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Background Biological drugs have improved the management of immune-mediated inflammatory diseases (IMIDs) despite being associated with important safety issues such as immunogenicity, infections, and malignancies in real-world settings. Objective The aim of this study was to explore the potential of a large Italian multi-database distributed network for use in the postmarketing surveillance of biological drugs, including biosimilars, in patients with IMID. Methods A retrospective cohort study was conducted using 13 Italian regional claims databases during 2010–2019. A tailor-made R-based tool developed for distributed analysis of claims data using a study-specific common data model was customized for this study. We measured the yearly prevalence of biological drug users and the frequency of switches between originator and biosimilars for infliximab, etanercept, and adalimumab separately and stratified them by calendar year and region. We then calculated the cumulative number of users and person-years (PYs) of exposure to individual biological drugs approved for IMIDs. For a number of safety outcomes (e.g., severe acute respiratory syndrome coronavirus 2 [SARS-COV-2] infection), we conducted a sample power calculation to estimate the PYs of exposure required to investigate their association with individual biological drugs approved for IMIDs, considering different strengths of association. Results From a total underlying population of almost 50 million inhabitants from 13 Italian regions, we identified 143,602 (0.3%) biological drug users, with a cumulative exposure of 507,745 PYs during the entire follow-up. The mean age ± standard deviation of biological drug users was 49.3 ± 16.3, with a female-to-male ratio of 1.2. The age-adjusted yearly prevalence of biological drug users increased threefold from 0.7 per 1000 in 2010 to 2.1 per 1000 in 2019. Overall, we identified 40,996 users of biosimilars of tumor necrosis factor (TNF)-α inhibitors (i.e., etanercept, adalimumab, and infliximab) in the years 2015–2019. Of these, 46% (N = 18,845) switched at any time between originator and biosimilars or vice versa. To investigate a moderate association (incidence rate ratio 2) between biological drugs approved for IMIDs and safety events of interest, such as optic neuritis (lowest background incidence rate 10.4/100,000 PYs) or severe infection (highest background incidence rate 4312/100,000 PYs), a total of 43,311 PYs and 104 PYs of exposure to individual biological drugs, respectively, would be required. As such, using this network, of 15 individual biological drugs approved for IMIDs, the association with those adverse events could be investigated for four (27%) and 14 (93%), respectively. Conclusion The VALORE project multi-database network has access to data on more than 140,000 biological drug users (and > 0.5 million PYs) from 13 Italian regions during the years 2010–2019, which will be further expanded with the inclusion of data from other regions and more recent calendar years. Overall, the cumulated amount of person-time of exposure to biological drugs approved for IMIDs provides enough statistical power to investigate weak/moderate associations of almost all individual compounds and the most relevant safety outcomes. Moreover, this network may offer the opportunity to investigate the interchangeability of originator and biosimilars of several TNFα inhibitors in different therapeutic areas in real-world settings. Supplementary Information The online version contains supplementary material available at 10.1007/s40259-021-00498-3.

Background: Magna Graecia is the ancient name for the modern geopolitical region of South Italy extensively populated by Greek colonizers, shown by archeological and historical evidence to be the oldest wine growing region of Italy, crucial for the spread of specialized viticulture around Mediterranean shores. Here, the genetic diversity of Magna Graecia grape germplasm was assessed and its role in grapevine propagation around the Mediterranean basin was underlined. Results: A large collection of grapevines from Magna Graecia was compared with germplasm from Georgia to the Iberian Peninsula using the 18 K SNP array. A high level of genetic diversity of the analyzed germplasm was determined; clustering, structure analysis and DAPC (Discriminant Analysis of Principal Components) highlighted the genetic relationships among genotypes from South Italy and the Eastern Mediterranean (Greece). Gene flow from east (Georgia) to west (Iberian Peninsula) was identified throughout the large number of detected admixed samples. Pedigree analysis showed a complex and well-structured network of first degree relationships, where the cultivars from Magna Graecia were mainly involved. Conclusions: This study provided evidence that Magna Graecia germplasm was shaped by historical events that occurred in the area due to the robust link between South Italian and Greek genotypes, as well as, by the availability of different thermal resources for cultivars growing in such different winegrowing areas. The uniqueness of this ampelographic platform was mainly an outcome of complex natural or human-driven crosses involving elite cultivars.

