New inter-correlated genes targeted by diatom-derived polyunsaturated aldehydes in the sea urchin Paracentrotus lividus

Abstract The marine environment is continually subjected to the action of stressors (including natural toxins), which represent a constant danger for benthic communities. In the present work using network analysis we identified ten genes on the basis of associated functions ( FOXA , FoxG, GFI-1 , nodal, JNK, OneCut/Hnf6, TAK1 , tcf4, TCF7, VEGF ) in the sea urchin Paracentrotus lividus , having key roles in different processes, such as embryonic development and asymmetry, cell fate specification, cell differentiation and morphogenesis, and skeletogenesis. These genes are correlated with three HUB genes, Foxo , Jun and HIF1A . Real Time qPCR revealed that during sea urchin embryonic development the expression levels of these genes were modulated by three diatom-derived polyunsaturated aldehydes (PUAs), decadienal, heptadienal and octadienal. Our findings show how changes in gene expression levels may be used as an early indicator of stressful conditions in the marine environment. The identification of key genes and the molecular pathways in which they are involved represents a fundamental tool in understanding how marine organisms try to afford protection against toxicants, to avoid deleterious consequences and irreversible damages. The genes identified in this work as targets for PUAs can be considered as possible biomarkers to detect exposure to different environmental pollutants

Environmental sustainability and service quality beyond economic and financial indicators: A performance evaluation of Italian water utilities

As water utilities operate as natural monopolists and they provide essential services for human life, their activities are regulated by public authorities. The sustainable use of water resources and a specific attention on social needs should be essential goals for this kind of firms, so that the evaluation of their business should go beyond their profitability and their financial solvency. Keeping pace with the new Circular Economy paradigm and the evolution of the water regulatory framework, in this paper we suggest a global composite indicator apt to evaluate in a novel way the water utilities performance, encompassing financial and economic indexes together with environmental sustainability and service quality measures. To show its empirical implementation we evaluate the performance of Italian water utilities. The operating context is also under scrutiny focusing on specific water utility features such as size, geographical location, degree of diversification and ownership. In this light, operating in the Center and being large are considered favourable background conditions, while the South and the medium size display a significant unfavourable influence on the water utility performance. Multi-utilities are more advantaged with respect to the mono-utilities and no significant distinction can be made among the different ownership models. ispartof: Socio-Economic Planning Sciences status: Published onlin

Fondazioni di partecipazione e gestione di beni e servizi culturali: problemi giuridici

The so called “participatory cultural foundation” (P.F.) is an innovative legal entity which can be used in the management of cultural Arts and public services. It is considered as a no-profit private Institution. In order to examine the characteristics of P.F., in the first part of the dissertation I will analyze the fundamentals of the constitutional division of competences in the cultural field, the theoretical notion of “cultural Arts”, the functions of "protection" and "enhancement" and the role of private individuals in this specific sector. The second part of the dissertation focuses on the "management models” of cultural Arts and services, within the framework of the subsidiarity principle and the Public-Private Partnership (P.P.P.), with a special emphasis on the EU Law. The last part of the thesis deepens the doctrinal elaboration of the P.F., with special regard to its peculiarities in the cultural field. Special attention is paid to the recent legal provisions which apparently restrict the uses of the institution, having been adopted as part of the public spending review

An eighteen-month follow-up study on the effects of intravitreal dexamethasone implant in diabetic macular edema refractory to anti-VEGF therapy

Abstract AIM: To evaluate the long-term efficacy and safety of dexamethasone implants in subjects affected by diabetic macular edema (DME) resistant to anti-vascular endothelial growth factor (VEGF) therapy. METHODS: Thirty-two DME patients were enrolled. A 700 microgram slow release Intravitreal Dexamethasone Implant (Ozurdex®) was placed in the vitreous cavity. All patients were followed for 18mo. Best-corrected visual acuity (BCVA) measured with Early Treatment Diabetic Retinopathy Study (ETDRS) and central macular thickness (CMT) exams were carried out at baseline (T0) and after 1 (T1), 3 (T3), 4 (T4), 6 (T6), 9 (T9), 12 (T12), 15 (T15), and 18mo (T18) post injection. RESULTS: Repeated measures ANOVA showed an effect of treatment on ETDRS (P<0.0001). Post hoc analyses revealed that ETDRS values were significantly increased at T1, T3, T4, T9, and T15 (P<0.001) as compared to baseline value (T0). At T6, T12, and T18, ETDRS values were still statistically higher than baseline (P<0.001 vs T0). However, at these time points, we observed a trend to return to baseline conditions. ANOVA also showed an effect of treatment (P<0.0001). CMT decreased significantly at T1, T3, T4, T9, and T15 (P<0.001). At T6 (P<0.01), T12 and T18 (P<0.001) CMT was also significantly lower than T0 although a trend to return to the baseline conditions was also observed. CONCLUSION: Our findings demonstrate that Intravitreal Dexamethasone Implant is a good option to improve BCVA and CMT in DME patients resistant to anti-VEGF therapy. Our data also show that the use of drugs administered directly into the vitreous allows achieving appropriate and long-lasting concentration at the site of disease without systemic side effects

