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In the pork production chain, the control at slaughterhouse aims to ensure safe food thanks to proper hygienic conditions during all steps of the slaughtering. Salmonella is one of the main foodborne pathogens in the EU causing a great number of human cases, and pigs also contribute to its spreading. Pig is the main reservoir of the zoonotic hepatitis E virus (HEV) that can be present in liver, bile, feces and even rarely in blood and muscle. The aim of this study was to assess the presence of both Salmonella and HEV in several points of the slaughtering chain, including pig trucks. Other viruses hosted in the gut flora of pigs and shed in feces were also assayed (porcine adenovirus PAdV, rotavirus, norovirus, and mammalian orthoreovirus MRV). Torque teno sus virus (TTSuV) present in both feces, liver and blood was also considered. Four Italian pig abattoirs were sampled in 12 critical points, 5 of which were the outer surface of carcasses before processing. HEV and rotavirus (RVA) were not detected. Norovirus was detected once. Salmonella was detected in two of the 4 abattoirs: in the two lairage pens, in the site of evisceration and on one carcass, indicating the presence of Salmonella if carcass is improper handled. The sampling sites positive for Salmonella were also positive for PAdV. MRV was detected in 10 swabs, from only two abattoirs, mainly in outer surface of carcasses. TTSuV was also detected in all abattoirs. Our study has revealed a diverse group of viruses, each serving as indicator of either fecal (NoV, RVA, PAdV, MRV) or blood contamination (TTSuV). TTSuV could be relevant as blood contamination indicators, crucial for viruses with a viremic stage, such as HEV. The simultaneous presence of PAdV with Salmonella is relevant, suggesting PAdV as a promising indicator for fecal contamination for both bacterial and viruses. In conclusion, even in the absence of HEV, the widespread presence of Salmonella at various points in the chain, underscores the need for vigilant monitoring and mitigation strategies which could be achieved by testing not only bacteria indicators as expected by current regulation, but also some viruses (PAdV, TTSuV, MRV) which could represent other sources of fecal contamination., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

Zinc oxide and polylactic-co-glycolic acid (ZnO-PLGA) nanocomposites are known to exhibit different biomedical applications and antibacterial activity, which could be beneficial for adding to wound dressings after different surgeries. However, possible cytotoxic effects along with various unexpected activities could reduce the use of these prominent systems. This is correlated to the property of ZnO, which exhibits different polymeric forms, in particular, wurtzite, zinc-blende, and rocksalt. In this study, we propose a computational approach based on the density functional theory to investigate the properties of ZnO-PLGA systems in detail. First, three different stable polymorphs of ZnO were considered. Subsequently, the abilities of each system to absorb the PLGA copolymer were thoroughly investigated, taking into account the modulation of electrical, optical, and mechanical properties. Significant differences between ZnO and PLGA systems have been found; in this study, we remark on the potential use of these models and the necessity to describe crucial surface aspects that might be challenging to observe with experimental approaches but which can modulate the performance of nanocomposites.

Self-consistent charge density functional tight-binding (DFTB) calculations have been performed to investigate the electrical properties and transport behavior of asymmetric graphene devices (AGDs). Three different nanodevices constructed of different necks of 8 nm, 6 nm and 4 nm, named Graphene-N8, Graphene-N6 and Graphene-N4, respectively, have been proposed. All devices have been tested under two conditions of zero gate voltage and an applied gate voltage of +20 V using a dielectric medium of 3.9 epsilon interposed between the graphene and the metallic gate. As expected, the results of AGD diodes exhibited strong asymmetric I ( V ) characteristic curves in good agreement with the available experimental data. Our predictions implied that Graphene-N4 would achieve great asymmetry ( A ) of 1.40 at | V | = 0.2 V with maximum transmittance ( DS | = 0.2 V with maximum transmittance ( T ) of 6.72 in the energy range 1.30 eV. More importantly, while the A of Graphene-N4 was slightly changed by applying the gate voltage, Graphene-N6/Graphene-N8 showed a significant effect with their A increased from 1.20/1.03 under no gate voltage (NGV) to 1.30/1.16 under gate voltage (WGV) conditions. Our results open up unprecedented numerical prospects for designing tailored geometric diodes., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Hepatitis E virus (HEV) genotype 3 is a prevalent zoonotic pathogen in European pig farms, posing a significant public health risk primarily through the foodborne route. The study aimed to identify effective biosecurity measures for controlling HEV transmission on pig farms, addressing a critical gap in current knowledge. Utilizing a cross-sectional design, fecal samples from gilts, dry sows, and fatteners were collected on 231 pig farms of all farm types across nine European countries. Real-time RT-PCR was employed to test these samples for HEV. Simultaneously, a comprehensive biosecurity questionnaire captured data on various potential measures to control HEV. The dependent variable was HEV risk, categorized as lower or higher based on the percentage of positive pooled fecal samples on each farm (25% cut-off). The data were analyzed using generalized linear models (one for finisher samples and one for all samples) with a logit link function with country and farm type as a priori fixed factors. The results of the final multivariable models identified key biosecurity measures associated with lower HEV risk, which were the use of a hygienogram in the breeding (OR: 0.06, p = 0.001) and/or fattening area after cleaning (OR: 0.21, p = 0.019), the presence of a quarantine area (OR: 0.29, p = 0.025), testing and/or treating purchased feed against Salmonella (OR: 0.35, p = 0.021), the presence of other livestock species on the farm, and having five or fewer persons in charge of the pigs. Contrary to expectations, some biosecurity measures were associated with higher HEV risk, e.g., downtime of 3 days or longer after cleaning in the fattening area (OR: 3.49, p = 0.005) or mandatory handwashing for farm personnel when changing barn sections (OR: 3.4, p = 0.026). This novel study unveils critical insights into biosecurity measures effective in controlling HEV on European pig farms. The identification of both protective and risk-associated measures contributes to improving strategies for managing HEV and underscores the complexity of biosecurity in pig farming., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Dubbert, Meester, Smith, Tobias, Di Bartolo, Johne, Pavoni, Krumova-Valcheva, Sassu, Prigge, Aprea, May, Althof, Ianiro, Żmudzki, Dimitrova, Alborali, D'Angelantonio, Scattolini, Battistelli and Burow.)

The COVID-19 pandemic, once a global crisis, is now largely under control, a testament to the extraordinary global efforts involving vaccination and public health measures. However, the relentless evolution of SARS-CoV-2, leading to the emergence of new variants, continues to underscore the importance of remaining vigilant and adaptable. Monoclonal antibodies (mAbs) have stood out as a powerful and immediate therapeutic response to COVID-19. Despite the success of mAbs, the evolution of SARS-CoV-2 continues to pose challenges and the available antibodies are no longer effective. New variants require the ongoing development of effective antibodies. In the present study, we describe the generation and characterization of neutralizing mAbs against the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein by combining plasmid DNA and recombinant protein vaccination. By integrating genetic immunization for rapid antibody production and the potent immune stimulation enabled by protein vaccination, we produced a rich pool of antibodies, each with unique binding and neutralizing specificities, tested with the ELISA, BLI and FACS assays and the pseudovirus assay, respectively. Here, we present a panel of mAbs effective against the SARS-CoV-2 variants up to Omicron BA.1 and BA.5, with the flexibility to target emerging variants. This approach ensures the preparedness principle is in place to address SARS-CoV-2 actual and future infections.

