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Background: Although digital mobility outcomes (DMOs) can be readily calculated from real-world data collected with wearable devices (WD) and ad-hoc algorithms, technical validation is still required. The aim of this paper is to comparatively assess and validate DMOs estimated using real-world gait data from six different cohorts, focusing on gait sequence detection (GSD), foot initial contact detection (ICD), cadence (CAD) and stride length (SL) estimates. Methods: Twenty healthy older adults, 20 people with Parkinson’s disease, 20 with multiple sclerosis, 19 with proximal femoral fracture (PFF), 17 with chronic obstructive pulmonary disease and 12 with congestive heart failure were monitored for 2.5 hours in the real-world, using a WD worn on the lower back. A reference system combining inertial modules with distance sensors and pressure insoles was used for comparison of DMOs from the WD. We assessed and validated three algorithms for GSD, four for ICD, three for CAD and four for SL by concurrently comparing their performances (e.g., accuracy, specificity, sensitivity, absolute and relative errors). Additionally, the effects of walking bout (WB) speed and duration on algorithm performance were investigated. Results: We identified two cohort-specific top performing algorithms for GSD and CAD, and a single best for ICD and SL. GSD best algorithms showed good performances (sensitivity > 0.73, positive predictive values > 0.75, specificity > 0.95, accuracy > 0.94). ICD and CAD algorithms presented excellent results, with sensitivity > 0.79, positive predictive values > 0.89 and relative errors < 11% for ICD and < 8.5% for CAD. The best identified SL algorithm showed lower performances than other DMOs (absolute error < 0.21 m). Lower performances across all DMOs were found for the cohort with most severe gait impairments (PFF). Algorithms’ performances were lower for short WBs; slower gait speeds (Conclusions: Overall, the identified algorithms enabled a robust estimation of key DMOs. Our findingsshowed that the choice of algorithm for estimation of GSD and CAD DMOs should be cohort-specific (e.g., slow walkers and with gait impairments). Short WB length and slow walking speed worsened algorithms’ performances. Trial registration: ISRCTN – 12246987.

Nature-based solutions are sustainable building materials produced recovering and enhancing agricultural biomasses which are by-products or waste of crops as, for example, rice, flax or hemp. Specifically, this research investigates the properties of hempcrete which is produced mixing lime, which acts as binder, and hemp shives, as vegetal aggregate. Hempcrete is characterized by breathability and excellent insulating properties, moreover it is a sustainable material due to the introduction of vegetal material and due to the carbonation of lime which gives further carbon dioxide sequestration. The mechanical properties of the material are largely variable and, in this research, triaxial tests have been performed to evaluate this experimental methodology as a technique applicable to evaluate the mechanical behavior of this material. The tests have been performed on samples produced with the same mix design developed by an Italian manufacturer for the production of prefabricated hempcrete blocks. These building components are used as non-loadbearing blocks, they are introduced in building envelopes or in indoor partition walls as insulating elements.

Accurately assessing people’s gait, especially in real-world conditions and in case of impaired mobility, is still a challenge due to intrinsic and extrinsic factors resulting in gait complexity. To improve the estimation of gait-related digital mobility outcomes (DMOs) in real-world scenarios, this study presents a wearable multi-sensor system (INDIP), integrating complementary sensing approaches (two plantar pressure insoles, three inertial units and two distance sensors). The INDIP technical validity was assessed against stereophotogrammetry during a laboratory experimental protocol comprising structured tests (including continuous curvilinear and rectilinear walking and steps) and a simulation of daily-life activities (SDA, including intermittent gait and short walking bouts). To evaluate its performance on various gait patterns, data were collected on 128 participants from seven cohorts: healthy young and older adults, patients with Parkinson’s disease, multiple sclerosis, chronic obstructive pulmonary disease, congestive heart failure, and proximal femur fracture. Moreover, INDIP usability was evaluated by recording 2.5-hours of real-world unsupervised activity. Excellent absolute agreement (ICC > 0.95) and very limited mean absolute errors were observed for all cohorts and DMOs (cadence ≤ 0.61 steps/min, stride length ≤ 0.02 m, walking speed ≤ 0.02 m/s) in the structured tests. Larger, but limited, errors were observed during the SDA (cadence 2.72–4.87 steps/min, stride length 0.04–0.06 m, walking speed 0.03–0.05 m/s). Neither major technical nor usability issues were declared during the 2.5-hours acquisitions. Therefore, the INDIP system can be considered a valid and feasible solution to collect reference data for analyzing gait in real-world conditions.

Inherited retinal diseases (IRDs) encompass a large heterogeneous group of rare blinding disorders whose etiology originates from mutations in the 280 genes identified to date. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems represent a promising avenue for the treatment of IRDs, as exemplified by FDA clinical trial approval of EDIT-101 (AGN-151587), which removes a deep intronic variant in the CEP290 gene that causes Leber congenital amaurosis (LCA) type 10. Prime editing is a novel double-strand break (DSB) independent CRISPR/Cas system which has the potential to correct all 12 possible transition and transversion mutations in addition to small deletions and insertions. Here, as a proof-of-concept study, we describe a methodology using prime editing for the in vitro installation and correction of the classical Pde6b