BACKGROUND Thyroid nodules diagnosed as Thy3B at fine-needle aspiration biopsy have a relevant risk of malignancy (15-30%) and are usually addressed to surgery. However surgery will result unnecessary in most cases. The present study aims at evaluating the possible increase of diagnostic accuracy for predicting malignancy using novel sonographic and elastographic parameters. METHODS In fifty patients undergoing thyroidectomy because of a Thy3B thyroid nodule, sonographic and elastosonographic evaluation was carried out by single operator before surgery. Five sonographic parameters (echogenicity, irregular margins, microcalcifications, intra-nodule blood flow and its irregularity) and two elastosonographic parameters (intra-nodule stiffness and its extension to adjacent tissue) were considered. After obtaining histological diagnosis, diagnostic accuracy was calculated. RESULTS When the two procedures were analyzed separately, sensitivity, specificity, positive (PPV) and negative (NPV) predictive values were 100%, 85%, 63% and 100% for ultrasonography and 60%, 92.5%, 67%, 90% for elastrosonography, respectively. The newly introduced evaluation procedures increased sensitivity. When a combined sonographic and elastosonographic evaluation was introduced, diagnostic accuracy was significantly improved: when ≥4 out of the seven parameters indicated were present, the risk of malignancy was very high (sensitivity 100%, specificity 92.55%, PPV 77%, NPV 100%). CONCLUSIONS A novel combined sonographic and elastosonographic parameter evaluation improved diagnostic accuracy for identifying thyroid nodules suspicious of malignancy.

Pyrethrum (Chrysanthemum cinerariifolium (Trev.) Vis. = Tanacetum cineariaefolium (Trev.) Schultz-Bip.) is a perennial herbaceous plant belonging to the family Asteraceae, native to Albania and the area of former Yugoslavia. It is the only species in the genus Tanacetum having an agronomic importance, although the genus consists of several species producing similar types of bioactive metabolites. The species is grown in order to obtain the insecticidal compounds collectively termed pyrethrins, which are found primarily in the flower heads. In this work we discuss the results found from a worldwide literature review about the micropropagation techniques followed on Pyrethrum, the in vitro culture conditions, and the ex vitro establishment trials under Mediterranean environmental conditions. Many technical problems concerning the propagation of the species seem to have been solved, and detailed protocols are available for an easy and fast propagation by seeds, vegetative splits, stem cuttings (rooted under mist or not), and tissue culture. The first attempts to introduce its cultivation into the semi-arid Mediterranean environments have brought to satisfactory results, and the species may be suggested as a valuable opportunity for the development strategies in new Mediterranean farming systems.

In Italy, grapevines are extensively cultivated, with Sicily representing one of the most significant wine regions. The high number of autochthonous grapevine varieties represents an important source of genetic diversity, and the many Sicilian varieties have anatomical and physiological traits that allow them to resist to different levels of drought stress. We investigated the water relation parameters of four cultivars of Vitis vinifera L. (Catarratto, Corinto, Nero d'Avola and Zibibbo) and characterized their leaf hydraulics. Measurements were conducted during summer on plants growing in the experimental field of the IBBR-CNR near Palermo. Daily patterns of leaf water potential (Yleaf) and stomatal conductance (gs) were measured in the field. Pressure-volume curves were constructed by the bench dehydration method to obtain leaf water potential at turgor loss point (Ytlp), osmotic potential at full rehydration (p0) and bulk modulus of elasticity (εmax). Leaf samples were collected to determine vein density using ImageJ. Major vein density was measured on digitally scanned leaves, while minor vein density was measured on photomicrographs of cleared and stained leaf portions.

