Evaluación del estado de los estocs costeros explotados por la flota de Viareggio (Mar Ligur meridional)

The coastal demersal fish assemblage exploited commercially by the Viareggio fleet was assessed in order to define its exploitation status and sustainability. A production model was used provided management benchmarks for the species for which available data are limited. The ASPIC Surplus production model was used. The results showed a depleted population for most of the species involved (B2008/B0 between 0.05 and 0.35) with high relative fishing mortality (F2008/FMSY between 1.18 and 1.64). Population projections using ASPIC-P allowed the exploitation strategies to be evaluated for a 10-year period. None of the populations are predicted to recover to BMSY if fishing effort remains at the 2008 levels. A reduction in effort of about 40% should increase the biomass in the medium-term of most of the species to BMSY or over, with a fairly good increase in yields of the most valuable species.El conjunto de especies costeras explotado comercialmente por la flota de Viareggio fue evaluado para definir su estado de explotación y sostenibilidad. El uso de un modelo de producción permitió definir pautas de manejo pesquero para estas especies para las que los datos disponibles son limitados. Se utilizó el modelo de producción excedente ASPIC. Los resultados mostraron para la mayoría de las especies, biomasas drásticamente reducidas (B2008/B0 entre 0.05 y 0.35) con altos valores relativos de mortalidad por pesca (F2008/FMSY entre 1.18 y 1.64). Proyecciones realizadas con ASPIC permitieron evaluar estrategias de explotación para un periodo de diez años. Manteniendo el esfuerzo a nivel del 2008, para ninguna especie se prevé que la recuperación de la población alcance el nivel de BMSY. Una reducción del esfuerzo del 40% debería conducir a un incremento, a medio plazo, de la biomasa de casi todas las especies por encima del BMSY, incrementándose asimismo sustancialmente los rendimientos de las especies más importantes

Antitumor immunization of mothers delays tumor development in cancer-prone offspring

Maternal immunization is successfully applied against some life-threatening infectious diseases as it can protect the mother and her offspring through the passive transfer of maternal antibodies. Here, we sought to evaluate whether the concept of maternal immunization could also be applied to cancer immune-prevention. We have previously shown that antibodies induced by DNA vaccination against rat Her2 (neu) protect heterozygous neu-transgenic female (BALB-neuT) mice from autochthonous mammary tumor development. We, herein, seek to evaluate whether a similar maternal immunization can confer antitumor protection to BALB-neuT offspring. Significantly extended tumor-free survival was observed in BALB-neuT offspring born and fed by mothers vaccinated against neu, as compared to controls. Maternally derived anti-neu immunoglobulin G (IgG) was successfully transferred from mothers to newborns and was responsible for the protective effect. Vaccinated mothers and offspring also developed active immunity against neu as revealed by the presence of T–cell-mediated cytotoxicity against the neu immunodominant peptide. This active response was due to the milk transfer of immune complexes that were formed between the neu extracellular domain, shed from vaccine-transfected muscle cells, and the anti-neu IgG induced by the vaccine. These findings show that maternal immunization has the potential to hamper mammary cancer in genetically predestinated offspring and to develop into applications against lethal neonatal cancer diseases for which therapeutic options are currently unavailable

Impact of pe_pgrs33 gene polymorphisms on mycobacterium tuberculosis infection and pathogenesis

PE_PGRS33 is a surface-exposed protein of Mycobacterium tuberculosis (Mtb) which exerts its role in macrophages entry and immunomodulation. In this study, we aimed to investigate the polymorphisms in the pe_pgrs33 gene of Mtb clinical isolates and evaluate their impact on protein functions. We sequenced pe_pgrs33 in a collection of 135 clinical strains, genotyped by 15-loci MIRU-VNTR and spoligotyping and belonging to the Mtb complex (MTBC). Overall, an association between pe_pgrs33 alleles and MTBC genotypes was observed and a dN/dS ratio of 0.64 was obtained, suggesting that a purifying selective pressure is acting on pe_pgrs33 against deleterious SNPs. Among a total of 19 pe_pgrs33 alleles identified in this study, 5 were cloned and used to complement the pe_pgrs33 knock-out mutant strain of Mtb H37Rv (Mtb\uce\u9433) to assess the functional impact of the respective polymorphisms in in vitro infections of primary macrophages. In human monocyte-derived macrophages (MDMs) infection, large in-frame and frameshift mutations were unable to restore the phenotype of Mtb H37Rv, impairing the cell entry capacity of Mtb, but neither its intracellular replication rate nor its immunomodulatory properties. In vivo studies performed in the murine model of tuberculosis (TB) demonstrated that the Mtb\uce\u9433 mutant strain was not impaired in the ability to infect and replicate in the lung tissue compared to the parental strain. Interestingly, Mtb\uce\u9433 showed an enhanced virulence during the chronic steps of infection compared to Mtb H37Rv. Similarly, the complementation of Mtb\uce\u9433 with a frameshift allele also resulted in a Mtb strain capable of causing a surprisingly enhanced tissue damage in murine lungs, during the chronic steps of infection. Together, these results further support the role of PE_PGRS33 in the pathogenesis and virulence of Mtb