Interest towards cellulose nanofibers obtained from virgin and waste sources has seen a significant growth, mainly thanks to the increasing sensitivity towards the concept of circular economy and the high levels of paper recycling achieved in recent years. Inspired by the guidelines of the green building industry, this study proposes the production and characterization of TEMPO-oxidized and homogenized cellulose nanofibers (TOHO CNF) from different sources and their use as additives for earth plasters on two different raw earth samples, characterized by geotechnical laboratory tests and mineralogical analysis: a high-plasticity clay (T2) and a medium-compressibility silt (ABS). Original sources, including those derived from waste (recycled cardboard and paper mill sludge), were characterized by determining chemical content (cellulose versus ashes and lignin) and fiber morphology. TOHO CNF derived from the different sources were compared in terms of nanofibers medium diameter, crystallinity degree, thermal decomposition and oxidation degree, that is the content of carboxylic groups per gram of sample. Then, a preliminary analysis of the influence of CNF on earth plasters is examined. Adhesion and capillary absorption tests highlighted the effect of such nanofibers on blends in function of two factors, namely the cellulose original source and the oxidation degree of the fibers. In particular, for both earth samples, T2 and ABS, a significant increase in adhesion strength was observed in the presence of some TOHO CNF additives. As far as capillary sorption tests, while an undesired increase in water adsorption was detected for T2 compared to the control, in the case of ABS, a significant reduction in water content was measured by adding TOHO CNF derived from recycled sources. These results pave the way for further in-depth investigation on the role of TOHO CNF as additives for earth plasters.

Aim Percutaneous coronary intervention with stent implantation (PCI-S) in patients requiring chronic oral anticoagulant therapy (OAC) is associated with an increased risk of bleeding and ischemic complications. Different randomized studies showed a significant advantage of a double antithrombotic therapy and superiority of direct oral anticoagulant (DOAC) compared with warfarin, but real-world data are limited. Aim is to evaluate the antithrombotic management and clinical outcome of patients with an indication for OAC who undergo PCI-S in a 'real-world' setting. Methods The multicentre prospective observational PERSEO (PERcutaneouS coronary intErventions in patients treated with Oral anticoagulant therapy) Registry (ClinicalTrials.gov Identifier: NCT03392948) has been designed to enrol patients requiring OAC treated by PCI-S in 25 Italian centres. A target of at least 1080 patients will be followed for 1 year and data on thromboembolic and bleeding events and changes in antithrombotic therapy will be registered. The primary end point is a combined measure of efficacy and safety outcome (NACE), including major bleeding events and major adverse cardiac and cerebral events at 1-year follow-up in patients treated with DOAC (and dual or triple antiplatelet therapy) compared with the corresponding strategies with vitamin K antagonists. A secondary prespecified analysis has been defined to evaluate NACE in dual versus triple antithrombotic therapy after hospital discharge at 1-year follow-up. Conclusion The PERSEO Registry will investigate in a 'real world' setting the safety and efficacy of DOAC versus warfarin and dual versus triple antithrombotic therapy in patients with indication for oral anticoagulant therapy who undergo PCI-S.

Background: Measuring mobility in daily life entails dealing with confounding factors arising from multiple sources, including pathological characteristics, patient specific walking strategies, environment/context, and purpose of the task. The primary aim of this study is to propose and validate a multi-task and multi-phase protocol for simulating real-world gait accounting for all these factors within a single set of observations carried out within a limited laboratory space, while ensuring minimisation of participant burden and safety.Methods: The protocol included eight motor tasks at varying speed, incline/steps, surface, path shape, cognitive demand, and included postures that may abruptly alter the participants’ strategy of walking. It was deployed in a convenience sample of 108 participants recruited from six cohort groups that included older healthy adults (HA) and participants with potentially altered mobility due to Parkinson’s Disease (PD), Multiple Sclerosis (MS), Proximal Femoral Fracture (PFF), Chronic Obstructive Pulmonary Disease (COPD) or Congestive Heart Failure (CHF). A novelty introduced in the protocol was the tiered approach to increase difficulty both within the same task (e.g., by allowing use of aids or armrests) and across tasks.Results: The protocol proved to be safe and feasible and the addition of the more complex tasks allowed a much greater spread in walking speeds to be achieved compared to standard straight walking trials. Furthermore, it allowed a relatively realistic representation of daily life relevant mobility aspects and can therefore be used for the validation of monitoring devices used in real life.Conclusions: The suitability of the protocol for measuring gait in a variety of pathological conditions suggests that it can also be used to detect changes in gait due to, for example, the onset or progression of a disease, or due to therapy.Trial registration: ISRCTN – 12246987.

Background and aims Novel genetic determinants associated with coronary artery disease (CAD) have been discovered by genome wide association studies. Variants encompassing the CELSR2- PSRC1-SORT1 gene cluster have been associated with CAD. This study is aimed to investigate the rs629301 polymorphism association with the extent of CAD evaluated by coronary angiography (CAG), and to evaluate its associations with an extensive panel of lipid and lipoprotein measurements in a large Italian cohort of 2429 patients. Methods and results The patients were collected by four Intensive Care Units located in Palermo and Verona (Italy). Clinical Records were filed, blood samples were collected, lipids and apolipoproteins (apo) were measured in separate laboratories. CAD was defined by the presence of stenotic arteries (>50% lumen diameter) by CAG. The presence of CAD was associated with the rs629301 genotype. Patients with CAD were 78% and 73% (p = 0.007) of the T/T vs. T/G + G/G genotype carriers respectively. T/T genotype was also correlated with the number of stenotic arteries, with a 1.29 (1.04–1.61) risk to have a three-arteries disease. T/T genotype correlated with higher levels of LDL-, non-HDL cholesterol, apoB, apoE and apoCIII, and lower HDL-cholesterol. Logistic Regression confirmed that rs629301was associated with CAD independently from the common risk factors, with a risk similar to that conferred by ten years of age [odds ratios were 1.43 (1.04–1.96) and 1.39 (1.22–1.58) respectively]. Conclusions rs629301 risk allele was independently associated with the extension and severity of CAD and positively with apoE and apoB containing lipoproteins.

