The Role of Spatial Exploration and Territoriality in Establishing Gilthead Seabream (Sparus aurata) Hierarchies, and Their Effects upon Underlying Stress Physiology

Territoriality, spatial exploration and social hierarchy are strictly related behaviors in gregarious fishes, and are often under-appreciated in farms where the individuals are confined within crowded spaces. In this study, we investigated the role of spatial exploration, elucidating the importance of time upon forming the social organization, and the role of the territoriality in gilthead seabream (Sparus aurata), using two experimental approaches. In the first approach, three fish were placed sequentially in the aquarium with an interval of two days (sequential model), while in the second (simultaneous model), two fish were simultaneously placed in an aquarium divided by a barrier which was removed after a certain period of time. To study the effect of social stress and spatial perception in the two models, we monitored behavior (aggressive acts and feeding priority), integrated with the evaluation of physiological and cellular stress parameters, such as phagocytosis, cortisol, glucose, and blood osmolarity levels. After the establishment of the social hierarchy in the "sequential model", we observed that the levels of cortisol and an immunological cell-mediated marker were higher in subordinate individuals than in the dominant ones. We observed a different modulation of phagocytic activity in peritoneal cavity cells between dominant and subordinates, demonstrating that social stress acts upon immune response. Differently from the first model, no behavioral, physiological, or phagocytic differences were found between the two fish involved in the simultaneous model, where both fish acted as co-dominants, defending their territory. The study achieved a deeper understanding of the role of spatial exploration, territorial dominance and intraspecific interaction in gilthead seabream, and elucidated the link between them and physiological stress indicators. The results highlight aspects of interest to the aquaculture industry, showing the importance of a greater focus on rearing conditions, finding solutions to mitigate crowding effects and promoting the quality of aquacultural products

Wnt3a neutralization enhances T-cell responses through indirect mechanisms and restrains tumor growth

The Wnt/beta-catenin pathway regulates T-cell functions, including the repression of effector functions to the advantage of memory development via Tcf1. In a companion study, we demonstrate that, in human cancers, Wnt3a/beta-catenin signaling maintains tumor-infiltrating T cells in a partially exhausted status. Here, we have investigated the effects of Wnt3a neutralization in vivo in a mouse tumor model. Abundant Wnt3a was released, mostly by stromal cells, in the tumor microenvironment. We tested whether Wnt3a neutralization in vivo could rescue the effector capacity of tumor-infiltrating T cells, by administering an antibody to Wnt3a to tumor-bearing mice. This therapy restrained tumor growth and favored the expansion of tumor antigen-specific CD8(+) effector memory T cells with increased expression of Tbet and IFN gamma and reduced expression of Tcf1. However, the effect was not attributable to the interruption of T-cell-intrinsic beta-catenin signaling, because Wnt3a/beta-catenin activation correlated with enhanced, not reduced, T-cell effector functions both ex vivo and in vitro. Adoptively transferred CD8(+) T cells, not directly exposed to the anti-Wnt3a antibody but infiltrating previously Wnt3a-neutralized tumors, also showed improved functions. The rescue of T-cell response was thus secondary to T-cell-extrinsic changes that likely involved dendritic cells. Indeed, tumor-derived Wnt3a strongly suppressed dendritic cell maturation in vitro, and anti-Wnt3a treatment rescued dendritic cell activities in vivo. Our results clarify the function of the Wnt3a/beta-catenin pathway in antitumor effector T cells and suggest that Wnt3a neutralization might be a promising immunotherapy for rescuing dendritic cell activities. (C) 2018 AACR

Molecular and clinical studies in five index cases with novel mutations in the GLA gene

Fabry disease is a metabolic and lysosomal storage disorder caused by the functional defect of the α-galactosidase A enzyme; this defect is due to mutations in the GLA gene, that is composed of seven exons and is located on the long arm of the X-chromosome (Xq21–22). The enzymatic deficit is responsible for the accumulation of glycosphingolipids in lysosomes of different cellular types, mainly in those ones of vascular endothelium. It consequently causes a cellular and microvascular dysfunction. In this paper, we described five novel mutations in the GLA gene, related to absent enzymatic activity and typical manifestations of Fabry disease. We identified three mutations (c.846_847delTC, p.E341X and p.C382X) that lead to the introduction of a stop codon in positions 297, 341 and 382. Moreover we found a missense mutation (p.R227P) in the exon 5 of the GLA gene and a single point mutation (c.639 + 5 G > T) occurring five base pairs beyond the end of the exon 4. These mutations have never been found in our group of healthy control subjects > 2300. The studied patients presented some clinical manifestations, such as cornea verticillata, hypo-anhidrosis, left ventricular hypertrophy, cerebrovascular disorders and renal failure, that, considering the null enzymatic activity, suggest that the new mutations reported here are related to the classic form of Fabry disease. The identification of novel mutations in patients with symptomatology referable to FD increases the molecular knowledge of the GLA gene and it gives clinicians an important support for the proper diagnosis of the disease