Abundance and characterization of floating microplastics along the Tuscany coast (Italy): the first aplication of the MSFD monitoring proticol

Due to the increasing use of plastic and its dispersion in the marine environment andaccumulation in all habitats, the issue of plastic debris needs to be deeply investigated [1]. In particular, despite the Mediterranean sea is one of the hot spot area in the world for plastic debris accumulation [2], the knowledge on distribution and occurrence of floating microplastics is still lacking. Microplastics can affect marine biota increasing the likelihoodof ingestion of plastics by marine organisms entering the food web. For this reason the European Union has promoted the Marine Strategy Framework Directive (MSFD) with the aim of reaching the "Good Environmental Status" by 2020. This work has been carried out as part of implementation of the Descriptor 10.1.3 [3] of the MSFD in Tuscany (Italy), with the aim to gain information on abundance and distribution of microplastics. Sampling has been realized in two seasons (winter and spring) using a manta trawl (330 μm mesh size). Samples were collected along 4 transects 100 km far from each other, located from the estuary of Arno river to the promontory of Argentario. Each transect was divided into 4 stations located at increasing distance from the coast (20 km, 10 km, 5 km, 0.5 km). The analysis were performed according to the MSFD protocol. All data was normalized to the total volume of water filtered and expressed as items/m3 and microplastics were characterized by colour, shape and size. A total of 2670 microplastics were isolated in the 72 samples, white was the predominant color; the majority of items are fragments and the most of microplastics fall in the measured from 1 to 2.5mm size class. In the winter, the highest values of microplastics have been found in the station at 20 km from the coast with a gradient decreasing in the stations closer to coast; whereas in spring, the highest was found in the station at 10 km. This work represents the first application of the MSFD protocol in the monitoring of microplastics in Tuscany and will allow to understand the distribution and abundance of microplastics in the Tuscany coastal waters

Erbb2 DNA Vaccine Combined with Regulatory T Cell Deletion Enhances Antibody Response and Reveals Latent Low-Avidity T Cells: Potential and Limits of Its Therapeutic Efficacy

Rat (r)Erbb2 transgenic BALB-neuT mice genetically predestined to develop multiple invasive carcinomas allow an assessment of the potential of a vaccine against the stages of cancer progression. Because of rErbb2 expression in the thymus and its overexpression in the mammary gland, CD8(+) T cell clones reacting at high avidity with dominant rErbb2 epitopes are deleted in these mice. In BALB-neuT mice with diffuse and invasive in situ lesions and almost palpable carcinomas, a temporary regulatory T cells depletion combined with anti-rErbb2 vaccine markedly enhanced the anti-rErbb2 Ab response and allowed the expansion of latent pools of low-avidity CD8(+) T cells bearing TCRs repertoire reacting with the rErbb2 dominant peptide. This combination of a higher Ab response and activation of a low-avidity cytotoxic response persistently blocked tumor progression at stages in which the vaccine alone was ineffective. However, when diffuse and invasive microscopic cancers become almost palpable, this combination was no longer able to secure a significant extension of mice survival

Surface Expression of MPT64 as a Fusion with the PE Domain of PE_PGRS33 Enhances Mycobacterium bovis BCG Protective Activity against Mycobacterium tuberculosis in Mice▿