Autosomal dominant disorders present unique challenges, as therapeutics must often distinguish between healthy and diseased alleles while maintaining high efficiency, specificity, and safety. For this task, CRISPR/Cas remains particularly promising. Various CRISPR/Cas systems, like homology-directed repair, base editors, and prime editors, have been demonstrated to selectively edit mutant alleles either by incorporating these mutations into sgRNA sequences (near the protospacer-adjacent motif ["near the PAM"]) or by targeting a novel PAM generated by the mutation ("in the PAM"). However, these probability-based designs are not always assured, necessitating generalized, mutation-agnostic strategies like ablate-and-replace and single-nucleotide polymorphism editing. Here, we detail recent advancements in CRISPR therapeutics to treat a wide range of autosomal dominant disorders and discuss how they are altering the landscape for future therapies.

The causal agent of mal secco disease is the fungus

Mutations in rhodopsin (RHO) are the most common causes of autosomal dominant retinitis pigmentosa (adRP), accounting for 20% to 30% of all cases worldwide. However, the high degree of genetic heterogeneity makes development of effective therapies cumbersome. To provide a universal solution to RHO-related adRP, we devised a CRISPR-based, mutation-independent gene ablation and replacement (AR) compound therapy carried by a dual AAV2/8 system. Moreover, we developed a novel hRHO(C110R)/hRHO(WT) humanized mouse model to assess the AR treatment in vivo. Results show that this humanized RHO mouse model exhibits progressive rod-cone degeneration that phenocopies hRHO(C110R)/hRHO(WT) patients. In vivo transduction of AR AAV8 dual vectors remarkably ablates endogenous RHO expression and overexpresses exogenous WT hRHO. Furthermore, the administration of AR during adulthood significantly hampers photoreceptor degeneration both histologically and functionally for at least 6 months compared with sole gene replacement or surgical trauma control. This study demonstrates the effectiveness of AR treatment of adRP in the human genomic context while revealing the feasibility of its application for other autosomal dominant disorders.

According to the different classification methods, the genus Opuntia belongs to the Cactaceae family native to the Americas, and encompasses several species ranging from 75 to 250. Many of these species are diffused, and some of them are cultivated in arid and semi-arid areas. The most economically important species is O. ficus-indica L. (Mill.), but the genus includes other important edible species (from diploid to octoploid), which are spread, wild, or cultivated in many regions worldwide. Opuntia species are gaining popularity in different areas, mainly arid and semi-arid, both for fruit production for human consumption and mainly for animal feeding using the tender and mature green part of the whole plant (cladodes). The taxonomic classification of the genus is hampered by several factors: the lack of reliable morphological descriptors, the recurrent intra- and intergeneric hybridization, the frequent polyploidy, and the phenotypic variation displayed by the genotypes under different ecological conditions. Due to the ambiguity of the different taxonomic hypotheses formulated based on morphological traits, in the last years, several DNA-based studies, relying on different categories of molecular markers, have been carried out in order to better assess variability level within the main cultivated species (O. ficus-indica) and to assess relationships and origin of the main species.

Mal secco is one of the most severe diseases of citrus, caused by the necrotrophic fungus Plenodomus tracheiphilus. With the main aim of identifying candidate genes involved in the response of citrus plants to “Mal secco”, we performed a de novo transcriptome analysis of rough lemon seedlings subjected to inoculation of P. tracheiphilus. The analysis of differential expressed genes (DEGs) highlighted a sharp response triggered by the pathogen as a total of 4986 significant DEGs (2865 genes up-regulated and 2121 down-regulated) have been revealed. The analysis of the most significantly enriched KEGG pathways indicated that a crucial role is played by genes involved in “Plant hormone signal transduction”, “Phenylpropanoid biosynthesis”, and “Carbon metabolism”. The main findings of this work are that under fungus challenge, the rough lemon genes involved both in the light harvesting and the photosynthetic electron flow were significantly down-regulated, thus probably inducing a shortage of energy for cellular functions. Moreover, the systemic acquired resistance (SAR) was activated through the induced salicylic acid cascade. Interestingly, RPM1 interacting protein 4, an essential positive regulator of plant defense, and BIR2, which is a negative regulator of basal level of immunity, have been identified thus representing useful targets for molecular breeding.

The causal agent of mal secco disease is the fungus Plenodomus tracheiphilus, mainly affecting lemon tree survival in the Mediterranean area. Using a fully compatible host-pathogen interaction, the aim of our work was to retrieve the fungus transcriptome by an RNA seq approach during infection of rough lemon (Citrus jambhiri Lush.) to identify crucial transcripts for pathogenesis establishment and progression. A total of 2438 clusters belonging to P. tracheiphilus were retrieved and classified into the GO and KEGG categories. Transcripts were categorized mainly within the “membrane”, “catalytic activity”, and “primary metabolic process” GO terms. Moreover, most of the transcripts are included in the “ribosome”, “carbon metabolism”, and “oxidative phosphorylation” KEGG categories. By focusing our attention on transcripts with FPKM values higher than the median, we were able to identify four main transcript groups functioning in (a) fungus cell wall remodeling and protection, (b) destroying plant defensive secondary metabolites, (c) optimizing fungus development and pathogenesis, and (d) toxin biosynthesis, thus indicating that a multifaceted strategy to subdue the host was executed.