The BIOPIGEE project (part of the One Health European Joint Programme under Horizon 2020) aimed to identify relevant measures to effectively control Salmonella, and another zoonotic pathogen, hepatitis E virus (HEV) within the pig meat food chain. The aim of this study was to identify biosecurity measures or management practices that are relevant for limiting Salmonella and/or HEV occurrence and spread within pig slaughterhouses. This was with the final goal of compiling a list of biosecurity measures for different processes and operations along the slaughter line with evidence of their effectiveness. To achieve this, a literature review was conducted on studies estimating the effectiveness of measures applied in slaughterhouses to reduce the microbial contamination of pig carcasses. Results of this literature search are discussed and presented in summary tables that could be used as a source of information for the pig slaughter industry to further develop their guidelines on hygienic slaughter., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Salmonella spp. is a common zoonotic pathogen, causing gastrointestinal infections in people. Pigs and pig meat are a major source of infection. Although farm biosecurity is believed to be important for controlling Salmonella transmission, robust evidence is lacking on which measures are most effective. This study enrolled 250 pig farms across nine European countries. From each farm, 20 pooled faecal samples (or similar information) were collected and analysed for Salmonella presence. Based on the proportion of positive results, farms were categorised as at higher or lower Salmonella risk, and associations with variables from a comprehensive questionnaire investigated. Multivariable analysis indicated that farms were less likely to be in the higher-risk category if they had '<400 sows'; used rodent baits close to pig enclosures; isolated stay-behind (sick) pigs; did not answer that the hygiene lock/ anteroom was easy to clean; did not have a full perimeter fence; did apply downtime of at least 3 days between farrowing batches; and had fully slatted flooring in all fattener buildings. A principal components analysis assessed the sources of variation between farms, and correlation between variables. The study results suggest simple control measures that could be prioritised on European pig farms to control Salmonella.

The decommissioned fahlore Cu-Sb(-Ag) mine at Mt. Avanza (Carnic Alps, Italy) is a rare example of exploited ore deposits, as the tetrahedrite (Cu 6 [Cu 4 (Fe,Zn) 2 ]Sb 4 S 13 ) is the main ore mineral found. This multi-compartmental geochemical characterisation approach provides one of the first case studies regarding the geochemical behaviour and fate of Hg, Sb, As, Cu, and other elements in solid and water matrices and of Hg in the atmosphere in an environment affected by the mining activity of a fahlore ore deposit. Elevated concentrations of the elements (Cu, Sb, As, Pb, Zn, Hg) associated with both (Zn-Hg)-tetrahedrite and to other minor ore minerals in mine wastes, soils, and stream sediments were observed. Concentrations in waters and stream sediments greatly decreased with increasing distance from the mining area and the I geo index values testify the highest levels of sediment contamination inside the mine area. Thallium and Ge were associated with the "lithogenic component" and not to sulfosalt/sulphide minerals. Although mine drainage water often slightly exceeded the national regulatory limits for Sb and As, with Sb being more mobile than As, the relatively low dissolved concentrations indicate a moderate stability of the tetrahedrite. The fate of Hg at the investigated fahlore mining district appeared similar to cinnabar mining sites around the world. Weak solubility but the potential evasion of gaseous elemental mercury (GEM) into the atmosphere also appear to be characteristics of Hg in fahlore ores. Although GEM concentrations are such that they do not present a pressing concern, real-time field surveys allowed for the easy identification of Hg sources, proving to be an effective, suitable high-resolution indirect approach for optimising soil sampling surveys and detecting mine wastes and mine adits., (© 2023. The Author(s).)

Foodborne transmission is considered the main way of spreading zoonotic hepatitis E virus (HEV) infection in Europe. In recent years, the human cases of hepatitis E in subjects without history of travel in endemic areas have raised, suggesting that domestic HEV transmission is increasing. Pork products with or without liver, are often indicated as the source of many human foodborne HEV cases as well as small outbreaks. Pigs are recognized as the main reservoir of the zoonotic HEV-3 genotype, the most frequently detected in human cases in the EU. In the absence of a harmonized surveillance of HEV circulation, data on prevalence are heterogeneous but confirm a widespread circulation of HEV-3 in pig herds across EU. HEV-3 can pass through the food chain from farm to fork when infected animals are slaughtered. In Italy, several studies reported the circulation of HEV-3 in pig farms, but results are heterogeneous due to different methodologies applied. In the present study, we performed a survey over 51 pig herds belonging to three main types of farms: breeding, fattening and farrow-to-finish. HEV-RNA was analyzed by broad range Real-time RT-PCR on 20 samples for each farm, obtained by pooling together feces from 10 individuals. Overall, HEV RNA was confirmed on 150 fecal pooled samples out of 1,032 (14.5%). At least one positive pooled sample was detected from 18 farms out of 51 tested (35.3%). By lowering the number of infected pigs at primary production, the risk of HEV-3 entering into the food chain can be reduced. Hence, information on HEV circulation in herds is highly relevant for choosing preventive measures and deserves development of a monitoring program and further investigations., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Ianiro, Pavoni, Aprea, Romantini, Alborali, D'Angelantonio, Garofolo, Scattolini, De Sabato, Magistrali, Burow, Ostanello, Smith and Di Bartolo.)

MoO 3 and MoO 2 systems have attracted particular attention for many widespread applications thanks to their electronic and optical peculiarities; from the crystallographic point of view, MoO 3 adopts a thermodynamically stable orthorhombic phase (α-MoO 3 ) belonging to the space group Pbmn , while MoO 2 assumes a monoclinic arrangement characterized by space group P 2 1 / c. In the present paper, we investigated the electronic and optical properties of both MoO 3 and MoO 2 by using Density Functional Theory calculations, in particular, the Meta Generalized Gradient Approximation (MGGA) SCAN functional together with the PseudoDojo pseudopotential, which were used for the first time to obtain a deeper insight into the nature of different Mo-O bonds in these materials. The calculated density of states, the band gap, and the band structure were confirmed and validated by comparison with already available experimental results, while the optical properties were validated by recording optical spectra. Furthermore, the calculated band-gap energy value for the orthorhombic MoO 3 showed the best match to the experimental value reported in the literature. All these findings suggest that the newly proposed theoretical techniques reproduce the experimental evidence of both MoO 2 and MoO 3 systems with high accuracy.

Metal-insulator-metal (MIM) diodes are very interesting in many different applications exploiting environment-friendly renewable energy solutions. Moreover, since the dimensions of such devices are at the nanoscale, the size and the characteristics of their constitutive elements can drastically influence their macroscale performance. As it could be difficult to describe in detail the physical phenomena occurring among materials in nanoscale systems, in this work first-principles calculations have been used to study the structural and electrical properties of three different hafnium oxide (HfO 2 )-MIM diodes. These devices have been simulated at the atomistic level by interposing 3 nm of HfO 2 between drain and source electrodes made of gold and platinum, respectively. The monoclinic and orthorhombic polymorphs of HfO 2 have been considered to model different types of MIM diodes, and the interface geometries have been optimized to compute the current-voltage characteristics, reflecting the tunneling mechanisms occurring in such devices. The calculation of the transmission pathways has also been carried out to investigate the effects of atomistic coordinates despite the use of the same material. The results demonstrate the role of the Miller indices of metals and the influence of the HfO 2 polymorphs on the MIM properties. In this study, the importance of interface phenomena on the measurable properties of the proposed devices has been investigated in detail., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

In this work, we present the design, atomistic/circuit/electromagnetic simulations, and the experimental results for graphene monolayer/zirconium-doped hafnium oxide (HfZrO) ultra-thin ferroelectric-based field effect transistors fabricated at the wafer scale, regarding the pyroelectricity generation directly from microwave signals, at room temperature and below it, namely at 218 K and at 100 K. The transistors work like energy harvesters, i.e. they collect low-power microwave energy and transform it into DC voltages with a maximum amplitude between 20 and 30 mV. The same devices function as microwave detectors in the band 1-10.4 GHz and at very low input power levels not exceeding 80 μ W when they are biased by using a drain voltage, with average responsivity values in the range 200-400 mV mW -1 ., (© 2023 IOP Publishing Ltd.)

