curie Hospital were obtained in accordance with institutional ethical guidelines. Samples were analyzed by multi-color flow cytometry. For statistical analysis, Wilcoxon matched paired test or Mann-Whitney test was performed using Prism (GraphPadSoftware)ResultsWe studied the distribution of 6 different DC subpopulations and observed in INV TDLNs a significant decrease in the percentage of BDCA1+ DCs (P<0.05) and a significant increase in the percentage of CD11c+HLADR+CD14+cells (P<0.01), including macrophages and inflammatory DCs, compared to NI TDLNs (P<0.05). We also found a significant lower frequency of naïve conventional and regulatory T cells in INV TDLNs (P<0.05). Both, in NI and INV TDLNs, memory conventional and regulatory T cells were highly polarized, mainly to the Th1 phenotype, but also to the Th2, Th17, Tfh and Th22 phenotypes, as determined by the expression of a panel of chemokine receptors and transcription factors. Notably, in INV TDLNs, a significantly higher proportion of regulatory and conventional T cells were Th1-polarized (P<0.05). Further functional analysis showed that after ex-vivo PMA/Iono stimulation, the Th1-polarized conventional T cells, but not the Th1-polarized regulatory T cells produced high amounts of IFN-γ, being the IFN-γ production significantly higher in INV TDLNs (P<0.05).ConclusionOverall, we observed that immune cells from metastatic TDLNs show evidence of high activation (increased proportion of inflammatory DCs and of Th1-polarized memory T cells) highlighting their potential role in the anti-tumor immune response.