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Urban trees are crucial in delivering essential ecosystem services, including air pollution mitigation. This service is influenced by plant associated microbiomes, which can degrade hydrocarbons, support tree health, and influence ecological processes. Yet, our understanding of tree microbiomes remains limited, thus affecting our ability to assess and quantify the ecosystem services provided by trees as complex systems. The main hypothesis of this work was that tree microbiomes concur to hydrocarbon biodegradation, and was tested through three case studies, which collectively investigated two tree micro-habitats (phyllosphere and tree cavity organic soil-TCOS) under various conditions representing diverse ecological scenarios, by applying different culture-based and molecular techniques and at different scales. The integration of all results provided a more comprehensive understanding of the role of microbiomes in urban trees. Firstly, bacterial strains isolated from the phyllosphere of Quercus ilex were characterized, indicating the presence of Plant-Growth Promoting bacteria and strains able to catabolize PAHs, particularly naphthalene and phenanthrene. Secondly, naphthalene biodegradation on artificially spiked Hedera helix leaves was quantified in greenhouse experiments on inoculated and untreated plants. The persistence of the inoculated strain and community structure of epiphytic bacteria were assessed by Illumina sequencing of V5-V6 hypervariable regions of 16S rRNA gene. Results showed that naphthalene degradation was initially faster on inoculated plants but later the degradation rates became similar, probably because bacterial populations with hydrocarbon-degrading abilities gradually developed also on non-inoculated plants. Finally, we explored bacterial and fungal biodiversity hosted by TCOS samples, collected from six large trees located in an urban park and belonging to different species. Microbial communities were characterized by Illumina sequencing of V5-V6 hypervariable regions of bacterial gene 16S rRNA and of fungal ITS1. Results indicated TCOS as a distinct substrate, whose microbiome is determined both by the host tree and by canopy environmental conditions and has a pronounced aerobic hydrocarbon degradation potential. Overall, a better assessment of biodiversity associated with trees and the subsequent provision of ecosystem services constitute a first step toward developing future new microbe-driven sustainable solutions, especially in terms of support for urban green planning and management policy., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Gandolfi, Canedoli, Rosatelli, Covino, Cappelletti, Sebastiani, Tatangelo, Corengia, Pittino, Padoa-Schioppa, Báez-Matus, Hernández, Seeger, Saati-Santamaría, García-Fraile, López-Mondéjar, Ambrosini, Papacchini and Franzetti.)

In this study, the anthropogenic contamination in Trasimeno lake (Central Italy) was investigated using three sediment cores spanning over the last 150 years (Anthropocene) to identify the primary sources of pollution and quantify the level of contaminant enrichment in the basin. First, based on the relative cumulative frequency and linear regression methods, we obtained a geochemical baseline for the lake using the deeper parts of the sediment cores. The geochemical baseline allowed us to determine the values of trace elements enrichment factors. On this knowledge, as a second result, we were able to reconstruct the natural sources and the anthropogenic impact on the lake with a biennial resolution. This goal has been obtained by combining different inorganic and organic chemical proxies such as trace elements, polycyclic aromatic hydrocarbons, and lead isotope ratios and exploiting both principal component and factor analysis to associate chemical proxies to human-driven contamination processes. Five different groups of elements have been identified, one of which is of natural origin and four of anthropogenic origin. In particular, it was possible to identify the times and impacts of the industrial activities during the Second World War, which dispersed heavy metals in sediments. Moreover, we found evidence of the recent human activities that have characterized the surroundings of the basin, such as Pb inputs related to the use of gasoline and the enrichment of certain elements generally used in agricultural activities (such as P, Cu, and Mn) due to the development of this sector in the last 40 years., (© 2022. The Author(s).)

Defining optimal nutrition in animals and humans remains a main scientific challenge. The objective of the work was to develop a dynamic model of reactive oxygen species (ROS)-polyunsaturated fatty acid (PUFA)-antioxidant homeostasis using the rabbit as a model. The problem entity was to evaluate the main metabolites generated from interactions between traits included in the conceptual model and identified by three main sub-models: (i) ROS generation, (ii) PUFA oxidation and (iii) antioxidant defence. A mathematical model (VENSIM software) that consisted of molecular stocks (INPUTs, OUTPUTs), exchange flows (intermediate OUTPUTs) and process rates was developed. The calibration was performed by using standard experimental data (Experiment 1), whereas the validation was carried out in Experiments 2 and 3 by using supra-nutritional dietary inputs (VIT E+ and PUFA+). The accuracy of the models was measured using 95% confidence intervals. Analytical OUTPUTs (ROS, PUFA, Vit E, Ascorbic acid, Iso-/NeuroProstanes, Aldehydes) were well described by the standard model. There was also good accuracy for the VIT E+ scenario, whereas some compensatory rates (K c1 -K c4 ) were added to assess body compensation when high levels of dietary PUFA were administered (Experiment 3). In conclusion, the model can be very useful for predicting the effects of dietary treatments on the redox homeostasis of rabbits.

