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Bioplastics are promoted as safer alternatives to tackle the long-term persistence of conventional plastics. However, information on the potential release of additives and non-intentionally added substances (NIAS) in the surrounding environment is limited, and biological effects of the leachates have been little studied. Leachates produced from three bioplastics, i.e. compostable bags (CB), bio-polyethylene terephthalate bottles (bioPET) and polylactic acid cups (PLA), and a control polymeric material, i.e. rubber tire (TR), were examined. The chemical nature of bioplastic polyesters PET, PLA and poly (butylene adipate-co-terephthalate) (PBAT) in CB, was confirmed by analytical pyrolysis. Fragments were incubated in artificial sea water for 14 days at 20 °C in darkness and leachate contents examined by GC-MS and HPLC-MS/MS. Catalysts and stabilizers represented the majority of chemicals in TR, while NIAS (e.g. 1,6-dioxacyclododecane-7,12-dione) were the main components of CB. Bisphenol A occurred in all leachates at a concentration range 0.3-4.8 μg/L. Trace metals at concentrations higher than control water were found in all leachates, albeit more represented in leachates from CB and TR. A dose response to 11 dilutions of leachates (in the range 0.6-100%) was tested for biological effects on early embryo stages of Mytilus galloprovincialis. Embryotoxicity was observed in the whole range of tested concentrations, the magnitude of effect depending on the polymers. The highest concentrations caused reduction of egg fertilization (CB, bioPET, TR) and of larvae motility (CB, PLA, TR). TR leachates also provoked larvae mortality in the range 10-100%. Effects on adult mussel physiology were evaluated after a 7-day in vivo exposure to the different leachates at 0.6% concentration. Nine biomarkers concerning lysosomal functionality, neurotransmission, antioxidant and immune responses were assessed. All lysosomal parameters were affected, and serum lysozyme activity inhibited. Harmonized chemical and biological approaches are recommended to assess bioplastic safety and support production of sustainable bioplastics.

Bisphenol A (BPA) is one of the most investigated compound as a suspected endocrine disrupting chemical. It has been found at nM concentrations in the maternal serum, cord serum, and amniotic fluid and also permeates placental tissues. Attempts are being made to replace BPA with the analog Bisphenol S (BPS). Also BPS was found in maternal and umbilical cord serum, and urine samples from a large population of pregnant women. A few studies investigated BPA impact on the placentation process, and even less are available for BPS. This work aimed to elucidate and compare the effects of BPA and BPS on physiological functions of HTR-8/SVneo cells, derived from extravillous trophoblast of first-trimester pregnancy. Proliferation and migration ability of trophoblast cells were assessed in vitro after exposure to BPA or BPS (10–13–10–3 M). Further, induction of the inflammatory response by the bisphenols was studied. To provide insight into the molecular pathways implicated in the responses, experiments were carried out in the presence or absence of tamoxifen as estrogen receptors (ERs) blocker, and U0126 as ERK1/2 phosphorylation inhibitor. Data indicate that BPA significantly affects both proliferation and migration of HTR-8/SVneo cells, through ER and ERK1/2 mediated processes. Differently, BPS only acts on proliferation, again through ER and ERK1/2 mediated processes. BPS, but not BPA, induces secretion of interleukins 6 and 8. Such effect is inhibited by blocking ERK1/2 phosphorylation. To the best of our knowledge, these are the first data showing that BPS affects trophoblast functions through ER/MAPK modulation. Summary sentence Bisphenol A and its analog bisphenol S, emerging contaminants founded in human serum and placental fluids, significantly affected the biological functions of HTR-8/SVneo cells, through estrogen receptors and MAPK mediated processes. Graphical Abstract

We empirically investigate incumbents’ and entrants’ bids on an original dataset of 192 scoring rule auctions for canteen services in Italy. Our findings show that winning rebates are lower (i.e., prices paid by the public buyer are higher) when the contract is awarded to the incumbent supplier. This result is not explained by the observable characteristics of the auction or the service awarded. We develop a simple theoretical model showing that the result is consistent with a setting in which the buyer exploits specific information on the incumbent supplier’s production cost.

We experimentally study bidding behavior in sequential first-price procurement auctions where bidders’ capacity constraints are private information. Treatment differs in the ex-ante probability distribution of sellers’ capacities and in the (exogenous) probability that the second auction is actually implemented. Our results show that: (i) bidding behavior in the second auction conforms with sequential rationality; (ii) while first auction’s bids negatively depend on capacity, bidders seem unable to recognize this link when, at the end of the first auction, they state their beliefs on the opponent’s capacity. To rationalize this inconsistency between bids and beliefs, we conjecture that bidding in the first auction is also affected by a hidden, behavioral type – related to the strategic sophistication of bidders – that obfuscates the link between capacity and bids. Building on this intuition, we show that a simple level-k model may help explain the inconsistency.

This study evaluates the transcriptional profiles of genes related to physiological responses in digestive glands (DG) of Mytilus galloprovincialis under the influence of seasonal changes of environmental variables, gender bias, and gonadal development. Composition of the DG microbiome was also explored. Mussels were collected across 7 months encompassing 3 seasons from a farm in the Northwestern Adriatic Sea. All gene products showed complex transcriptional patterns across seasons. Salinity, surface oxygen and transparency significantly correlate with transcriptional profiles of males, whereas in females temperature and gonadal maturation mostly explained the observed transcriptional changes. Seasonal variations and gender-specific differences were observed in DG microbiome composition, with variations resembling metabolic accommodations likely facing season progression and reproductive cycle. Results provide baseline information to improve actual monitoring strategies of mussel farming conditions and forecast potential detrimental impacts of climatological/environmental changes in the study area.