The Gulf of Trieste (Northern Adriatic Sea, Italy) is the coastal area of the Mediterranean Sea most highly contaminated by mercury (Hg) due to fluvial inputs from the Isonzo/Soča River system, draining over 500 years' worth of cinnabar extraction activity from the Idrija mining district (Western Slovenia). The aim of this research is to investigate the concentration of Hg in hair samples taken from the general population of the Friuli Venezia Giulia (FVG) Region coastal area, as a marker of chronic exposure to Hg. Three hundred and one individuals - 119 males and 182 females - were recruited by convenience sampling in Trieste in September 2021. An amount of approximately 100 mg of hair was collected from the occipital scalp of each participant to measure the respective Hg concentrations (expressed as mg/kg). Moreover, participants completed a self-report questionnaire collecting extensive socio-demographic and life-style information. A multiple linear regression analysis was employed to investigate factors associated with increased levels of Hg concentration in hair. A mean Hg concentration in hair of 1.63 mg/kg was found, slightly above the 1.0 mg/kg threshold recommended by the WHO for pregnant women and children, although still well below the no observed adverse effects level (NOAEL) of 10 mg/kg. Among respondents, 55.6% showed a Hg concentration in hair > 1 mg/kg, 22.9% > 2 mg/kg, and 2 participants exhibited Hg levels > 10 mg/kg. The adjusted mean hair Hg level increased in those subjects who reported a preference for shellfish/crayfish/mollusks (RC = 0.35; 95%CI: 0.16; 0.55), whereas it decreased in those who reported a preference for frozen fish (RC = -0.23; 95%CI: - 0.39; - 0.06). Though a risk alert for Hg exposure for coastal residents from FVG is deemed unnecessary at this time, it is recommended that pregnant women limit their ingestion of locally caught fish to < 4 servings/month., (© 2022. The Author(s).)

Reliable point-of-care (POC) rapid tests are crucial to detect infection and contain the spread of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). The emergence of several variants of concern (VOC) can reduce binding affinity to diagnostic antibodies, limiting the efficacy of the currently adopted tests, while showing unaltered or increased affinity for the host receptor, angiotensin converting enzyme 2 (ACE2). We present a graphene field-effect transistor (gFET) biosensor design, which exploits the Spike-ACE2 interaction, the crucial step for SARS-CoV-2 infection. Extensive computational analyses show that a chimeric ACE2-Fragment crystallizable (ACE2-Fc) construct mimics the native receptor dimeric conformation. ACE2-Fc functionalized gFET allows in vitro detection of the trimeric Spike protein, outperforming functionalization with a diagnostic antibody or with the soluble ACE2 portion, resulting in a sensitivity of 20 pg/mL. Our miniaturized POC biosensor successfully detects B.1.610 (pre-VOC), Alpha, Beta, Gamma, Delta, Omicron ( i.e. , BA.1, BA.2, BA.4, BA.5, BA.2.75 and BQ.1) variants in isolated viruses and patient's clinical nasopharyngeal swabs. The biosensor reached a Limit Of Detection (LOD) of 65 cps/mL in swab specimens of Omicron BA.5. Our approach paves the way for a new and reusable class of highly sensitive, rapid and variant-robust SARS-CoV-2 detection systems., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. The ACE2-Fc functionalization of gFET and the design of the POC device are under patent pending, applied by Polytechnic University of Marche. D.DM., M.DA., C.A., A.R., I.DA., D.M., E.P., P.C., G.B., L.P., M.F. are the inventors of the patent application N. 102021000000533 filed in 01/13/2021. All other authors declare they have no competing interests., (© 2022 The Authors. Published by Elsevier Ltd.)

The increasing use of graphene-related materials (GRMs) in everyday-life products raises concerns for their possible release into the environment and consequent impact on organisms. GRMs have widely varying effects on plants and, according to recent evidences, graphene oxide (GO) has the potential to interfere with the sexual reproduction owing to its acidic properties and production residues. Here, stigmas of the model plant Cucurbita pepo (summer squash) were subjected to simulated dry depositions of GO and GO purified from production residues (PGO). Stigmas were then hand-pollinated and GRM deposition was checked by ESEM and confocal microscopy. Analysis of stigma integrity, pH homeostasis and pollen-stigma interactions did not reveal negative effects. Fruit and seed production were not affected, but GO depositions of 22.1 ± 7.2 ng mm -2 affected the normal development of seeds, decreasing seed dimensions, seed germination and germination speed. The elemental analysis revealed that GO has significant quantities of production residues, such as strong acids and oxidants, while PGO has only traces, which justifies the differences observed in the effects caused by the two materials. Our results show that GO depositions of up to 11.1 ± 3.6 ng mm -2 , which fall within the variation range of total dry particulate matter depositions reported in the literature, are safe for reproduction of C. pepo. This is the first "safety" limit ever recorded for depositions of "out-of-the-box" GO concerning the reproduction of a seed plant. If confirmed for wind-pollinated species, it might be considered for policymaking of GRMs emissions in the air., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Scanning microwave microscopy (SMM) is a novel metrological tool that advances the quantitative, nanometric, high-frequency, electrical characterization of a broad range of materials of technological importance. In this work, we report an inverted near-field scanning microwave microscopy (iSMM) investigation of a graphene oxide-based epoxy nanocomposite material at a nanoscopic level. The high-resolution spatial mapping of local conductance provides a quantitative analysis of the sample's electrical properties. In particular, the electrical conductivity in the order of ∼10-1 S/m as well as the mapping of the dielectric constant with a value of ∼4.7 ± 0.2 are reported and validated by the full-wave electromagnetic modeling of the tip-sample interaction.

HfO 2 can assume different crystalline structures, such as monoclinic, orthorhombic, and cubic polymorphs, each one characterized by unical properties. The peculiarities of this material are also strongly related to the presence of doping elements in the unit cell. Thus, the present paper has the main purpose of studying and comparing twelve different systems characterized by diverse polymorphs and doping percentages. In particular, three different crystalline structures were considered: the monoclinic P2 1 /c , the orthorhombic Pca2 1 , and the cubic Fm3¯m phases of HfO 2 . Each one has been studied by using Y as a doping agent with three different contents: 0% Y:HfO 2 , 8% Y:HfO 2 , 12% Y:HfO 2 , and 16% Y:HfO 2 . For all the systems, density functional theory (DFT) methods based on PBE/GGA, and on the HSE hybrid functionals were used to optimize the geometry as well as to study their optical properties. Depending on the polymorphs, Y affects the formation energy in different ways and causes changes in the optical properties. When the percentage of Y did not exceed 12%, a stabilization of the cubic phase fraction and an increase of the dielectric constant was observed. Additionally, the calculated optical bandgap energies and the refractive index are examined to provide an overview of the systems and are compared with experimental data. The bandgaps obtained are in perfect agreement with the experimental values and show a slight increase as the doping percentage grows, while only minor differences are found between the three polymorphs in terms of both refractive index and optical band gap. The adopted first principles study generates a reasonable prediction of the physical-chemical properties of all the systems, thus identifying the effects of doping phenomena., Competing Interests: The authors declare no conflict of interest.