In the present study, we carried out a quantitative analysis of the monoterpenes composition in different tissues of the non-model conifer Pinus nigra J.F. Arnold subsp. laricio Palib. ex Maire ( P. laricio , in short). All the P. laricio tissues examined showed the presence of the same fourteen monoterpenes, among which the most abundant were β-phellandrene, α-pinene, and β-pinene, whose distribution was markedly tissue-specific. In parallel, from the same plant tissues, we isolated seven full-length cDNA transcripts coding for as many monoterpene synthases, each of which was found to be attributable to one of the seven phylogenetic groups in which the d1-clade of the canonical classification of plants' terpene synthases can be subdivided. The amino acid sequences deduced from the above cDNA transcripts allowed to predict their putative involvement in the biosynthesis of five of the monoterpenes identified. Transcripts profiling revealed a differential gene expression across the different tissues examined, and was found to be consistent with the corresponding metabolites profiles. The genomic organization of the seven isolated monoterpene synthase genes was also determined.

Sample manipulation for storage and storage itself, interfere with the stability of labile lipids in human plasma, including vitamin E (α-tocopherol), polyunsaturated fatty acids (PUFAs), and their enzymatic and free radical-derived oxidation metabolites. This remains a main limit of lipidomics studies that often lack of sufficient standardization and validation at the pre-analytical level. In order to characterize the stability of these lipids in human plasma and to develop a standardized pre-analytical protocol for lipidomics methods, the oxidation metabolites of α-tocopherol, the free form of ω3 and ω6 PUFAs, and some arachidonic acid (AA)-derived eicosanoids were investigated in human plasma during storage at different freezing temperatures. The effect of a protection/defense cocktail of antioxidants and lipoxygenase inhibitors (PD solution) on these lipid parameters was also evaluated. The temperature of storage markedly affected the formation of α-tocopheryl quinone (α-TQ), the main lipoperoxyl radical-derived oxidation metabolite of vitamin E, with the lowest production rate observed in samples stored at -80 °C or in liquid nitrogen. A similar effect of the storage temperature was observed for the free form of the ω-3 species eicosapentaenoic and docosahexaenoic acid, and for the ω-6 AA. Freezing samples at -20 °C resulted in a time-dependent formation of the pro-inflammatory eicosanoid LTB4. The PD solution prevents non-specific alterations of these lipid parameters in samples that are processed for direct analysis and protects from the temperature-dependent modifications of free PUFAs. Combining PD solution and preservation at -80 °C or in liquid nitrogen, resulted in levels of α-TQ and PUFAs that remained stable over 1 month and up to 8 months of storage, respectively. This method paper provides indications for the optimal processing and storage of human plasma utilized in lipidomics studies., (Copyright © 2021 Elsevier Inc. All rights reserved.)

A quali-quantitative analysis of diterpenoid composition in tissues obtained from different organs of Pinus nigra subsp. laricio (Poiret) Maire (Calabrian pine) was carried out. Diterpene resin acids were the most abundant diterpenoids across all the examined tissues. The same nine diterpene resin acids were always found, with the abietane type prevailing on the pimarane type, although their quantitative distribution was found to be remarkably tissue-specific. The scrutiny of the available literature revealed species specificity as well. A phylogeny-based approach allowed us to isolate four cDNAs coding for diterpene synthases in Calabrian pine, each of which belonging to one of the four groups into which the d3 clade of the plants' terpene synthases family can be divided. The deduced amino acid sequences allowed predicting that both monofunctional and bifunctional diterpene synthases are involved in the biosynthesis of diterpene resin acids in Calabrian pine. Transcript profiling revealed differential expression across the different tissues and was found to be consistent with the corresponding diterpenoid profiles. The isolation of the complete genomic sequences and the determination of their exon/intron structures allowed us to place the diterpene synthase genes from Calabrian pine on the background of current ideas on the functional evolution of diterpene synthases in Gymnosperms.