The sorption of organic pollutants to marine plastic litter may pose risks to marine organisms, notably for what concerns their intake and transfer through microplastic (MP) ingestion. This study investigated the effects of polystyrene MP loaded with chrysene (CHR) on early-stage and physiological endpoints measured in the Mediterranean mussel Mytilus galloprovincialis. The same concentrations of virgin microplastics (MP) and MP loaded with 10.8 µg CHR/mg (CHR-MP) were administered to mussel gametes/embryos (25 × 103 items/mL) and adults (5⋅× 103 items/L), further treatments included 0.1 mg/L of freely dissolved CHR and a second CHR concentration corresponding to that vehiculated by CHR-MP during exposure (3.78 µg/L and 0.73 ng/L for gamete/embryos and adults, respectively). None of the treatments affected gamete fertilization, while 0.1 mg/L CHR induced embryotoxicity. In adults, CHR-MP and MP similarly affected lysosomal membrane stability and neutral lipids and induced slight effects on oxidative stress endpoints. CHR affected tested endpoints only at 0.1 mg/L, with lysosomal, oxidative stress and neurotoxicity biomarkers generally showing greater alterations than those induced by CHR-MP and MP. This study shows that the CHR sorption on MP does not alter the impact of virgin MP on mussels and may pose limited risks compared to other routes of exposure.

International audience; We use a statistical micro matching procedure to obtain a synthetic dataset to investigate the risk of fuel poverty in small areas. Specifically, we link 19,174 homes with Energy Performance Certificates (EPCs) in the Italian province of Treviso with information - from the General Census of Population and Housing and Household Budget Survey - about the socio-economic features of the families that most likely inhabit them. Based on this original dataset, we find that poor housing is as important as low income and the use of natural gas in determining fuel poverty. In particular, the risk of fuel poverty is increased by i) the energy inefficiency of the home; ii) the size of the home in relation to household size; iii) the lower total household expenditure. Conversely, the risk decreases when the main heating is natural gas and/or the home is endowed with renewable energy sources. Our results show that information in EPCs, matched at the micro level with the socio-economic data usually available from administrative sources or surveys, can be used to identify - on the one hand - the individual households and the municipal areas risking fuel poverty, and - on the other - the most effective policy options to tackle the phenomenon.

We empirically compare bids (i.e. prices) from temporary partnerships (TPs), that outsource part of the contract before the auction, and firms that outsource afterwards. Using a comprehensive dataset on procurement auctions for public works in Valle d’Aosta (Italy), we find that the timing of outsourcing affects the bids and the probability of winning the auction. Specifically, TPs bid closer to the payoff maximizing offer and are more likely to win. Hence, the price paid by the public buyer is lower. These results are supported by a simple theoretical setting showing that, by pre-committing to a TP, suppliers have a lower risk of being “held up” by subcontractors than firms that outsource part of the work after the bidding phase. Our results show the advantage for TPs of freely choosing partners, size and boundaries before the auction, highlighting their potential in fostering the effective participation in public procurement procedures of Small and Medium-sized Enterprises (SMEs).

This paper investigates the price variability of standardized medical devices purchased by Italian Public Buyers (PBs). A semiparametric approach is used to recover the marginal cost of each device. Average prices vary substantially between PBs; we show that most of the difference between the purchase prices and estimated costs is associated with a PB fixed effect, which, in turn, is related to the institutional characteristics and size of the PB. Repeating the main estimation using device fixed effects yields similar results. Finally, an exogenous policy change, i.e. the termination of the mandatory reference price regime, is used to assess how discretion affects medical device procurement given the skills of each PB. Our results show that less PB discretion — i.e. when mandatory reference prices apply — determines efficiency gains and losses for low- and high-skilled PBs, respectively.

Ocean contamination by synthetic polymers can represent a risk for the fitness of marine species due to the leaching of chemical additives. This study evaluated the sub-lethal effects of plastic and rubber leachates on the mussel Mytilus galloprovincialis through a battery of biomarkers encompassing lysosomal endpoints, oxidative stress/detoxification parameters, and specific responses to metals/neurotoxicants. Mussels were exposed for 7 days to leachates from car tire rubber (CTR), polypropylene (PP), polyethylene terephthalate (PET), polystyrene (PS) and polyvinyl chloride (PVC), containing organic additives and metals in the ng-μg/L range. The leachate exposure affected general stress parameters, including the neutral lipid content (all leachates), the lysosomal membrane stability (PS, PP, PVC and CTR leachates) and lysosomal volume (PP, PVC and TR leachates). An increased content of the lipid peroxidation products malondialdehyde and lipofuscin was observed in mussels exposed to PET, PS and PP leachates, and PP, PVC and CTR leachates, respectively. PET and PP leachates increased the activity of the phase-II metabolism enzyme glutathione S-transferase, while a decreased acetylcholinesterase activity was induced by PVC leachates. Data were integrated in the mussel expert system (MES), which categorizes the organisms’ health status based on biomarker responses. The MES assigned healthy status to mussels exposed to PET leachates, low stress to PS leachates, and moderate stress to PP, CTR and PVC leachates. This study shows that additives leached from selected plastic/rubber polymers cause sub-lethal effects in mussels and that the magnitude of these effects may be higher for CTR, PVC and PP due to a higher content and release of metals and organic compounds.

