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A four-year survey was conducted to monitor the presence of multiple pesticide residues contaminating tomatoes, with the aim of evaluating the potential health and environmental risks. A multiresidue liquid chromatography–triple mass spectrometry with a multiple reaction monitoring (LC-MS/MS-MRM) method was fully validated and used to test 252 pesticides in 360 samples analysed. According to SANTE guidelines, the proposed method was considered suitable for the purpose. Dietary risk assessment was conducted using the Hazard Quotient (HQ) approach and the European Food Safety Authority (EFSA) Pesticide Residue Intake Model; meanwhile, the cumulative environmental risk assessment was conducted using the Concentration Addition (CA) and Independent Action (IA) methods. Data obtained revealed multiple contaminations in most fields examined over the years. Twenty-two pesticide residues were identified, comprising 68.2% fungicides, 27.3% insecticides, and the remaining 4.5% acaricides. Higher levels were detected for Boscalid in 2022 in three fields, with an average value of 0.42 mg/kg. Multi-residue contamination occurred each year; the lowest abundance was detected in 2023 (3.9%), and the highest in 2022 (12.3%), with 5 pesticide residues as the maximum number of compounds detected in one sample in 2022. The consumer risk assessment identified no potential health concerns for adults or toddlers, and the combined risk was considered acceptable. The environmental assessment showed maximum cumulative ratio (MCR) values that were always ≥1, indicating a contribution to the toxicity of the mixture, only slightly higher than that of the single compound with the highest toxicity. The results of this study highlight the critical need to include cumulative dietary exposure assessments in pesticide risk evaluations, especially for food products that are susceptible to contamination by multiple residues.

Social insects employ venom as an external immune defence against pathogens and parasites. Like other Hymenopterans, the venom gland of honey bee ( Apis mellifera ) serves as a reservoir of antimicrobial substances, primarily melittin. This study investigated the presence and origin of venom on honey bee body that could act as an external immune defence in honey bee workers infested by the ectoparasite mite, Varroa destructor . Using a multi-step approach, we first confirmed the presence of venom on bees’ bodies using melittin as a marker. We then examined how grooming could facilitate the distribution of venom on the bee’s body through behavioural observations. Further assays were used to compare melittin levels on the bodies of Varroa -free and Varroa -infested workers and assess the effects of bee-venom on mite activity. Our findings confirmed the occurrence of “venom bathing” in A. mellifera , excluding social components or environmental contamination, with bees likely coating their bodies with antimicrobial substances through self-grooming. Our results further suggest that infested bees spread higher amount of venom on their bodies compared to uninfested bees, and bee-venom significantly reduced mite activity, suggesting that the venom functions as an external defence. However, Varroa negatively impacted melittin production. Our study reveals a previously unknown negative effect of V. destructor : impairment of honey bees’ external immune defence through reduced melittin production.

Purpose Here we describe a patient admitted for a stroke that was unexpectedly correlated with subclinical infective endocarditis attributable to a rarely opportunistic pathogen, Abiotrophia defectiva . Case report A 75-year-old man presented with a stroke. Transesophageal echocardiography suggested vegetation on all aortic valve cusps, despite the absence of clinical or laboratory signs of infection. Surprisingly, three sets of blood cultures collected without fever were positive for A. defectiva . Although the patient did not exhibit classic signs of infection during hospitalization, the severity of the valve condition necessitated replacement with a bioprosthesis. Conclusions This clinical case underscores the importance of investigating the infective origin of endocarditis, even in the absence of clinical or laboratory evidence. Physicians should maintain a high level of suspicion, especially in patients with highly suggestive anamnestic characteristics.

Artichoke (Cynara cardunculus L.) is an herbaceous perennial plant from the Mediterranean Basin, cultivated as a poly-annual crop in different countries. Artichoke produces a considerable amount of waste at the end of the harvesting season in the field (5.2 tons/ha/year, DW) and from the industrial processing of fresh-cut products during the harvesting time (800 tons/year). The qualitative and quantitative phenolic profile and inulin content of artichoke samples from the field and industrial processing waste have been investigated after green extraction. The best operative conditions were achieved using the dried biomass extracted with water at 80 °C for 120 min and a matrix-to-solvent ratio of 1:30. The data obtained showed that the concentration of total polyphenols in fresh artichokes followed this order: stems > heads > leaves > outer bracts. Chlorogenic acid and 3,4 di-O-caffeoylquinic acid were the most concentrated caffeoylquinic derivates, whereas luteolin 7-O-malonyglucoside, luteolin 7-O-glucoside and 7-O-rutinoside were the most abundant flavonoids. The artichoke by-products showed high polyphenolic and inulin values, thus representing an important source of health-promoting biomolecules for application in pharmaceutical and cosmeceutical fields. According to the principles of circular economy, the work scheme proposed in this article, the use of waste and its processing into useful products such as nutraceutical supplies could provide a practical and economic opportunity for companies and farmers involved in the cultivation and industrial processing of artichokes.

