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The myeloma overexpressed gene (MYEOV) has been proposed to be a proto-oncogene due to high RNA transcript levels found in multiple cancers, including myeloma, breast, lung, pancreas and esophageal cancer. The presence of an open reading frame (ORF) in humans and other primates suggests protein-coding potential. Yet, we still lack evidence of a functional MYEOV protein. It remains undetermined how MYEOV overexpression affects cancerous tissues. In this work, we show that MYEOV has likely originated and may still function as an enhancer, regulating CCND1 and LTO1. Firstly, MYEOV 3′ enhancer activity was confirmed in humans using publicly available ATAC-STARR-seq data, performed on B-cell-derived GM12878 cells. We detected enhancer histone marks H3K4me1 and H3K27ac overlapping MYEOV in multiple healthy human tissues, which include B cells, liver and lung tissue. The analysis of 3D genome datasets revealed chromatin interactions between a MYEOV-3′-putative enhancer and the proto-oncogene CCND1. BLAST searches and multi-sequence alignment results showed that DNA sequence from this human enhancer element is conserved from the amphibians/amniotes divergence, with a 273 bp conserved region also found in all mammals, and even in chickens, where it is consistently located near the corresponding CCND1 orthologues. Furthermore, we observed conservation of an active enhancer state in the MYEOV orthologues of four non-human primates, dogs, rats, and mice. When studying this homologous region in mice, where the ORF of MYEOV is absent, we not only observed an enhancer chromatin state but also found interactions between the mouse enhancer homolog and Ccnd1 using 3D-genome interaction data. This is similar to the interaction observed in humans and, interestingly, coincides with CTCF binding sites in both species. Taken together, this suggests that MYEOV is a primate-specific gene with a de novo ORF that originated at an evolutionarily older enhancer region. This deeply conserved putative enhancer element could regulate CCND1 in both humans and mice, opening the possibility of studying MYEOV regulatory functions in cancer using non-primate animal models.

Despite the plasma proteome being able to provide a unique insight into the health and disease status of individuals, holding singular promise as a source of protein biomarkers that could be pivotal in the context of personalized medicine, only around 100 proteins covering a few human conditions have been approved as biomarkers by the US Food and Drug Administration (FDA) so far. Mass spectrometry (MS) currently has enormous potential for high-throughput analysis in clinical research; however, plasma proteomics remains challenging mainly due to the wide dynamic range of plasma protein abundances and the time-consuming procedures required. We applied a new MS-based multiplexed proteomics workflow to quantitate proteins, encompassing 67 FDA-approved biomarkers, in >1300 human plasma samples from a clinical cohort. Our results indicate that this workflow is suitable for large-scale clinical studies, showing good accuracy and reproducibility (coefficient of variation (CV) < 20 for 90% of the proteins). Furthermore, we identified plasma signature proteins (stable in time on an individual basis), stable proteins (exhibiting low biological variability and high temporal stability), and highly variable proteins (with low temporal stability) that can be used for personalized health monitoring and medicine.

Summary: Topologically associated domains (TADs) are interaction subnetworks of chromosomal regions in 3D genomes. TAD boundaries frequently coincide with genome breaks while boundary deletion is under negative selection, suggesting that TADs may facilitate genome rearrangements and evolution. We show that genes co-localize by evolutionary age in humans and mice, resulting in TADs having different proportions of younger and older genes. We observe a major transition in the age co-localization patterns between the genes born during vertebrate whole-genome duplications (WGDs) or before and those born afterward. We also find that genes recently duplicated in primates and rodents are more frequently essential when they are located in old-enriched TADs and interact with genes that last duplicated during the WGD. Therefore, the evolutionary relevance of recent genes may increase when located in TADs with established regulatory networks. Our data suggest that TADs could play a role in organizing ancestral functions and evolutionary novelty.

An estimate of the environmental impact of dairy farms in Northern Italy producing milk for hard cheese (protected designation of origin) has been obtained through a comprehensive life cycle assessment. The estimate focused on climate change (CC) and photochemical ozone creation potential (POCP) indicators, which were evaluated according to the Intergovernmental Panel on Climate Change (IPCC) guidelines and interpreted with the aid of the feeds’ composition evaluated using near-infrared reflectance spectroscopy (Foss NIR-System 5000) as well as with a diet evaluation according to the NRC (National Research Council) or the CNCPS (Cornell Net Carbohydrate and Protein System) nutrient requirement modeling. Herds were classified into high-, mid-, and low-performing based on the daily milk yield per cow. A lower impact on indicators was observed as herd performance increased. The high-performing herds had a lower contribution from enteric fermentation (6.30 × 10−1 kgCO2-eq), and the more milk that they produced allowed for a differentiation of CC from land use and transformation (2.39 × 10−1 kgCO2-eq), compared to low-performing herds (3.66 × 10−1 kgCO2-eq). Compared to the IPCC approach, the CC and POCP indicator estimates were reduced when addressing the feed’s quality, particularly in mid- and high-performing herds. The results could be helpful in the dairy sector as they provide an insight into how diet quality affects the environmental impact of milk.

Additive manufacturing workflows generate large amounts of data in each phase, which can be very useful for monitoring process performance and predicting the quality of the finished part if used correctly. In this paper, a framework is presented that utilises machine learning methods to predict porosity defects in printed parts. Data from process settings, in-process sensor readings, and post-process computed tomography scans are first aligned and discretised using a voxelisation approach to create a training dataset. A multi-step classification system is then proposed to classify the presence and type of porosity in a voxel, which can then be utilised to find the distribution of porosity within the build volume. Titanium parts were printed using a laser powder bed fusion system. Two discretisation techniques based on voxelisation were utilised: a defect-centric and a uniform discretisation method. Different machine learning models, feature sets, and other parameters were also tested. Promising results were achieved in identifying porous voxels; however, the accuracy of the classification requires improvement before being applied industrially. The potential of the voxelisation-based framework for this application and its ability to incorporate data from different stages of the additive manufacturing workflow as well as different machine learning models was clearly demonstrated.