Potassium availability affects drought responses in plants through several metabolic roles, among which stomatal regulation, cell growth and xylem hydraulics. Vitis vinifera L. is a highly valuable crop and several genotypes have been selected during its millennial cultivation. Varieties show differences in their adaptability to stress conditions, making them more or less suitable to certain climatic and edaphic conditions. The varieties cultivated in Sicily are characterized by high variability. We investigated the response of two Sicilian cultivars (Nero d'Avola and Catarratto) to potassium deficiency and drought stress. Two-year-old grafted plants were grown in agriperlite, with or without potassium in the fertigation solution, and subjected to moderate drought stress by suspending irrigation for 6-8 days. Potassium content of xylem sap, leaf and root tissues were measured with an ion-selective electrode. Changes in stomatal conductance, plant transpiration and hydraulic conductance were compared between genotypes and treatments, in order to gain information for the development of optimal fertigation practices and the selection of the most drought tolerant varieties.

The current rates of biodiversity decrease are estimated from 100 to 1,000 times the natural background levels. Among the initiatives to stop this trend, IUCN launched the campaign "Countdown 2010" to encourage and enable decision makers and stakeholders to uphold their commitments and achieve the goal to "halt the loss of biodiversity by 2010". Accordingly, the Italian Ministry of Environment funded in 2006 a project to save from the extinction the tree species Zelkova sicula, a very rare endemism from Sicily. Zelkova sicula is a relic of the Tertiary flora and belongs to a genus which became extinct in the whole continental Europe during the Quaternary glacial age. Currently the entire population consists of about 230 individuals and the species is included in the IUCN Red List as "Critically Endangered" of extinction. Several investigations showed that its current habitat is somewhat far away from a bioclimatic optimum and the most recent trends in climate change certainly give rise to discouraging scenarios. Summer water stress, exclusive vegetative regeneration, heavy anthropic disturbances and lack of active protection measures are the main points of criticism in the conservation of this species. Furthermore, the long geographic isolation and the limited population size might have caused a rapid loss of intra-specific genetic variability. The project to save Zelkova sicula consists of specific actions to promote in-situ and ex-situ conservation. It entails research activities, on one side, focusing on the multiplication and the genetic structure of the residual population, and, on the other side, education and awareness campaigning. CNR-IGV and Legambiente, respectively, are the subjects involved. Several steps have been performed. Numbering, labelling and positioning of each single tree through a GPS device were first carried out. Leaf material and current year twigs were then sampled from all trees in order to perform genetic analyses and vegetative multiplication. Different protocols for the vegetative propagation were developed. Cuttings were collected at three periods of the year (February, May and October) from three types of plant stocks (herbaceous, semi-woody and woody) and then treated with several concentrations of indole-3-butyric acid (IBA) and naphtalene acetic acid (NAA). Introduction in vitro and micropropagation were performed using newly flushed nodal explants as starting material. Explants were firstly sterilized under several conditions and then introduced in vitro on WPM medium supplemented with 6-benzylaminopurine (BAP) at different concentrations. The genetic diversity was investigated by molecular markers. Preliminary screening of genetic variation was performed using three ISSR primers. Then, codominant and potentially highly polymorphic nuclear SSRs were screened using primers designed in Zelkova serrata. Eleven SSR primers produced amplification products of the expected sizes in Zelkova sicula. Near absence of variation was observed in this species.