Honey, a natural food with a rich history, is produced by honeybees and other species of bees from nectar, other plant fluids, and honeydew of sap-sucking insects. During foraging, these bees may be exposed to plant protection products (PPPs), metals, and metalloids, potentially leading to residues in honey and hive products that could have a negative impact on human safety. Recognizing the lack of an appropriate methodology for pesticide contamination of honey and other hive products, this research aims to support the need for studies on residues in pollen and bee products for human consumption to establish safe maximum residue levels (MRLs) for consumers. A UHPLC-MS/MS residues method and a modified QuEChERS extraction were applied to simultaneously determine 237 pesticide residues in honey and pollen. The study in North Sardinia analyzed honey and pollen samples from six areas for pesticide residues and verified 27 heavy metals and metalloid residues using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES). The data obtained showed residues at levels close to the LOQ of the method, cycloate in a few samples of pollen, and dichlorvos, zoxamide, cycloate, and chlorantraniliprole in honey samples. All samples showed the absence of heavy metal contamination. Overall, no risk to human health was identified. The results of this study confirm that honey and pollen may be a good bioindicator of environmental contamination of a wide area surrounding honeybee hives.

Fourth-range products are those types of fresh fruit and vegetables that are ready for raw consumption or after cooking, and belong to organic or integrated cultivations. These products are subject to mild post-harvesting processing procedures (selection, sorting, husking, cutting, and washing), and are afterwards packaged in packets or closed food plates, with an average shelf life of 5–10 days. Artichokes are stripped of the leaves, stems and outer bracts, and the remaining heads are washed with acidifying solutions. The A LC-MS/MS analytical method was developed and validated following SANTE guidelines for the detection of 220 pesticides. This work evaluated the distribution of pesticide residues among the fraction of artichokes obtained during the industrial processing, and the residues of their wastes left on the field were also investigated. The results showed quantifiable residues of one herbicide (pendimethalin) and four fungicides (azoxystrobin, propyzamide, tebuconazole, and pyraclostrobin). Pendimethalin was found in all samples, with the higher values in leaves 0.046 ± 8.2 mg/kg and in field waste 0.30 ± 6.7 mg/kg. Azoxystrobin was the most concentrated in the outer bracts (0.18 ± 2.9 mg/kg). The outer bracts showed the highest number of residues. The industrial waste showed a significant decrease in the number of residues and their concentration.

This work aimed to investigate the bio-distribution and the persistence of fipronil and its primary metabolite fipronil sulfone after oral and dermal administration by simulating natural farming conditions. Fipronil and fipronil sulfone detection and quantification were performed in different poultry matrices using an LC-MS/MS method coupled with modified QuEChERS extraction. After oral administration, fipronil was detected in feathers at each sampling time, in eggs for 28 days, and in the internal organs at the end of the experiment. After dermal administration, high levels of fipronil were detected in feathers, accounting for 195.85 ± 8.54 mg/kg, which were reduced by a third after 60 days. No traces of fipronil were detected in the eggs or internal organs. In addition, fipronil sulfone showed remarkable residues in all samples in trial 2. The data obtained confirmed that inappropriate use of unauthorized pesticides can lead to severe contamination of entire poultry farms. The contemporary presence of fipronil sulfone in feathers and eggs associated with the lack of fipronil in eggs suggests recent dermal contamination or past oral contamination. Moreover, simultaneous analysis of hens’ feathers and eggs could represent a new method to improve large-scale monitoring programs and animal welfare, limiting their slaughter.