International audience; For the period 2003–2014, we investigate unexplored effects of fiscal consolidation in decentralized public finance on a large dataset of Italian municipalities. Based on a simple, realistic theoretical model, we show that municipalities increase arrears on committed investment expenditure as a response to intergovernmental transfer cuts. Then, we test our predictions controlling for potential sources of endogeneity, and find that a reduction in intergovernmental transfers causes a significant increase in arrears, in addition to other common adjustments to local fiscal policies (e.g., tax revenues). Our results highlight a perverse effect of fiscal consolidation packages implemented by centrally imposed fiscal restraints.

The possible impact of carbamazepine (CBZ) and propranolol (PROP), two widespread pharmaceuticals in the aquatic environment, were investigated on morphology and gene transcription of early larvae of Mytilus galloprovincialis . Pharmaceuticals were first tested in a wide concentration range (from 0.01 to 1000 μg/L) through the 48-hpf embryotoxicity assay. The results showed that both compounds significantly affected embryo development from environmental concentrations. Although similar EC 50 were obtained, (≅ 1 μg/L) CBZ induced a progressive increase in embryo malformations, whereas PROP apparently showed greater impacts in terms of arrested development and embryo mortality at higher concentrations (>10 μg/L). Transcriptional analyses of 17 genes involved in different physiological functions in mussels and/or in their response to environmental contaminants, were performed at 24 and 48 h pf at two selected concentrations of CBZ and PROP (0.01 and 1 μg/L). Both compounds induced down-regulation of shell-specific and neuroendocrine related transcripts, while distinct effects were observed on antioxidant, lysosomal, and immune-related transcripts, also depending on the larval stage investigated. The results demonstrate that CBZ and PROP can affect development and gene transcription in mussel early larvae at environmental concentrations.

The emerging paradigm on plastic pollution in marine environments is that microsize particles (MPs) have far more subtle effects than bigger fragments, given their size range overlapping with that of particles ingested by filter-feeders. The impacts include gut blockage, altered feeding and energy allocation, with knock-on effects on widespread physiological processes. This study investigated whether ingestion of polystyrene MPs (PS-MPs) triggers protective processes in marine mussels. The Multixenobiotic resistance (MXR) system is a cytoprotective mechanism acting as an active barrier against harmful xenobiotics and a route of metabolite detoxification. Both larvae and adults were employed in laboratory experiments with different concentrations of 3-μm PS-MPs (larvae), and 3-μm and 45-μm PS-MPs (adults) matching size range of planktonic food through the mussel lifecycle. Embryos grown in the presence of 3-μm PS-MPs showed significant reduction of MXR activity and down-regulation of ABCB and ABCC transcripts encoding the two main MXR-related transporters P-glycoprotein and the Multidrug resistance-related protein, respectively. In adults, effects of PS-MPs were assessed in haemocytes and gills, which showed different modulation of MXR activity and ABCB/ABCC expression according to MP size (haemocyte and gills) or particle concentration (haemocyte). These data showed that modulation of MXR activity is part of a generalized response triggered by particle ingestion.

This study investigated functional and transcriptional modulation of the Multixenobiotic resistance (MXR) system as a cytoprotective mechanism contributing to the physiological chemoresistance of haemocytes in the Mediterranean mussel. Basal transport activity was assessed using the model substrate rhodamine 123 and specific inhibitors for the MXR-related transporters P-glycoprotein (ABCB mRNA) and Multidrug resistance-related protein (ABCC mRNA). Results showed that MXR activity in mussel haemocytes was mainly supported by the Mrp-mediated efflux. In agreement, ABCC was expressed at higher levels than ABCB. Activation of the cyclic-AMP (cAMP) dependent protein kinase A (PKA) resulted in increased rhodamine efflux, which was counteracted by the selective PKA inhibitor H89. Although serotonin, a physiological modulator of cAMP/PKA signaling and ABCB transcription in haemocytes, did not affect basal MXR transport, the environmental pharmaceuticals fluoxetine, propranolol, and carbamazepine, which interact in different ways with the adrenergic and serotoninergic pathways, were showed to act as modulators and substrates of MXR-related transporters and to affect cell viability. While the increased MXR activity may have lowered the cytotoxic effects of propranolol and carbamazepine, the lack of MXR efflux induction by fluoxetine may play a role in the observed cytotoxicity of the compound.

In this paper, we study bidding behavior by firms in beauty-contest auctions, i.e. auctions in which the winning bid is the one which gets closest to some function (average) of all submitted bids. Using a dataset on public procurement beauty-contest auctions in Italy and exploiting a change in the auction format, we show that firmsÕ observed bidding behavior departs from equilibrium and can be predicted by an index of sophistication, which captures the firmsÕ accumulated capacity of bidding well (i.e., close to optimality) in the past. We show that our empirical evidence is consistent with a Cognitive Hierarchy model of biddersÕ behavior. We also investigate whether and how firms learn to think and bid strategically through experience.

