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Climate change significantly contributes to water scarcity in various regions worldwide. While wastewater reuse is a crucial strategy for mitigating water scarcity, it also carries potential risks for human health due to the presence of pathogenic and antibiotic resistant bacteria (ARB). Antibiotic resistance represents a Public Health concern and, according to the global action plan on antimicrobial resistance, wastewater role in selecting and spreading ARB must be monitored. Our aim was to assess the occurrence of ARB, antibiotic resistance genes (ARGs), and potential pathogenic bacteria throughout a wastewater treatment plant (WWTP) designed for water reuse. Furthermore, we aimed to evaluate potential association between ARB and ARGs with antibiotics and heavy metals. The results obtained revealed the presence of ARB, ARGs and pathogenic bacteria at every stage of the WWTP. Notably, the most prevalent ARB and ARG were sulfamethoxazole-resistant bacteria (up to 7.20 log CFU mL -1 ) and sulII gene (up to 5.91 log gene copies mL -1 ), respectively. The dominant pathogenic bacteria included Arcobacter, Flavobacterium and Aeromonas. Although the abundance of these elements significantly decreased during treatment (influent vs. effluent, p < 0.05), they were still present in the effluent designated for reuse. Additionally, significant correlations were observed between heavy metal concentrations (copper, nickel and selenium) and antibiotic resistance elements (ampicillin-resistant bacteria, tetracycline-resistant bacteria, ARB total abundance and sulII) (p < 0.05). These results underscore the importance of monitoring the role of WWTP in spreading antibiotic resistance, in line with the One Health approach. Additionally, our findings suggest the need of interventions to reduce human health risks associated with the reuse of wastewater for agricultural purposes., 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 © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Extracts from locally grown aromatic plants can enhance the geographical characteristics and microbial stability of craft beers, which are often not pasteurized or filtered. Here, the chemical and antimicrobial properties of aqueous extracts from leaves of Myrtus communis L., Pistacia lentiscus L., Artemisia arborescens L., and floral wastes of Crocus sativus L., all cultivated in Sardinia (Italy), were assessed. P. lentiscus extract had the highest polyphenol content (111.20 mg GAE/g), followed by M. communis (56.80 mg GAE/g), C. sativus (32.80 mg GAE/g), and A. arborescens (8.80 mg GAE/g). Notably, only the M. communis extract demonstrated significant inhibitory activity against pathogenic and spoilage microorganisms, with minimum inhibitory concentrations of 0.18, 0.71, and 1.42 mg GAE/mL against Staphylococcus aureus , Lactiplantibacillus plantarum , and Lacticaseibacillus casei , respectively. Additionally, it reduced the growth of Levilactobacillus brevis and Fructilactobacillus lindneri at concentrations of 0.35 and 0.71 mg GAE/mL, respectively. Based on its significant antimicrobial activity, the M. communis extract was further characterized using high-resolution mass spectrometry, revealing high abundances of nonprenylated phloroglucinols, flavonoid derivatives (myricetin), and quinic acids. Lastly, adding M. communis extract (2.84 mg GAE/mL) to commercial beer effectively prevented the growth of L. brevis and F. lindneri , showing its potential to avoid beer's microbial spoilage.

The World Health Organization (WHO) promotes research aimed at developing new drugs from natural compounds. Fungi are important producers of bioactive molecules, and they are often effective against other fungi and/or bacteria and are thus a potential source of new antibiotics. Basidiomycota crude extracts, which have previously been proven to be active against Pseudomonas aeruginosa ATCC27853, were subjected to liquid chromatographic separation by RP-18, leading to six macro-fractions for each fungal extract. The various fractions were tested for their bioactivities against P. aeruginosa ATCC27853, and ten of them were characterized by HPLC-HRMS and NMR. Further chromatographic separations were performed for a few selected macro-fractions, yielding seven pure compounds. Bioactivity was mainly found in the lipophilic fractions containing fatty acids and their derivatives, such as hydroxy or keto C-18 unsaturated acids, and in various complex lipids, such as glycolipids and related compounds. More hydrophilic molecules, such as GABA, phenethylamine, two chromogenic anthraquinoids and pistillarin, were also isolated, and their antibacterial activities were recorded. The novelties of this research are as follows: (i) the genera Cortinarius and Mycena have never been investigated before for the synthesis of antibiotic compounds; (ii) the molecules produced by these genera are known, but their production has never been reported in the investigated fungi; (iii) the determination of bacterial siderophore synthesis inhibition by certain compounds from Cortinarius and Mycena .

Introduction: The lack of functional hepatocytes poses a significant challenge for drug safety testing and therapeutic applications due to the inability of mature hepatocytes to expand and their tendency to lose functionality in vitro . Previous studies have demonstrated the potential of Human Liver Stem Cells (HLSCs) to differentiate into hepatocyte-like cells within an in vitro rotary cell culture system, guided by a combination of growth factors and molecules known to regulate hepatocyte maturation. In this study, we employed a matrix multi-assay approach to comprehensively characterize HLSC differentiation. Methods: We evaluated the expression of hepatic markers using qRT-PCR, immunofluorescence, and Western blot analysis. Additionally, we measured urea and FVIII secretion into the supernatant and developed an updated indocyanine green in vitro assay to assess hepatocyte functionality. Results: Molecular analyses of differentiated HLSC aggregates revealed significant upregulation of hepatic genes, including CYP450, urea cycle enzymes, and uptake transporters exclusively expressed on the sinusoidal side of mature hepatocytes, evident as early as 1 day post-differentiation. Interestingly, HLSCs transiently upregulated stem cell markers during differentiation, followed by downregulation after 7 days. Furthermore, differentiated aggregates demonstrated the ability to release urea and FVIII into the supernatant as early as the first 24 h, with accumulation over time. Discussion: These findings suggest that a 3D rotation culture system may facilitate rapid hepatic differentiation of HLSCs. Despite the limitations of this rotary culture system, its unique advantages hold promise for characterizing HLSC GMP batches for clinical applications., Competing Interests: GC is a member of the Scientific Advisory Board of Unicyte AG. MT, GS, VF, GC, and MBHS are named inventors in related patents. The remaining 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 Tapparo, Saccu, Pasquino, Fonsato, Medana, Schiavo, Mecarelli, Maccagno, Silengo, Bruno, Camussi and Herrera Sanchez.)

A healthy vascular endothelium plays an essential role in modulating vascular tone by producing and releasing vasoactive factors such as nitric oxide (NO). Endothelial dysfunction (ED), the loss of the endothelium physiological functions, results in the inability to properly regulate vascular tone, leading to hypertension and other cardiovascular risk factors. Alongside NO, the gasotransmitter hydrogen sulfide (H 2 S) has emerged as a key molecule with vasodilatory and antioxidant activities. Since a reduction in H 2 S bioavailability is related to ED pathogenesis, natural H 2 S donors are very attractive. In particular, we focused on the sulfur-containing amino acid S -allyl cysteine (SAC), a bioactive metabolite, of which black garlic is particularly rich, with antioxidant activity and, among others, anti-diabetic and anti-hypertensive properties. In this study, we analyzed the protective effect of SAC against ED by evaluating reactive oxygen species level, H 2 S release, eNOS phosphorylation, and NO production (by fluorescence imaging and western blot analysis) in Bovine Aortic Endothelial cells (BAE-1). Furthermore, we chemically characterized a Black Garlic Extract (BGE) for its content in SAC and other sulfur-containing amino acids. BGE was used to carry out an analysis on H 2 S release on BAE-1 cells. Our results show that both SAC and BGE significantly increase H 2 S release. Moreover, SAC reduces ROS production and enhances eNOS phosphorylation and the consequent NO release in our cellular model. In this scenario, a natural extract enriched in SAC could represent a novel therapeutic approach to prevent the onset of ED-related diseases.