Pesticides are broadly used to improve food safety, although they can lead to adverse health effects on consumers. Various food processing approaches, at the industrial or domestic level, have been found to highly reduce the amount of pesticide residues in most food materials. In this work, samples of raw tomatoes were collected directly from the field and processed at the industrial level to produce purée, triple concentrated paste, fine pulp, and diced tomatoes. A multiresidue method based on a modified QuEChERS (Quick, Easy, Cheap, Effective, Rugged e Safe) sample preparation, followed by liquid chromatography-tandem mass spectrometry analysis (LC-MS/MS) for the assessment of 116 pesticides residues, was used. The analytical method has been validated according to SANTE indications. The recovery yields ranged from 75.5% to 115.3%, repeatability (RSDr) ranged from 3.4% to 18.3%, while reproducibility (RSDwR) ranged from 5.4% to 19.8%. The limit of quantifications (LOQs) ranged from 2.35 µg kg−1 for benthiavalicarb to 6.49 µg kg−1 for allethrin. A total of 159 raw tomato samples were collected from the field. The analysis showed the presence of 46 pesticides with azoxystrobin and chlorantraniliprole the most represented. On the other hand, all industrially processed samples showed values ≤ LOD, confirming that post-harvest processes can lead to a decrease in pesticide residues from agricultural commodities.

Homeostasis of the host immune system is regulated by white blood cells with a variety of cell surface receptors for cytokines. Chemotactic cytokines (chemokines) activate their receptors to evoke the chemotaxis of immune cells in homeostatic migrations or inflammatory conditions towards inflamed tissue or pathogens. Dysregulation of the immune system leading to disorders such as allergies, autoimmune diseases, or cancer requires efficient, fast-acting drugs to minimize the long-term effects of chronic inflammation. Here, we performed structure-based virtual screening (SBVS) assisted by the Keras/TensorFlow neural network (NN) to find novel compound scaffolds acting on three chemokine receptors: CCR2, CCR3, and one CXC receptor, CXCR3. Keras/TensorFlow NN was used here not as a typically used binary classifier but as an efficient multi-class classifier that can discard not only inactive compounds but also low- or medium-activity compounds. Several compounds proposed by SBVS and NN were tested in 100 ns all-atom molecular dynamics simulations to confirm their binding affinity. To improve the basic binding affinity of the compounds, new chemical modifications were proposed. The modified compounds were compared with known antagonists of these three chemokine receptors. Known CXCR3 compounds were among the top predicted compounds; thus, the benefits of using Keras/TensorFlow in drug discovery have been shown in addition to structure-based approaches. Furthermore, we showed that Keras/TensorFlow NN can accurately predict the receptor subtype selectivity of compounds, for which SBVS often fails. We cross-tested chemokine receptor datasets retrieved from ChEMBL and curated datasets for cannabinoid receptors. The NN model trained on the cannabinoid receptor datasets retrieved from ChEMBL was the most accurate in the receptor subtype selectivity prediction. Among NN models trained on the chemokine receptor datasets, the CXCR3 model showed the highest accuracy in differentiating the receptor subtype for a given compound dataset.

Humans are exposed daily to complex mixtures of chemical substances via food intake, inhalation, and dermal contact. Developmental neurotoxicity is an understudied area and entails one of the most complex areas in toxicology. Animal studies for developmental neurotoxicity (DNT) are hardly performed in the context of regular hazard studies, as they are costly and time consuming and provide only limited information as to human relevance. There is a need for a combination of in vitro and in silico tests for the assessment of chemically induced DNT in humans. The zebrafish (Danio rerio) embryo (ZFE) provides a powerful model to study DNT because it shows fast neurodevelopment with a large resemblance to the higher vertebrate, including the human system. One of the suitable readouts for DNT testing in the zebrafish is neurobehaviour (stimulus-provoked locomotion) since this provides integrated information on the functionality and status of the entire nervous system of the embryo. In the current study, environmentally relevant pharmaceuticals and their mixtures were investigated using the zebrafish light-dark transition test. Zebrafish embryos were exposed to three neuroactive compounds of concern, carbamazepine (CBZ), fluoxetine (FLX), and venlafaxine (VNX), as well as their main metabolites, carbamazepine 10,11-epoxide (CBZ 10,11E), norfluoxetine (norFLX), and desvenlafaxine (desVNX). All the studied compounds, except CBZ 10,11E, dose-dependently inhibited zebrafish locomotor activity, providing a distinct behavioural phenotype. Mixture experiments with these pharmaceuticals identified that dose addition was confirmed for all the studied binary mixtures (CBZ-FLX, CBZ-VNX, and VNX-FLX), thereby supporting the zebrafish embryo as a model for studying the cumulative effect of chemical mixtures in DNT. This study shows that pharmaceuticals and a mixture thereof affect locomotor activity in zebrafish. The test is directly applicable in environmental risk assessment; however, further studies are required to assess the relevance of these findings for developmental neurotoxicity in humans.