Airborne bacteria were characterized over a 2-y period via high-throughput massive sequencing of 16S rRNA gene in aerosol samples collected at a background mountain European Monitoring and Evaluation Programme (EMEP) Network site (Monte Martano, Italy) located in the Central Mediterranean area. The air mass origin of nineteen samples was identified by air mass modelling and a detailed chemical analysis was performed. Four main origins (Saharan, North-western, North-eastern, and Regional) were identified, and distinct microbial communities were associated with these air masses. Samples featured a great bacterial diversity with Protobacteria being the most abundant phylum, and Sphingomonas followed by Acidovorax, Acinetobacter and Stenotrophomonas the most abundant genera of the dataset. Bacterial genera including potential human and animal pathogens were more abundant in European and in Regional samples compared to Saharan samples; this stressed the relevance of anthropic impact on bacterial populations transported by air masses that cross densely populated areas. The principal aerosol chemical characteristics and the airborne bacterial communities were correlated by cluster analysis, similarity tests and non-metric multidimensional scaling analysis, explaining most of the variability observed. However, the strong correlation between bacterial community structure and air mass origin hampered the possibility to disentangle the effects of variations in bacterial populations and in dust provenance on variations in chemical variables., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The metabolism of α-tocopherol (α-TOH, vitamin E) shows marked interindividual variability, which may influence the response to nutritional and therapeutic interventions with this vitamin. Recently, new metabolomics protocols have fostered the possibility to explore such variability for the different metabolites of α-TOH so far identified in human blood, i.e., the "vitamin E metabolome", some of which have been reported to promote important biological functions. Such advances prompt the definition of reference values and degree of interindividual variability for these metabolites at different levels of α-TOH intake. To this end, a one-week oral administration protocol with 800 U RRR-α-TOH/day was performed in 17 healthy volunteers, and α-TOH metabolites were measured in plasma before and at the end of the intervention utilizing a recently validated LC-MS/MS procedure; the expression of two target genes of α-TOH with possible a role in the metabolism and function of this vitamin, namely pregnane X receptor (PXR) and the isoform 4F2 of cytochrome P450 (CYP4F2) was assessed by immunoblot in peripheral blood leukocytes. The levels of enzymatic metabolites showed marked interindividual variability that characteristically increased upon supplementation. With the exception of α-CEHC (carboxy-ethyl-hydroxychroman) and the long-chain metabolites M1 and α-13'OH, such variability was found to interfere with the possibility to utilize them as sensitive indicators of α-TOH intake. On the contrary, the free radical-derived metabolite α-tocopheryl quinone significantly correlated with the post-supplementation levels of α-TOH. The supplementation stimulated PXR, but not CYP4F2, expression of leucocytes, and significant correlations were observed between the baseline levels of α-TOH and both the baseline and post-supplementation levels of PXR. These findings provide original analytical and molecular information regarding the human metabolism of α-TOH and its intrinsic variability, which is worth considering in future nutrigenomics and interventions studies.

The present paper describes the assessment of the atmospheric deposition processes in a basin valley through a multidisciplinary approach based on the data collected within an extensive physico-chemical characterization of the soils, combined with the local meteorology. Surface soil cores were collected on a NNW-SSE transect across the Terni basin (Central Italy), between the Monti Martani and the Monti Sabini chains (956 m a.s.l.), featuring the heavily polluted urban and industrial enclave of Terni on its bottom. Airborne radiotracers, namely 210 Pb and 137 Cs, have been used to highlight atmospheric deposition. We observed an increased deposition flux of 210 Pb and 137 Cs at sites located at the highest altitudes, and the associated concentration profiles in soil allowed to evaluate the role of atmospheric deposition. We also obtained a comprehensive dataset of stable anthropogenic pollutants of atmospheric origin that showed heterogeneity along the transect. The behavior has been explained by the local characteristic of the soil, by seeder-feeder processes promoted by the atmospheric circulation, and was reconciled with the concentration profile of radiotracers by factor analysis. Finally, the substantial impact of the local industrial activities on soil profiles and the role of the planetary boundary layer has been discussed and supported by simulations employing a Lagrangian dispersion model., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Garcinoic acid (GA or δ-T3-13'COOH), is a natural vitamin E metabolite that has preliminarily been identified as a modulator of nuclear receptors involved in β-amyloid (Aβ) metabolism and progression of Alzheimer's disease (AD). In this study, we investigated GA's effects on Aβ oligomer formation and deposition. Specifically, we compared them with those of other vitamin E analogs and the soy isoflavone genistein, a natural agonist of peroxisome proliferator-activated receptor γ (PPARγ) that has therapeutic potential for managing AD. GA significantly reduced Aβ aggregation and accumulation in mouse cortical astrocytes. Similarly to genistein, GA up-regulated PPARγ expression and apolipoprotein E (ApoE) efflux in these cells with an efficacy that was comparable with that of its metabolic precursor δ-tocotrienol and higher than those of α-tocopherol metabolites. Unlike for genistein and the other vitamin E compounds, the GA-induced restoration of ApoE efflux was not affected by pharmacological inhibition of PPARγ activity, and specific activation of pregnane X receptor (PXR) was observed together with ApoE and multidrug resistance protein 1 (MDR1) membrane transporter up-regulation in both the mouse astrocytes and brain tissue. These effects of GA were associated with reduced Aβ deposition in the brain of TgCRND8 mice, a transgenic AD model. In conclusion, GA holds potential for preventing Aβ oligomerization and deposition in the brain. The mechanistic aspects of GA's properties appear to be distinct from those of other vitamin E metabolites and of genistein., Competing Interests: Conflict of interest—The authors declare that they have no conflicts of interest., (© 2020 Marinelli et al.)