Citrus limon (L.) Burm. F. is an important evergreen fruit crop whose rhizosphere and phyllosphere microbiota have been characterized, while seed microbiota is still unknown. Bacterial and fungal endophytes were isolated from C. limon surface-sterilized seeds. The isolated fungi—belonging to Aspergillus, Quambalaria and Bjerkandera genera—and bacteria—belonging to Staphylococcus genus—were characterized for indoleacetic acid production and phosphate solubilization. Next Generation Sequencing based approaches were then used to characterize the endophytic bacterial and fungal microbiota structures of surface-sterilized C. limon seeds and of shoots obtained under aseptic conditions from in vitro growing seedlings regenerated from surface-sterilized seeds. This analysis highlighted that Cutibacterium and Acinetobacter were the most abundant bacterial genera in both seeds and shoots, while Cladosporium and Debaryomyces were the most abundant fungal genera in seeds and shoots, respectively. The localization of bacterial endophytes in seed and shoot tissues was revealed by Fluorescence In Situ Hybridization coupled with Confocal Laser Scanning Microscopy revealing vascular bundle colonization. Thus, these results highlighted for the first time the structures of endophytic microbiota of C. limon seeds and the transmission to shoots, corroborating the idea of a vertical transmission of plant microbiota and suggesting its crucial role in seed germination and plant development.

Sansevieria Thunb. species are traditionally known as succulent ornamental plants worldwide. They are also cultivated for medicinal, fodder, soil conservation and fiber uses, and for their capacity to reduce environmental pollution. Sansevieria sexual propagation is limited by the lack of viable seeds, and reproduction is largely made via vegetative propagation by suckers or cuttings. For these reasons, genetic improvement by conventional breeding is limited. To overcome this problem and to address the increasing demand from customers for novel Sansevieria varieties, many commercial companies regularly use in vitro propagation, as is the case in the breeding process of several ornamental plants. In this paper, for the first time, we report a procedure for in vitro somatic embryogenesis and plant regeneration starting from three flower explants for seven different Sansevieria genotypes. Regeneration was attempted using stigmas/styles, anther/filament, and ovary which were cultured on a Murashige and Skoog solidified medium under three different plant growth regulator combinations. A good regeneration rate was obtained with all genotypes used under all culture conditions tested from every explant type, with percentages ranging from 0 to 73.3%. “Genetic stability” assessment of regenerated plants in respect to their mother plants was verified through flow cytometry analysis showing a high degree of uniformity, with only S. parva exhibiting a different level of DNA fluorescence among in vitro regenerated plants. This is an interesting achievement in the aim to produce true-to-type plants and new variants with desirable characteristics, both of which are desired features in ornamentals improvement.

The genus Calendula (Asteraceae) includes several species that are renowned for their therapeutic properties and/or use as ingredients in the preparation of cosmetics. The rare and critically endangered sea marigold, Calendula maritima Guss., an endemic species from Western Sicily, has also been recognized as a potential “farm plant species” for several important compounds used in cosmetics. However, the few remnant populations of this species are currently threatened with extinction because of several factors, such as hybridization with the congeneric species Calendula suffruticosa subsp. fulgida (Raf.) Guadagno and anthropogenic disturbance of its habitat. Therefore, in order to preserve the genetic integrity from pure genetic lineages, seed-based propagation and seed storage are not recommended for either conservation or massive production purposes. In this paper, we describe a protocol adopted for mass propagation of C. maritima from selected genotypes. Nodal segments collected from selected plants growing in the field were used as starting explants and were cultured for micropropagation on MS medium with and without phloroglucinol. New shoots produced were cultured for rooting under several conditions with the aim of finding the best procedure favoring root induction. The best rooting performance was obtained with zeolite and rooted plants were successfully acclimatized outdoors. The technique described allowed the multiplication of genotypes of interest as well as to overcome the problems of hybridization of this species, hence contributing concretely to the conservation of the sea marigold.