Irinotecan, a widely prescribed anticancer drug, is an emerging contaminant of concern that has been detected in various aquatic environments due to ineffective removal by traditional wastewater treatment systems. Solar photodegradation is a viable approach that can effectively eradicate the drug from aqueous systems. In this study, we used the design of experiment (DOE) approach to explore the robustness of irinotecan photodegradation under simulated solar irradiation. A full factorial design, including a star design, was applied to study the effects of three parameters: initial concentration of irinotecan (1.0-9.0 mg/L), pH (5.0-9.0), and irradiance (450-750 W/m 2 ). A high-performance liquid chromatography coupled with a high-resolution mass spectrometry (HPLC-HRMS) system was used to determine irinotecan and identify transformation products. The photodegradation of irinotecan followed a pseudo-first order kinetics. In the best-fitted linear model determined by the stepwise model fitting approach, pH was found to have about 100-fold greater effect than either irinotecan concentration or solar irradiance. Under optimal conditions (irradiance of 750 W/m 2 , 1.0 mg/L irinotecan concentration, and pH 9.0), more than 98% of irinotecan was degraded in 60 min. With respect to irradiance and irinotecan concentration, the degradation process was robust in the studied range, implying that it may be effectively applied in locations and/or seasons with solar irradiance as low as 450 W/m 2 . However, pH needs to be strictly controlled and kept between 7.0 and 9.0 to maintain the degradation process robust. Considerations about the behavior of degradation products were also drawn., (© 2022. The Author(s), under exclusive licence to European Photochemistry Association, European Society for Photobiology.)

Background: Polycystic ovary syndrome (PCOS) is the most common reproductive-endocrine disorder affecting between 5 and 18% of women worldwide. An elevated frequency of pulsatile luteinizing hormone (LH) secretion and higher serum levels of anti-Müllerian hormone (AMH) are frequently observed in women with PCOS. The origin of these abnormalities is, however, not well understood., Methods: We studied brain structure and function in women with and without PCOS using proton magnetic resonance spectroscopy (MRS) and diffusion tensor imaging combined with fiber tractography. Then, using a mouse model of PCOS, we investigated by electron microscopy whether AMH played a role on the regulation of hypothalamic structural plasticity., Findings: Increased AMH serum levels are associated with increased hypothalamic activity/axonal-glial signalling in PCOS patients. Furthermore, we demonstrate that AMH promotes profound micro-structural changes in the murine hypothalamic median eminence (ME), creating a permissive environment for GnRH secretion. These include the retraction of the processes of specialized AMH-sensitive ependymo-glial cells called tanycytes, allowing more GnRH neuron terminals to approach ME blood capillaries both during the run-up to ovulation and in a mouse model of PCOS., Interpretation: We uncovered a central function for AMH in the regulation of fertility by remodeling GnRH terminals and their tanycytic sheaths, and provided insights into the pivotal role of the brain in the establishment and maintenance of neuroendocrine dysfunction in PCOS., Funding: INSERM (U1172), European Research Council (ERC) under the European Union's Horizon 2020 research and innovation program (grant agreement n° 725149), CHU de Lille, France (Bonus H)., Competing Interests: Declaration of interests A.-L.B. received a honorarium from Merck-Serono for a booklet writing on the use of AMH in Assisted Reproductive Technology; G.K. was a recipient of a Research Grant from Société Française de Neuroradiologie that covered part of his salary. All the other authors have declared that no conflict of interest exists., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

Introduction: Despite major advances in the field of dialysis, there are still some unmet needs such as reducing inflammation through adequate depuration. It is well known that the wide spectrum of pro-inflammatory and pro-atherosclerotic uremic toxins are inefficiently removed by current dialysis techniques. Adsorption seems to be an extra tool to remove toxins, but its effect and optimization have not been widely studied. The aim of this report was to present preliminary results regarding the possibility of performing hemodiafiltration with a highly adsorptive polymethylmethacrylate membrane., Methods: The study was first conducted in 10 patients in which the safety and feasibility of hemodiafiltration with PMMA BG-U 2.1 membrane were tested through measurement of hemolysis indices, transmembrane pressures, and dialysis adequacy. Twenty patients were prospectively observed for 18-month period in which they consecutively underwent standard hemodialysis, standard post-dilution hemodiafiltration, and polymethylmethacrylate-based post-dilution hemodiafiltration. Protein-bound uremic toxins concentrations and inflammatory markers were measured throughout the observed period., Results: HDF-PMMA was inferior to HDF in convective volume, but KT/V was similar, and no differences were noted in operating pressures during the two treatments. During HDF-PMMA period of treatment, we observed a significant reduction of CPR levels, and HDF-PMMA was superior to all other treatments in hepcidin removal even if this did not significantly affect hemoglobin levels. HDF-PMMA could significantly reduce indoxyl sulfate (indoxyl) concentration over a period of 6 months but not for p-cresyl sulfate (p-cresyl)., Conclusion: PMMA BG-U 2.1 membrane can be safely and efficiently used in hemodiafiltration. Moreover, as these preliminary results show, adding adsorption properties to convection and diffusion enabled an increased removal of indoxyl uremic toxin associated to a reduction in inflammation markers as CRP and hepcidin without any negative impact on albumin levels., (© 2024 The Author(s). Published by S. Karger AG, Basel.)

Gonadotropin-releasing hormone isoform I (GnRH), a neuro-deca-peptide, plays a fundamental role in development and maintenance of the reproductive system in vertebrates. The anomalous release of GnRH is observed in reproductive disorder such as hypogonadotropic hypogonadism, polycystic ovary syndrome (PCOS), or following prenatal exposure to elevated androgen levels. Quantitation of GnRH plasma levels could help to diagnose and better understand these pathologies. Here, a validated nano-high-performance liquid chromatography-high-resolution mass spectrometry (HPLC-HRMS) method to quantify GnRH in ewe plasma samples is presented. Protein precipitation and solid-phase extraction (SPE) pre-treatment steps were required to purify and enrich GnRH and internal standard (lamprey-luteinizing hormone-releasing hormone-III, l-LHRH-III). For the validation process, a surrogate matrix approach was chosen following the International Council for Harmonisation (ICH) and FDA guidelines. Before the validation study, the validation model using the surrogate matrix was compared with those using a real matrix such as human plasma. All the tested parameters were analogous confirming the use of the surrogate matrix as a standard calibration medium. From the validation study, limit of detection (LOD) and limit of quantitation (LOQ) values of 0.008 and 0.024 ng/mL were obtained, respectively. Selectivity, accuracy, precision, recovery, and matrix effect were assessed with quality control samples in human plasma and all values were acceptable. Sixteen samples belonging to healthy and prenatal androgen (PNA) exposed ewes were collected and analyzed, and the GnRH levels ranged between 0.05 and 3.26 ng/mL. The nano-HPLC-HRMS developed here was successful in measuring GnRH, representing therefore a suitable technique to quantify GnRH in ewe plasma and to detect it in other matrices and species., (© 2022. The Author(s).)

In this study, the phytochemical profile and the antioxidative properties of Eugenia involucrata fruits were evaluated. Spectrophotometric assays indicated that these berries are a rich source of polyphenols with very high radical-scavenging and metal-reducing activities. High-performance liquid chromatography-Orbitrap analysis was able to carry out the annotation of 36 different compounds, mainly belonging to the flavonol, flavan-3-ol, and anthocyanin families. Antioxidant activity of the fruit extract was evaluated in a cell-based lipid peroxidation model. Obtained data showed that the extract, at very low concentration, was able to prevent oxidative damage in HepG2 cells exposed to oxidative stimuli. Moreover, the evaluation of the gene expression of the most important antioxidant enzymes suggested that the observed antioxidant protection in cells also involves an improvement in enzymatic antioxidant defenses. Finally, the collected data show that E. involucrata fruits are a good source of natural antioxidant molecules and provide evidence of their potential application in the nutraceutical field.