To evaluate the safety and efficacy of the Orsiro sirolimus-eluting stent (Biotronik) in patients with ST-segment elevation myocardial infarction undergoing primary percutaneous coronary intervention (pPCI). Specific drug-eluting stent (DES) platforms might influence pPCI success rate in the mid-to-long term. Orsiro, a hybrid sirolimus DES with thin struts and a biodegradable polymer, may potentially cause less stent malapposition, stent-induced inflammation, and mechanical damage, improving clinical outcomes.We retrospectively enrolled all patients who received 1 or more Orsiro DES in the target vessel of pPCI at 9 Italian centers from January 2012 to March 2016. The primary endpoint was a device-oriented composite endpoint (DOCE) of cardiac death, any myocardial infarction clearly attributable to the intervention culprit vessel (TVMI), and ischemic-driven target-lesion revascularization (ID-TLR) at 1-year follow-up. Secondary endpoints were: (1) DOCE at 6-month and 3-year follow-up; (2) any definite/probable stent thrombosis; and (3) any major bleeding.The study cohort comprised 353 patients. At 1-year follow-up, we observed a 3.7% cumulative incidence of DOCE, consisting of 11 cardiac deaths (3.1%), 2 TVMIs (0.6%), and 2 ID-TLRs (0.6%). There was only 1 definite stent thrombosis (0.3%) and 8 bleedings (2.4%). Kaplan-Meier analysis showed DOCE-free survival rates of 96.6% at 6 months, 96.3% at 1 year, and 93.8% at 3 years.Our findings support the real-world safety and efficacy of the Orsiro stent for pPCI.

Background: Anthocyanin pigmentation characterizes a number of tissues of Citrus and its relatives. The gain and loss of pigmentation is intriguing and is inherited variously among species. Methods: Citrus germplasm was used to investigate the anthocyanin pigmentation of tissues never before considered, including stamen, style and stigma, and of young leaves, petals, rind and flesh of 28 genotypes belonging to 14 species. Citrus genotypes encompassed citron, lemon, sweet orange, lime, and Citrus relatives included Microcitrus, Murraya, and Severinia. A relative qRT-PCR analysis was carried out on the structural and regulatory genes: phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3′-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), anthocyanidin synthase (ANS), uridine diphosphate glucose flavonoid glucosyl-transferase (UFGT), glutathione S-transferase (GST), Ruby and Noemi. Image analysis and a genomic approach were employed to evaluate how the red pigmentation is inherited among tissues and species. Results: Pigmentation of young leaves and petals is specific to citron and its hybrids. Ruby controls the pigmentation of petals, but not of leaves. The red color of the rind and flesh is a trait that particularly characterizes a diversity of sweet oranges, citron hybrids and Citrus relatives. Color expression depends on external factors and also on developmental stage. The coloration of stamen and style is citron-specific, while a red stigma is exclusive to Moro orange and its hybrids. Conclusion: It is hypothesized that there is a relationship among Citrus species and genes controlling anthocyanin pigmentation.

The origin, development, and characteristics of the principal citrus scion varieties in commercial production are described. As citrus moved from its site of origin, the primitive, ancestral forms were selected resulting in the wide range of varieties and selections of varieties currently available today. Selection criteria differed according to local preference (traditions, customs, beliefs, etc.), and the perception of fruit quality differs according to regional preferences. Most citrus varieties in commercial production were developed through the selection of natural mutations, e.g., pummelos, grapefruit, lemons, limes, oranges, and many mandarin types, although induced mutations by irradiation have resulted in several seedless citrus varieties, whereas numerous mandarin hybrids are either man-made or naturally occurring.

This chapter describes traditional strategies used by breeding programs around the world, and how they have resulted in improved citrus varieties. The first part examines scion breeding objectives and the advantages/disadvantages of important techniques including somatic mutants, nucellar selection, diploid and polyploid hybridization, and mutation breeding. Seedlessness remains an enormous challenge for breeders and the available methods are discussed within the context of having to simultaneously maintain or improve a host of other critical traits. The second part describes the main steps leading to the release of improved rootstocks. The importance of hybridization with citrus relatives to introgress useful genes both in scions and rootstocks breeding programs is also emphasized. New diseases and increasing consumer expectations present additional challenges for breeding programs and more efficient phenotyping and molecular markers are required.

The recent discovery of a Roman ceramics manufacturing workshop at Montelabate (Perugia, Italy), in use from the first century BC until the late-fourth to fifth centuries AD, offers a unique opportunity to study the technical processes for producing Roman amphorae. Ancient and modern clays were sampled and analysed; they do not differ significantly, supporting the hypothesis of the exploitation of the rich local clay source that allowed a continuity of production. Characterization of the clays was performed using geotechnical methods (Atterberg limits and size distribution) and by thermogravimetric and differential thermogravimetric analysis, Fourier-transform infrared spectroscopy, X-ray diffraction and X-ray fluorescence analyses. The material was suitable for pottery making with the addition of calcite and quartz sand temper. Production waste and discarded materials as well as good-quality products were also analysed with the same methodology. It is therefore possible to reconstruct the ancient technology by defining the recipe for the production of the amphorae and their firing temperature on the basis of the decomposition of clay materials and the presence of newly formed minerals.