Disputes on the penalties enforceable for breach of contract are often solved in court. Using a large dataset on Italian public procurement contracts, we study the effects of the inefficiency of the local law courts on the delay with which contractors deliver public works. First we sketch a simple model to see how inefficient law courts - i.e. those characterized by longer average duration of trials - may induce public buyers to refrain from enforcing penalties for late delivery in order to avoid costly disputes in court. Then our empirical findings show that, where the local law courts are inefficient: i) public works are delivered with longer delays, and this applies particularly to higher-value contracts - i.e. complex projects; ii) contracts are more often awarded to larger firms; and iii) on average, a higher share of final payment in contracts is adopted. These results are not driven by omitted environmental variables, since we show that delays in the completion of contracted works are still a selected by the efficiency of the local law courts after including province-related fixed effects in the model and considering other possible explanations for our findings.

The widespread occurrence of microplastics (MP) in the marine environment is cause of increasing concerns about the safety of the exposed ecosystems. Although the effects associated to the MP uptake have been studied in most marine taxa, the knowledge about their sub-lethal impacts on early life stages of marine species is still limited. Here, we investigated the uptake/retention of 3-μm polystyrene MP by early stages of the Mediterranean mussel Mytilus galloprovincialis, and the related effects on gut clearance, feeding efficiency, morphological and transcriptional parameters involved in embryo-larval development. Uptake measurements were performed on larvae at 48 h, 3, 6 and 9 days post fertilization (pf) after exposure to a range of 50-10,000 particles mL-1. At all tested pf periods, treatments resulted in a significant and linear increase of MP uptake with increasing concentrations, though levels measured at 48 h pf were significantly lower compared to 3-9 d pf. Ingested MP were retained up to 192 h in larvae's gut, suggesting a physical impact on digestive functions. No change was noted between the consumption of microalgae Nannochloropsis oculata by larvae when administered alone or in the presence of an identical concentration (2000 items mL-1) of MP. The exposure to 50-10,000 MP mL-1 did not alter the morphological development of mussel embryos; however, transcriptional alterations were observed at 50 and 500 MP mL-1, including the up-regulation of genes involved in shell biogenesis (extrapallial protein; carbonic anhydrase; chitin synthase) and immunomodulation (myticin C; mytilin B), and the inhibition of those coding for lysosomal enzymes (hexosaminidase; β-glucorinidase; catepsin-L). In conclusion, though not highlighting morphological or feeding abnormalities, data from this study revealed the onset of physical and transcriptional impairments induced by MP in mussel larvae, indicating sub-lethal impacts which could increase their vulnerability toward further environmental stressors.

We discuss public procurement instruments for acquiring innovation, focusing on the European Pre-commercial Procurement, Procurement of Innovative Solutions and Innovation Partnerships. We analyse, in particular, how firms’ innovation incentives are affected by: (i) economies of scope and externalities between RD (ii) the degree of specificity of the innovation; (iii) the presence of Small and Medium Enterprises in the market and the level of market competition; (iv) the risk of market foreclosure and supplier lock-in. Our study contributes to the literature on incentives in demand-side innovation policy by tapping into the contractual design features and by offering relevant implications for academics and policy-makers.

Using a dataset of medical devices purchased by Italian Public Buyers (PBs), for each purchase, we measure the difference between each item's price and its marginal cost. We define PBs' ability in purchasing as PBs' fixed effect (FE) on that difference. We find that average prices vary substantially amongst PBs, and this variation is largely captured by PBs' FE. We then exploit the exogenous termination of the mandatory reference price regime to assess how discretion affects procurement performance, given each PB's ability. Our results highlight that reduced PBs' discretion - in presence of mandatory prices - determines efficiency gains and losses for low- and high-ability PBs, respectively.

This study aimed at evaluating the environmental quality of a coastal lagoon (Pialassa Piomboni, NW-Adriatic, Italy) by combining analyses of biomarkers of environmental stress and bioaccumulation of contaminants in marine mussels (Mytilus galloprovincialis) transplanted for 28 days to six selected sites. Assessed biomarkers encompassed lysosomal endpoints, oxidative stress and detoxification parameters, specific responses to metals, neuro- and genotoxic substances; chemical analyses focused on PAHs, metals, pesticide and pharmaceuticals. Results showed up to a 67-fold bioaccumulation of 4- to 6-ring PAHs, including pyrene, fluoranthene, chrysene and benzo(ghi)perylene in transplanted mussels compared to reference conditions (T0). A 10-fold increase of Fe, Cr and Mn was observed, while pesticides and pharmaceuticals were not or slightly detected. The onset of a significant (p < 0.05) general stress syndrome occurred in exposed mussels, as outlined by a 50–57.7% decrease in haemocytes lysosomal membrane stability and an increased lysosomal volume (22.6–26.9%) and neutral lipid storage (18.9–48.8%) observed in digestive gland. Data also revealed a diffuse lipofuscin accumulation (86.5–139.3%; p < 0.05) in digestive gland, occasionally associated to a catalase activity inhibition in gill, indicating an increased vulnerability toward pro-oxidant factors. Higher levels of primary DNA damage (258%; p < 0.05) and PAH accumulation were found in mussels exposed along the eastern shoreline, hosting a petrochemical settlement. Bioaccumulated metals showed a positive correlation with increased metallothionein content (85–208%; p < 0.05) observed in mussels from most sites. Overall, the use of physiological and chemical analyses detected chronic alterations of the mussel health status induced by specific toxicological pathways, proving a suitable approach in the framework of biomonitoring programs of coastal lagoons.