Tornillos at Vulcano: Clues to the dynamics of the hydrothermal system

The number of tornillo events has recently increased at the Vulcano Island, Italy. While only 15 tornillos were recorded during 2004–2006, 584 events occurred in 2007–2008. They were located just below La Fossa Crater at depths ranging between 0.1 and 1 km b.s.l. During two intervals in 2007–2008 increases in the number of tornillos took place at the same time as temperature and geochemical anomalies were observed. The spectral content of the tornillos, generally characterized by one–two dominant spectral peaks near 6 and 10 Hz, varied over time, with changes also noted in the quality factors. The simplest source mechanism proposed for tornillos is the free eigenvibration of a fluid volume within a crack or a conduit. Based on this model, we propose a causal relationship between the temperature and geochemical anomalies and the increases in numbers of tornillos. As the amount of hydrothermal fluids increases during the anomalies, the upward flux of fluids grows. The consequent changes in the pressure, temperature and dynamics of the system of cracks and conduits result in the generation of tornillos. Based on the fluid-filled crack/conduit model, the shallow depths of the sources and the values of the quality factors, the fluid within the resonant crack/conduit was inferred to be an ash–gas or water droplet–gas mixture. Moreover, the observed variations in the wavefield can be caused by small changes in the location of the source, in the source mechanism, or in the medium in between the source and the seismic station. Finally, another peculiar feature of tornillos is the amplitude modulation that can be explained as a result of a beating phenomenon.Published377-3933V. Proprietà chimico-fisiche dei magmi e dei prodotti vulcaniciJCR Journalreserve

Deletion of the miR172 target site in a TOE-type gene is a strong candidate variant for dominant double-flower trait in Rosaceae

Double flowers with supernumerary petals have been selected by humans for their attractive appearance and commercial value in several ornamental plants, including Prunus persica (peach), a recognized model for Rosaceae genetics and genomics. Despite the relevance of this trait, knowledge of the underlying genes is limited. Of two distinct loci controlling the double-flower phenotype in peach, we focused on the dominant Di2 locus. High-resolution linkage mapping in five segregating progenies delimited Di2 to an interval spanning 150858bp and 22 genes, including Prupe.6G242400 encoding an euAP2 transcription factor. Analyzing genomic resequencing data from single- and double-flower accessions, we identified a deletion spanning the binding site for miR172 in Prupe.6G242400 as a candidate variant for the double-flower trait, and we showed transcript expression for both wild-type and deleted alleles. Consistent with the proposed role in controlling petal number, Prupe.6G242400 is expressed in buds at critical times for floral development. The indelDi2 molecular marker designed on this sequence variant co-segregated with the phenotype in 621 progenies, accounting for the dominant inheritance of the Di2 locus. Further corroborating the results in peach, we identified a distinct but similar mutation in the ortholog of Prupe.6G242400 in double-flower roses. Phylogenetic analysis showed that these two genes belong to a TARGET OF EAT (TOE)-type clade not represented in Arabidopsis, indicating a divergence of gene functions between AP2-type and TOE-type factors in Arabidopsis and other species. The identification of orthologous candidate genes for the double-flower phenotype in two important Rosaceae species provides valuable information to understand the genetic control of this trait in other major ornamental plants. Significance Statement We used peach as a model to gain insight into the molecular basis of double flowers, an important trait in many ornamental plants. We propose that a deletion causes a TOE-type transcription factor to escape miR172-mediated repression, in turn resulting in an increased number of petals, as corroborated by results on the orthologous gene in rose

Preliminary study of novel SRC tyrosine kinase inhibitor and proton therapy combined effect on glioblastoma multiforme cell line: In vitro evaluation of target therapy for the enhancement of protons effectiveness

The aim of this work was to evaluate proton therapy effectiveness in combination with a molecule SRC protein inhibitor for glioblastoma multiforme treatment. The role of this novel compound, Si306, is to interfere with glioblastoma carcinogenesis and progression, creating a radiosensitivity condition. The experiments were performed on U87 human glioblastoma multiforme cell line. Molecule concentrations of 10 μM and 20μM were tested in combination with proton irradiation doses of 2, 4, 10 and 21Gy. Cell survival evaluation was performed by clonogenic assay. The results showed that Si306 increases the efficacy of proton therapy reducing the surviving cells fraction significantly compared to treatment with protons only. These studies will support the preclinical phase realization, in order to evaluate proton therapy effects and molecularly targeted drug combined treatments

Proton-irradiated breast cells: molecular points of view

Breast cancer (BC) is the most common cancer in women, highly heterogeneous at both the clinical and molecular level. Radiation therapy (RT) represents an efficient modality to treat localized tumor in BC care, although the choice of a unique treatment plan for all BC patients, including RT, may not be the best option. Technological advances in RT are evolving with the use of charged particle beams (i.e. protons) which, due to a more localized delivery of the radiation dose, reduce the dose administered to the heart compared with conventional RT. However, few data regarding proton-induced molecular changes are currently available. The aim of this study was to investigate and describe the production of immunological molecules and gene expression profiles induced by proton irradiation. We performed Luminex assay and cDNA microarray analyses to study the biological processes activated following irradiation with proton beams, both in the non-tumorigenic MCF10A cell line and in two tumorigenic BC cell lines, MCF7 and MDA-MB-231. The immunological signatures were dose dependent in MCF10A and MCF7 cell lines, whereas MDA-MB-231 cells show a strong pro-inflammatory profile regardless of the dose delivered. Clonogenic assay revealed different surviving fractions according to the breast cell lines analyzed. We found the involvement of genes related to cell response to proton irradiation and reported specific cell line- and dose-dependent gene signatures, able to drive cell fate after radiation exposure. Our data could represent a useful tool to better understand the molecular mechanisms elicited by proton irradiation and to predict treatment outcome