The use of microbial biostimulants in agriculture is recognized as a sustainable approach to promoting crop productivity and quality due to improved nutrient uptake, enhanced stress tolerance, and improved ability to cope with non-optimal environments. The present paper aimed to comparatively investigate the effect of seven different commercial mycorrhizal-based treatments in terms of yield, phytochemical components, and technological traits of Malvasia di Candia Aromatica grape (Vitis vinifera L.) plants. Metabolomic analysis and photosynthetic performance were first investigated in leaves to point out biochemical differences related to plant growth. Higher photosynthetic efficiency and better PSII functioning were found in biostimulant-treated vines, reflecting an overall decrease in photoinhibition compared to untreated plants. Untargeted metabolomics followed by multivariate statistics highlighted a robust reprogramming of primary (lipids) and secondary (alkaloids and terpenoids) metabolites in treated plants. The analysis of berry yield and chemical components exhibited significant differences depending on the biostimulant product. Generally, berries obtained from treated plants displayed improved contents of polyphenols and sugars, while yield remained unchanged. These results elucidated the significant role of microbial biostimulants in determining the quality of grape berries and eliciting biochemical changes in vines.

Italy is Europe's leading rice producer, with over half of total production almost totally concentrated in the north-western part of the Padana plain. In this area, rice irrigation has traditionally been carried out by wet seeding and continuous flooding. The replacement of this technique with the dry seeding and delayed flooding in the last 15 years brought economic benefits to farmers but also strong impacts to the water resource system of the area, characterized by a strong interaction between irrigation and shallow aquifer levels. Wet seeding and Alternate Wetting and Drying (AWD) could be seen as an alternative irrigation strategy; however, its environmental, economic and social sustainability must be investigated before supporting the dissemination of this “water-saving” irrigation technique over the territory in cooperation with decision-makers and water resource managers. In this paper, the results of an experimental activity carried out in a platform established in the main Italian rice district to compare three irrigation strategies (wet seeding and traditional flooding—WFL, dry seeding and delayed flooding from around the 3-leaf stage—DFL, wet seeding and alternated wetting and drying from the tillering stage—AWD) and data further provided by the farm holder (agronomic inputs and economic costs for the three options) were used to assess their economic and environmental sustainability through an assessment procedure based on quantitative indicators. Interviews with rice growers in the area were used to identify barriers to the adoption of AWD and subsequently propose ways to overcome them. AWD proved to be economically viable and to reduce the irrigation need in the peak month (June) without significantly affecting rice yield or quality (cadmium and arsenic content in rice grain). Additionally, it guarantees the recharging of the phreatic aquifer in the first period of the irrigation season, while reducing the methane (CH4) emissions, facing another significant environmental challenge of rice cultivation. Rice growers in the region stated their willingness to adopt AWD, although they would need technical, technological and financial support. The assessment procedure adopted in this paper proved to be effective for comparing rice irrigation techniques and is available for further studies.

The environmental impacts of the dairy industry, particularly global warming, are heavily influenced by milk production. Thus, there is an urgent need for farm-level actions and opportunities for improvement, implying mitigation strategies. The aim of this paper is to investigate five possible mitigation actions at the dairy farm and which one the farmers were willing to adopt: management and distribution of livestock manure and fertilizers, anaerobic manure treatment, optimization of the herd composition, feed quality, and heat recovery. A life cycle assessment was conducted on 63 farms using the product environmental footprint approach. The latter was divided into four quartiles, from which four representative farms were selected. For each farm, three scenarios have been analyzed considering the reference impact (reference scenario), the application of the mitigation actions (best-case scenario), and what farmers would implement (realistic scenario). Overall, the most effective mitigation actions in the best-case scenario were anaerobic manure treatment and the management and distribution of livestock manure and fertilizers, showing a potential reduction in total environmental impacts of 7–9% and 6–7%, respectively. Farmers’ responses indicated a willingness to implement the latter mitigation strategy better. The optimization of the herd composition, feed quality, and heat recovery reported a range impact reduction between 0.01–5%.

The application of microbial biostimulants to plants has revealed positive effects related to nutrients uptake, stress tolerance, root development and phenological growth. However, little information is available exploiting the potential synergistic biostimulant action of microbes on the functional quality of the yields. The current research elucidated the effect of single or coupled action of biostimulants, associated with either optimal or reduced nitrogen application, on the functional quality of tomato fruits. Chemical assays and untargeted metabolomics were applied to investigate Rhizoglomus irregulare and Funneliformis mosseae administration (both being arbuscular mycorrhiza, AMF), under optimal or low N input conditions, alone or coupled to Trichoderma atroviride application. The coupling of AMF and Trichoderma fungal inoculations resulted in a synergistic biostimulant effect on tomato fruits under sub-optimal fertility, revealing improved concentrations of carotenoid compounds—B-carotene (0.647 ± 0.243 mg/100 g), Z-carotene (0.021 ± 0.021 mg/100 g), 13-z-lycopene (0.145 ± 0.052 mg/100 g) and all-trans-lycopene (12.586 ± 1.511 mg/100 g), and increased values for total phenolic content (12.9 ± 2.9 mgGAE/g), total antioxidant activity (phosphomolybdenum, 0.9 ± 0.2 mmolTE/g), radical scavenging activity (DPPH, 3.4 ± 3.7 mgTE/g), reducing power (FRAP, 23.6 ± 6.3 mgTE/g and CUPRAC, 37.4 ± 7.6 mg TE/g), and enzyme inhibitory activity (AChE, 2.4 ± 0.1 mg GALAE/g), when compared to control. However, evidence of carotenoid and bioactive compounds were exclusively observed under the sub-optimal fertility and no significant differences could be observed between the biostimulant treatment and control under optimal fertility.