Health and safety of occupations entailing extensive skin contact with cinnabar-enriched sand in beaches of Friuli-Venezia Giulia (FVG) Region (North-eastern Italy) have been questioned for possible skin absorption of mercury (Hg). One hundred mg hair was collected from the occipital scalp of 50 male workers of Grado beach and 121 males from FVG general population. Factors associated with hair Hg content were investigated by multivariable logistic (considering Hg levels >1 vs ≤1 mg/kg) and log-transformed linear regression. The median hair concentration of Hg in male beach workers was 0.70 (IQR = 0.42; 1.34) mg/kg, lower than FVG general population's [1.29 (IQR = 0.87-2.06) mg/kg (p < 0.001)]. In both regression models the hair Hg increased with fish consumption, both among beach workers of Grado and FVG general population. The mean Hg levels in beach workers of Grado fell within an acceptable range, not requiring restrictions of their occupational activities., Competing Interests: Declaration of competing interest None to declare., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

A VO 2 (B) polymorph has been thoroughly investigated by means of density functional theory (DFT) calculations to evaluate the structure, Raman spectrum, cohesive energy, phonon band structure, an delectronic and optical properties. Among the computed Raman modes, eight of them have been assigned to the VO 2 (B) structure in full agreement with the corresponding experimental spectra. The minimized structure of the VO 2 (B) polymorph indicated the presence of negative frequencies in its phonon dispersion curves, confirming the dynamic instability of this material. Herein, the combination of generalized gradient approximation (GGA)/PBEsol with a hybrid HSE functional has been employed to perform ab initio calculations on VO 2 (B), since the conventional semi-local DFT calculations are believed to underestimate the band gap of materials. By considering the electronic structure calculations, for the first time, we found that the calibration of the PBEsol functional can efficiently model the metallic-like properties of VO 2 (B) with a band gap of 0.26 eV, while the corresponding electronic bandgap of VO 2 (B) based on the HSE functional possesses a larger band gap of 0.67 eV. The prediction of optical characteristics of VO 2 (B) indicated that the optical conductivity of VO 2 (B) lies in the infrared region of light. This work strongly suggests the combination of GGA/PBEsol with HSE hybrid functionals to carefully describe the physical properties of smart materials exploitable in electronics and optoelectronics applications. The nanostructure of VO 2 (B) looks promising for IR photodetectors and smart windows applications as a semiconductor material with excellent optical features. It is predicted that in the future VO 2 (B) will continue to expand the envelope of its capabilities because of its remarkable properties., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

We have studied the structural, electronic, magnetic, and optical properties of the VO 2 (B) polymorph using first-principles calculations based on density functional theory (DFT). This polymorph was found to display four optimized structures namely VO 2 (B) PP , VO 2 (B) LP , VO 2 (B) PPD , and VO 2 (B) LPD using the generalized gradient approximation (GGA) PBE exchange-correlation functional by including/excluding van der Waals interaction. Our derivation provides a theoretical justification for adding an on-site Coulomb U value in the conventional DFT calculations to allow a direct comparison of the two methods. We predicted a zero bandgap of the VO 2 (B) structure based on GGA/PBE. However, by GGA/PBE + U, we found accurate bandgap values of 0.76, 0.66, and 0.70 eV for VO 2 (B) PP , VO 2 (B) LP , and VO 2 (B) PPD , respectively. The results obtained from DFT + U were accompanied by a structural transition from the metallic to semiconductor property. Here, we verified the non-magnetic characteristic of the monoclinic VO 2 (B) phase with some available experimental and theoretical data. However, the debate on the magnetic property of this polymorph remains unresolved. Imaginary and real parts of the dielectric function, as computed with the GGA/PBE functional and the GGA/PBE + U functional, were also reported. The first absorption peaks of all considered geometries in the imaginary part of the dielectric constants indicated that the VO 2 (B) structure could perfectly absorb infrared light. The computed static dielectric constants with positive values, as derived from the optical properties, confirmed the conductivity of this material. Among the four proposed geometries of VO 2 (B) in this study, the outcomes obtained by VO 2 (B) PPD reveal good results owing to the excellent consistency of its bandgap, magnetic and optical properties with other experimental and theoretical observations. The theoretical framework in our study will provide useful insight for future practical applications of the VO 2 (B) polymorph in electronics and optoelectronics., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Products containing graphene-related materials (GRMs) are becoming increasingly common, allowing GRM nanoparticles (NPs) to enter the environment during their life cycle. Thanks to their lightness and bidimensional geometry, GRM NPs can be easily dispersed in the air and travel very long distances. The flowers of wind-pollinated plants may be exposed to airborne GRMs, being apt to intercept pollen from the air and, inevitably, other airborne particles. Here, stigmas of four wind-pollinated plants (Corylus avellana, common hazel; Juglans regia, walnut; Quercus ilex, holm oak; Zea mays, maize) were exposed to airborne graphene oxide (GO) and GO purified from production residues (PGO) at a concentration of 3.7 ng m -3 . Subsequently, the stigmas were pollinated and the adhesion of GOs and their effects on stigma integrity and pollen-stigma interaction were examined. The effect of GO NPs in presence of liquid water on the stigma of C. avellana was also investigated. GOs NPs were intercepted by all species, but their effect varied among them. GO reduced pollen adhesion in J. regia and Q. ilex, whereas pollen germination was unaffected in all four species. The presence of a film of water neither completely removed GO NPs from the stigma, nor it enhanced the toxic effect of GO acidity. PGO never affected pollen-stigma interaction, indicating that the phytotoxic substances used for the production of GO, still in traces in commercial GO, are the main cause of GO toxicity. These results reconfirm the need to verify GRMs effects also on key biological processes beside single model organisms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

HfO 2 shows different polymorphs, including monoclinic and orthorhombic ones, that exhibit singular properties. Moreover, the character of HfO 2 is also influenced by the Zr atoms as a doping agent. Here, an extensive study of the monoclinic P2 1 /c and the orthorhombic Pca2 1 polymorphs of HfO 2 , Hf 0.75 Zr 0.25 O 2 , and Hf 0.5 Zr 0.5 O 2 is reported. For all six systems, density functional theory (DFT) methods based on generalized gradient approximations (GGAs) were first used; then the GGA + U method was settled and calibrated to describe the electrical and optical properties of polymorphs and the responses to the oxygen vacancies. Zr had different effects in relation to the polymorph; moreover, the amount of Zr led to important differences in the optical properties of the Pca2 1 polymorph. Finally, oxygen vacancies were investigated, showing an important modulation of the properties of Hf x Zr y O 2 nanostructures. The combined GGA and GGA + U methods adopted in this work generate a reasonable prediction of the physicochemical properties of o- and m-Hf x Zr y O 2 , identifying the effects of doping phenomena.