We experimentally study the effects of allotment—the division of an item into homogeneous units—in independent private value auctions. We compare a bundling first-price auction with two equivalent treatments where allotment is implemented: a two-unit discriminatory auction and two simultaneous single-unit first-price auctions. We find that allotment in the form of a discriminatory auction generates a loss of efficiency with respect to bundling. In the allotment treatments, we observe large and persistent bid spread, and the discriminatory auction is less efficient than simultaneous auctions. We provide a unified interpretation of our results that is based on both a non-equilibrium response to the coordination problem characterizing the simultaneous auction format and a general class of behavioral preferences that includes risk aversion, joy of winning and loser’s regret as specific cases.

We consider the supplier’s strategic choice on delivery time in a public procurement setting as the result of the firm’s opportunistic behavior on the optimal investment timing when production costs are uncertain. We model the supplier’s trade-off between the option value to defer the contract execution and the penalty payment in the event of delays. We also take into account the issue of penalty enforcement, which in turn depends on both the discretion of the court of law in voiding contractual clauses and the “efficiency” of the judicial system (i.e. the average length of civil trials). We test our main results on Italian public procurement data showing that the supplier’s incentive to delay is greater the higher the volatility of production costs and the lower the “efficiency” of the judicial system. We then calibrate the model using parameters that mimic the Italian scenario on public works procurement and calculate the maximum amount that a supplier is “willing to pay” (per day) to postpone the delivery date and infringe the contract provisions. Our calibration results are consistent with the theoretical model’s predictions and the empirical findings.

We test the robustness of recent findings on the benefits of penalty contracts to the environments typical of B2B (and B2G) procurement, where buyers and sellers interact repeatedly, matching is endogenous and competitive, there are contractible and non-contractible tasks, and reputation-based relationships can emerge. Both bonuses and penalties boost efficiency, strongly increasing effort in the contractible task while only mildly crowding it out in the non-contractible one. However, sellers grab a higher fraction of surplus with bonuses, as buyers’ offers become more generous. Consequently, buyers prefer penalties, which may explain why they are so widespread in procurement.

Bank of Italy Working Paper no. 1076, https://www.bancaditalia.it/pubblicazioni/temi-discussione/2016/2016-1076/en_tema_1076.pdf; Local governments may increase expenditure arrears to relax the financial constraints induced by intergovernmental transfer cuts. We assess this hypothesis using information from accounting and financial reports from Italian municipalities for the period 2003-2010. By exploiting the long-lasting effect of the 1977-1978 structural reform of Italian local public finance, we employ an instrumental variable approach to address endogeneity concerns. We find robust empirical evidence that the lower the intergovernmental grants, the larger the use of arrears in public investment expenditures by municipalities. We argue that, when local governments are not subject to effective controls on the formation of arrears but fiscal rules impose binding budget constraints, a cut in intergovernmental transfers can partially diminish the effectiveness of fiscal consolidation at local level.

Purpose Due to its role in learning, memory and in many neurodegenerative diseases, acetylcholinesterase (AChE) represents an interesting endpoint to assess possible targets of exposure to radiofrequency electromagnetic fields (RF-EMF) generated by mobile phones. We investigated possible alterations of enzymatic activity, gene and protein expression of AChE in neuronal-like cells exposed to a 1.8 GHz Global System for Mobile Communication (GSM) modulated signal (217-GSM).Materials and methods Rat PC12 cells were exposed for 24 h to 1.8 GHz 217-GSM signal. Specific adsorption rate (SAR) was 2 W/kg. AChE enzyme activity was assessed spectrophotometrically by Ellmans method, mRNA expression level was evaluated by real time polymerase chain reaction, and protein expression was assessed by Western blotting.Results AChE enzymatic activity increased of 1.4-fold in PC12 cells exposed to 217-GSM signal for 24 h, whilst AChE transcriptional or translational pathways were not affected.Conclusion Our results provide the first evidence of effects on AChE activity after in vitro exposure of mammalian cells to the RF-EMF generated by GSM mobile phones, at the SAR value 2 W/kg. The obtained evidence promotes further investigations on AChE as a possible target of RF-EMF and confirm the ability of 1.8 GHz 217-GSM signal to induce biological effects in different mammalian cells.

We consider alternative European public procurement mechanisms for acquiring RD (ii) degree of specificity of the innovation; (iii) role of SMEs in the market and level of market competition; (iv) risk of market foreclosure and supplier lock-in. This article contributes to the literature on incentives in demand-side innovation policy by tapping into the contractual design features and by offering relevant implications for academics and policy makers.