In this work, the impact of terroir, cultivar, seasonality, and farming systems on functional traits of tomato was hierarchically investigated. Untargeted metabolomics, antioxidant capacity, colorimetric assays, and enzyme inhibition were determined. The total phenolic and carotenoid contents significantly varied between growing years, whereas an interaction between the farming system and growing year (p < 0.01) was observed for total phenolics, carotenoids, and flavonoids, and for acetylcholinesterase inhibition. Hierarchical clustering showed that geographical origin and growing year were the major contributors to the differences in phytochemical profiles. Nonetheless, supervised modeling allowed highlighting the effect of the farming system. Several antioxidants (L-ascorbic acid, α-tocopherol, and 7,3′,4′-trihydroxyflavone) decreased, whereas the alkaloid emetine and phytoalexin phenolics increased under organic farming. Taken together, our findings indicate that cultivar and pedo-climatic conditions are the main determinants for the functional quality of tomato, whereas the farming system plays a detectable but hierarchically lower.

The present study was designed to evaluate the functional potential of common duckweed (Lemna minor L.) as a source of bioactive compounds of nutraceutical interest. The untargeted profiling of the bioactive components of common duckweed was carried out through ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS), in parallel with assessing in vitro antioxidant and enzymatic inhibition properties. The optimization of extraction parameters was determined using the response surface methodology (RSM) through a 3-factor central composite design. The process parameters included extraction temperature, % of ethanol, and ultrasound power, while the response variables were the phenolic content (considering each main phenolic class), total glucosinolates, total carotenoids, the antioxidant potential, and enzyme inhibition activities. The results revealed that common duckweed was a rich source of carotenoids and total flavonoids (mainly flavones and flavonols), followed by phenolic acids, low-molecular-weight phenolics, and glucosinolates. Interestingly, the total flavones, total flavonols and total carotenoid equivalents showed the highest and most positive correlation values with the bioactive properties measured. Finally, the combined RSM approach and unsupervised statistics allowed us to point out the pivotal impact of ethanol percentage in the extraction solvent to recover the highest amounts of bioactive compounds efficiently.

Changes in melanocytic naevi and development of new naevi have been reported in pregnant women. The association between pregnancy and melanoma is a controversial topic. We conducted this review to identify the dermatoscopic changes that occur in naevi during pregnancy that could facilitate in distinguishing benign from suspicious lesions. Medline, Scopus, and Embase datasets were reviewed for clinical studies on dermatoscopic characteristics of melanoma and naevus in pregnancy. Six cohort studies with a total of 258 patients with 1,167 skin lesions that were examined fulfilled the conditions to be included in the review. None of the patients developed melanoma. Development of new naevi, when reported, was observed in less than half of the participants. The most frequent observed dermatoscopic change among the studies was the increase in the number of dots. Development of new vessels, hypo- and hyperpigmentations and changes in the pigment network were common described changes. The included studies were heterogeneous not allowing head-to-head comparisons between them. Robust and larger studies of dermatoscopic evaluation of naevi in pregnant women are needed to determine high-risk dermatoscopic characteristics.

Metal trace elements (MTE) can damage crops if present in excessive amounts in the environment. This research investigated the effect of a plant extract of an aquatic species, Lemna minor L. (duckweed) (LE), on the ability of maize to cope with copper (Cu) toxicity. LE reversed the effects of Cu2+ on photosynthetic activity (Pn), evapotranspiration (E), stomatal conductance (gs), sub-stomatal CO2 concentration (Ci) and biomass which did not differ from the untreated controls. LE did not regulate the amount of copper in maize leaves, but compared to Cu-treated samples, the extract decreased the hydrogen peroxide (H2O2; −26% on average) and malondialdehyde (MDA; −47% on average) content, regardless of the dosage applied. Furthermore, the activity of antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX) and catalase (CAT) was significantly increased by LE compared to samples treated with Cu alone. Untargeted metabolomic profiling revealed that LE activated maize secondary metabolism, eliciting the content of non-enzymatic antioxidants (flavonoids, glutathione and glutathione-related compounds, tocopherols and tocotrienols) and modulating plant stress-related hormones (brassinosteroids and ABA derivatives). The results of this study are promising and pave the way for using duckweed as a biostimulant to trigger beneficial effects in maize and increase its resistance to MTEs.

In severe viral pneumonia, including Coronavirus disease 2019 (COVID-19), the viral replication phase is often followed by hyperinflammation, which can lead to acute respiratory distress syndrome, multi-organ failure, and death. We previously demonstrated that alpha-1 adrenergic receptor (⍺1-AR) antagonists can prevent hyperinflammation and death in mice. Here, we conducted retrospective analyses in two cohorts of patients with acute respiratory distress (ARD, n = 18,547) and three cohorts with pneumonia (n = 400,907). Federated across two ARD cohorts, we find that patients exposed to ⍺1-AR antagonists, as compared to unexposed patients, had a 34% relative risk reduction for mechanical ventilation and death (OR = 0.70, p = 0.021). We replicated these methods on three pneumonia cohorts, all with similar effects on both outcomes. All results were robust to sensitivity analyses. These results highlight the urgent need for prospective trials testing whether prophylactic use of ⍺1-AR antagonists ameliorates lower respiratory tract infection-associated hyperinflammation and death, as observed in COVID-19.