Breakthrough SARS-CoV-2 infections in fully vaccinated individuals are considered a consequence of waning immunity. Serum antibodies represent the most measurable outcome of vaccine-induced B cell memory. When antibodies decline, memory B cells are expected to persist and perform their function, preventing clinical disease. We investigated whether BNT162b2 mRNA vaccine induces durable and functional B cell memory in vivo against SARS-CoV-2 3, 6, and 9 months after the second dose in a cohort of health care workers (HCWs). While we observed physiological decline of SARS-CoV-2-specific antibodies, memory B cells persist and increase until 9 months after immunization. HCWs with breakthrough infections had no signs of waning immunity. In 3-4 days, memory B cells responded to SARS-CoV-2 infection by producing high levels of specific antibodies in the serum and anti-Spike IgA in the saliva. Antibodies to the viral nucleoprotein were produced with the slow kinetics typical of the response to a novel antigen., Competing Interests: Declaration of interests All authors declare no competing interests., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

To observe the prevalence of contamination by hepatitis A virus (HAV) and norovirus (NoV) in different food types, 9242 samples were analyzed over a 6-year period (January 2014-December 2019). Samples were from routine official activities by Competent Authorities (CAs) and Food Business Operators, according to Hazard Analysis and Critical Control Points plans. Analyses were performed in accordance with European and Italian regulations. Food types were obtained from different production areas of Italy, and ranged from mollusks, ready-to-eat (RTE) and packaged vegetables, frozen berries, tap water, fruit and RTE fruit salads, and processed and preserved foods. No risk management plans were set by the authors' laboratory, because they were still adopted by conferring customers. Analyses were conducted according to ISO/TS 15216-2:2013 (ISO in Part 2: Method for Qualitative Detection. International Organization for Standardization, Geneva, 2013). The data showed that 2.25% (95% CI: 2.0-2.6) of samples were contaminated by at least one virus type, and that the most detected pathogen was NoV GII (89.50% of all positives). Mollusks (filter-feeding animals) were the most contaminated category (92.31% of all positives) not only by NoV or HAV individually, but also by multiple HAV/NoV contaminations consisting of 22.59% of all positives. For NoV, there was a significant correlation between shellfish positivity and season, with the autumn-winter period being the most associated with risk. Conversely, berries, drinking water and RTE vegetables, previously linked to several outbreaks, showed a low rate of contamination. These results from data collection have implications for the improvement of sampling plans for HAV and NoV by Italian CAs, and by food-producing and distribution operators. Moreover, these findings obtained by a standardized qualitative method contribute the collection of data aimed at establishing new microbiological criteria not yet foreseen (but advocated) by current European rules., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Torque teno sus virus (TTSuV) is a non-enveloped circular ssDNA virus which frequently infects swine and has been associated with hepatic, respiratory, and autoimmune disorders. TTSuV's pathogenic role is still uncertain, and clear data in the literature on virus reservoirs are lacking. The aims of this study were to investigate the presence of potentially zoonotic TTSuV in wild animals in Northern Italy and to evaluate their role as reservoirs. Liver samples were collected between 2016 and 2020 during four hunting seasons from wild boars ( Sus scrofa ), red deer ( Cervus elaphus ), roe deer ( Capreolus capreolus ), and chamois ( Rupicapra rupicapra ). Samples originated from areas in Northern Italy characterized by different traits, i.e., mountains and flatland with, respectively low and high farm density and anthropization. Viral identification was carried out by end-point PCR with specific primers for TTSuV1a and TTSuVk2a species. TTSuV prevalence in wild boars was higher in the mountains than in the flatland (prevalence of 6.2% and 2.3%, respectively). In wild ruminants only TTSuVk2a was detected (with a prevalence of 9.4%). Our findings shed light on the occurrence and distribution of TTSuV in some wild animal species, investigating their possible role as reservoirs.

The COVID-19 pandemic caused by SARS-CoV-2 has made the development of safe and effective vaccines a critical priority. To date, four vaccines have been approved by European and American authorities for preventing COVID-19, but the development of additional vaccine platforms with improved supply and logistics profiles remains a pressing need. Here we report the preclinical evaluation of a novel COVID-19 vaccine candidate based on the electroporation of engineered, synthetic cDNA encoding a viral antigen in the skeletal muscle. We constructed a set of prototype DNA vaccines expressing various forms of the SARS-CoV-2 spike (S) protein and assessed their immunogenicity in animal models. Among them, COVID-eVax-a DNA plasmid encoding a secreted monomeric form of SARS-CoV-2 S protein receptor-binding domain (RBD)-induced the most potent anti-SARS-CoV-2 neutralizing antibody responses (including against the current most common variants of concern) and a robust T cell response. Upon challenge with SARS-CoV-2, immunized K18-hACE2 transgenic mice showed reduced weight loss, improved pulmonary function, and lower viral replication in the lungs and brain. COVID-eVax conferred significant protection to ferrets upon SARS-CoV-2 challenge. In summary, this study identifies COVID-eVax as an ideal COVID-19 vaccine candidate suitable for clinical development. Accordingly, a combined phase I-II trial has recently started., Competing Interests: Declaration of interests A.C. and M.M. are Evvivax employees. E.M., F.P., G.R., A.M., L.L., L.L., E. Salvatori, M. Cappalletti, F.F.F., E.D., V.C., and L.A. are Takis employees. G. Caselli and L.R. are Rottapharm Biotech employees. Takis and Rottapharm Biotech are jointly developing COVID-eVax. M.I. participates in advisory boards/consultancies for or receives funding from Gilead Sciences, Roche, Third Rock Ventures, Amgen, Allovir, Asher Bio. L.G.G is a member of the board of directors at Genenta Science and Epsilon Bio and participates in advisory boards/consultancies for Gilead Sciences, Roche, and Arbutus Biopharma., (Copyright © 2021 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.)

The skin constitutes a protective barrier to external physical and chemical aggressions. Although it is constantly exposed to various xenobiotics, it is generally considered poorly permeable to them, as for example metal ions, becoming unfortunately an entry route of such substances. Metals may penetrate inside the skin inducing more or less local effects such as skin sensitization and potential metals diffusion into the bloodstream. The objective of the study was to investigate the percutaneous penetration of metals in vitro - ex vivo in Franz cell with intact as well damaged skin applying a road dust powder. Moreover, porcine and human skins were compared. This study demonstrated that, after the application of a road dust powder on the skin, metals can penetrate and permeate this cutaneous membrane. From this experimental analysis, in intact skin lead (Pb) achieved the highest skin absorption in both human and porcine skin, while skin absorption profile of cobalt (Co) was the lowest in human skin than the one in porcine model. The concentrations of Ni present in receiving solution were higher compared to other metals in all experiments performed. The present work, definitely shows that metals permeation through damaged skin is accelerated than intact skin, as a result of the weaker cutaneous barrier function. According to published data, pig skin appeared as a suitable model for human skin. Our results confirmed that skin absorption of metals can be relevant in environmental exposures., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Children are potentially exposed to products that contain nanoparticles (NPs). In particular, silver NPs are commonly present both in products used by and around children, primarily due to their antibacterial properties. However, very few data are available regarding the ability of silver NPs to penetrate through the oral mucosa in children. In the present work, we used baby porcine buccal mucosa mounted on vertical Franz diffusion cells, as an in vitro model to investigate penetration of silver NPs (19 ± 5 nm). Permeability experiments were performed using pristine physiologically-relevant saline solution in the receiver chamber and known concentrations of NPs or ions in the donor chamber; conditions mimicked the in vivo physiological pH conditions. After physicochemical characterization of silver nanoparticles dispersed in physiological solution, we evaluated the passage of ions and NPs through the mucosa, using single particle inductively coupled plasma mass spectrometry. A flux of 4.1 ± 1.7 ng cm -2  min -1 and a lag time of 159 ± 17 min were observed through mucosa exposed to silver nanoparticles. The latter suggests nanoparticle penetration through the baby porcine mucosa and release Ag + ions in the receptor fluid, as confirmed by computational model. Due to physiological similarity between human and pig membranes it is reasonable to assume that a trans-oral mucosa penetration could occur in children upon contact with silver nanoparticles.