The geochemical evolution of olivine from primitive magmatic skarn environments has been studied by atmospheric pressure experiments carried out at 1250, 1200, and 1150 °C under QFM oxygen buffering conditions. The starting materials were three synthetic basalts (i.e., meltMg#78, meltMg#75, and meltMg#72) doped with variable amounts of CaCO3, in order to reproduce the natural concentration levels of CaO-rich magmas interacting with the skarn rock shells. Results from decarbonation experiments evidence that the crystallization of Fo-CaO-rich, NiO-poor olivines is more favored at higher temperatures when primitive basaltic magmas assimilate increasing amounts of carbonate materials. The number of large size Ca cations entering olivine crystal lattice is proportional to the amount of Ca-O-Si bonds available in the melt. Due to differences between Fe2+ and Mg cation radii, the Ca-Fe2+ substitutions into M2 crystallographic site are more facilitated than Ca Mg ones, thus enhancing the forsterite component in olivine. The partitioning behavior of Ni, Mg, Fe2+, Mn, and Ca between olivine and melt has been also investigated to better understand cation redistribution mechanisms at the magma‑carbonate reaction zone. In this context, some partitioning models from the literature have been refined to more accurately quantify the geochemical evolution of primitive skarn systems. Under the effect of CaCO3 assimilation, the partitioning of divalent cations, can be parameterized as a function of temperature, bulk composition (mostly, CaO and MgO contents in both olivine and melt) and melt structure (expressed as the number of non-bridging oxygens per tetrahedrally coordinated cations). Conversely, the exchange partition coefficients between Fe2+/Ca/Mn/Ni and Mg do not vary significantly as a function of temperature and meltMg#, due to the limited influence of these parameters on the melt structure. In turn, cation exchange reactions are primarily controlled by the strong depolymerizing effect of CaCO3 assimilation that increases the number of structural sites critically important to accommodating network-modifying cations in the melt phase. The comparison between cumulates and magmatic skarns from the Colli Albani Volcanic District (Italy) and experiments from this study provides quantitative constraints on the geochemical evolution of olivine phenocrysts and their melt inclusions as a function of carbonate assimilation.

Nowadays, the installation of efficient lighting sources and Information and Communications Technologies can provide economic benefits, energy efficiency, and visual comfort requirements. More advantages can be derived if the public lighting infrastructure integrates a smart grid. This study presents an experimental multifunctional infrastructure for public lighting, installed in Palermo. The system is able to provide smart lighting functions (hotspot Wi-Fi, video-surveillances, car and pedestrian access control, car parking monitoring and support for environmental monitoring). A remote control and monitoring platform called “Centro Servizi” processes the information coming from different installations as well as their status in real time, and sends commands to the devices (e.g. to control the luminous flux), each one provided with a machine to machine interface. Data can be reported either on the web or on a customised app. The study has shown the efficient operation of such new infrastructure and its capability to provide new functions and benefits to citizens, tourists, and public administration. Thus, this system represents a starting point for the implementation of many other lighting infrastructure features typical of a “smart city.&rdquo

Fruits of sixteen Tarocco sweet orange (Citrus sinensis [L.] Osbeck) clonal selections were compared for their long-period cold storage aptitude, for fresh fruit consumption. Selections were evaluated, for two consecutive seasons, at harvest (T0) and after 30 (T30), 60 (T60) days of storage at 5 ± 1 °C and 85–90% relative humidity (RH), followed by 7 days of shelf-life at 20±2 °C (T60+7). The following parameters were evaluated: fruit weight, peel and pulp color, initial and residual deformation, elasticity, firmness, juice yield, TSS, pH, TA, total anthocyanin content, and for each control: weight loss and physiological disorders (chilling injury and aging). The selections showed significant variability regarding the physiological disorders, especially at T60+7 with a range of 0.0–0.6 for chilling injury index and a range of values of 0–63% for aging. At T60+7 the weight loss range was between 10.0–14.3%. Regarding overall qualitative parameters, relevant changes were observed for firmness which decreased during the storage, with values ranging from 2.58-5.28 (T0) to 2.56–4.41 (T60+7), and for the anthocyanins which increased from a range of 1.67–39.71 (T0) to 30.47–114.86 mg/L (T60+7), with different values depending on the clonal selections. Based on our results, the clonal selections that showed a better aptitude to long-period cold storage were Gangi, 2B-12–9, Pedalino and Amantea, together with the reference cultivars Meli and Messina. These clones resulted particularly tolerant to chilling injury and aging, they maintained the firmness and the most relevant qualitative traits (TSS and TA) and showed the best fruit external appearance after one week of shelf-life (T60+7).

Sweet oranges are an important source of ascorbic acid (AsA). In this study, the content of AsA in the juice and leaves of four orange clonal selections, different in terms of maturity time and the presence/absence of anthocyanins, was correlated with the transcription levels of the main genes involved in the biosynthesis, recycling, and degradation pathways. Within each variety, differences in the above pathways and the AsA amount were found between the analysed tissues. Variations were also observed at different stages of fruit development and maturation. At the beginning of fruit development, AsA accumulation was attributable to the synergic action of l-galactose and Myo-inositol, while the l-gulose pathway was predominant between the end of fruit development and the beginning of ripening. In leaves, the l-galactose pathway appeared to play a major role in AsA accumulation, even though higher GalUr isoform expression suggests a synergistic contribution of both pathways in this tissue. In juice, the trend of the AsA content may be related to the decrease in the transcription levels of the GME, GDH, MyoOx, and GalUr12 genes. Newhall was the genotype that accumulated the most AsA. The difference between Newhall and the other varieties seems to be attributable to the GLDH, GalUr12, APX2, and DHAR3 genes.