Baron Antonio Mendola was devoted to the study of grapevine, applying ampelography and dabbling in crosses between cultivars in order to select new ones, of which Moscato Cerletti, obtained in 1869, was the most interesting. Grillo, one of the most important white cultivars in Sicily, was ascertained to be an offspring of Catarratto Comune and Zibibbo, the same parents which Mendola claimed he used to obtain Moscato Cerletti. Thus the hypothesis of synonymy between Moscato Cerletti and Grillo or the same parentage for both sets of parents needs to be verified.In the present study, historical documents were consulted and genetic analyses and ampelographic, agronomic and qualitative characterisation carried out to determine the distinctiveness of each cultivars. These were also compared with Catarratto Comune and Zibibbo in order to establish the Moscato Cerletti pedigree. Due to their different SSR profiles, Grillo and Moscato Cerletti were confirmed as two distinct cultivars; they also differed in ripening times and sugar storage ability, as well as in the aromatic grape produced by Moscato Cerletti only. The trio genotype genetic analysis confirmed that Zibibbo is a parent of Moscato Cerletti (justifying the aromatic grape), whilst the SSR profiles did not show Catarratto Comune to be a second parent.Moscato Cerletti was found to have oenological potential in the production of sparkling muscat wines due to its ability to adapt to a changing climate in warm and dry environments and in different winegrowing regions.

Nowadays network is the preferred governance form to conduct economic transactions. Network solution allows to reach flexibility maintaining cost and quality level. Since network concept refers to a great variety of organizational hybrids it is possible to choose the one that fits better market requirements. The new trends in inter-organization relationships push towards network solutions: companies are interested in relationships with partners and customers to overcome resource dependence, to enter too risky market or simply differentiate their business portfolio. The proposed research focuses on the network concept aiming at highlighting threats and opportunities to investigate the double nature of the risk concept. Network structures offer flexibility and higher profit as a consequence and business risk sharing opportunity. These two aspects (profit and risk) are strictly related and have to be considered together to depict a complete scenario; this implies that risk assessment and management in network environment cannot neglect profit sharing or, in other words, that profit sharing mechanisms should use risk as driver. In this context our research proposes a methodology to measure risk taking into account network peculiarities; risk estimation is a basic step to evaluate the opportunity cost of capital needed to compute the network Net Present Value (NPV) that is assumed as base in the profit sharing process. The profit sharing process has been tackled using the Shapley value approach that is inspired to the fairness principle while the opportunity cost of capital is assessed using the Capital Asset Pricing Model (CAPM).

Background: The European post-authorisation study (EU PAS) register is a repository launched in 2010 by the European Medicines Agency (EMA). All EMA-requested PAS, commonly observational studies, must be recorded in this register. Multi-database studies (MDS) leveraging secondary data have become an important strategy to conduct PAS in recent years, as reflected by the type of studies registered in the EU PAS register. Objectives: To analyse and describe PAS in the EU PAS register, with focus on MDS. Methods: Studies in the EU PAS register from inception to 31st December 2018 were described concerning transparency, regulatory obligations, scope, study type (e.g., observational study, clinical trial, survey, systematic review/meta-analysis), study design, type of data collection and target population. MDS were defined as studies conducted through secondary use of >1 data source not linked at patient-level. Data extraction was carried out independently by 14 centres with expertise in pharmacoepidemiology, using publicly available information in the EU PAS register including study protocol, whenever available, using a standardised data collection form. For validation purposes, a second revision of key fields for a 15% random sample of studies was carried out by a different centre. The inter-rater reliability (IRR) was then calculated. Finally, to identify predictors of primary data collection-based studies/versus those based on secondary use of healthcare databases) or MDS (vs. non-MDS), odds ratios (OR) and 95% confidence intervals (CI) were calculated fitting univariate logistic regression models. Results: Overall, 1426 studies were identified. Clinical trials (N = 30; 2%), systematic reviews/meta-analyses (N = 16; 1%) and miscellaneous study designs (N = 46; 3%) were much less common than observational studies (N = 1227; 86%). The protocol was available for 63% (N = 360) of 572 observational studies requested by a competent authority. Overall, 36% (N = 446) of observational studies were based fully or partially on primary data collection. Of 757 observational studies based on secondary use of data alone, 282 (37%) were MDS. Drug utilisation was significantly more common as a study scope in MDS compared to non-MDS studies. The overall percentage agreement among collaborating centres that collected the data concerning study variables was highest for study type (93.5%) and lowest for type of secondary data (67.8%). Conclusions: Observational studies were the most common type of studies in the EU PAS register, but 30% used primary data, which is more resource-intensive. Almost half of observational studies using secondary data were MDS. Data recording in the EU PAS register may be improved further, including more widespread availability of study protocols to improve transparency.