Mercury (Hg) contamination in the Gulf of Trieste (northern Adriatic Sea) due to mining activity in Idrija (Slovenia) still represents an issue of environmental concern. The Isonzo/Soča River's freshwater inputs have been identified as the main source of Hg into the Gulf, especially following periods of medium-high discharge. This research aims to evaluate the occurrence and distribution of dissolved (DHg) and particulate (PHg) Hg along the water column in the northernmost sector of the Gulf, a shallow and sheltered embayment suitable for the accumulation of fine sediments. Sediment and water samples were collected under unperturbed and perturbed environmental conditions induced by natural and anthropogenic factors. Mercury in the sediments (0.77-6.39 µg g -1 ) and its relationship to grain size were found to be consistent with previous research focused on the entire Gulf, testifying to the common origin of the sediment. Results showed a notable variability of DHg (-1 ) and PHg (0.39-12.5 ng L -1 ) depending on the interaction between riverine and marine hydrological conditions. Mercury was found to be mainly partitioned in the suspended particles, especially following periods of high discharge, thus confirming the crucial role of the river inputs in regulating PHg distribution in the Gulf.

Specific memory B cells and antibodies are a reliable read-out of vaccine efficacy. We analysed these biomarkers after one and two doses of BNT162b2 vaccine. The second dose significantly increases the level of highly specific memory B cells and antibodies. Two months after the second dose, specific antibody levels decline, but highly specific memory B cells continue to increase, thus predicting a sustained protection from COVID-19. We show that although mucosal IgA is not induced by the vaccination, memory B cells migrate in response to inflammation and secrete IgA at mucosal sites. We show that the first vaccine dose may lead to an insufficient number of highly specific memory B cells and low concentration of serum antibodies, thus leaving vaccinees without the immune robustness needed to ensure viral elimination and herd immunity. We also clarify that the reduction of serum antibodies does not diminish the force and duration of the immune protection induced by vaccination. The vaccine does not induce sterilizing immunity. Infection after vaccination may be caused by the lack of local preventive immunity because of the absence of mucosal IgA.

Electrochemical microscopy techniques have extended the understanding of surface chemistry to the micrometer and even sub-micrometer level. However, fundamental questions related to charge transport at the solid-electrolyte interface, such as catalytic reactions or operation of individual ion channels, require improved spatial resolutions down to the nanoscale. A prerequisite for single-molecule electrochemical sensitivity is the reliable detection of a few electrons per second, that is, currents in the atto-Ampere (10 -18 A) range, 1000 times below today's electrochemical microscopes. This work reports local cyclic voltammetry (CV) measurements at the solid-liquid interface on ferrocene self-assembled monolayer (SAM) with sub-atto-Ampere sensitivity and simultaneous spatial resolution < 80 nm. Such sensitivity is obtained through measurements of the charging of the local faradaic interface capacitance at GHz frequencies. Nanometer-scale details of different molecular organizations with a 19% packing density difference are resolved, with an extremely small dispersion of the molecular electrical properties. This is predicted previously based on weak electrostatic interactions between neighboring redox molecules in a SAM configuration. These results open new perspectives for nano-electrochemistry like the study of quantum mechanical resonance in complex molecules and a wide range of applications from electrochemical catalysis to biophysics., (© 2021 The Authors. Small published by Wiley-VCH GmbH.)

The sediments in the Gulf of Trieste (northern Adriatic Sea, Italy) are contaminated by mercury (Hg) due to historic mining which took place in Idrija (Slovenia). Despite many studies having been done regarding Hg, no information is available on the potential impact of dredging required along the main channel approaching the Port of Monfalcone. Sixteen surface sediment samples were collected along the channel to determine both total Hg concentration and chemical species using the thermo-desorption (TD) technique. Six samples were also chosen to apply a selective sequential extraction (SSE). The TD technique showed the maximum Hg release approximately at 260 and 335°C, corresponding to metacinnabar (β-HgS) and cinnabar (α-HgS), respectively. The SSE demonstrated that Hg was mainly associated with poorly soluble or insoluble compounds (98.7%). A resuspension event over a limited period of time can be considered of negligible impact to the water column due to the scarce Hg mobility from sediments.

Hepatitis E virus (HEV) is the causative agent of hepatitis E, an emerging public health infection which has an increasing incidence across Europe. Because of the apparent lack of species barriers, HEV was characterized as a zoonotic agent. Swine are recognized as the main reservoir, but HEV is also found in wild animals such as ungulates, lagomorphs, and bats. Our work aimed at detecting the HEV presence in wild fauna in two hunting areas of Northern Italy (Parma and Sondrio areas) with different environmental and anthropic characteristics to investigate its possible role as reservoir. Liver samples were collected from wild boars, red deer, roe deer and chamois, and viral identification was carried out by One-Step RT Real-time PCR. Positive samples were genotyped, and phylogenetic analysis was performed. The virus was found only in the wild boar population, with different prevalence and subtypes in the two areas (14% HEV3a and 1.2% close to HEV3f in Parma and Sondrio, respectively). Wild ruminants seem otherwise to pose a marginal risk. Given the high pig farm density in the Parma area, and expansion of the wild boar population, continuous monitoring of the strains circulating in wildlife is crucial.

Mammalian orthoreoviruses (MRVs) are emerging infectious agents that may affect wild animals. MRVs are usually associated with asymptomatic or mild respiratory and enteric infections. However, severe clinical manifestations have been occasionally reported in human and animal hosts. An insight into their circulation is essential to minimize the risk of diffusion to farmed animals and possibly to humans. The aim of this study was to investigate the presence of likely zoonotic MRVs in wild ungulates. Liver samples were collected from wild boar, red deer, roe deer, and chamois. Samples originated from two areas (Sondrio and Parma provinces) in Northern Italy with different environmental characteristics. MRV detection was carried out by PCR; confirmation by sequencing and typing for MRV type 3, which has been frequently associated with disease in pigs, were carried out for positive samples. MRV prevalence was as high as 45.3% in wild boars and 40.6% in red deer in the Sondrio area, with lower prevalence in the Parma area (15.4% in wild boars). Our findings shed light on MRV occurrence and distribution in some wild species and posed the issue of their possible role as reservoir.

Heparanase is a validated target in cancer therapy and a potential target for several inflammatory pathologies. A ligand-based virtual screening of commercial libraries was performed to expand the chemical space of small-molecule inhibitors. The screening was based on similarity with known inhibitors and was performed in several runs, starting from literature compounds and progressing through newly discovered inhibitors. Among the fifty-five tested compounds, nineteen had IC 50 values lower than 5 µM and some showed remarkable potencies. Importantly, tere- and isophthalamides derivatives belong to new structural classes of heparanase inhibitors and some of them showed enzyme affinities ( 61 and 63 , IC 50 = 0.32 and 0.12 µM, respectively) similar to those of the most potent small-molecule inhibitors reported so far. Docking studies provided a comprehensive binding hypothesis shared by compounds with significant structural diversity. The most potent inhibitors reduced cell invasiveness and inhibited the expression of proangiogenic factors in tumour cell lines.