The present study aimed to investigate the effects of alcoholic fermentation on the physicochemical and phytochemical characteristics of the tissues flavedo and albedo from five citrus fruits, namely grapefruit (Citrus paradisi), lemon (Citrus limon), and sweet oranges (Citrus sinensis) cvs. ‘Tarocco’, ‘Washington navel’, and ‘Moro’. The tissues were fermented with pure and mixed cultures of Saccharomyces cerevisiae, Brettanomyces bruxellensis, and Torulaspora delbrueckii. Flavedo and albedo increased the water holding capacity (WHC) and oil binding capacity (OBC) when subjected to fermentation. The treatment with B. bruxellensis resulted the most effective at enhancing WHC and OBC, e.g., lemon flavedo achieved the highest WHC (9.67 ± 0.15 g/g; P

The taxonomic classification of the genus Opuntia, which includes several cultivated species, is complicated mainly because of inadequate morphological descriptors, the common intra- and inter-generic hybridization and the relationships between phenotypic variation and ecological conditions. The phylogenetic relationships among 72 cultivated genotypes, either selected for fruit or forage production, and wild accessions belonging to approximately 15 different species of Opuntia were inferred using cytoplasmic markers. Previous studies indicated as the most important cultivated accessions for fruit production have a polyphyletic origin but polyploidy hampered their clear phylogenetic assignment based on nuclear markers. Cytoplasmic markers are considered helpful for their maternal inheritance and for overcoming the multiple gene copy problem in polyploid phylogenetics already reported in the Opuntia genus. In particular, we combined in this work capillary electrophoresis for newly designed cpSSRs and high resolution melting for SNV analyses to identify chloroplast (ndhF-rpl32, psbJ-petA, atpB-rbcL, matK, ycf1) and mitochondrial (rpl5) DNA markers in a selected group of genotypes. The results revealed the presence of polymorphisms in the predicted cpSSRs and SNVs and clearly evidenced that most of the studied genotypes were closely related to Opuntia ficus-indica (L.) Miller. Plastid markers identified 11 chlorotypes and 8 unique genotypes. Interestingly the analysis evidenced a multiple maternal phylogeny for the fleshy fruit varieties classified as O. ficus-indica. These results allow questioning of the reliability of the current classification based on morphological parameters and reveal the narrow genetic base of the most common cultivated opuntias for fruit production, while forage genotypes evidenced greater variability.

In spite of the increasing diffusion of tunnel boring machines, conventional tunnelling is still preferred for economic reasons in case of short tunnels, unconventional cross sections or irregular tunnel trajectories. In conventional tunnelling, the mechanical response of the tunnel front is a main concern and, when tunnels are excavated in cohesive soils, this is dominated by the time factor, related to geometry, to the mean excavation rate and to the hydro-mechanical properties of the materials involved. This is particularly evident during excavation standstill: front displacements progressively increase with time and, in many cases, the system response under long-term conditions becomes unstable. In conventional tunnelling, a common technique employed to improve the system response (under both short- and long-term conditions) is the installation of fibreglass tubes within the advance core. In this paper, the mechanical response of both unreinforced and reinforced deep tunnel fronts in cohesive soils is experimentally analysed. In particular, the results of a series of 1g small-scale tests, taking into account both the influence of the excavation rate (the unloading time) on the system response and the evolution with time of the tunnel face displacements, induced by a rapid reduction in the horizontal stress applied on the tunnel face, are reported.

Blood oranges have been cultivated in Italy since the early 19th century. Growers and breeders were able to identify and collect many somatic mutants, which differed in their major pomological traits, such as fruit size and firmness, pulp and peel pigmentation, and ripening period. Recently, in the framework of a genetic resource conservation program, most of the accessions collected at CREA in recent decades were planted in a single block to evaluate their characteristics. The presence of many selections in the same block, of the same age and grafted on the same rootstock is extremely important for a reliable characterization of the pomological characters, particularly pigmentation, which is strongly influenced by both the environment and the rootstock. The collection consists of 88 genotypes, including old lines and nucellar selections. Some commercial Tarocco, Moro and Sanguinello varieties were also included in the collection to facilitate comparison among clones. Moreover, the pomological features of 20 elite Tarocco clones cultivated nearby in the same experimental station were used as a further reference. The pomological characterization, based on 13 traits and performed in two consecutive years, revealed significant variability. Some genotypes showed strong year-to-year variability, mainly in fruit pigmentation, probably due to different climatic conditions during winter. Similarly, other traits such as fruit size, total soluble solids and acidity also varied among and within the varietal groups. These findings have a potential utility in identifying promising clones, which might fulfil the growing market needs such as higher anthocyanin content and longer shelf life.

Pain management is one of the main challenges in addressing the improved care of hospitalized newborns. The administration of oral sucrose with and without non-nutritive suction has been proposed as a nonpharmacological intervention to relieve procedural pain in newborns. The effects have not yet been well characterized. The aim of this study is to investigate, using skin conductance algesimeter (SCA) pain monitor index, the effects of 24% sucrose solution on pain perception during capillary and arterial blood sampling. It is a prospective, randomized controlled study: sucrose versus placebo. Sucrose was given orally to infants who were submitted to arterial or capillary sampling. The SCA was measured during, and for 3 min before and after the intervention. Fifty-six infants were enrolled: 31 in the sucrose group and 25 in the placebo group. SCA showed that the measurement of peaks per second of pain during and 3 min after the procedures was lower in the sucrose group than the placebo group and that this difference was statistically significant (

This study aimed to evaluate the effects of fermentation with lactic acid bacteria (LAB) on organic acids, ascorbic acid, phenolic compounds, anthocyanins, and carotenoids of sweet orange (Citrus sinensis) juices. Juices from ‘Tarocco’ and ‘Washington navel’ oranges were fermented at 30 °C for 48 h with L. plantarum, L. rhamnosus, L. paracasei, and L. brevis without supplementation. The juice from ‘Washington navel’ resulted the best medium to grow LAB, especially L. brevis (1.98 ± 0.36 log CFU mL−1 at 48 h). Malic acid was thoroughly converted into lactic acid. Ascorbic acid increased in ‘Tarocco’ juice, peaking to 295 ± 25.3 g L−1 when fermented by L. brevis. The effects of LAB on phenolic compounds were matrix-specific, e.g., L. rhamnosus increased the levels of phenolics in ‘Washington navel’ juice (36.8 ± 1.51 mg L−1; p