Water pollution in wastewater treatment plants: An efficiency analysis with undesirable output

The environmental efficiency of 96 Tuscan (Italian) wastewater treatment plants (WWTPs) is investigated taking into account the quality of the outgoing water in terms of pollutant. In this regard, the presence of the residual nitrogen in the outgoing treated water is considered as undesirable output. The efficiency analysis is performed by applying a novel integrated Analytic Hierarchy Process/Non-radial Directional Distance Function (AHP/NDDF) approach, combining the benefits of the two techniques. Similarly to the standard NDDF approach, the suggested model allows to include simultaneously inputs, desirable and undesirable outputs and not to overestimate the efficiency scores. At the same time, the AHP inclusion gives the possibility to directly take into account the decision maker preferences in the weighting system and to encompass some existing directional distance function models as special cases.The obtained results are then used to identify the efficiency explanatory variables: among them, the facilities' capacity, the percentage of wastewater discharged by the industrial and agricultural activities and the level of compliance with the pollutant concentration threshold set by the legislator have a significant impact on the WWTP performance. The integrated performance assessment allows the water authorities to combine the WWTP efficiency together with the environmental sustainability issue and it has the potential for further promising environmental inspections

Nitric Oxide Mediates the Stress Response Induced by Diatom Aldehydes in the Sea Urchin Paracentrotus lividus

Diatoms are ubiquitous and abundant primary producers that have been traditionally considered as a beneficial food source for grazers and for the transfer of carbon through marine food webs. However, many diatom species produce polyunsaturated aldehydes that disrupt development in the offspring of grazers that feed on these unicellular algae. Here we provide evidence that production of the physiological messenger nitric oxide increases after treatment with the polyunsaturated aldehyde decadienal in embryos of the sea urchin Paracentrotus lividus. At high decadienal concentrations, nitric oxide mediates initial apoptotic events leading to loss of mitochondrial functionality through the generation of peroxynitrite. At low decadienal concentrations, nitric oxide contributes to the activation of hsp70 gene expression thereby protecting embryos against the toxic effects of this aldehyde. When nitric oxide levels were lowered by inhibiting nitric oxide synthase activity, the expression of hsp70 in swimming blastula decreased and the proportion of abnormal plutei increased. However, in later pluteus stages nitric oxide was no longer able to exert this protective function: hsp70 and nitric oxide synthase expression decreased with a consequent increase in the expression of caspase-8. Our findings that nitric oxide production increases rapidly in response to a toxic exogenous stimulus opens new perspectives on the possible role of this gas as an important messenger to environmental stress in sea urchins and for understanding the cellular mechanisms underlying toxicity during diatom blooms

Molecular response to toxic diatom-derived aldehydes in the sea urchin Paracentrotus lividus.

Diatoms are dominant photosynthetic organisms in the world's oceans and represent a major food source for zooplankton and benthic filter-feeders. However, their beneficial role in sustaining marine food webs has been challenged after the discovery that they produce secondary metabolites, such as polyunsaturated aldehydes (PUAs), which negatively affect the reproductive success of many invertebrates. Here, we report the effects of two common diatom PUAs, heptadienal and octadienal, which have never been tested before at the molecular level, using the sea urchin, Paracentrotus lividus, as a model organism. We show that both PUAs are able to induce teratogenesis (i.e., malformations), as already reported for decadienal, the better-studied PUA of this group. Moreover, post-recovery experiments show that embryos can recover after treatment with all three PUAs, indicating that negative effects depend both on PUA concentrations and the exposure time of the embryos to these metabolites. We also identify the time range during which PUAs exert the greatest effect on sea urchin embryogenesis. Finally, we report the expression levels of thirty one genes (having a key role in a broad range of functional responses, such as stress, development, differentiation, skeletogenesis and detoxification processes) in order to identify the common targets affected by PUAs and their correlation with morphological abnormalities. This study opens new perspectives for understanding how marine organisms afford protection from environmental toxicants through an integrated network of genes

The effect of FK506 on transforming growth factor beta signaling and apoptosis in chronic lymphocytic leukemia B cells.