Antifungal azoles are the most frequently used fungicides worldwide and occur as active ingredients in many antifungal pharmaceuticals, biocides, and pesticides. Azole fungicides are frequent environmental contaminants and can affect the quality of surface waters, groundwater, and drinking water. This study examined the potential of combined vacuum UV (185 nm) and UVC (254 nm) irradiation (VUV/UVC) of the azole fungicide tebuconazole and the transformation product 1,2,4-trizole on degradation and changes in ecotoxicity. In vivo ecotoxicity was examined before and after UV treatment using bioassays with test organisms from different trophic levels to integrate changes in biological effect of the parent compound and the degradation products. The test battery included the luminescent bacterium Aliivibrio fischeri, the Gram-positive bacterium Bacillus subtilis, the fungus Fusarium graminearum, the green microalga Raphidocelis subcapitata, and the crustacean Daphnia magna. The combined VUV/UVC treatment of tebuconazole in drinking water efficiently degraded the parent compound at the µg/L-mg/L level and resulted in transformation products with lower toxicity than the parent compound. A direct positive correlation was observed between the applied UV dose (fluence, J/cm 2 ), the disappearance of tebuconazole, and the decrease in ecotoxicity. The combined VUV/UVC process does not require addition of supplementary oxidants or catalysts and our study suggests that VUV/UVC-mediated photolysis of azole fungicides in water can decrease the overall toxicity and represent a potentially environmentally friendly treatment method., (© 2022. The Author(s).)

During a survey for isolating sourdough lactic acid bacteria (LAB), 20 dough samples produced at the bakery level (BL) or home-made (HM) were collected. An enzyme-based colorimetric method revealed a total biogenic amines (BAs) concentration in the range 41.4-251.8 ppm for six (three BL and three HM) sourdoughs characterised by unpleasant odours. Eight BAs generally investigated in foods were identified and quantified from these six samples by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). Only one HM sample contained almost all analysed BAs. Tryptamine was exclusively detected in HM sourdoughs (0.71-24.1 ppm). Putrescine, tryptamine, spermidine, and spermine were the only BAs detected in BL sourdoughs. MiSeq Illumina analysis was applied to study the total bacterial community of sourdoughs. LAB accounted from 67.89 to 92.17% of total bacterial diversity, and Levilactobacillus brevis was identified in all six sourdoughs. Leuconostoc , Pediococcus , and Weissella were also dominant. Plate counts detected neither the presence of Pseudomonas nor members of the Enterobacteriaceae family, and LAB levels were, on average, barely 5.89 Log CFU/g for BL, and 7.33 Log CFU/g for HM sourdoughs. Data suggested that the microorganisms mainly imputable of BAs formation in sourdough are members of the LAB community.

Terpenes are among the major causes of pleasant or unpleasant odors close to active or inactive landfills. We studied R-limonene and p-cymene environmental degradation products using the heterogeneous photocatalysis mediated by titanium dioxide to explore the odor pollution. The aim of the study was the development of mass spectrometry based methods both hyphenated with GC and HPLC to identify and characterize transformation products (TPs) derived from photodegradation of R-limonene and p-cymene. With the GC-MS method we identified three TPs for R-limonene and two for p-cymene comparing the obtained mass spectra with those in the NIST library. While with HPLC-MS method, thanks to the use of the high resolution of MS tool, we recognized four and five TPs for R-limonene and p-cymene respectively. No p-cymene was detected as R-limonene transformation product. The methods developed were then applied to real environmental samples coming from landfills active (Lan1) or inactive (Lan2 and Lan3) located in northern Italy. R-limonene was detected in the active landfill (Lan1 at the concentration of 2.35 μg/mL) together with one of its TPs and one TP derived from p-cymene. p-Cymene was detected in the other two inactive landfills (Lan2 and Lan3 concentrations 0.025 and 0.15 μg/mL, respectively) together with one of its TP and two TPs coming from R-limonene photodegradation. The finding of TPs together with R-limonene and p-cymene both in active and inactive landfills point out the attention on the reduction of these molecules in the environment to reduce pollution and human risks., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

We searched for five neonicotenoids (namely acetamiprid, clothianidin, imidacloprid, thiacloprid and thiamethoxam) in 67 samples of fruits, leaves, pollen and honey via HPLC-MS by employing QueChERs for extraction and purification. Clothianidin was never detected, while imidacloprid was identified in apple (9.2 µg/kg) and pollen (18-28 µg/Kg), thiacloprid in peaches (21-35 µg/kg) and acetamiprid was identified in the hazel leaves (1266 µg/kg), honey (13-26 µg /Kg) and pollen (11-24 µg/kg). Since the levels found of acetamiprid in hazel, honey and pollen were concerning, we accomplished a study to identify and characterize the possible transformation products via a laboratory simulation. The methodology exploited the analysis by HPLC-HRMS and its application in all matrices. We identify twelve transformation products, whose formation involved dimerization, hydroxylation, oxidation, demethylation and cleavage of the molecule. Three of them were also detected in hazel leaves., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Aliskiren was selected as a compound of potential concern among a suspect screening list of more than 40,000 substances on a basis of high occurrence, potential risk and the absence of information about its environmental fate. This study investigated the photoinduced degradation of aliskiren in river water samples spiked at trace levels exposed to simulated sunlight. A half-life time of 24 h was observed with both direct and indirect photolysis playing a role on pollutant degradation. Its photo-induced transformation involved the formation of six transformation products (TPs), elucidated by LC-HRMS - resulted from the drug hydroxylation, oxidation and moieties loss with subsequent cyclization structurally. The retrospective suspected analysis performed on a total of 754 environmental matrices evidenced the environmental occurrence of aliskiren and two TPs in surface waters (river and seawater), fresh water, sediments and biota. In silico bioassays suggested that aliskiren degradation undergoes thought the formation of TPs with distinct toxicity comparing with the parent compound., 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 © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

This study investigated the direct and indirect photochemical degradation of citalopram (CIT), a selective serotonin reuptake inhibitor (SSRI), under natural and artificial solar radiation. Experiments were conducted in a variety of different operating conditions including Milli-Q (MQ) water and natural waters (lake water and municipal WWT effluent), as well as in the presence of natural water constituents (organic matter, nitrate and bicarbonate). Results showed that indirect photolysis can be an important degradation process in the aquatic environment since citalopram photo-transformation in the natural waters was accelerated in comparison to MQ water both under natural and simulated solar irradiation. In addition, to investigate the decontamination of water from citalopram, TiO 2 -mediated photocatalytic degradation was carried out and the attention was given to mineralization and toxicity evaluation together with the identification of by-products. The photocatalytic process gave rise to the formation of transformation products, and 11 of them were identified by HPLC-HRMS, whereas the complete mineralization was almost achieved after 5 h of irradiation. The assessment of toxicity of the treated solutions was performed by Microtox bioassay ( Vibrio fischeri ) and in silico tests showing that citalopram photo-transformation involved the formation of harmful compounds.