Plants and phyllosphere microorganisms may effectively contribute to reducing air pollution in cities through the adsorption and biodegradation of pollutants onto leaves. In this work, during all seasons, we sampled atmospheric particulate matter (PM 10 ) and leaves of southern magnolia Magnolia grandiflora and deodar cedar Cedrus deodara, two evergreen plant species widespread in the urban area of Milan where the study was carried out. We then quantified Polycyclic Aromatic Hydrocarbons (PAHs) both in PM 10 and on leaves and used sequencing of 16S rRNA gene, shotgun metagenomics and qPCR analyses to investigate the microbial communities hosted by the sampled leaves. Taxonomic and functional profiles of epiphytic bacterial communities differed between host plant species and seasons and the microbial communities on leaves harboured genes involved in the degradation of hydrocarbons. Evidence collected in this work also suggested that the abundance of hydrocarbon-degrading microorganisms on evergreen leaves increased with the concentration of hydrocarbons when atmospheric pollutants were deposited at high concentration on leaves, and that the biodegradation on the phyllosphere can contribute to the removal of PAHs from the urban air., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This study investigated the oxidative stability of meat from lambs fed silages in which timothy grass was totally or partially replaced by sainfoin and/or red clover. Five groups of 8 lambs were fed the following silages: timothy grass (T), mixture (50:50) of timothy with either sainfoin (T-SF) or red clover (T-RC), mixture of timothy, sainfoin and red clover (50:25:25; T-SF-RC), or mixture (50:50) of sainfoin and red clover (SF-RC). Feeding the silages containing red clover (T-RC, T-SF-RC and SF-RC) decreased the deposition of vitamin E in muscle relative to the highly unsaturated fatty acids (P < .001), did not affect the oxidative stability of fresh meat, but reduced oxidative deterioration in cooked meat and in meat homogenates incubated with pro-oxidant catalysts (P < .001). The results of this study demonstrated that feeding lambs with silages containing sainfoin and red clover improve the oxidative stability of meat subjected to strong pro-oxidant conditions and suggest that these effects should be further clarified., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Lipid peroxidation is one of the earliest pathogenic events of non-alcoholic fatty liver disease (NAFLD). In this context, an increased oxidation of the lipoperoxyl radical scavenger α-tocopherol (α-TOH) should occur already in the subclinical phases of the disease to compensate for the increase oxidation of the lipid excess of liver and possibly of other tissues. However, this assumption remains unsupported by direct analytical evidence. In this study, GC-MS/MS and LC-MS/MS procedures have been developed and applied for the first time to measure the vitamin E oxidation metabolite α-tocopheryl quinone (α-TQ) in plasma of fatty liver (FL) subjects that were compared in a pilot cross-sectional study with healthy controls. The protein adducts of 4-hydroxynonenal (4-HNE) and the free form of polyunsaturated free fatty acids (PUFA) were measured as surrogate indicators of lipid peroxidation. α-TQ formation was also investigated in human liver cells after supplementation with α-TOH and/or fatty acids (to induce steatosis). Compared with controls, FL subjects showed increased (absolute and α-TOH-corrected) levels of plasma α-TQ and 4-HNE, and decreased concentrations of PUFA. α-TQ levels positively correlated with indices of liver damage and metabolic dysfunction, such as alanine aminotransferase, bilirubin and triglycerides, and negatively correlated with HDL cholesterol. Fatty acid supplementation in human hepatocytes stimulated the generation of cellular oxidants and α-TOH uptake leading to increased α-TQ formation and secretion in the extracellular medium - both were markedly stimulated by α-TOH supplementation. In conclusion, plasma α-TQ represents an early biomarker of the lipoperoxyl radical-induced oxidation of vitamin E and lipotoxicity of the fatty liver., (Copyright © 2018 Elsevier Inc. All rights reserved.)