The occurrence of caffeine (CF), a biologically active drug, has widely been documented in coastal waters, and whether its environmental concentrations do represent a threat for marine organisms is unclear. The present study aimed at assessing sub-lethal effects induced by a 7-day exposure to environmentally relevant concentrations of CF (5, 50 and 500 ng/L) in the Mediterranean mussel, Mytilus galloprovincialis. CF in water and mussel tissues, and a battery of biomarkers, including lysosomal parameters of general stress, oxidative stress responses and endpoints of neurological and genetic damages, were evaluated and tested for significance vs controls (p

We investigate how the general principles of the Treaty have been applied to the car sector in the EU, given the specific soft law provisions which are typical of the sector. A detailed quantitative analysis from 1990 to 2008 highlights a reduction of aid over time. A shift from sectoral to Òregional developmentÓ motives in granting aid to the sector is also observed in the last ten years. However, sector specific aid is now less explicit but it remains important. Large amounts of public money are spent without a consistent strategy, reducing capacity in some cases, expanding it in others. The scarcity of public funds calls for a more focussed European policy for this industry.

The focus of the present study was to set a methodological approach for evaluating molecular mechanisms underlying silvering transformation in the European eel, Anguilla anguilla. Silvering is a tightly controlled process during which eels undergo significant morphological, physiological and behavioral changes, pre-adapting for the oceanic spawning migration. Female eels showing different silver indexes were caught in different seasons in the Comacchio Lagoon (North Adriatic Sea, Italy). Isolated hepatocytes from these eels were selected as the experimental model given the relevant role of these cells in metabolic functions potentially altered during silvering. Expression profiles of 7 candidate reference transcripts were analyzed seeking the most viable and robust strategies for accurate qPCR data normalization during silvering. Stability analysis and further statistical validation identified transcripts encoding the ribosomal proteins L13 and ARP as the appropriate reference genes in studies on A. anguilla through silvering. The identified reference transcripts were further used to evaluate expression profiles of target transcripts encoding the thyroid hormone receptor β (THRβ) and vitellogenin (vtg), known to be involved in silvering processes. To the best of our knowledge, this is the first study comparing THRβ expression in European eels across silvering.

We investigated the effect of high-frequency electromagnetic fields (HF-EMFs) and 17-β-estradiol on connexins (Cxs), integrins (Ints), and estrogen receptor (ER) expression, as well as on ultrastructure of trophoblast-derived HTR-8/SVneo cells. HF-EMF, 17-β-estradiol, and their combination induced an increase of Cx40 and Cx43 mRNA expression. HF-EMF decreased Int alpha1 andβ1 mRNA levels but enhanced Int alpha5 mRNA expression. All the Ints mRNA expressions were increased by 17-β-estradiol and exposure to both stimuli. ER-βmRNA was reduced by HF-EMF but augmented by 17-β-estradiol alone or with HF-EMF. ER-βimmunofluorescence showed a cytoplasmic localization in sham and HF-EMF exposed cells which became nuclear after treatment with hormone or both stimuli. Electron microscopy evidenced a loss of cellular contact in exposed cells which appeared counteracted by 17-β-estradiol. We demonstrate that 17-β-estradiol modulates Cxs and Ints as well as ER-βexpression induced by HF-EMF, suggesting an influence of both stimuli on trophoblast differentiation and migration.

Aquatic organisms are unintentionally exposed to a large number of pharmaceutical residues in their natural habitats. Ecotoxicological studies have agreed that these compounds are not harmful to aquatic organisms, as their environmental concentrations are typically too low. However, recent reports have shown biological effects at such low concentrations when biological endpoints related to the therapeutic effects are assessed. Therefore, conservation of molecular targets is now addressed as a key aspect for the development of more efficient test strategies for pharmaceutical environmental risk assessment, providing the rationale for the mode of action (MOA) approach. In the present study the MOA approach was used to investigate the interactive effects of fluoxetine (FX) and propranolol (PROP) on the Mediterranean mussels (Mytilus galloprovincialis). Indeed, organisms in the environment are exposed to pharmaceutical mixtures throughout their lifetime, and particular combinations may be of concern. The antidepressant FX increases serotonin (5-HT) levels in the synaptic cleft by inhibiting 5-HT reuptake. PROP, a prototypical β-adrenoceptor antagonist, also blocks 5-HT1 receptors, which are negatively coupled to cAMP-mediated signaling. Cell signaling alterations potentially triggered by 5-HT1 receptor occupation were therefore assessed after a 7-day mussel exposure to FX or PROP, alone or in combination, each at 0.3 ng/L concentration. FX decreased cAMP levels and PKA activities in digestive gland and mantle/gonads, in agreement with an increased occupation of 5-HT1 receptors. PROP caused a decrease in cAMP levels and PKA activities in digestive gland and an increase in cAMP levels in mantle/gonads, consistent with a differential expression of adrenergic and 5-HT receptors in the two tissues. Co-exposure to FX and PROP provides significant indications for antagonistic effects of the pharmaceuticals, consistent with a direct (PROP) and indirect (FX) action on the same molecular target. Interestingly, FX induced over-expression of a 5-HT1 gene product, and PROP counteracted such increase when the mixture was administered, while having per se no effect. Finally, mRNA expression of the ABCB gene encoding the MXR-related transporter P-glycoprotein was reduced by both pharmaceuticals in the digestive gland, while decreased by FX, increased by PROP, and not affected by the mixture in mantle/gonads. Since transcription of this gene product is under cAMP/PKA modulation, the impairment of regulatory pathways triggered by low concentrations of pharmaceuticals have the potential to affect the ability of animals to elaborate strategies of defense or adaptation toward further stress factors. In this specific case, the pharmaceutical mixture limits the detrimental effects of the single compounds.