Decision support systems (DSSs) have been traditionally identified as useful information technology tools in a variety of fields, including the context of cultural heritage. However, to the best of our knowledge, no prior study has developed a DSS framework that incorporates all the main decision areas simultaneously in the context of cultural heritage. We fill this gap by focusing on design-science research and specifically by developing a DSS framework whose features support all the main decision areas for the sustainable management of cultural assets in a comprehensive manner. The main decision-making areas considered in our study encompass demand management, segmentation and communication, pricing, space management, and services management. For these areas, we select appropriate decision-making supporting techniques and data management solutions. The development of our framework, in the form of a web-based system, results in an architectural solution that is able to satisfy critical requirements such as ease of use and response time. We present an application of the innovative DSS framework to a museum and discuss the main managerial implications and future improvements.

This study aimed at surveying lower secondary schools in southern Italy, in a highly polluted area. A community close to an industrial area and three villages in rural areas was investigated. Indoor temperature, relative humidity (RH), gaseous pollutants (CO 2 and NO 2 ), selected biological pollutants in indoor dust, and the indoor/outdoor mass concentration and elemental composition of PM 2.5 were ascertained. Temperature and RH were within, or close to, the comfort range, while CO 2 frequently exceeded the threshold of 1000ppm, indicating inadequate air exchange rate. In all the classrooms, median NO 2 levels were above the WHO threshold value. Dermatophagoides p. allergen concentration was below the sensitizing threshold, while high endotoxin levels were detected in the classrooms, suggesting schools may produce significant risks of endotoxin exposure. Concentration and solubility of PM 2.5 elements were used to identify the sources of indoor particles. Indoor concentration of most elements was higher than outdoors. Resuspension was responsible for the indoor increase in soil components. For elements from industrial emission (Cd, Co, Ni, Pb, Sb, Tl, V), the indoor concentration depended on penetration from the outside. For these elements, differences in rural vs industrial concentrations were found, suggesting industrial sources may influence indoor air quality nearby schools.

Polyploidy plays an important role in plant adaptation to biotic and abiotic stresses. Alterations of the ploidy in grapevine plants regenerated via somatic embryogenesis (SE) may provide a source of genetic variability useful for the improvement of agronomic characteristics of crops. In the grapevine, the SE induction process may cause ploidy changes without alterations in DNA profile. In the present research, tetraploid plants were observed for 9.3% of ‘Frappato’ grapevine somatic embryos regenerated in medium supplemented with the growth regulators β-naphthoxyacetic acid (10 µM) and N6-benzylaminopurine (4.4 µM). Autotetraploid plants regenerated via SE without detectable changes in the DNA profiles were transferred in field conditions to analyze the effect of polyploidization. Different ploidy levels induced several anatomical and morphological changes of the shoots and mature leaves. Alterations have been also observed in stomata. The length and width of stomata of tetraploid leaves were 39.9 and 18.6% higher than diploids, respectively. The chloroplast number per guard cell pair was higher (5.2%) in tetraploid leaves. On the contrary, the stomatal index was markedly decreased (12%) in tetraploid leaves. The observed morphological alterations might be useful traits for breeding of grapevine varieties in a changing environment.