Estuarine sediments must be dredged to allow for navigation, and where these sediments are placed after dredging depends upon guidelines based only on the total concentration of contaminants. However, resuspension events could seriously affect the mobility and speciation of contaminants, including potentially toxic trace elements stored in sediments. The effects of resuspension on the cycling of mercury (Hg) and arsenic (As) between the sediment and water column was investigated in a mesocosm study. Four experiments were conducted in three estuaries in northern Spain based on samples collected from sites which have been impacted by decommissioned Hg and As mines and periodically subjected to dredging activities. Designed to mimic the resuspension of particles, each of the experiments revealed that the release of Hg and As species does not only depend on the total concentration in the sediments (16.3-50.9 mg kg -1 , for As and 0.52-5.01 mg kg -1 for Hg). The contribution from porewaters and the subsequent reductive dissolution and/or desorption appear to be the main processes responsible for the abrupt increase in dissolved Hg and As species (maximum release of 427% and 125%, respectively). In some cases, As and Hg continued to remain at high concentrations in the water column even after the experiments were completed, thus testifying to their critical persistence in the dissolved form. Conversely, at the other sites, the restoration of pre-resuspension conditions was observed only a few hours after resuspension, mainly due to the role of Fe oxy-hydroxides which provides suitable surfaces for adsorption and/or co-precipitation involving dissolved Hg (maximum removal of -58%) and As (maximum removal of -25%) species. The results of this research could be helpful to take appropriate decisions regarding dredging especially at the Nalòn estuary, where the release of dissolved As(V) and MeHg appeared to be favoured by sediment resuspension., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Estuaries are transitional water systems where the hydrodynamic processes governing water circulation actively influence suspended particle transport and deposition. In the estuarine mixing zone, the strong physico-chemical gradients resulting from the interaction between river freshwater and seawater may affect the distribution, mobility and fate of several potentially toxic compounds, among which trace elements are of major concern. Knowledge regarding the partitioning behaviour of trace elements would provide essential scientific support for the environmental management of estuaries. In this study, trace element occurrence and phase partitioning among suspended particulate matter, colloidal material and the truly dissolved fraction were investigated in the main Italian and Slovenian estuarine environments of the Gulf of Trieste (northern Adriatic Sea). Further information about the water quality at the river mouths was provided and, in addition to the traditional evaluation of single chemical parameters, a multi-way principal component analysis was employed in order to depict disparities among sampling sites, water layers and seasonal conditions with the final aim of evaluating trace element phase partitioning. Results indicated that the suspended particulate matter acts as the main effective vehicle for Cu, Cr, Fe, Ni and Pb, and enhanced adsorption processes resulted in elevated partitioning coefficients, especially for Fe and Pb. Although disparities occurred between sampling sites and seasons, trace elements showing affinity for the solid phase appeared to be partially bound to the colloidal material. Conversely, As and Cs prevailed in the truly dissolved fraction, especially in seawater and showed scarce affinity for both the suspended particles and colloids., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Hepatitis E virus (HEV) is a singlestrand RNA virus that causes an acute viral hepatitis in humans. Among its eight recognized genotypes, HEV-3 and HEV-4 are zoonotic, infecting humans, pigs and wild boars. Recently, HEV-3 has been also detected in red deer, which represents another reservoir of HEV. Consumption of raw pork products (mainly liver sausages), undercooked wild boar meat, raw wild boar liver and deer meat has been responsible for foodborne HEV human worldwide. From November 2018 to March 2019, liver samples collected from 97 wild boars hunted in Emilia-Romagna region (Northern Italy) were tested for HEV RNA. The hunting area included two territories for an extension of 33 km 2 , named A (about 13 km 2 ,natural park, deciduous wood) and B (about 20 km 2 , cultivated fields in proximity of a river) areas. Distance between the two areas ranged between 8 to 10 km. A total of 73 wild boars were hunted in area A, and 24 in area B. HEV RNA was detected by Real-time RT- PCR in 23/73 liver samples of wild boars living in area A only (31.5% - 95% CI: 22.0-42.8%). The HEV sequences (n=13) clustered within genotype 3. The majority of positives belonged to animals < 12 months (12/25; 48%), followed by subadults (13-24 months) (7/16; 43.8%) and adults (4/32; 12.5%). This difference was found to be statistically significant (p=0.0024). In absence of pig farms, the restriction of HEV-positive animals to a well-defined territory of 13 km 2 (Boschi di Carrega Regional Park) could hypothetically be related to the presence of red deer ( Cervus elaphus ), which lived in area A at the beginning of the hunting season. Further studies are needed to confirm or deny our hypothesis., Competing Interests: Conflict of interests: The authors declare no potential conflict of interests., (©Copyright: the Author(s).)

A series of molecules in which 9H-carbazole (electron donor, D) and 2,2':6',2''-terpyridine (electron acceptor, A) are connected through rigid π-conjugated bridges (D-π-A systems) have been synthesized and their photophysical properties examined in detail, with the support of DFT calculations. The bridges are made of different sequences of ethynylene, phenylene, and anthracene groups. The synthetic strategies involve condensation of 2-acetylpyridine with the aromatic aldehyde moiety on different functionalized π-conjugated bridges and couplings with carbazole derivatives. The system incorporating anthracene in the bridge shows the typical absorption and emission fingerprints of this polycyclic hydrocarbon. The other systems have HOMOs and LUMOs centred, respectively, over the carbazole and the bridge and exhibit solvatochromic charge-transfer (CT) luminescence with high photoluminescence yield up to 70 %, except when an ethynylene unit is directly attached to the carbazole ring, due to a trans-bent non-emissive π-σ* excited state., (© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Scanning microwave microscopy (SMM) is based on the interaction between a sample and an electromagnetic evanescent field, in the microwave frequency range. SMM is usually coupled with a scanning probe microscopy (SPM) technique such as in our case, a scanning tunneling microscope (STM). In this way, the STM tip is used to control the distance between the probe and the sample while acting as an antenna for the microwave field. Thanks to the peculiarity of our home-made setup, the SMM is a suitable method to study blisters formed on HOPG surface as consequence of an electrochemical treatment. Our system has a "broad-band" approach that opens the way to perform local microwave spectroscopy over a broad frequency range. Moreover, microwaves have the ability to penetrate into the sample allowing the sub-surface characterization of materials. The application of the SMM to characterize blisters formed on the HOPG surface provides information on the sub-layer structures., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Berries have repeatedly been associated with outbreaks of hepatitis A virus (HAV) infection. The fruits are usually minimally processed in the food industry due to their delicate nature. While washing treatments partially remove enteric viruses, the commonly used chemical additives produce toxic by-products. A valid alternative to preserve the food safety of these products could be the use of essential oils (EOs). EOs exert antimicrobial activity and do not interfere with the nutritional characteristics of food products. We investigated the efficacy of four essential oils, lemon (Citrus limon), sweet orange (Citrus sinensis), grapefruit (Citrus paradisi), and rosemary cineole (Rosmarinus officinalis chemotype 1.8 cineole) in reducing viral loads of HAV in soft fruits. Mixed fruit berries were inoculated with 10 6.74 TCID 50 /ml of HAV, and treated with four different EOs (0.5% lemon, 0.1% sweet orange and grapefruit, and 0.05% rosemary) for 1 h at room temperature. Virus infectivity was then assessed by titration assays for its ability to grow on cell cultures. A statistically significant reduction in HAV titer on the fruit surface was observed after treating the berries with EOs of lemon (2.84 log TCID 50 /ml), grapefruit (2.89 log TCID 50 /ml), and rosemary cineole (2.94 log TCID 50 /ml). Rosemary cineole was the most effective EO in reducing viral titer on berries, followed by grapefruit EO. These results improve our knowledge about the antiviral activity of these EOs and highlight their potential use in fresh produce sanitation.