The reconstitution is a pedotecnique producing environmental proper and fertile Technosols, applying chemical-mechanical treatment to alluvial sediments, degraded soils and pedomaterials included waste by different productive processes. By means of reconstitution, the environmental restoration of the covering degraded soil of a closed landfill near Piacenza is made (LIFE10 ENV/IT/000400 NEW LIFE). In order to assess this environmental restoration, LCC and FCC are calculated on 5 soil samples before and after reconstitution. The results, which highlight the transition from worst to best LCC and FCC classes show how reconstitution was able to convert the environmental and agronomic conditions from soil have very severe limitations that restrict the choice of plants or require very careful management, and that limit or restrict its use mainly to pasture, range, forestland, or wildlife food and cover, to soil have moderate limitations that restrict the choice of plants or that require moderate conservation practices or have optimum fertility.

Plant growth promoting rhizobacteria (PGPR) can display several plant-beneficial properties, including support to plant nutrition, regulation of plant growth, and biocontrol of pests. Mechanisms behind these effects are directly related to the presence and expression of specific genes, and different PGPR strains can be differentiated by the presence of different genes. In this study we reported a comprehensive evaluation of a novel PGPR Klebsiella variicola UC4115 from the field to the lab, and from the lab to the plant. The isolate from tomato field was screened in-vitro for different activities related to plant nutrition and growth regulation as well as for antifungal traits. We performed a functional annotation of genes contributing to plant-beneficial functions previously tested in-vitro. Furthermore, the in-vitro characterization, the whole genome sequencing and annotation of K. variicola UC4115, were compared with the well-known PGPR Azospirillum brasilense strain Sp7. This novel comparative analysis revealed different accumulation of plant-beneficial functions contributing genes, and the presence of different genes that accomplished the same functions. Greenhouse assays on tomato seedlings from BBCH 11–12 to BBCH > 14 were performed under either organic or conventional management. In each of them, three PGPR inoculations (control, K. variicola UC4115, A. brasilense Sp7) were applied at either seed-, root-, and seed plus root level. Results confirmed the PGP potential of K. variicola UC4115; in particular, its high value potential as indole-3-acetic acid producer was observed in increasing of root length density and diameter class length parameters. While, in general, A. brasilense Sp7 had a greater effect on biomass, probably due to its high ability as nitrogen-fixing bacteria. For K. variicola UC4115, the most consistent data were noticed under organic management, with application at seed level. While, A. brasilense Sp7 showed the greatest performance under conventional management. Our data highlight the necessity to tailor the selected PGPR, with the mode of inoculation and the crop-soil combination.

Current farm systems rely on the use of Plant Protection Products (PPP) to secure high productivity and control threats to the quality of the crops. However, PPP use may have considerable impacts on human health and the environment. A study protocol is presented aiming to determine the occurrence and levels of PPP residues in plants (crops), animals (livestock), humans and other non-target species (ecosystem representatives) for exposure modelling and impact assessment. To achieve this, we designed a cross-sectional study to compare conventional and organic farm systems across Europe. Environmental and biological samples were/are being/will be collected during the 2021 growing season, at 10 case study sites in Europe covering a range of climate zones and crops. An additional study site in Argentina will inform the impact of PPP use on growing soybean which is an important European protein-source in animal feed. We will study the impact of PPP mixtures using an integrated risk assessment methodology. The fate of PPP in environmental media (soil, water and air) and in the homes of farmers will be monitored. This will be complemented by biomonitoring to estimate PPP uptake by humans and farm animals (cow, goat, sheep and chicken), and by collection of samples from non-target species (earthworms, fish, aquatic and terrestrial macroinvertebrates, bats, and farm cats). We will use data on PPP residues in environmental and biological matrices to estimate exposures by modelling. These exposure estimates together with health and toxicity data will be used to predict the impact of PPP use on environment, plant, animal and human health. The outcome of this study will then be integrated with socio-economic information leading to an overall assessment used to identify transition pathways towards more sustainable plant protection and inform decision makers, practitioners and other stakeholders regarding farming practices and land use policy.

Sweet basil (Ocimum basilicum L.) is one of the most produced aromatic herbs in the world, exploiting hydroponic systems. It has been widely assessed that macronutrients, like nitrogen (N) and sulfur (S), can strongly affect the organoleptic qualities of agricultural products, thus influencing their nutraceutical value. In addition, plant-growth-promoting rhizobacteria (PGPR) have been shown to affect plant growth and quality. Azospirillum brasilense is a PGPR able to colonize the root system of different crops, promoting their growth and development and influencing the acquisition of mineral nutrients. On the bases of these observations, we aimed at investigating the impact of both mineral nutrients supply and rhizobacteria inoculation on the nutraceutical value on two different sweet basil varieties, i.e., Genovese and Red Rubin. To these objectives, basil plants have been grown in hydroponics, with nutrient solutions fortified for the concentration of either S or N, supplied as SO42– or NO3–, respectively. In addition, plants were either non-inoculated or inoculated with A. brasilense. At harvest, basil plants were assessed for the yield and the nutraceutical properties of the edible parts. The cultivation of basil plants in the fortified nutrient solutions showed a general increasing trend in the accumulation of the fresh biomass, albeit the inoculation with A. brasilense did not further promote the growth. The metabolomic analyses disclosed a strong effect of treatments on the differential accumulation of metabolites in basil leaves, producing the modulation of more than 400 compounds belonging to the secondary metabolism, as phenylpropanoids, isoprenoids, alkaloids, several flavonoids, and terpenoids. The primary metabolism that resulted was also influenced by the treatments showing changes in the fatty acid, carbohydrates, and amino acids metabolism. The amino acid analysis revealed that the treatments induced an increase in arginine (Arg) content in the leaves, which has been shown to have beneficial effects on human health. In conclusion, between the two cultivars studied, Red Rubin displayed the most positive effect in terms of nutritional value, which was further enhanced following A. brasilense inoculation.