In this paper we investigate how tightening fiscal constraints (e.g., through intergovernmental transfer cuts) can lead local governments to postpone investments’ payments. We first provide a simple model showing how local governments can use arrears to relax their short-run financial constraints. We then empirically assess our theoretical prediction, using information from accounting and financial reports of all Italian municipalities for the period 2003-2010. Exploiting the long-lasting effect of 1979 structural reform of Italian local public finance, we employ an instrumental variable approach to face endogeneity concerns. We find robust evidence that tighter fiscal and financial conditions of the local governments determine larger arrears for public investment expenditures.

We present results from an experiment with multiple public goods, where each good produces benefits only if total contributions to it reach a minimum threshold. The presence of multiple public goods makes coordination among participants more difficult, discouraging donor participation and decreasing the likelihood of any public good being effectively funded. Applied to the case of fundraising, the results show how overall donations and the number of effectively funded projects may both decrease as the total number of projects vying for funding increases. The analysis considers whether making one of the contribution options salient, either through its merits or by arbitrarily choosing one to feature during the experiment, helps overcome the increased coordination problem. The results have implications for the growing popularity of crowdfunding websites, and suggest benefits to these sites from helping donors compare and identify the most promising projects.

Using a newly assembled dataset, we empirically investigate the effects of subcontracting on procurement auction prices in Italy. In this setting, the pre-qualifications required for firms aiming to bid on public contracts determine the firms’ different subcontracting formats. We find that fully qualified firms in a position to choose whether to subcontract generally offer lower prices than partially qualified firms, which must proceed with mandatory subcontracts. This result indicates that the firms’ voluntary arrangements tend to improve market performance, while imposed arrangements tend to worsen market performance, in the public procurement supply-chain. (JEL H57, L23, L24, D44)

Adenyl cyclase (AC) plays a pivotal role in cell signaling. The AC system of bivalves has received little attention so far, and our study has been addressed to the characterization of AC properties in the gills of T. philippinarum. The enzyme showed a Km value of 0.77 mM for ATP in the presence of 5 mM Mg2+; in the absence of agonists, it was poorly affected by GTP, while it was stimulated by GTPgammaS and GppNHp up to 14-fold and 4-fold, respectively. Similarly to other invertebrates, the enzyme activity was scarcely stimulated by forskolin. The receptor agonist serotonin (5-HT) significantly stimulated the AC activity, and the pharmacological profile of the 5-HT receptor/s was as follows: (+)butaclamol > dihydroergocryptine > methysergide > prazosin > yohimbine. The AC activity was assessed in vitro in the presence of tributyltin chloride and HgCl2, which reduced the AC activity only at the highest dose tested (10-100 microM). Our data indicate the presence of a membrane-bound AC in gill membranes of T. philippinarum, coupled to Gs proteins and to a specific class of 5-HT receptors. Such receptors show a pharmacological profile slightly different from that reported for 5-HT invertebrate receptors cloned so far.

G proteins are members of a highly conserved superfamily of GTPases, which includes heterotrimeric (alpha, beta, gamma) proteins acting as critical control points for transmembrane signaling. In ectothermal vertebrates, knowledge about these proteins is scarce, and our work provides the first demonstration that G(s), G(q), and G(i) proteins are all present in the liver of a fish. G(q)alpha subunits of about 42 kDa have been identified in European eel (Anguilla anguilla) liver membranes, supporting previous reports about the existence of hormone transduction pathways coupled to inositol 1,4,5-trisphosphate/Ca(2+) enhancement in fish hepatocytes. Although two G(s)alpha proteins of about 45 and 52 kDa have been reported in mammals, a single isoform of approximately 45 kDa has been recognized in eel liver. G(s)alpha and G(q)alpha proteins are involved in the epinephrine transduction pathway, leading to cAMP and Ca(2+) intracellular increments, respectively. Interestingly, both messengers significantly stimulated glucose release from eel hepatocytes but with a different time course. In fact, the Ca(2+)-dependent glucose output preceded the cAMP-mediated release by about 7 min. G(i)alpha subunits of about 40 kDa were also immunodetected, suggesting the presence of hormone receptors leading to adenylyl cyclase inhibition in eel liver; however, alpha(2)- adrenoreceptor ligands were ineffective on both enzyme activity and glucose release.