Background Loss of response to transforming growth factor-beta (TGF-beta) is thought to contribute to the progression of chronic lymphocytic leukemia. Recent findings of over-activation of the TGF-beta signal in FKBP12-knockout mouse prompted us to investigate whether FK506, the canonical ligand of FKBP, can activate the TGF-beta signal in chronic lymphocytic leukemia. Design and Methods We studied 62 chronic lymphocytic leukemia samples from patients with Rai/Binet stage 0 to 4 disease. The TGF-beta signal was investigated by western blotting and flow cytometry. The levels of Bcl2-family members and death-associated-protein kinase were also investigated by western blotting, whereas apoptosis was studied in flow cytometry. Down-modulation of FKBP12 was obtained by gene silencing with short interfering RNA. Results Twenty-two out of 62 chronic lymphocytic leukemia samples were sensitive to TGF-beta-induced apoptosis. All but two of the responsive samples underwent apoptosis also when cultured with FK506, but not with cyclosporine. Thirteen samples that were not sensitive to TGF-beta were sensitive to FK506. Overall, response to FK506 occurred in 33 samples. FK506 induced Smad2 phosphorylation and nuclear translocation. Accordingly, death-associated-protein kinase, a transcriptional target of Smad, was induced. At the same time, Bcl-2 and Bcl-xL levels decreased whereas the levels of Bim and Bmf increased. A loss of mitochondrial membrane potential preceded caspase activation and cell death. FK506 removed FKBP12 from its binding to the TGF-beta-receptor. FKBP12 release activated the receptor-kinase activity as suggested by the enhanced levels of phospho-Smad found in cells depleted of FKBP12. Conclusions Our study shows that most chronic lymphocytic leukemia cells escape the homeostatic control of TGF-beta and that FK506 restores the TGF-beta signal in a proportion of non-responsive samples. We demonstrated that FK506 activates TGF-beta receptor I kinase activity in chronic lymphocytic leukemia, which transduces apoptosis by a mitochondrial-dependent pathway

Microtubule Organization during the Early Development of the Parthenogenetic Egg of the HymenopteranMuscidifurax uniraptor

AbstractThe origin of the zygotic centrosome is an important step in developmental biology. It is generally thought that sperm at fertilization plays a central role in forming the functional centrosome which subsequently organizes the first mitotic spindle. However, this view is not applicable in the case of parthenogenetic eggs which develop without the sperm contribution. To clarify the problem of the origin of the zygotic centrosome during parthenogenetic development, we studied a hymenopteran,Muscidifurax uniraptor.Antitubulin antibody revealed that after activation several asters assembled in the egg cytoplasm. The number of asters varied in relation to the cell cycle. They became visible from anaphase of the first meiotic division and increased in number as meiosis progressed, reaching a maximum at the first mitosis. From anaphase–telophase of the first mitosis they decreased in number and were no longer found during the third mitotic division. To elucidate the nature of these asters we performed an ultrastructural study with transmission electron microscopy and immunofluorescence with antibodies against anti-γ-tubulin and CP190. In this way we showed the presence in these asters of centrosomal components and centrioles. Our observations suggest that the cytoplasm ofMuscidifuraxeggs contains a pool of inactive centrosomal precursor proteins becoming able to nucleate microtubules into well-defined asters containing centrioles after activation

UV-protective compounds in marine organisms from the Southern Ocean

Solar radiation represents a key abiotic factor in the evolution of life in the oceans. In general, marine, biota particularly in euphotic and dysphotic zones depends directly or indirectly on light, but ultraviolet radiation (UV-R) can damage vital molecular machineries. UV-R induces the formation of reactive oxygen species (ROS) and impairs intracellular structures and enzymatic reactions. It can also affect organismal physiologies and eventually alter trophic chains at the ecosystem level. In Antarctica, physical drivers, such as sunlight, sea-ice, seasonality and low temperature are particularly influencing as compared to other regions. The springtime ozone depletion over the Southern Ocean makes organisms be more vulnerable to UV-R. Nonetheless, Antarctic species seem to possess analogous UV photoprotection and repair mechanisms as those found in organisms from other latitudes. The lack of data on species-specific responses towards increased UV-B still limits the understanding about the ecological impact and the tolerance levels related to ozone depletion in this region. The photobiology of Antarctic biota is largely unknown, in spite of representing a highly promising reservoir in the discovery of novel cosmeceutical products. This review compiles the most relevant information on photoprotection and UV-repair processes described in organisms from the Southern Ocean, in the context of this unique marine polar environment