Friedreich ataxia (FRDA) is caused by the reduced expression of the mitochondrial protein frataxin (FXN) due to an intronic GAA trinucleotide repeat expansion in the FXN gene. Although FRDA has no cure and few treatment options, there is research dedicated to finding an agent that can curb disease progression and address symptoms as neurobehavioral deficits, muscle endurance, and heart contractile dysfunctions. Because oxidative stress and mitochondrial dysfunctions are implicated in FRDA, we demonstrated the systemic delivery of catalysts activity of gold cluster superstructures (Au 8 -pXs) to improve cell response to mitochondrial reactive oxygen species and thereby alleviate FRDA-related pathology in mesenchymal stem cells from patients with FRDA. We also found that systemic injection of Au 8 -pXs ameliorated motor function and cardiac contractility of YG8sR mouse model that recapitulates the FRDA phenotype. These effects were associated to long-term improvement of mitochondrial functions and antioxidant cell responses. We related these events to an increased expression of frataxin, which was sustained by reduced autophagy. Overall, these results encourage further optimization of Au 8 -pXs in experimental clinical strategies for the treatment of FRDA., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

High plasma levels of the sphingolipid intermediates ceramide (Cer) and sphingosine-1-phosphate (S1P) are suggested to be involved in the development of insulin resistance (IR). Recent evidence indicates that advanced glycation endproducts (AGEs) can alter the sphingolipids metabolism equilibrium. Since enzymes responsible for sphingolipid rheostat maintenance are highly expressed in liver, we thus investigated whether AGEs accumulation can affect hepatic sphingolipids metabolism in insulin resistant mice. Two different models of IR were examined: genetically diabetic LeptrDb-/- (DbDb) and diet-induced insulin resistant C57Bl/6J mice fed a 60% trans-fat diet (HFD). In addition, a group of HFD mice was supplemented with the anti-AGEs compound pyridoxamine. AGEs were evaluated in the liver by western blotting. Cer and S1P were measured by UHPLC-MS/MS. The expression of RAGE and of enzymes involved in sphingolipid metabolism were assessed by RT-PCR and western blotting. HepG2 cells were used to study the effect of the major AGE Nε-(carboxymethyl)lysine (CML)-albumin on sphingolipid metabolism and the role of the receptor of AGEs (RAGE). High levels of AGEs and RAGE were detected in the liver of both DbDb and HFD mice in comparison to controls. The expression of enzymes of sphingolipid metabolism was altered in both models, accompanied by increased levels of Cer and S1P. Specifically, ceramide synthase 5 and sphingosine kinase 1 were increased, while neutral ceramidase was reduced. Pyridoxamine supplementation to HFD mice diminished hepatic AGEs and prevented alterations of sphingolipid metabolism and the development of IR. CML administration to HepG2 cells evoked alterations similar to those observed in vivo, that were in part mediated by the binding to RAGE. The present study shows a direct involvement of AGEs in alterations of sphingolipid metabolism associated to the development of IR. The modulation of sphingolipids metabolism through the prevention of AGEs accumulation by pyridoxamine may reduce the development of IR., (Copyright © 2021. Published by Elsevier Inc.)

The presence of residues from fining agents in wines may represent a risk for allergic consumers and a source of discomfort for others, such as vegans. Even though ELISA is the official detection method for such residues, this technique may be hindered by cross-reactivity issues, or by matrix-molecule interference due to a high polyphenol content, especially in red wines. An HRMS-based method has been developed to detect pig gelatin and egg white in experimental five-year aged Nebbiolo-based red wine. Biomarker peptides were selected, after tryptic digestion, and quantified by multitarget nanoHPLC-HRMS analysis. The method resulted in an LLOQs of 5 µg/mL in the experimental wine, and between 1 and 2 µg/mL in the buffer. This method allowed both gelatin and egg white proteins to be detected and quantified in aged red wine, while whereas the commercial ELISA kit was instead unable to detect egg white in the same samples., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Rationale: Bisphenol E (BPE) and bisphenol S (BPS) have recently replaced bisphenol A as monomers for producing polycarbonates. However, BPE and BPS can pose hazards as they are known to be endocrine disruptors. Despite the huge increase in their use, there is a lack of data regarding the toxicity and effects of BPE and BPS., Methods: We investigated the photoinduced transformation of BPE and BPS when subjected to sun-simulated radiation and using TiO 2 as a photocatalyst. Analyses of BPE, BPS and their by-products were performed by high-performance liquid chromatography/high-resolution mass spectrometry (HPLC/HRMS) using an orbitrap mass analyzer in negative electrospray ionisation (ESI) mode. The chromatographic separations were achieved by employing a C18 reversed-phase column, and the transformation products (TPs) were elucidated structurally using HRMS and multistage MS experiments performed in collision-induced dissociation (CID) mode., Results: The transformation of bisphenol S involved the formation of twelve by-products, while ten TPs were detected following BPE degradation. For bisphenol S, the cleavage of the molecule is a very important transformation route, together with the hydroxylation of the substrate to provide mono- and poly-hydroxylated TPs. For bisphenol E, the two main routes were hydroxylation and ring opening. Acute toxicity for BPS, BPE and their TPs was assessed using the Vibrio fischeri assay, highlighting that their initial transformation involved the formation of TPs that were more toxic than the parent compound., Conclusions: The HPLC/HRMS method developed was useful for characterising and identifying newly formed TPs from bisphenol E and bisphenol S. This study aimed to examine the structure of twenty by-products identified during TiO 2 -mediated photolysis and to evaluate acute toxicity over time., (© 2021 John Wiley & Sons Ltd.)

Protium heptaphyllum (Aubl.) Marchand (PH) trees are endemic to the tropical region of South America, mostly Brazil. Antibacterial, antinociceptive, anti-inflammatory, anxiolytic, antidepressant and anti-hyperlipidemic/anti-hypercholesterolemic effects were reported for its resinous exudate Protium heptaphyllum resin (PHR). This work aims to provide a qualitative and quantitative consistent chemical profiling of the major constituents of this resin and two extracts enriched in acid (acidic triterpene concentrated extract, ATCE) and neutral triterpenes (α and β-amyrin concentrated extract, AMCE). GC-MS/GC-FID was used for volatile terpene fraction, a validated GC-MS method was developed for quantification of neutral α and β-amyrin and HPLC-APCI HRMS 2 was used for acidic triterpenes analysis. The chemical investigation reported 29 molecules, including 14 volatile terpenes, 6 neutral triterpenes and 11 acid triterpenes. The most abundant compounds were α-amyrin (251.28 g kg -1 , 123.98 g kg -1 and 556.82 g kg -1 in PHR, ATCE and AMCE, respectively), β-amyrin (172.66 g kg -1 , 95.39 g kg -1 and 385.58 g kg -1 in PHR, ATCE and AMCE, respectively), 3-oxo-tirucalla-7,24-dien-21-oic acid (80.64 g kg -1 , 157.10 g kg -1 and 15.31 g kg -1 in PHR, ATCE and AMCE, respectively) and 3α-hydroxy-tirucalla-8,24-dien-21-oic acid (77.71 g kg -1 , 130.40 g kg -1 and 11.64 g kg -1 in PHR, ATCE and AMCE, respectively). Results showed specific enrichment of acidic and neutral triterpenoids in the two respective extracts.

Hypercholesterolemia is one of the major causes of cardiovascular disease, the risk of which is further increased if other forms of dyslipidemia occur. Current therapeutic strategies include changes in lifestyle coupled with drug administration. Statins represent the most common therapeutic approach, but they may be insufficient due to the onset of resistance mechanisms and side effects. Consequently, patients with mild hypercholesterolemia prefer the use of food supplements since these are perceived to be safer. Here, we investigate the phytochemical profile and cholesterol-lowering potential of Protium heptaphyllum gum resin extract (PHE). Chemical characterization via HPLC-APCI-HRMS 2 and GC-FID/MS identified 13 compounds mainly belonging to ursane, oleanane, and tirucallane groups. Studies on human hepatocytes have revealed how PHE is able to reduce cholesterol production and regulate the expression of proteins involved in its metabolism. ( HMGCR , PCSK9 , LDLR , FXR , IDOL , and PPAR ). Moreover, measuring the inhibitory activity of PHE against HMGR, moderate inhibition was recorded. Finally, molecular docking studies identified acidic tetra- and pentacyclic triterpenoids as the main compounds responsible for this action. In conclusion, our study demonstrates how PHE may be a useful alternative to contrast hypercholesterolemia, highlighting its potential as a sustainable multitarget natural extract for the nutraceutical industry that is rapidly gaining acceptance as a source of health-promoting compounds.