In this paper, we present a comprehensive taxonomic survey of the bacterial community and accurate quantification of polycyclic aromatic hydrocarbons (PAHs) associated with an intense Saharan dust advection, which impacted Central Mediterranean area in the whole 2014-2015 period. This work is part of an intensive field campaign at the EMEP regional background site of Monte Martano (Central Italy), considered well representative of long-range transport in the Central Mediterranean area. 22 samples have been characterized in their provenance region and have been considered for the chemical and biological characterization. The event described in the present paper was exceptionally intense at the sampling site allowing a detailed evaluation of the dust load on a regional scale, an estimation of the impact of PAH based on the Toxic Equivalency Factor methodology and a thorough characterization of the airborne bacterial fraction performed by High Throughput Sequencing approach. Afterward, we cultured viable bacteria and evaluated several enzymatic activities and conducted UV survival tests. Principal findings include: (i) the striking evidence that, during the Saharan dust event, a highly diverse and abundant bacterial community was associated with PAH concentrations higher than the yearly mean; (ii) the tangible presence of cultivable microbes; (iii) the proof that the isolates recovered from Saharan dust had the potential to be metabolically active and that almost all of them were able to persist following UV radiation exposure. Comparisons of results for the present case study with mean values for the 2014-2015 experimental campaign are presented. The bacterial community and chemical speciation associated with the Saharan dust advection were specific and very different from those associated with other air masses. The particular case of North-Western Atlantic, which represents one of the most typical advection route reaching the sampling site is discussed in detail., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Sterol intermediates of the cholesterol biosynthetic pathway have drawn attention for novel biological activities. Follicular fluid meiosis activating sterol (FF-MAS) is a LXRα ligand and a potential modulator of physiologic processes regulated by nuclear receptors, such as lipid homeostasis and cell proliferation. In this work, we established a model to selectively accumulate FF-MAS in HepG2 cells, by using a combination of the inhibitors AY9944 and 17-hydroxyprogesterone to block C14-sterol reductases and the downstream C4-demethylase complex. We investigated the effects produced by altered levels of cholesterol biosynthesis intermediates, in order to dissect their influence on LXRα signaling. In particular, endogenously accumulated FF-MAS was able to modulate the expression of key genes in cholesterol metabolism, to activate LXRα nuclear signaling resulting in increased lipogenesis, and to inhibit HepG2 cells proliferation. Moreover, a fluorescent ester derivative of FF-MAS localized in nuclear lipid droplets, suggesting a role for these organelles in the storage of signaling lipids interacting with nuclear partners., (Copyright © 2017 Elsevier B.V. All rights reserved.)

In the present study, we report the attempt to characterize the chemical composition of fruit kernel of Lodoicea maldivica coco nucifera palm (commonly named as 'Coco de mer') by gas chromatographic method. The analysis was performed by HS-SPME and GC/MS techniques to determine volatile aroma, sterol, and fatty acid composition profiles in the internal and external pulp of two distinct coconuts. Although no qualitative differences in flavour composition were observed between the two analysed coconuts and the relative two pulp parts, variations in the abundance levels of the prominent compounds have been recorded. The averaged quantity of total phytosterols, resulting from the two analysed 'Coco de mer' samples, was almost constant in both kernels coconut, being 24.5 μg/g (of dry net matter) for the external, and 26.9 μg/g (of dry net matter) for the internal portion. In both coconuts, the fatty acid pattern composition was characterized by seven saturated acids ranged from C14:0 (myristic) to C20:0 (arachidic) and two monounsaturated acids, the palmitoleic (C16:1, ω7) and the oleic (C18:1, ω9). Palmitic acid (C16:0) was the predominant one with an average contribution of about 49.0%, followed by pentadecanoic 16.5%, stearic (C18:0) 11.6%, and myristic (C14:0) 9.9% acids in all two examined kernel portions., (© 2017 Wiley-VHCA AG, Zurich, Switzerland.)