The elicitation of plant secondary metabolism may offer interesting opportunities in the framework of sustainable approaches in plant science and in terms of their ability to prime resistance to biotic and abiotic stressors. The broad metabolic reprogramming triggered by different molecular elicitors, namely salicylate (SA), polyamines (PAs), and chitosan, was comprehensively investigated using a metabolomics approach and the tomato (Solanum lycopersicum L.) as the model crop. Six different treatments were compared: a negative control (no treatments), a second negative control treated with 1 M acetic acid (the reference for chitosan, since chitosan was solubilized in acetic acid), and four molecular elicitors, 1 mM 2,1,3-benzothiadiazole (BTH, a positive control), 10 mg/mL chitosan, 0.01 mM SA, and a 0.1 mM PA (putrescine, spermidine, and spermine). All treatments determined a slight increase in biomass, in particular following PA treatment. A broad reprogramming of secondary metabolism could be observed, including membrane lipid remodeling, phenylpropanoid antioxidants, and phytohormone crosstalk. Overall, our results suggest that PAs, SA, and BTH shared a systemic acquired resistance (SAR)-related response, whereas chitosan induced a more distinct induced systemic resistance (ISR)-like jasmonate-related response. These results pave the way towards the possible use of elicitors as a sustainable tool in plant science and agriculture by increasing crop resilience to biotic and abiotic stressors without detrimental effects on plant biomass.

The two alkaloids gramine and hordenine have been known for playing a role in the allelopathic ability in barley (Hordeum vulgare L.). These allelochemicals can be both found in leaves and roots in some barley cultivars whereas in others one seems to exclude the other. In this study eighteen accessions of barley from the Middle-East area, one accession from Tibet and the modern spring cultivar Barke, already used as parental donor in a nested associated mapping (NAM) population, were screened for their gramine, hordenine and N-methyltyramine (the direct precursor of hordenine) content in leaves, roots and exudates. Moreover, the toxicity of the three allelochemicals on root growth inhibition on lettuce (Lactuca sativa L.) was evaluated. Results of this study showed the preferential production of gramine and hordenine in leaves and roots, respectively, in the nineteen barley accessions. On the other hand, in the modern barley cultivar Barke, the highest content of hordenine in roots and the general lack of gramine suggests a favored biosynthesis of the former. Gramine was not detected in the root exudates. In additions, different metabolomic profiles were observed in wild relatives compared to modern barley genotypes. The results also showed the phytotoxic effects of the three compounds on root growth of lettuce seedlings, with a reduction in root length and an increase of root surface area and diameter. In conclusion, this study highlighted the impact of the domestication effects on the production and distribution of the two allelopathic alkaloids gramine and hordenine in barley.

Abstract Background Sewage sludge, the inevitable byproduct of municipal wastewater-treatment plant operation, is a key issue in many countries due to its increasing volume and the impacts associated with its disposal. According to the report of European Commission published in 2010, 39% of sewage sludge produced in the European Union is recycled into agriculture. Management options require extensive waste characterization, since many of them may contain compounds, which could be harmful to the ecosystem, such as heavy metals, organic pollutants, etc. The present study aims to show the results of 2 years’ sampling of sewage sludge—based on 130 samples collected from 35 wastewater-treatment plants situated in the North of Italy—and to assess its suitability as soil fertilizer regarding contents of nonylphenol (NP), nonylphenolethoxylates (NPnEOs), and phthalates (DEHP). Results An effective analytic method for organic pollutants detection in the sewage sludge has been developed, showing an excellent repeatability and recoveries. Ecotoxicological risk assessment was evaluated using risk quotients (RQs) for sludge-amended soil. Most of the analyzed samples do not contain NP, NPnEOs, and DEHP at levels higher than the limit established by the draft-working document of the European Commission on Sludge. The assessment using RQs reports that NP and NPnEOs never give values higher than 1, and for DEHP the obtained RQs exceed the value of 1 just three times. Data obtained were compared to the data from other European and Asiatic countries, showing a huge variability for all the compounds considered. Conclusions Based on the obtained results, it appears that the proposed EU limits for the selected substances on sewage sludge intended to be used as soil fertilizer in agriculture are sufficiently conservative to avoid negative effects on soil fauna.

The phenolic profile of Goji berries from Italy and China was investigated through ultra-high-pressure liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry. Hydroxycinnamic acids, followed by flavonols and flavanols, were the most representative phenolics, whereas glycoside forms of anthocyanins and flavonols were the compounds best discriminating between Chinese and Italian berries.A mix of dried berries was then subjected to in vitro gastro-intestinal digestion and fermentation to investigate the fate and bioaccessibility of phenolic compounds. The total phenolic content and the oxygen radical absorbance capacity recorded during the large intestine fermentation increased at 24 h, up to 1.56 mg gallic acid equivalents g−1 and 1.76 μmol trolox equivalents g−1, respectively. Multivariate statistics applied to phenolic profiling during the in vitro fermentation depicted those compounds having the highest bioaccessibility; stilbenes, alkyl-resorcinols and lignans were the best markers of Goji berries during in vitro fermentation.