Heparanase is the only mammalian endo-β-d-glucuronidase involved in a variety of major diseases. The up-regulation of heparanase expression increases tumor size, angiogenesis, and metastasis, representing a validated target in the anti-cancer field. To date, only a few small-molecule inhibitors have been described, but none have gotten through pre-clinical development. Previously, we explored 2-(4-(4-(bromo-methoxybenzamido)benzylamino)phenyl) benzazole derivatives as anti-heparanase agents, proposing this scaffold for development of broadly effective heparanase inhibitors. Herein, we report an extended investigation of new symmetrical 2-aminophenyl-benzazolyl-5-acetate derivatives, proving that symmetrical compounds are more effective than asymmetrical analogues, with the most-potent compound, 7g, being active at nanomolar concentration against heparanase. Molecular docking studies were performed on the best-acting compounds 5c and 7g to rationalize their interaction with the enzyme. Moreover, invasion assay confirmed the anti-metastatic potential of compounds 5c, 7a, and 7g, proving the inhibition of the expression of proangiogenic factors in tumor cells.

We have fabricated and electrically characterized at the wafer scale tens of metal-ferroelectric (HfZrO)-semiconductor capacitors and metal-graphene monolayer-ferroelectric (HfZrO)-semiconductor capacitors with the same top electrode dimensions. We have found that the memory windows of the capacitors containing graphene are 3-4 times larger than the ferroelectric capacitors without graphene, and increase even more after annealing. This physical effect can be attributed to the additional electric field exerted by the graphene monolayer on the HfZrO ferroelectric semiconductor capacitor, and to the negative thermal extension coefficient of graphene, respectively.

The recent outbreak of Salmonella Agona linked to the consumption of infant formula (powdered formula) has rekindled the attention about the correct procedures for preparation and use of these products. International guidelines have already been published so far, particularly in association with Cronobacter sakazakii in early 2000s. FAO/WHO suggested to reconstitute formula with water at no less than 70°C. We therefore contaminated powdered formula with low levels of Salmonella spp and C sakazakii to evaluate the pathogens inactivation during the formula preparation using water at 70°C. In these conditions we observed a survival of both pathogens, indicating that the suggested recommendations may be not enough to guarantee the safety of this product. Higher temperatures are needed to reduce the biological risk, even if it may be not easily realized in actual domestic conditions. Moreover, the impact on the nutritional value of reconstituted formulas should be evaluated.

One of the main environmental issues affecting coastal marine environments is the accumulation of contaminants in sediments and their potential mobility. In situ benthic chamber experiments were conducted at two tourist ports (marinas) located in the Gulf of Trieste, one in Slovenia and one in Italy. The aim was to understand if and where recycling at the sediment-water interface (SWI) may affect metal(loid)s. Short sediment cores were also collected near the chamber to investigate the solid (sediments) and dissolved phases (porewaters). Both diffusive and benthic fluxes were estimated to elucidate the release of metal(loid)s at the SWI. Total element concentrations and their labile fractions were determined in sediments to quantify their potential mobility. The total element contents were found to be two orders of magnitude higher in the Italian marina than in the Slovenian one, especially for Hg (up to 1000 mg kg -1 ), whereas the labile fraction was scarce or null. The opposite occurred in the Slovenian marina. Metal(loid)s in porewaters showed a clear diagenetic sequence and a close dependence upon the suboxic/anoxic conditions of sediments. The results suggest that although the sediments of the Italian marina exhibit the highest total metal(loid) concentration, these elements are scarcely remobilisable. Conversely, in the Slovenian marina, sediments seem to be comparatively more prone to release metal(loid)s at the SWI.

Heparanase is the sole mammalian enzyme capable of cleaving glycosaminoglycan heparan sulfate side chains of heparan sulfate proteoglycans. Its altered activity is intimately associated with tumor growth, angiogenesis, and metastasis. Thus, its implication in cancer progression makes it an attractive target in anticancer therapy. Herein, we describe the design, synthesis, and biological evaluation of new benzazoles as heparanase inhibitors. Most of the designed derivatives were active at micromolar or submicromolar concentration, and the most promising compounds are fluorinated and/or amino acids derivatives 13a, 14d, and 15 that showed IC 50 0.16-0.82 μM. Molecular docking studies were performed to rationalize their interaction with the enzyme catalytic site. Importantly, invasion assay confirmed the antimetastatic potential of compounds 14d and 15. Consistently with its ability to inhibit heparanase, compound 15 proved to decrease expression of genes encoding for proangiogenic factors such as MMP-9, VEGF, and FGFs in tumor cells.

Exosomes are nanovesicles known to mediate intercellular communication. Although it is established that zinc ions can act as intracellular signaling factors, the measurement of zinc in circulating nanovesicles has not yet been attempted. Providing evidence of the existence of this zinc fraction and methods for its measurement might be important to advance our knowledge of zinc status and its relevance in diseases. Exosomes from 0.5 ml of either fresh or frozen human plasma were isolated by differential centrifugation. A morphological and dimensional evaluation at the nanoscale level was performed by atomic force microscopy (AFM) and Transmission Electron Microscopy (TEM). Energy Dispersive X-Ray Microanalysis (EDX) revealed the elemental composition of exosomes and their respective total Zinc content on a quantitative basis. The zinc mole fraction (in at%) was correlated to the phosphorous mole fraction, which is indicative for exosomal membrane material. Both fresh (Zn/P 0.09 ± 0.01) and frozen exosomes (Zn/P 0.08 ± 0.02) had a significant zinc content, which increased up to 1.09 ± 0.12 for frozen exosomes when treated with increasing amounts of zinc (100-500 μM; each p < 0.05). Interestingly, after zinc addition, the Calcium mole fractions decreased accordingly suggesting a possible exchange by zinc. In order to estimate the intra-exosomal labile zinc content, an Imaging Flow Cytometry approach was developed by using the specific membrane permeable zinc-probe Fluozin-3AM. A labile zinc content of 0.59 ± 0.27 nM was calculated but it is likely that the measurement may be affected by purification and isolation conditions. This study suggests that circulating nano-vesicular-zinc can represent a newly discovered zinc fraction in the blood plasma whose functional and biological properties will have to be further investigated in future studies., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

The extracellular guanosine 5'-triphosphate, GTP, has been demonstrated to be an enhancer of myogenic cell differentiation in a murine cell line, not yet in human muscle cells. Our hypothesis was that GTP could influence also human skeletal muscle regeneration, specifically in the first phases. We tested GTP stimulus on human muscle precursor cells established in culture by human satellite cells derived from Vastus Lateralis of three young male. Our data show that extracellular GTP (a) up-regulated miRNA (specifically miR133a and miR133b) and myogenic regulator factor and (b) induces human myogenic precursor cells to release exosomes stuffed with guanosine based molecules (mainly guanosine) in the extracellular milieu. We think that probably these exosomes could be addressed to influence by means of their content (mainly guanosine) in paracrine or autocrine manner the surrounding cells and/or at distance other muscles or tissues.

Biolox ® delta has been widely used in joint replacements thanks to its high strength and wear resistance. In this study, eleven Biolox ® delta femoral head retrievals affected by metal transfer (MT) were analysed by Raman spectroscopy to estimate the tetragonal to monoclinic zirconia phase transformation, whose occurrence may compromise ceramic chemical stability and mechanical strength. The residual stress state was evaluated by both Raman and photoemission spectroscopy. V m monoclinic zirconia contents were higher near the centre of the articulating surface and in the MT area than in the border control area of the retrievals. In only one retrieval, stress related to MT appeared a more severe condition, able to induce zirconia phase transformation; for all the others, stresses related to loading in the central region and related to MT, were conducive to a zirconia phase transformation of nearly the same extent. V m depth profiling analyses showed that the transformation involved different thicknesses in different samples. Raman data allowed for the investigation of the mechanism of zirconia phase transformation and confirmed that the growth stage was absent and the nucleation stage was not occurring as freely as it would in unconstrained zirconia., Competing Interests: The authors declare no conflict of interest. The founding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in the decision to publish the results.