Evidence on treatments for early-stage COVID-19 in outpatient setting is sparse. We explored the pattern of use of drugs prescribed for COVID-19 outpatients’ management in Southern Italy in the period February 2020–January 2021. This population-based cohort study was conducted using COVID-19 surveillance registry from Caserta Local Health Unit, which was linked to claims databases from the same catchment area. The date of SARS-CoV-2 infection diagnosis was the index date (ID). We evaluated demographic and clinical characteristics of the study drug users and the pattern of use of drugs prescribed for outpatient COVID-19 management. Overall, 40,030 patients were included in the analyses, with a median (IQR) age of 44 (27–58) years. More than half of the included patients were asymptomatic at the ID. Overall, during the study period, 720 (1.8%) patients died due to COVID-19. Azithromycin and glucocorticoids were the most frequently prescribed drugs, while oxygen was the less frequently prescribed therapy. The cumulative rate of recovery from COVID-19 was 84.2% at 30 days from ID and it was lower among older patients. In this study we documented that the drug prescribing patterns for COVID-19 treatment in an outpatient setting from Southern Italy was not supported from current evidence on beneficial therapies for early treatment of COVID-19, thus highlighting the need to implement strategies for improving appropriate drug prescribing in general practice.

Supply chain finance aims at finding the best financing arrangements within a given buyer-supplier dyad. The source of capital can be internal (buyer or supplier) or external (financial institution) to the supply chain. So far, many studies have investigated the optimal mix of the sources of capital; our study aims at contributing to the recent literature that explores the interface of operations and finance extending the supplier-based financing models. As the Covid-19 pandemic hits economic activity, the financial constraints have ever greater importance; knock-on effects of the Covid-19 crisis urges on the critical role of a supply chain that should provide financial resources, along to the flow of goods, in the more efficient way. The proposed model considers a supply chain formed by a supplier and a retailer, both capital-constrained that can ask for a loan to a financial institution or resort on their internal reciprocal sources. Demand is uncertain, retailer, that acts as a price taker, may also affect her product demand applying some effort in increasing sales. Both retailer and supplier can fail. The model optimizes the supplier and retailer's’ profit varying their contract parameters; doing this, the research allows to understand the interplay between supply chain operational and financial issues when retailer's sales effort is at work, and its findings can support retailer in the suppliers selection basing on their credit rating. Our findings show how operational (retailer's effort) and financial (trade credit conditions) issues can synergically interact also in supply chain with low working capital or conversely how retailer with high working capital can perform better working with low rating supplier, however boosting the chance to successfully compete in the Covid-19 pandemic era.

In Sicily, small differences exist between wild and cultivated rosemary biotypes; VOCs and genetic profiles may be a useful tool to distinguish them. A germplasm collection of Rosmarinus officinalis L. was harvested from 15 locations in Sicily. Eleven wild and four cultivated populations were collected and, due to the surveyed area covered, they can be considered as a representative panel of Sicilian genetic background of the species. Ex situ plant collection was transferred to the field cultivation in homogeneous conditions for characterizing through a multidisciplinary approach. The study included morphological traits observations (growth habitus, flower color, number and size of leaves, length and number of internodes), VOC profiles using HS-SPME, genome size by flow cytometry analysis, and genetic characterization by means of DNA and nuclear microsatellite (nSSR) investigation. To detect any pattern within- and among-populations variability, all morphological and chemical data were submitted to ANOVA, while clustering and structure population analysis were carried out using genetic profiles. The present work allowed us to distinguish rather well between wild and cultivated genotypes and to underline the biodiversity richness among rosemary Sicilian germplasm, never highlighted, useful for future breeding programs addressed to exploit this important resource.

The cosmopolitan genus Fraxinus comprises about 40 species occupying several habitats in the Northern Hemisphere. With some species hybridizing and sharing genetic variants, questions remain on the species assignment of germplasm within the genus Fraxinus despite numerous species-specific assessments. A multidisciplinary approach was employed to provide a definitive insight into the genetics of an endangered Fraxinus &ldquo, manna ash&rdquo, collection, located in a rich plant biodiversity hotspot of the Madonie Mountains (Sicily). Although the collection size was small, genetic diversity, assessed by chloroplast (cpSSR) and nuclear (nSSR) microsatellites (SSR&mdash, Simple Sequence Repeats), allowed identifying three different chloroplast haplotypes, with one (H5) dominant, and several polymorphic loci, able to discriminate most of the local accessions studied. Molecular data were linked to cytofluorimetric and phenotypic evaluations and, contrary to popular belief that manna ash is Fraxinus ornus L., the germplasm currently used for manna production belongs to Fraxinus angustifolia Vahl. Interestingly, joint analysis of our genetic panel with a large European dataset of Fraxinus spp. suggested the presence of a possible glacial refuge in Sicily, confirming its importance as biodiversity source. Our results will be helpful for the design of long-term conservation programs for genetic resources, such as in situ and ex situ conservation, seed collection and tree reintroduction.