To improve the current vaccine against tuberculosis, a recombinant strain of Mycobacterium bovis bacillus Calmette-Guérin (rBCG) expressing a Mycobacterium tuberculosis vaccine candidate antigen (MPT64) in strong association with the mycobacterial cell wall was developed. To deliver the candidate antigen on the surface, we fused the mpt64 gene to the sequence encoding the PE domain of the PE_PGRS33 protein of M. tuberculosis (to create strain HPE-ΔMPT64-BCG), which we have previously shown to transport proteins to the bacterial surface. In a series of protection experiments in the mouse model of tuberculosis, we showed that (i) immunization of mice with HPE-ΔMPT64-BCG provides levels of protection significantly higher than those afforded by the parental BCG strain, as assessed by bacterial colonization in lungs and spleens and by lung involvement (at both 28 and 70 days postchallenge), (ii) rBCG strains expressing MPT64 provide better protection than the parental BCG strain only when this antigen is surface expressed, and (iii) the HPE-ΔMPT64-BCG-induced MPT64-specific T cell repertoire when characterized by β chain variable region-β chain joining region (BV-BJ) spectratyping indicates that protection is correlated with the ability to recruit gamma interferon (IFN-γ)-secreting T cells carrying the BV8.3-BJ1.5 (172 bp) shared rearrangement. These results demonstrate that HPE-ΔMPT64-BCG is one of the most effective new vaccines tested so far in the mouse model of tuberculosis and underscore the impact of antigen cellular localization on the induction of the specific immune response induced by rBCG

Qualitative and quantitative comparison of intracellular markers expressed by neural cells.

<p>(<b>A</b>) Flow cytometry for the intracellular antigens TUJ1, MAP2 and doublecortin (DCX) analyzed on various cell sources including neuroblastoma BE(2)-M17 and SH-SY5Y as well as human iPS cell-derived differentiated neural stem cell cultures (hiPS-NSC). Grey boxes in flow cytometry plots specify gates set on negative control samples to capture 0.4%. Green boxes specify positive fraction on stained samples. Error bars in (<b>B</b>) indicate standard deviation.</p

Combinatorial detection of known neural intracellular markers plus a panel of candidate surface antigens in human cell lines.

<p>(<b>A</b>) In SH-SY5Y cells CD49f, CD90^HIGH and CD29 expression clustered distinct from doublecortin (DCX)-positivity, while subsets of CD56, CD24 and CD200 closely correlated with neuronal differentiation (DCX+). Patterns of exclusive positivity on either the y- or the x-axis (upper left and lower right quadrant) yield CD marker candidates for negative selection strategies. Patterns of shared intracellular and surface expression (upper right quadrant) are suitable for positive selection strategies. Blue outlines illustrate co-expression patterns. (<b>B</b>) Co-localization analysis of the catecholaminergic intracellular marker tyrosine hydroxylase (TH) is shown on BE(2)-M17 cells. The TH-positive subset stained negative for the putative proliferative indicator CD49f but was colocalized with CD90, CD24 and CD200 in this cell line. (<b>C</b>) Co-localization analysis of the glial intracellular marker glial fibrillary acidic protein (GFAP) is shown on SNB-19 cells. In contrast to the other neural cell lines analyzed, GFAP-positive subsets stained positive for CD49f, and also CD90. CD24 was present to a lower degree and the putative neuronal marker CD200 in this context was virtually absent.</p

Application for neural cell isolation in human neural cancer lines.

<p>(<b>A</b>) Mixed SH-SY5Y/SNB-19 neuro-glial cell suspensions (without fixation and permeabilization) analyzed by flow cytometry for CD49f-PE and CD200-APC. Polygon gates within the left dot plot indicate selection of CD49f⁺/CD200^- (red) versus CD49f ^-/CD200^HIGH (green) populations. (<b>B</b>) Corresponding microscopic analysis of unsorted (left panel) vs. sorted conditions (mid and right panels) 1 div post-sort. Arrows indicate SH-SY5Y cells, arrowhead SNB-19 [phase contrast; scale bars: 50µm]. (<b>C</b>) Immunofluorescence for GFAP (red) and doublecortin (green) of surface marker code-based cell suspensions. Left panel: unsorted; mid panel: CD49f⁺/CD200^- population post-sort; right panel: CD49f ^-/CD200^HIGH population. [1 div post-FACS; scale bars: 20µm].</p