The aim of the present study was to assess the effects of bisphenol (BP) exposure on pregnancy and neonatal life. We have (a) determined BP (BPA and BPS) concentration levels in a group of newborns and their mothers; (b) identified factors, habits, and devices possibly responsible for BP uptake; and (c) determined the effect of BP exposure. No significant correlations were detected between maternal and neonatal BP concentration levels. In newborns, positive correlations between pacifier use and BPS total ( p = 0.04) and free BPS ( p = 0.03) concentrations were detected. A significant correlation was also found between oral glucose administration and concentration levels of free BPA ( p < 0.05). Our study points to a central role of lifestyle, hospital procedures, and neonatal devices in inducing BP exposure, especially during the perinatal period. This is the first report of BP contamination in newborns due to widely non-alimentary products designed for newborn care, such as glucose-solution containers for BPA and pacifiers for BPS. Further studies are advocated in order to clarify both the impact of other BP forms on human health and development, as well as potential BPA exposure sources during neonatal and childhood life.

Scope: A high fructose and galactose intake show adverse metabolic effects in animal models and in humans, but it is yet unknown if addition of fermentable dietary fiber can mitigate such effects. This study investigate the effects of high intakes of fructose and galactose, with/without added fructooligosaccharides (FOS), on metabolic factors, inflammation, and gut integrity markers in rats., Methods and Results: Rats (n = 6/group) receive different carbohydrates at isocaloric conditions for 12 weeks as follows: 1) starch (control), 2) fructose, 3) galactose, 4) starch + FOS (FOS control), 5) fructose + FOS, and 6) galactose + FOS, together with a high amount of n-6 polyunsaturated fatty acids (n-6 PUFA) in all diets except for in 7) starch + olive oil (negative control). The rats fed the galactose and galactose + FOS diets exhibit lower body weight than other groups. High-galactose diets has more pronounced effects on metabolic factors and gut permeability than high-fructose diets. High-fructose diets show less pronounced effect on these selected markers. No differences in inflammatory markers are detected for any of the diets., Conclusions: The results suggest potential adverse effects of high galactose and fructose on metabolic factors and gut integrity markers, but not on inflammation. However, several mechanisms are at play, and general net effects are difficult to determine conclusively for the conditions tested., (© 2021 The Authors. Molecular Nutrition & Food Research published by Wiley-VCH GmbH.)

In the present study, the photoinduced degradation of the antidepressant drug sertraline under artificial solar radiation was examined. Photolysis was studied under different experimental conditions to explore its photolytic fate in the aqueous environment. Photolytic degradation kinetics were carried out in ultrapure water, wastewater effluent, as well as in the presence of dissolved organic matter (humic acids), bicarbonate and nitrate ions which enabled their assessment on sertraline photo-transformation. The reaction of sertraline with photoactive compounds accelerated sertraline transformation in comparison with direct photolysis. Moreover, TiO 2 -mediated photocatalytic degradation of sertraline was investigated, and focus was placed on the identification of by-products. As expected, photocatalysis was extremely effective for sertraline degradation. Photocatalytic degradation proceeded through the formation of forty-four transformation products identified by HPLC-HRMS and after 240 min of irradiation total mineralization was achieved. Microtox bioassay (Vibrio fischeri) was employed to assess the ecotoxicity of the photocatalysis-treated solutions and results have indicated that sertraline photo-transformation proceeds through the formation of toxic compounds., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

Maprotiline was identified as a compound of potential interest further to a suspect screening test carried out for a list of more than 40,000 substances based on specific occurrence, hazard and risk indicators. Despite the high frequency of appearance of this drug in wastewater treatment stations, his environmental fate is still unknown. Herein, we investigated for the first time the maprotiline degradation pathways in river water spiked with the drug at a concentration close to those detected in natural waters. Preliminary photocatalytic experiments in ultrapure water produced 32 transformation products (TPs) resulted mainly from the multiple hydroxylation/oxidation in different positions of the drug molecule. From the river water experiments, 12 TPs were formed by photolysis matching with those observed in ultrapure water experiments, and 2 were also formed resulted from biotic degradation. Employing HPLC-HRMS, we were able to elucidate the chemical structures of TPs and assess the overall degradation mechanism. Preliminary bioassays suggested lower toxicity of TPs relatively to the parent compound., 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 The Authors. Published by Elsevier B.V. All rights reserved.)

Olive quick decline syndrome (OQDS) is a multifactorial disease affecting olive plants. The onset of this economically devastating disease has been associated with a Gram-negative plant pathogen called Xylella fastidiosa (Xf). Liquid chromatography separation coupled to high-resolution mass spectrometry detection is one the most widely applied technologies in metabolomics, as it provides a blend of rapid, sensitive, and selective qualitative and quantitative analyses with the ability to identify metabolites. The purpose of this work is the development of a global metabolomics mass spectrometry assay able to identify OQDS molecular markers that could discriminate between healthy (HP) and infected (OP) olive tree leaves. Results obtained via multivariate analysis through an HPLC-ESI HRMS platform (LTQ-Orbitrap from Thermo Scientific) show a clear separation between HP and OP samples. Among the differentially expressed metabolites, 18 different organic compounds highly expressed in the OP group were annotated; results obtained by this metabolomic approach could be used as a fast and reliable method for the biochemical characterization of OQDS and to develop targeted MS approaches for OQDS detection by foliage analysis.

Quorum sensing (QS) is the ability of some bacteria to detect and to respond to population density through signalling molecules. QS molecules are involved in motility and cell aggregation mechanisms in diseases such as sepsis. Few biomarkers are currently available to diagnose sepsis, especially in high-risk conditions. The aim of this study was the development of new analytical methods based on liquid chromatography-mass spectrometry for the detection and quantification of QS signalling molecules, including N-acyl homoserine lactones (AHL) and hydroxyquinolones (HQ), in biofluids. Biological samples used in the study were Pseudomonas aeruginosa bacterial cultures and plasma from patients with sepsis. We developed two MS analytical methods, based on neutral loss (NL) and product ion (PI) experiments, to identify and characterize unknown AHL and HQ molecules. We then established a multiple-reaction-monitoring (MRM) method to quantify specific QS compounds. We validated the HPLC-MS-based approaches (MRM-NL-PI), and data were in accord with the validation guidelines. With the NL and PI MS-based methods, we identified and characterized 3 and 13 unknown AHL and HQ compounds, respectively, in biological samples. One of the newly found AHL molecules was C12-AHL, first quantified in Pseudomonas aeruginosa bacterial cultures. The MRM quantitation of analytes in plasma from patients with sepsis confirmed the analytical ability of MRM for the quantification of virulence factors during sepsis. Graphical abstract.

Rationale: Gabapentin is a drug used to treat epilepsy and peripheral neuropathic pain. It is an analog of gamma-aminobutyric acid, and it is a selective blocker of voltage-gated calcium channels. The drug is excreted unmetabolized; it is stable in the environment and is classified as a persistent mobile organic contaminant. Because wastewater treatment plants (WWTPs) are not completely efficient, some bioactive molecules may be released unaltered into the environment. The aim of this study was to provide information about degradation pathways of gabapentin in water by studying its photoinduced transformation products (TPs) through laboratory simulation experiments. Gabapentin and its TPs were monitored in influent and effluent water samples from WWTPs in Germany and Italy., Methods: The laboratory simulation used heterogeneous photodegradation mediated by titanium dioxide (TiO 2 ). Chromatographic separation was achieved using a C18 reverse-phase column, and the structural identification of TPs was performed using high-resolution electrospray ionization high-resolution mass spectrometry (ESI-HRMS) and multistage MS n experiments., Results: Several TPs were observed during TiO 2 photodegradation. Nine new compounds were detected, and potential structures were assigned by studying the fragmentation pathways of the [M + H] + ions of these TPs and gabapentin. Gabapentin and some of the newly identified TPs were found in environmental samples from WWTPs., Conclusions: The developed high-performance liquid chromatography/high-resolution mass spectrometry method was used to identify TPs from gabapentin. It was then successfully applied to real environmental samples to monitor the TPs as potential environmental pollutants., (© 2020 John Wiley & Sons, Ltd.)