Background Plant seeds possess a complex microbiota which may play a crucial rolein a plethora of mechanisms, such as preservation, germination, seedling development, plant growth and health. In particular, seed endophytic microbiota is gaining more and more consideration due to the fact that it may be vertically transmitted to ensure individual competitive advantages. The Citrusspecies, including C.limon,areone of the most economically important evergreen fruit crops in the world. Objectives i)Identification and characterization ofmicrobial isolates from Citrus limon L. Burm. F. seed endosphere. ii) Structure of Citrus limon L. Burm. F. seed endosphytic microbiota. Methods -Surface-sterilization of Citrus limon L. Burm. F. seeds. - Strain isolation and phylogenetic characterization by 16S rDNA sequence. - Next Generation Sequence Technology analysis (NGS) of metagenomic DNA by pyrosequencing of 16S rDNA. - Fluorescence In Situ Hybridization coupled with Confocal Laser Scanning Microscopy (FISH-CLSM). Conclusions Culture-dependent analysis allowed the isolation of several Staphylococcus spp. and fungi belonging to genera Aspergillus, Quambalaria and Efibula from surface-sterilized seeds. These results weresupported by the detection of bacterial cells and micro–colonies in seed cryosections by FISH-CLSM. In fact, this finding highlighted the presence of Firmicutes and other bacteria colonizing intercellular spaces. NGS-basedcharacterization using metagenomic DNA from surface-sterilized seeds is being carried out in order to elucidatethe microbiota structure. Culture-dependent approaches allowed the isolation of several bacterial strains belonging to the genus Staphylococcus and several fungal strains belonging to the genera Aspergillus, Quambalaria and Efibulafrom seed endosphere. These results were supported by the detection of bacterial cells and micro–colonies in seed cryosections by FISH-CLSM. In particular, this analysis highlighted the presence of Firmicutes and other bacteria colonizing intercellular spaces. NGS-based characterization using metagenomic DNA from surface-sterilized seeds is being carried out in order to elucidate the microbiota structure.

In our study, we evaluated the feasibility of a new sampling method for splenic stiffness (SS) measurement by Quantitative Acoustic Radiation Force Impulse Elastography (Virtual Touch Tissue Quantification (VTTQ)).We measured SS in 54 patients with HCV-related cirrhosis of whom 28 with esophageal varices (EV), 27 with Chronic Hepatitis C (CHC) F1–F3, and 63 healthy controls. VTTQ-SS was significantly higher among cirrhotic patients with EV (3.37 m/s) in comparison with controls (2.19 m/s,P<0.001), CHC patients (2.37 m/s,P<0.001), and cirrhotic patients without EV (2.7 m/s,P<0.001). Moreover, VTTQ-SS was significantly higher among cirrhotic patients without EV in comparison with both controls (P<0.001) and CHC patients (P<0.01). The optimal VTTQ-SS cut-off value for predicting EV was 3.1 m/s (AUROC = 0.96, sensitivity 96.4%, specificity 88.5%, positive predictive value 90%, negative predictive value 96%, positive likelihood ratio 8.36, and negative likelihood ratio 0.04). In conclusion, VTTQ-SS is a promising noninvasive and reliable diagnostic tool to screen cirrhotic patients for EV and reduce the need for upper gastrointestinal endoscopy. By using our cut-off value of 3.1 m/s, we would avoid endoscopy in around 45% of cirrhotic subjects, with significant time and cost savings.