Citrus rootstocks are highly polyembryonic. Seeds usually produce many nucellar seedlings that are identical to the mother plant and sometimes one zygotic plant. Nucellar seedlings hamper the identification of hybrids if the morphological markers that distinguish sexual and nucellar plants are missing, and competition among embryos might impair zygotic embryo viability. Accurate morphological and/or molecular screening methods are needed to identify and discard the nucellar plants, which is labor-intensive and increases the costs of rootstock breeding programs. This study describes an affordable and reliable method for an early identification of hybrids in progenies derived from highly polyembryonic female parents based on DNA isolation from pooled leaf samples (2 to 4 plants per sample) coupled with a high resolution melting (HRM) analysis at two SNP loci. The method was validated on more than one thousand progeny derived from the crosses alemow ( Citrus macrophylla Wester) × Swingle citrumelo [ Citrus paradisi Macf. × Poncirus trifoliata (L.) Raf.] (A × S), and Cleopatra mandarin ( Citrus reshni Hort. ex Tan) × Swingle citrumelo (C × S). Bulking reduced the number of DNA isolations for population genotyping, while HRM unequivocally identified, in a single step, the bulks containing the unifoliate hybrids. A consistent difference was found between A × S and C × S populations in the frequency of uni- and multifoliate hybrids (69% unifoliate in A × S vs. 10% in C × S), suggesting that the combination of parents, rather than just the male parent carrying the trifoliate character, influenced the early multifoliate leaf expression. Moreover, several multifoliate nucellar seedlings were found among the C × S progenies, making the multifoliate leaf character inappropriate for the early identification of all hybrids.

Congenital anomalies of the coronary arteries, although uncommon, have the potential to cause serious myocardial damage, ischemic cardiomyopathy, and sudden cardiac death. We report a rare case of origin of all three coronaries from the right sinus of Valsalva.

Earthen dwellings are part of the vernacular architecture and cultural heritage of many countries in the Mediterranean region, and a renovated interest towards these types of structures is now widespread in the same countries mainly due to sustainability reasons. However, poor resistance to weathering compromises their durability and their popular acceptance. A fascinating option to improve the resistance of earthen structures while preserving their environmental sustainability is to use locally available waste materials as stabilisers. In this paper, the evolution over time of the mechanical resistance of rammed earth stabilised with residues from widespread industrial processes (i.e. fly ash from coal combustion and calcium carbide residue from acetylene production) was investigated. Waste-stabilisation prompted optimal long-term mechanical resistance; on the other hand, laboratory samples exhibited low compressive strengths in the short-term when cured under standard conditions. The addition of a supplementary industrial residue (i.e. gypsum from flue gas desulfurization) was explored to enhance the early-age strength. Results confirmed the short-term strength benefits induced by gypsum addition.

Italian germplasm is characterized by a wide diversity rapidly developed determining a massive genetic pool of cultivars in several growing areas. On the whole, regions of southern Italy, and in particular Sicily, were considered as one of the main trade routes along which almond was spread throughout the shores of the Mediterranean Sea. In this work, 9 SSR markers have been used to analyze 113 almond cultivars and accessions coming from ex-situ conservation, including most of almond genotypes spread in Sicily and Apulia and foreign cultivars from Mediterranean, American and Australian areas in order to determine the level of genetic diversity within Italian genotypes and elucidate phylogenetic and possible parentage relationships between Italian accessions and foreign germplasm. Distance and model-based analyses revealed a high level of variability both among and within geographical areas. In total, 159 alleles were detected altogether, which identified the average of 44 genotypes in the population investigated. The wide genetic diversity detected in the Italian almond cultivars represents a valuable source of variability to be applied for breeding. These data represent the first overview of genetic diversity among Italian almond cultivars using molecular markers.

Telomeres have been postulated as a universal clock that shortens in parallel with cellular aging. They are specialized DNA-protein structures at the ends of chromosome with remarkable functions--preventing their recognition as double-stranded DNA breaks, protecting their recombination and degradation, and avoiding a DNA damage cellular response. Telomere shortening is currently considered the best aging marker, but is also a predictor for age-related diseases, including cardiovascular diseases. Biological age clearly seems to be a better predictor of vascular risk rather than chronological age. This concept is supported by key assumptions that peripheral blood leukocyte telomere content accurately reflects that of the vascular wall and its decrease is associated with premature vascular disease. Thus, we are analyzing whether the mean of blood leukocyte telomere length might also be a predictor for sporadic thoracic aortic aneurysm (S-TAA). The preliminary results seem to be promising. Shorter telomeres were detected in patients than in controls. Thus, mean of blood leukocyte telomere length could contribute to identify individuals at S-TAA risk.