The active herbicide ingredient glyphosate [N-(phosphonomethyl)glycine] is frequently detected as a contaminant in groundwater and surface waters. This study investigated effects of UV-A (365 nm), UV-B (302 nm) and UV-C (254 nm) irradiation of glyphosate in water on photolysis and toxicity to aquatic organisms from different trophic levels. A test battery with bacteria (Bacillus subtilis, Aliivibrio fischeri), a green microalga (Raphidocelis subcapitata), and a crustacean (Daphnia magna) was used to assess biological effect of glyphosate and bioactive transformation products before and after UV irradiation (4.7-70 J/cm 2 ). UV-C irradiation at 20 J/cm 2 resulted in a 2-23-fold decrease in toxicity of glyphosate to aquatic test organisms. UV-B irradiation at 70 J/cm 2 caused a twofold decrease whereas UV-A did not affect glyphosate toxicity at doses ≤ 70 J/cm 2 . UV-C irradiation of glyphosate in drinking water and groundwater with naturally occurring organic and inorganic constituents showed comparable or greater reduction in toxicity compared to irradiation in deionized water. High-resolution mass spectrometry analyses of samples after UV-C irradiation showed > 90% decreases in glyphosate concentrations and the presence of multiple transformation products. The study suggests that UV mediated indirect photolysis can decrease concentrations of glyphosate and generate less toxic products with decreased overall toxicity to aquatic organisms.

Hazelnut skin (HS) was evaluated as a source of nutrients for dairy cows. In total, 26 Aosta Red Pied cows were divided into two balanced groups. All cows were fed hay ad libitum. The control group was also given 6 kg of concentrate, while the hazelnut skin group (HAZ) was given 1 kg of the same concentrate that was substituted by 1 kg of HS. The dry matter intake of the cows was reduced by the dietary inclusion of HS ( p ≤ 0.001). The milk yield and main constituents were unaffected by treatment. Milk from HAZ cows showed decreased concentrations of de novo saturated fatty acids (FAs), odd- and branched-chain FA, α-linolenic acid, and long-chain n-3 FAs, as well as increased concentrations of stearic acid, oleic acid, linoleic acid, total monounsaturated FAs, trans biohydrogenation intermediates, and α-tocopherol. Replacing the concentrate with HS increased the human-inedible feed quota in the diet and improved the sustainability of milk production in terms of the food-feed competition. Our results suggest that it is possible to add economic value to organic waste from the hazelnut industry using HS as a feed ingredient for dairy cows, enhancing the feed efficiency and milk antioxidant activity and having expected impacts on the nutraceutical quality of milk fat.

The popularity of edible wild fruits has increased in industrialized countries due to their composition and positive effects. The aim of this study has been to characterize the polyphenolics and anthocyanins of black mulberry (Morus nigra L.), cornelian cherry (Cornus mas L.), elderberry (Sambucus nigra L.), hawthorn (Crataegus monogyna L.), lingonberry (Vaccinium vitis-idaea L.) and rose hip (Rosa canina L.) harvested in the north-west of Italy by means of HPLC-DAD-ESI HRMS in positive ion mode. Although there is an abundant amount of literature related to the polyphenolics of cultivated fruit, a new type of comparison has here been conducted between wild and cultivated fruits on their polyphenolic content. The HPLC-DAD-ESI HRMS method has detected 64 different polyphenolic molecules and it can be used to perform qualitative and quantitative analyses. Furthermore, the cornelian cherry and elderberry samples showed the highest polyphenolic compound levels. The quercetin glycosylated compounds showed the highest percentage of flavonols in most of the analyzed wild fruits.

Inhibition of either P2Y12 receptor or the nucleotide-binding oligomerization domain- (NOD-) like receptor pyrin domain containing 3 (NLRP3) inflammasome provides cardioprotective effects. Here, we investigate whether direct NLRP3 inflammasome inhibition exerts additive effects on myocardial protection induced by the P2Y12 receptor antagonist Ticagrelor. Ticagrelor (150 mg/kg) was orally administered to rats for three consecutive days. Then, isolated hearts underwent an ischemia/reperfusion (30 min ischemia/60 min reperfusion; IR) protocol. The selective NLRP3 inflammasome inhibitor INF (50  μ M) was infused before the IR protocol to the hearts from untreated animals or pretreated with Ticagrelor. In parallel experiments, the hearts isolated from untreated animals were perfused with Ticagrelor (3.70  μ M) before ischemia and subjected to IR. The hearts of animals pretreated with Ticagrelor showed a significantly reduced infarct size (IS, 49 ± 3% of area at risk, AAR) when compared to control IR group (69 ± 2% of AAR). Similarly, ex vivo administration of INF before the IR injury resulted in significant IS reduction (38 ± 3% of AAR). Myocardial IR induced the NLRP3 inflammasome complex formation, which was attenuated by either INF pretreatment ex vivo , or by repeated oral treatment with Ticagrelor. The beneficial effects induced by either treatment were associated with the protective Reperfusion Injury Salvage Kinase (RISK) pathway activation and redox defence upregulation. In contrast, no protective effects nor NLRP3/RISK modulation were recorded when Ticagrelor was administered before ischemia in isolated heart, indicating that Ticagrelor direct target is not in the myocardium. Our results confirm that Ticagrelor conditioning effects are likely mediated through platelets, but are not additives to the ones achieved by directly inhibiting NLRP3., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2020 Claudia Penna et al.)

The popularity of small berries has rapidly increased in Western countries given their antioxidant, anti-inflammatory, and antimicrobial activities and health-promoting properties. The aim of this study was to compare the fatty acid (FA) profile, phenolic compounds, and antioxidant capacity of extracts of 11 berries cultivated in the North West of Italy. Berry samples were extracted and evaluated for FA profile and total anthocyanin (TAC), total flavonoid contents (TFC), ferric-reducing antioxidant power (FRAP), and for their radical scavenging activities against 2,2'-diphenyl-1-picrylhydrazyl (DPPH • ) radical. The main polyphenols of berry extracts were characterized by HPLC-DAD-UV-ESI HRMS in positive ion mode. Results showed that the highest TAC and TFC contents were recorded in black currants, blackberries, and blueberries. Maximum and minimum DPPH • radical scavenging activities, Trolox Equivalent Antioxidant Capacity, and FRAP measurements confirmed the same trend recorded for TAC and TFC values. HPLC-HRMS analyses highlight how blueberries and blackberries have the highest concentration in polyphenols. Palmitic, stearic, oleic, linoleic, α-linolenic, and γ-linolenic acids significantly differ between berries, with oleic and α-linolenic acid representing the most abundant FAs in raspberries. Among the berries investigated, results of phytochemical characterization suggest choosing black currants and blueberries as an excellent source of natural antioxidants for food and health purposes.

Fishery products are perishable due to the action of many enzymes, both endogenous and exogenous. The latter are produced by bacteria that may contaminate the products. When fishes age, there is a massive bacteria growth that causes the appearance of off-flavor . In order to obtain "false" freshness of fishery products, an illicit treatment with hydrogen peroxide is reported to be used. Residues of hydrogen peroxide in food may be of toxicology concern. We developed two mass spectrometry based methodologies to identify and quantify molecules related to the treatment of fishes with hydrogen peroxide. With ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS) we evaluated the concentration of trimethylamine-N-oxide (TMAO), trimethylamine (TMA), dimethylamine (DMA), and cadaverine (CAD) in fish products. After evaluating LOQ, we measured and validated the lower limits of quantification (LLOQs as first levels of calibration curves) values of 50 (TMAO), 70 (TMA), 45 (DMA), and 40 (CAD) ng/mL. A high ratio between TMAO and TMA species indicated the freshness of the food. With a GC-MS method we confirmed the illicit treatment measuring the levels of H 2 O 2 after an analytical reaction with anisole to give 2-hydroxyanisole as a marker. This latter product was detected in the headspace of the homogenized sample with simplification of the work-up. A LLOQ of 50 ng/mL was checked and validated. When fish products were whitened and refreshed with hydrogen peroxide, the detected amount of the product 2-hydroxyanisole could be very important, (larger than 100 mg/kg). The developed analytical methods were suitable to detect the illicit management of fishery products with hydrogen peroxide; they resulted as sensitive, selective, and robust.