Biostimulants are becoming increasingly popular in agriculture for their ability to induce beneficial effects in crops, paving the way towards the identification of new materials with biostimulant potential. This study evaluated the potential of different concentrations of an aqueous extract (0.25%, 0.50%, and 1.00%, dry weight/water volume, respectively) obtained from duckweed (Lemna minor L.) to stimulate olive plants. Leaf net photosynthesis (Pn), leaf transpiration rate (E), stomatal conductance (gs), sub-stomatal CO2 concentration (Ci), chlorophyll content and other plant growth parameters were investigated. As a result, the extract improved Pn, gs, Ci, chlorophyll content and plant biomass production (leaf fresh and dry weight). Furthermore, the duckweed extract generally increased the uptake of nitrogen (N), potassium (K), calcium (Ca), magnesium (Mg), iron (Fe) and zinc (Zn), while it did not influence the content of sodium (Na), manganese (Mn) and copper (Cu). The untargeted metabolomic profiling of the extract revealed the presence of signalling compounds (including phytohormones), phenolics and glutathione. Such broad diversity of bioactives may support the stimulatory potential observed in olive. In summary, this study revealed for the first time that duckweed could be seen as a promising species to obtain extracts with biostimulant properties in olive trees.

Despite being not essential to plants, Silicon (Si) has proven to have promoting effects on plants growth, yield, and resistance against biotic and abiotic stressors. The increase of concentration in specific minerals in plant tissues can also improve shelf-life, which, in fruits like strawberries, is also affected by the epiphytic microbial community. The present research was carried out to assess whether Si biofortification of strawberry plants, grown in soilless system, could affect plants yield and post-harvest feature of fruits during the storage period, carried out at three different temperatures (i.e., 1, 4 and 10 °C) for 7 and 14 days. Furthermore, we investigated whether the plant nutritional regime, specifically the Si fertilization, can impact the composition of microbial community. Our results showed that biofortification did not significantly affect fruits firmness, whereas, at the highest Si levels, an increase in titratable acidity was observed. The microbial community analysis highlighted for the first time the presence of probiotic bacteria, as Bacillus breve, which could present interesting technological features as strains adapted to the strawberry fruit-sphere. In addition, with the increasing levels of Si biofortification, the depletion of potentially pathogenic microorganisms, like Escherichia coli and Terrisporobacter glycolicus, was also observed. In conclusion, data here reported highlight for the first time the possible role played by the nutritional regimes of strawberry plants in shaping composition of the fruit epiphytic microbial community.

Arbuscular mycorrhiza and Trichoderma are well-known beneficial fungi whose plant growth promotion and defense elicitation effects are known. However, the molecular and biochemical processes underlying the beneficial effects of these priming microorganisms have not been fully elucidated yet. On this basis, the present work aimed to use metabolomics to dissect comprehensively the modulation of secondary metabolism induced by mycorrhiza and Trichoderma, using tomato as a model plant. To this aim, either mycorrhiza or Trichoderma were applied to tomato roots at transplanting using a commercial formulation and then harvested once the mutualistic relationship was well established. Shoots were analyzed using an MS-based untargeted metabolomics approach, and differential metabolites identified by multivariate statistics were subjected to pathway analysis. Together with promoting plant growth, the treatments induced a broad molecular reprogramming with the phenylpropanoid biosynthetic pathway (including defense phenolics like coumarins and glycosylated anthocyanins) being strongly elicited. An accumulation of auxins, cytokinins, and jasmonate (especially after treatment with Trichoderma) could be observed concerning phytohormone profiles. Overall, the broad and distinctive effects triggered by mycorrhiza and Trichoderma in tomato secondary metabolism supported both plant growth promotion and immunity.

The need to increase crop productivity and resistance directs interest in nanotechnology. Indeed, biogenic metal oxide nanoparticles can promote beneficial effects in plants, while their synthesis avoids the environmental impacts of conventional synthetic procedures. In this context, this research aimed to synthesize biogenic zinc oxide nanoparticles (ZnO-NPs) using, for the first time, an extract of a wild and spontaneous aquatic species, Lemna minor (duckweed). The effectiveness of this biogenic synthesis was evidenced for comparison with non-biogenic ZnO-NPs (obtained without using the plant extract), which have been synthesized in this research. XRD (X-ray diffraction), FE-SEM (field emission gun electron scanning microscopy), EDX (energy dispersive x-ray spectroscopy), TEM (transmission electron microscope) and UV-vis (ultraviolet-visible spectrophotometry) showed the biogenic approach effectiveness. The duckweed extract was subjected to UHPLC-ESI/QTOF-MS (ultra high-pressure liquid chromatography quadrupole time of flight mass spectrometry) phenolic profiling. This untargeted characterization highlighted a high and chemically diverse content in the duckweed extract of compounds potentially implicated in nanoparticulation. From an application standpoint, the effect of biogenic nanoparticles was investigated on some traits of maize subjected to seed priming with a wide range of biogenic ZnO-NPs concentrations. Inductive effects on the shoot and root biomass development were ascertained concerning the applied dosage. Furthermore, the biogenic ZnO-NPs stimulated the content of chlorophylls, carotenoids, and anthocyanin. Finally, the study of malondialdehyde content (MDA) as a marker of the oxidative status further highlighted the beneficial and positive action of the biogenic ZnO-NPs on maize.