Aim To compare the incidence, and risk factors, in-hospital and at the 18-month prognosis of contrast-induced nephropathy (CIN) according to the definition utilized: as an increase in serum creatinine (Scr) ≥0.5 mg/dL (CIN 1) or as an increase in Scr ≥25% above baseline values (CIN 2). Methods and Results We prospectively evaluated CIN according to two different definitions in 150 patients who underwent percutaneous coronary intervention (PCI) in simple lesions employing a low-medium dose of contrast media. Incidence of CIN was higher using the CIN 2 definition than CIN 1 (9.3% vs. 4%; p=0.0133). Patients with CIN 1 had a higher incidence of chronic kidney disease (CKD) (66.7% vs. 13.9%; p=0.006), higher mean serum creatinine levels (1.35±0.42 vs. 0.98±0.35; p=0.001) and lower mean eGFR levels (58.3±19.6 vs. 84±25.9; p=0.002). Patients with CIN 2 had a higher incidence of anemia (57.1% vs. 30.9%; p=0.049) and a higher mean contrast media volume was used (142.6±62.2 mL vs. 110.6±57.2 mL; p=0.05). In the multivariate analysis the only significant variable associated with CIN (CIN 2) was a higher volume of contrast media (OR=1.01; p=0.04). There were no differences in the major in-hospital events, but patients with CIN (both definitions) had a longer in-hospital stay. A persistent rise in serum creatinine at discharge occurred in only one patient. There were no differences between patients with and without CIN in events at the follow-up, with the exception of an increased risk of new hospitalization in patients with CIN 2. Conclusion After PCI employing low-medium dose of contrast media the incidence of CIN varied according to the definition used. Neither of the two definitions offers additional information compared with the other. Chronic kidney disease and baseline parameters of renal function are the risk factors associated with CIN 1; anemia and higher volume of contrast media are associated with CIN 2.

Opuntia spp. belong to the Cactaceae family and are native to Central America. The most economically important species is O. ficus indica, cultivated both for fruits and cladodes. The genus includes other important edible species (from diploid to octoploid) that occur worldwide as either wild or cultivated species in many arid or semiarid areas (e.g., the Mediterranean region). Several accessions are cultivated in different growing regions, but little is known about their ancestries and levels of genetic diversity. The aim of this study was to investigate the level of intraspecific genetic diversity among O. ficus indica cultivated varieties and some related species. Specifically, six highly polymorphic simple sequence repeats (SSR) and two expressed sequence tag (EST)-SSR loci were investigated in 62 wild and cultivated genotypes belonging to 16 Opuntia species. The clusters identified by the distance and model-based analyses clearly separated the wild opuntias from the cultivated ones. However, the O. ficus indica accessions did not cluster separately from other arborescent cactus pear species, such as O. amyclaea, O. megacantha, O. streptacantha, O. fusicaulis, and O. albicarpa, indicating that their current taxonomical classifications do not fit with their genetic variability. In general, the genotypes cultivated in Mexico showed high levels of diversity, whereas most of the spineless accessions collected in other countries had a very narrow genetic base. This study increases our knowledge of the variability among some of the most diffused Opuntia cultivated accessions. This study also points to the inconsistencies of previous taxonomical genotype assignments that were based solely on morphological characteristics.

Acute myocardial infarction (AMI) in young adult presents a typical pattern of risk factors, clinical, angiographic and prognostic characteristics. In the last years we demonstrated that hemorheological profile is altered in these patients in a persistent way and independently of the number of risk factors and of the extent of coronary lesions. Thus, the hyperviscosity syndrome following AMI could be considered an intrinsic characteristic of these patients. Consequently it is possible to hypothesise the presence of a genetic background at the origin of this predisposition. If this background is able to influence the risk of ischemic heart disease, this should be particularly evident in young subjects. Since inflammatory mechanisms play a central role in mediating all phases of atherosclerosis, genes encoding for inflammatory or anti-inflammatory molecules are candidates for the risk of developing atherosclerosis. As atherosclerosis is the first cause of mortality in Western countries and if pro-inflammatory genotypes contribute to risk of coronary heart disease, alleles associated to disease susceptibility should not be included in the genetic background favouring longevity: People genetically predisposed to a weak inflammatory activity have fewer chances to develop cardiovascular disease and, therefore, have better chance for a long-life. According to this hypothesis, we studied in our population of young patients with AMI, the distribution of some polymorphisms influencing a inflammation and found an higher prevalence of pro-inflammatory polymorphisms (SNP A2080G of pyrin gene, SNP Gly670Arg of PECAM gene, C1019T of Cx 37 gene, SNP G1059C of PCR gene) and a lower prevalence of anti-inflammatory polymorphisms (Asp299Gly of TLR4 gene, SNP -1082 G/A of IL10 gene, CCR5Δ32). Results of these studies show that early myocardial infarction could be associated with a genetic predisposition to an intense inflammatory response, associated also to an hyperviscosity syndrome.

Background Late survival and freedom from retreatment on the descending aorta was evaluated after ascending aortic repair for type A acute aortic dissection (TAAAD). Methods Between March 1992 and January 2006, 189 TAAAD patients (mean age, 52 ± 11; range, 17 to 83 years) were included; of these, 58 had a patent false lumen, and 49 had Marfan syndrome. The descending aorta was evaluated postoperatively with computed tomography (CT). Late outcomes were assessed by Cox regression analysis and actuarial survival and freedom from retreatment by the Kaplan-Meier method. Mean follow-up was 88 ± 44 months. Results There were 38 (20%) late deaths. At 10 years, survival was 89.8% ± 2.1% for patients with an occluded false lumen and 59.8% ± 3.5% for patients with a patent false lumen ( p = 0.001), and freedom from retreatment on the descending aorta was 94.2% ± 3.1% for an occluded false lumen and 63.7% ± 2.6% for a patent false lumen ( p p = 0.002) and a patent false lumen ( p = 0.001) were predictors for late death. Patent false lumen ( p = 0.0001), Marfan syndrome ( p = 0.03), and descending aortic diameter 4.5 cm or larger ( p = 0.002) were predictors for retreatment. Conclusions A patent false lumen was a predictor for late death and retreatment on the descending aorta. Marfan syndrome and aortic size exceeding 4.5 cm were predictors for late retreatment. These patients require very close follow-up and a plan for retreatment on the descending aorta to prevent sudden rupture and late death.