Purpose: We previously reported effects on heat shock protein 70 (HSP70) mRNA expression, a cytoprotective protein induced under stressful condition, in human trophoblast cells exposed to amplitude-modulated Global System for Mobile Communication (GSM) signals. In the present work the same experimental conditions were applied to the rat PC12 cells, in order to assess the stress responses mediated by HSP70 and by the Mitogen Activated Protein Kinases (MAPK) in neuronal-like cells, an interesting model to study possible effects of mobile phone frequencies exposure. Materials and methods: HSP70 gene expression level was evaluated by reverse transcriptase polymerase chain reaction, HSP70 protein expression and MAPK phosphorylation were assessed by Western blotting. PC12 cells were exposed for 4, 16 or 24 h to 1.8 GHz continuous wave signal (CW, carrier frequency without modulation) or to two different GSM modulation schemes, GSM-217Hz and GSM-Talk (which generates temporal changes between two different GSM signals, active during talking or listening phases, respectively, thus simulating a typical conversation). Specific adsorption rate (SAR) was 2 W/kg. Results: After PC12 cells exposure to the GSM-217Hz signal for 16 or 24 h, HSP70 transcription significantly increased, whereas no effect was observed in cells exposed to the CW or GSM-Talk signals. HSP70 protein expression and three different MAPK signaling pathways were not affected by the exposure to any of the three different 1.8 GHz signals. Conclusion: The positive effect on HSP70 mRNA expression, observed only in cells exposed to the GSM-217Hz signal, is a repeatable response previously reported in human trophoblast cells and now confirmed in PC12 cells. Further investigations towards a possible role of 1.8 GHz signal modulation are therefore advisable.

In this paper we discuss a number of ways to define and measure the affordability of energy consumption, and we examine the emergence of fuel poverty in Italy in the period from 1998 to 2011. The paper examines the eligibility criteria for claiming the benefits available to support energy consumption for vulnerable families and it identifies the potential beneficiaries. The study assesses the appropriateness of the eligibility criteria by comparing the population targeted by the policy with the population actually living in fuel poverty. A simulation exercise, using the hypothetical scenario most likely to result in energy benefits being made available, shows that, regardless of the affordability index adopted, the provision of state energy benefits has little impact on fuel poverty.

In this paper, we discuss a number of ways to define and measure the affordability of energy consumption, and we examine the emergence of energy poverty in Italy in the period from 1998 to 2011. The paper examines the eligibility criteria for claiming the benefits available to support energy consumption for vulnerable families and it identifies the potential beneficiaries. The study assesses the appropriateness of the eligibility criteria by comparing the population targeted by the policy with the population actually facing affordability problems. A simulation exercise, using the hypothetical scenario most likely to result in energy benefits being made available, shows that, regardless of the affordability index adopted, the provision of state energy benefits has little impact on fuel poverty.

The method for settling telecommunications payments between operators in different countries is the reciprocal accounting rate system. This is a discriminatory system, because different operators pay different prices to access the same national network and these price differences are not related to different costs of providing the service. Reforms of the accounting rate system are currently under discussion in international organizations. In this paper we study the effects of the existing regime and of the main alternative proposal, the international traffic terminating fee, on the retail price of international telephone calls. Our main result is that the current regime of reciprocal accounting rates may determine lower prices than the proposed alternative system.

Polynucleotide: adenosine glycosidases (PNAG) are a class of plant and bacterial enzymes commonly known as ribosome-inactivating proteins (RIP). They are presently classified as rRNA N-glycosidases in the enzyme nomenclature [EC 3.2.2.22]. Several activities on nucleic acids, other than depurination, have been attributed to PNAG: in particular modifications induced in circular plasmids, including linearisation and topological changes, and cleavage of guanidinic residues. Here we describe a chromatographic procedure to obtain nuclease-free PNAG by dye-chromatography onto Procion Red derivatized Sepharose((R)). Highly purified enzymes depurinate extensively pBR322 circular, supercoiled DNA at neutral pH and exhibit neither DNase nor DNA glycolyase activities, do not cause topological changes, and adenine is the only base released from DNA and rRNA, even at very high enzyme concentrations. A scanning force microscopy (SFM) study of pBR322 treated with saporin-S6 confirmed that (i) this PNAG binds extensively to the plasmid, (ii) the distribution of the bound saporin-S6 molecules along the DNA chain is markedly variable, (iii) plasmids already digested with saporin-S6 do not appear fragmented or topologically modified. The observations here described demonstrate that polynucleotide:adenosine glycosidase is the sole enzymatic activity of the four ribosome-inactivating proteins gelonin, momordin I, pokeweed antiviral protein from seeds and saporin-S6. These proteins belong to different families, suggesting that the findings here described may be generalized to all PNAG.

Momordin II, a ribosome-inactivating protein from Momordica charantia seeds, was purified by a procedure involving a series of chromatographies on S-Sepharose, Sephadex G-50, CM-Sepharose, and Red Sepharose columns. Highly purified momordin II inhibited cell-free protein synthesis, released adenine from rat liver ribosomes and from DNA, and had no RNase activity.

Protein synthesis in the thermoacidophilic archaeon Sulfolobus solfataricus (Ss) was inhibited by polynucleotide:adenosine glycosylase activity of some type 1 ribosome-inactivating proteins (RIP). The target of RIP was S. solfataricus rRNA that was depurinated thus producing inactive ribosomes. The amount of RIP required to half-inactivated Ss-ribosomes was comparable to that needed for eubacterial ribosomes, but two orders of magnitude higher than that required for mammalian ribosomes. In addition, RIP treated Ss-ribosomes were also less efficient in stimulating the ribosome dependent GTPase activity of the S. solfataricus elongation factor 2 (SsEF-2) thus suggesting that the inhibition of protein synthesis was probably due to the lack of the interaction between depurinated Ss-ribosomes and SsEF-2. Since SsEF-2 protects Ss-ribosomes against RIP activity it can be hypothesised that also on Ss-ribosomes the sites of interaction for the translocation factor 2 and the RIP are topographically close.