The aim of this work was to investigate the effect of meteorological conditions on resveratrol concentration of red wines produced in Piacenza viticultural region (Italy). In this regard, six representative estates producing Colli Piacentini Gutturnio DOC (a blend of V. vinifera L. cvs. Barbera and Croatina) vintage wines were analysed for trans- and cis-resveratrol over an 8-year period (1998–2005). Grapes were taken from the same vineyard in each estate by using the same enological practices over the entire investigated period. The meteorological conditions corresponding to the production areas were recorded, and bioclimatic indices were calculated as well. Overall, cis-resveratrol concentration was negatively correlated to Huglin index and August mean temperature, whilst positive correlation coefficients were found when considering the Selianinov index and the rainfall of September.

In this work, the comprehensive metabolomic changes in pork burgers treated with different antioxidants, namely, (a) a control without antioxidants, (b) 200 mg/kg butylated hydroxytoluene (BHT), and (c) 250 mg/kg pitanga leaf extract (PLE, from Eugenia uniflora L.), each one packaged under modified atmosphere (80% O2 and 20% CO2) for 18 days storage at 2 ± 1 °C, were deeply studied. In particular, untargeted metabolomics was used to evaluate the impact of the antioxidant extracts on meat quality. The PLE phytochemical profile revealed a wide variety of antioxidant compounds, such as polyphenols, alkaloids, and terpenoids. Multivariate statistics (both unsupervised and supervised) allowed to observe marked differences in BHT and PLE burgers metabolomic profiles during storage. Most of the differences could be attributed to hexanoylcarnitine, 4-hydroxy-2-nonenal, 6-hydroxypentadecanedioic acid, 9S,11S,15S,20-tetrahydroxy-5Z,13E-prostadienoic acid (20-hydroxy-PGF2a), sativic acid, followed by glycerophospholipids. In addition, significant correlations (p < 0.01) were observed between thiobarbituric acid reactive substances and metabolites related to lipid oxidation processes. Therefore, the approach used showed a clear modulation of lipid oxidation, likely promoted by the plant leaf extract, thus confirming the ability of PLE to delay lipid oxidative phenomena during storage.

Plant growth promoting rhizobacteria provide an innovative solution to address challenges in sustainable agro-ecosystems, improving plant growth as well as acting as agents of biocontrol. In this study autochthonous bacteria were isolated from the rhizosphere of processing tomato plants (Solanum lycopersicum L.) cultivated with conservation agriculture practices (i.e., reduced tillage and cover crops), and evaluated for both growth-promoting activities (PGPAs), and antagonistic potential against the phytopathogenic pest Sclerotinia sclerotiorum. Considering the several activities of PGPR, we decided to structure the screening with a hierarchic approach, starting from testing the capability of fixing nitrogen. The obtained bacteria were processed through the molecular typing technique rep-PCR (Repetitive Extragenic Palindromic) in order to discriminate microbial strains with the same profiles, and identified via 16S rDNA sequencing. Thirty-eight selected isolates were screened in vitro for different activities related to plant nutrition and plant growth regulation as well as for antifungal traits. Isolated bacteria were found to exhibit different efficiencies in indoleacetic acid production and siderophore production, phosphate solubilization and biocontrol activity against the widespread soil-borne plant pathogen S. sclerotiorum. All the 38 bacterial isolates showed at least one property tested. With a view to detect the suitable candidates to be developed as biofertilizers, the selected isolates were ranked by their potential ability to function as PGPR. Thus, consortium of native PGPR bacteria inoculants may represent a suitable solution to address the challenges in sustainable agriculture, to ensure crop yield and quality, lowering the application of chemicals input.

In this work, different commercial extra-virgin olive oils (EVOO) were subjected to in vitro gastrointestinal digestion and the changes in bioactive compounds were evaluated by ultra-high-pressure liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry, using untargeted metabolomics. As expected, raw EVOO samples were abundant in total sterols (on average: 3007.4 mg equivalents/kg) and tyrosol equivalents (on average: 334.1 mg equivalents/kg). However, the UHPLC-QTOF screening allowed us to annotate 309 compounds, with a large abundance of sterols (219 compounds), followed by polyphenols (67 compounds) and terpenoids. The in vitro gastrointestinal digestion was found to affect the phytochemical composition of the different EVOO samples. In particular, both unsupervised and supervised statistics depicted the modifications of the bioactive profile following gastric and pancreatic phases. Overall, the compounds which resulted as the most affected by the in vitro digestion were flavonoids (cyanidin and luteolin equivalents), whilst relatively high % bioaccessibility values were recorded for tyrosol equivalents during the pancreatic phase (on average, 66%). In this regard, oleuropein-aglycone (i.e., the major phenolic compound in EVOO) was converted to hydroxytyrosol, moving from an average value of 1.3 (prior to the in vitro digestion) up to 9.7 mg equivalents/kg during the pancreatic step. As proposed in the literature, the increase in hydroxytyrosol might be the result of the combined effect of lipase(s) activity and acidic conditions. Taken together, the present findings corroborate the suitability of untargeted metabolomics coupled to in vitro digestion methods to investigate the bioaccessibility of phenolic compounds. In this regard, a significant impact of in vitro gastrointestinal digestion on polyphenolic profiles has been detected, thus suggesting the need to account for actual bioaccessibility values rather than just considering the amounts in the raw commodity.