Background: Cytochrome P450 enzymes (CYPs) are monooxygenases present in every domain of life. In fungi CYPs are involved in virulence. Fusarium wilt of lettuce, caused by F. oxysporum f. sp. lactucae, is the most serious disease of lettuce. F. oxysporum f.sp. lactucae MSA35 is an antagonistic fungus. Pathogenic formae specialis of F. oxysporum possess a CYP belonging to the new family CYP505. This enzyme hydroxylates saturated fatty acids that play a role in plant defence., Methods: Molecular tools were adopted to search for cyp505 gene in MSA35 genome. cyp505 gene expression analysis in pathogenic and antagonistic Fusarium was performed. The enzyme was expressed in its recombinant form and used for catalytic reactions with fatty acids, the products of which were characterized by mass spectrometry analysis., Results: A novel MSA35 self-sufficient CYP505 is differentially expressed in antagonistic and pathogenic F. oxysporum. Its expression is induced by the host plant lettuce in both pathogenesis and antagonism during the early phase of the interaction, while it is silenced during the late phase only in antagonistic Fusarium. Mass-spectrometry investigations proved that CYP505A1 mono-hydroxylates lauric, palmitic and stearic acids., Conclusions: The ability of CYP505A1 to oxidize fatty acids present in the cortical cell membranes together with its differential expression in its Fusarium antagonistic form point out to the possibility that this enzyme is associated with Fusarium pathogenicity in lettuce., General Significance: The CYP505 clan is present in pathogenic fungal phyla, making CYP505A1 enzyme a putative candidate as a new target for the development of novel antifungal molecules., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In this research, the degradation of seven different micropollutants (MPs) and the formation of their transformation products (TPs) have been assessed during the application of different advanced oxidation processes: photolytic and photocatalytic activation of peroxymonosulfate (PMS) and persulfate (PS). The results were compared with those obtained from the photolytic experiments using hydrogen peroxide (H 2 O 2 ) as oxidant. A significant abatement of almost all MPs was achieved, even with very low UV-C contact time (9 and 28 s). The degradation of atenolol (ATN) and caffeine (CFN) ranged from 84 to 100% with a dose of 0.5 mM of any oxidant. The efficiencies for bisphenol-A (BPA), carbamazepine (CBZ), diclofenac (DCF), ibuprofen (IBP), and sulfamethoxazole (SMX) varied depending on the oxidation system and operating conditions (oxidant dose and UV-C contact time), leading to the photolysis of PMS to higher efficiencies than PS and H 2 O 2 . In all cases, the abatement of MPs ranged from 63 to 83%, even with the lowest PMS dosage. Moreover, the addition of Fe(II) as a catalyst enhanced the removal efficiency, reaching almost total removal, especially over CBZ, DCF, and IBP. The Dissolved Organic Carbon (DOC) removal ranged between 44 and 62%, suggesting the transformation of MPs in intermediate compounds. The identification of transformation products was carried out for each micropollutant and each oxidation treatment, being observed some transformation products specific of oxidation by sulfate radicals. For example, m/z 165.0432 only appeared after PMS/Fe(II)/UV-C on the degradation of BFA, m/z 251.082 appeared after photolytic activation of PMS and PS on CBZ removal, and m/z 128.0452 was observed after any sulfate radical oxidation treatment, but not after photolysis of H 2 O 2 ., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The extensive use of mollusc shell as a versatile raw material is testament to its importance in prehistoric times. The consistent choice of certain species for different purposes, including the making of ornaments, is a direct representation of how humans viewed and exploited their environment. The necessary taxonomic information, however, is often impossible to obtain from objects that are small, heavily worked or degraded. Here we propose a novel biogeochemical approach to track the biological origin of prehistoric mollusc shell. We conducted an in-depth study of archaeological ornaments using microstructural, geochemical and biomolecular analyses, including 'palaeoshellomics', the first application of palaeoproteomics to mollusc shells (and indeed to any invertebrate calcified tissue). We reveal the consistent use of locally-sourced freshwater mother-of-pearl for the standardized manufacture of 'double-buttons'. This craft is found throughout Europe between 4200-3800 BCE, highlighting the ornament-makers' profound knowledge of the biogeosphere and the existence of cross-cultural traditions., Competing Interests: JS, SA, PB, MB, TC, AC, FD, AG, MH, HK, GM, CM, KP, LP, HS, ST, CT, JT, JW, FM, BD No competing interests declared, (© 2019, Sakalauskaite et al.)

Here, the performances of two different catalysts, Ce-ZnO and TiO2 synthetized in our laboratories, were compared with the commercial TiO2-P25 for degradation of a mixture of seven emerging pollutants under UV irradiation. The investigation included monitoring pollutants abatement in Milli-Q water and in wastewater effluent and identifying their transformation products by HPLC-HRMS. Structural characterization of intermediates supported by data available from literature allowed elucidation of the transformation pathways occurring in the presence of all investigated catalysts in the wastewater effluent. Preliminary results showed a good removal efficacy for almost all examined contaminants, even in real water matrix (i.e. wastewater effluent). The type of matrix and catalyst affects the number and/or the abundance of transformation products, which suggests differences in their transformation routes.

A new class of foreign substances present in the unsaponifiable fraction of vegetable oils undergone to chemical interesterification was systematically investigated. Their chemical structure, corresponding to dialkyl ketones (DAK) molecules, was elucidated both by gas chromatography-mass spectrometry (GC-MS), and liquid chromatography-high resolution mass spectrometry (LC-HRMS). An analytical protocol aimed to qualitative and quantitative detection of DAK molecules in vegetable oils of confectionery industry interest was developed. Being the range of concentration levels to be evaluated dependent on the technological task of interesterification process, the quantitation step was thoroughly examined. All the validation parameters were satisfactory and particularly the concentration determinations were made more reliable by the contemporary use of several quantitation standards. GC-MS and LC-HRMS analytical techniques exhibited comparable performances even if the second one shown better detection sensitivity., (Copyright © 2018 Elsevier B.V. All rights reserved.)

PI3K activation plays a central role in the development of pulmonary inflammation and tissue remodeling. PI3K inhibitors may thus offer an improved therapeutic opportunity to treat non-resolving lung inflammation but their action is limited by unwanted on-target systemic toxicity. Here we present CL27c, a prodrug pan-PI3K inhibitor designed for local therapy, and investigate whether inhaled CL27c is effective in asthma and pulmonary fibrosis. Mice inhaling CL27c show reduced insulin-evoked Akt phosphorylation in lungs, but no change in other tissues and no increase in blood glycaemia, in line with a local action. In murine models of acute or glucocorticoid-resistant neutrophilic asthma, inhaled CL27c reduces inflammation and improves lung function. Finally, inhaled CL27c administered in a therapeutic setting protects from bleomycin-induced lung fibrosis, ultimately leading to significantly improved survival. Therefore, local delivery of a pan-PI3K inhibitor prodrug reduces systemic on-target side effects but effectively treats asthma and irreversible pulmonary fibrosis.