Pesticides are intentionally applied to agricultural fields for crop protection. They can harm non-target organisms such as soil microorganisms involved in important ecosystem functions with impacts at the global scale. Within the frame of the pesticide registration process, the ecotoxicological impact of pesticides on soil microorganisms is still based on carbon and nitrogen mineralization tests, despite the availability of more extensive approaches analyzing the abundance, activity or diversity of soil microorganisms. In this study, we used a high-density DNA microarray (PhyloChip) and 16S rDNA amplicon next-generation sequencing (NGS) to analyze the impact of the organophosphate insecticide chlorpyrifos (CHL), the phenyl-urea herbicide isoproturon (IPU), or the triazole fungicide tebuconazole (TCZ) on the diversity and composition of the soil bacterial community. To our knowledge, it is the first time that the combination of these approaches are applied to assess the impact of these three pesticides in a lab-to-field experimental design. The PhyloChip analysis revealed that although no significant changes in the composition of the bacterial community were observed in soil microcosms exposed to the pesticides, significant differences in detected operational taxonomic units (OTUs) were observed in the field experiment between pesticide treatments and control for all three tested pesticides after 70 days of exposure. NGS revealed that the bacterial diversity and composition varied over time. This trend was more marked in the microcosm than in the field study. Only slight but significant transient effects of CHL or TCZ were observed in the microcosm and the field study, respectively. IPU was not found to significantly modify the soil bacterial diversity or composition. Our results are in accordance with conclusions of the Environmental Food Safety Authority (EFSA), which concluded that these three pesticides may have a low risk toward soil microorganisms.

Spinal muscular atrophy (SMA) is a pediatric genetic disease, characterized by motor neuron (MN) death, leading to progressive muscle weakness, respiratory failure, and, in the most severe cases, to death. Abnormalities at the neuromuscular junction (NMJ) have been reported in SMA, including neurofilament (NF) accumulation at presynaptic terminals, immature and smaller than normal endplates, reduced transmitter release, and, finally, muscle denervation. Here we have studied the role of agrin in SMAΔ7 mice, the experimental model of SMAII. We observed a 50% reduction in agrin expression levels in quadriceps of P10 SMA mice compared to age-matched WT controls. To counteract such condition, we treated SMA mice from birth onwards with therapeutic agrin biological NT-1654, an active splice variant of agrin retaining synaptogenic properties, which is also resistant to proteolytic cleavage by neurotrypsin. Mice were analyzed for behavior, muscle and NMJ histology, and survival. Motor behavior was significantly improved and survival was extended by treatment of SMA mice with NT-1654. At P10, H/E-stained sections of the quadriceps, a proximal muscle early involved in SMA, showed that NT-1654 treatment strongly prevented the size decrease of muscle fibers. Studies of NMJ morphology on whole-mount diaphragm preparations revealed that NT-1654-treated SMA mice had more mature NMJs and reduced NF accumulation, compared to vehicle-treated SMA mice. We conclude that increasing agrin function in SMA has beneficial outcomes on muscle fibers and NMJs as the agrin biological NT-1654 restores the crosstalk between muscle and MNs, delaying muscular atrophy, improving motor performance and extending survival.

The novel multi-million read generating sequencing technologies are very promising for resolving the immense soil 16S rRNA gene bacterial diversity. Yet they have a limited maximum sequence length screening ability, restricting studies in screening DNA stretches of single 16S rRNA gene hypervariable (V) regions. The aim of the present study was to assess the effects of properties of four consecutive V regions (V3-6) on commonly applied analytical methodologies in bacterial ecology studies. Using an in silico approach, the performance of each V region was compared with the complete 16S rRNA gene stretch. We assessed related properties of the soil derived bacterial sequence collection of the Ribosomal Database Project (RDP) database and concomitantly performed simulations based on published datasets. Results indicate that overall the most prominent V region for soil bacterial diversity studies was V3, even though it was outperformed in some of the tests. Despite its high performance during most tests, V4 was less conserved along flanking sites, thus reducing its ability for bacterial diversity coverage. V5 performed well in the non-redundant RDP database based analysis. However V5 did not resemble the full-length 16S rRNA gene sequence results as well as V3 and V4 did when the natural sequence frequency and occurrence approximation was considered in the virtual experiment. Although, the highly conserved flanking sequence regions of V6 provide the ability to amplify partial 16S rRNA gene sequences from very diverse owners, it was demonstrated that V6 was the least informative compared to the rest examined V regions. Our results indicate that environment specific database exploration and theoretical assessment of the experimental approach are strongly suggested in 16S rRNA gene based bacterial diversity studies.

This study give a preliminary survey of pharmaceutical contamination and accumulation in surface waters and sediments along the river Po basin (74,000 km2, the largest in Italy), a strategic region for the Italian economy: it collects sewage from a vast industrialized area of Italy (Autorità di Baciono del fiume Po, 2006, 2009). 10 pharmaceuticals (atenolol, propanolol, metoprolol, nimesulide, furosemide, carbamazepine, ranitidine, metronidazole, paracetamol, and atorvastatin) from several therapeutic classes were searched in 54 sampling points along the river Po from the source to the delta, and at the mouth of its major effluents. In water samples were found pharmaceuticals in the range of 0.38–0.001 μg/L, except for furosemide (max conc. 0.605 μg/L), paracetamol (max conc. 3.59 μg/L), metoprolol (never detected) and for atenolol (not analysed). In sediment samples, only paracetamol was not detected, while the others were generally found in the range of 0.4–0.02 μg/kg ww with high concentrations for atenolol (max conc. 284 μg/kg ww) and furosemide (max conc. 98.4 μg/kg ww). The findings confirm also STPs as point sources of contamination. Despite of the much evidence for the adverse effects of pharmaceuticals in the aquatic environment, the observed low levels cannot be considered to pose a serious risk to human health; further studies are necessary for a comprehensive risk assessment.