Polycystic ovary syndrome (PCOS) is the main cause of female infertility worldwide and corresponds with a high degree of comorbidities and economic burden. How PCOS is passed on from one generation to the next is not clear, but it may be a developmental condition. Most women with PCOS exhibit higher levels of circulating luteinizing hormone, suggestive of heightened gonadotropin-releasing hormone (GnRH) release, and anti-Müllerian hormone (AMH) as compared to healthy women. Excess AMH in utero may affect the development of the female fetus. However, as AMH levels drop during pregnancy in women with normal fertility, it was unclear whether their levels were also elevated in pregnant women with PCOS. Here we measured AMH in a cohort of pregnant women with PCOS and control pregnant women and found that AMH is significantly more elevated in the former group versus the latter. To determine whether the elevation of AMH during pregnancy in women with PCOS is a bystander effect or a driver of the condition in the offspring, we modeled our clinical findings by treating pregnant mice with AMH and followed the neuroendocrine phenotype of their female progeny postnatally. This treatment resulted in maternal neuroendocrine-driven testosterone excess and diminished placental metabolism of testosterone to estradiol, resulting in a masculinization of the exposed female fetus and a PCOS-like reproductive and neuroendocrine phenotype in adulthood. We found that the affected females had persistently hyperactivated GnRH neurons and that GnRH antagonist treatment in the adult female offspring restored their neuroendocrine phenotype to a normal state. These findings highlight a critical role for excess prenatal AMH exposure and subsequent aberrant GnRH receptor signaling in the neuroendocrine dysfunctions of PCOS, while offering a new potential therapeutic avenue to treat the condition during adulthood.

Interesterification is an industrial transformation process aiming to change the physico-chemical properties of vegetable oils by redistributing fatty acid position within the original constituent of the triglycerides. In the confectionery industry, controlling formation degree of positional isomers is important in order to obtain fats with the desired properties. Silver ion HPLC (High Performance Liquid Chromatography) is the analytical technique usually adopted to separate triglycerides (TAGs) having different unsaturation degrees. However, separation of TAG positional isomers is a challenge when the number of double bonds is the same and the only difference is in their position within the triglyceride molecule. The TAG positional isomers involved in the present work have a structural specificity that require a separation method tailored to the needs of confectionery industry. The aim of this work was to obtain a chromatographic resolution that might allow reliable qualitative and quantitative evaluation of TAG positional isomers within reasonably rapid retention times and robust in respect of repeatability and reproducibility. The resulting analytical procedure was applied both to confectionery raw materials and final products.

We report a new High Performance Liquid Chromatography-Mass Spectrometry (HPLC-MS) method to rapidly detect and quantify meglumine by-products (specifically reducing sugar(s) and nitrogen impurities) that could be present in the meglumine samples. Meglumine is a secondary amine obtained from glucose and it is an excipient used as counter-ion in several pharmaceutical formulations, especially when the concentration of the active pharmaceutical ingredient (API) is so high that the sodium is not a suitable option. Moreover, the increased use of meglumine is related to its ability to improve solubility in aqueous solutions due to the presence of a large number of hydroxyl groups. Thus, even if meglumine is widely used as excipient in pharmaceutical formulations, its impurity profile has never been fully evaluated. Here, we propose the use of a commercial agent that specifically reacts with carbonyl compounds, 1-(4-aminobenzyl)-1,2,4-triazole, with the aim of improving the detection of reducing sugars, such as glucose, after an easy derivatization procedure. Finally, we describe the method validation and the analysis of the impurity profile of meglumine samples from different manufacturers., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The photocatalytic transformation of acesulfame K - an artificial sweetener that has gained popularity over the last decades for being a calorie-free additive in food, beverages and several pharmaceutical products - was studied using three different photocatalysts, the benchmark TiO 2 -P25 and two other forms of synthetized titanium oxides named TiO 2 -SG1 and TiO 2 -SG2. The two latter materials were synthesized by a sol gel process in which the hydrolysis rate of titanium n-butoxide was controlled by the water formed in situ through an esterification reaction between ethanol and acetic acid. The investigation included monitoring the sweetener disappearance, identifying its intermediate compounds, assessing mineralization and evaluating toxicity. The analyses were carried out using high-performance liquid chromatography (HPLC) coupled with a LTQ-Orbitrap analyzer via an electrospray ionization (ESI) in the negative ion mode. This is a powerful tool for the identification, characterization and measurement of the transformation products (TPs); overall 13 species were identified. The use of several semiconductors has pointed out differences in terms of both photocatalytic efficiency and mechanism: the assessment of the evolution kinetics of each species (TPs, total organic carbon and inorganic ions) has brought to the elaboration of a general transformation pathway of acesulfame K. TiO 2 -SG2 proved to be the most efficient material in degrading the artificial sweetener and leads to the complete mineralization within 6 h of irradiation, while up to 16 h are required for TiO 2 -P25., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

We studied some ionic liquids (ILs) belonging to the pyridinium class under photocatalytic treatment. In particularly, we analysed how the length of the alkyl chain, the kind of inorganic ion and the type of substituents could influence the disappearance rate, the mineralization extent, the acute toxicity and the transformation mechanism. For such, we selected some pyridinium derivatives with different alkyl chain but the same anion, namely tetrafluoroborate (1-ethylpyridinium, 1-butylpyridinium, 1-hexylpyridinium), with two alkyl substituents (4-methyl-1-butylpyridinium) and with a different substituent (1-cyanopropylpyridinium). Then, on a selected IL (1-butylpyridinium), we evaluate the role of different inorganic anions (bromine and chlorine). The results show that irrespective to the alkyl chain or the number of substituents, the transformation involved an attack to the alkyl chain, proceeded through the formation of harmless compounds and the mineralization was easily achieved within 4h. Nitrogen was mainly released as ammonium ion. When introducing a cyano group, the extent of nitrate ions and the number of possible transformation route increased. Conversely, the type of inorganic ion deeply affected the transformation pathways and the extent of mineralization. Actually, in the presence of bromide as anion, IL was only partially mineralized and the formation of highly persistent transformation products occurred., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The present study reports the photocatalytic transformation of stevioside, under simulated solar irradiation using TiO 2 as a photocatalyst. As a tool of investigating the effect of various aqueous matrices, as well as, the initial stevioside concentration on the variation of the photocatalytic efficiency, a fully nested experimental design was employed. A significant impact on the degradation rate of the sweetener was observed: degradation rate decreases in the order distilled water>river water>lake water, attributed to the increased natural organic matter content of the respective natural water samples. Moreover, the investigation has involved the identification of intermediate compounds, as well as the assessment of mineralization and toxicity evaluation. More than one hundred unknown transformation products, most of them in the form of several isobaric species, were identified. By employing accurate mass determination, we were able to attribute an empirical formula to each species and through MS n analyses we were capable to distinguish several isobaric species. The overall transformation mechanism was assessed and involved the hydroxylation/oxidation of the molecule and the subsequent loss of the glucose units bound to the parent compound., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Rationale: Ionic liquids (ILs) are a subject of active research in the field of alternative solvents. We studied the behaviour of a piperidine IL, 1-butyl-1-methylpiperidinium tetrafluoroborate (BMPA), through the elucidation of its transformation products (TPs) in water., Methods: The transformation pathways of BMPA were investigated using high-performance liquid chromatography (HPLC) combined with a hybrid LTQ-Orbitrap instrument on the basis of mass defect filtering. TPs of BMPA were identified by fragmentation patterns and accurate mass measurements., Results: The separation and identification of 32 TPs was achieved. BMPA can be oxidized at different positions in the alkyl chains. The ultimate products corresponds to N-methyl-piperidinium and some byproducts involving ring-opening. Tests of acute toxicity, evaluated with Vibrio Fischeri bacteria, show that BMPA transformation proceeds through the formation of slightly harmful compounds., Conclusions: Results showed that the main transformation pathways of BMPA were alkyl chain hydroxylation/shortening and de-alkylation, and that HPLC/LTQ-Orbitrap can serve as an important analytical platform to gather the unknown TPs of ILs., (Copyright © 2017 John Wiley & Sons, Ltd.)