Proteomica e metallomica: studio del tessuto placentare in presenza di diabete gestazionale

The present research work concerns the development of new analytical methods aimed to the study of the proteome and the metallome of placental tissue and the evaluation of its possible modifications and/or alterations due to gestational diabetes. Proteomics analysis was performed by different experimental approaches based on mass spectrometry (ESI-MS and MALDI-MS), and on one- and two-dimensional electrophoresis. These analytical techniques are presently the most suitable experimental approaches to be used in omics applications thanks to their high sensitivity and specificity. Results showed the presence of protein species differently expressed in the case of gestational diabetes; in particular, it was measured an increment of the chorionic somatomammotrophin level and a decrease in the concentration of alpha, beta and gamma chains of fibrinogen and tubulointerstitial nephritis antigen-like in the case of placental tissues impaired by GDM. Metallomics analysis was carried out by ICP-MS, an analytical technique characterized by high sensitivity and robustness. The study reported an increased concentration of Se and a decrease of Cd level in pathological placental tissues compared to healthy ones. Furthermore, this approach allowed the evaluation of the effectiveness of the sampling procedure, the influence of the blood on the obtained results and showed the importance of other parameters, such as the lifestyle of the patients, in the statistical evaluation of the experimental data. In conclusion, the obtained data might be useful for the study and the explanation of the biochemical processes that characterize the disease and for the determination of new biomarkers aimed to the development of innovative diagnostic tests for gestational diabetes

Ecotoxicological effects and bioaccumulation of BPA analogues and their mixture in the clam Ruditapes philippinarum

Bisphenol A is recognized as an endocrine disruptor that can affect several biological processes in marine species. Consequently, its use has been restricted and it has been replaced with other similar compounds named bisphenol A analogues (BPA analogues). BPA analogues are speculatively considered safer compounds than BPA and their usage is increasing with a consequent higher environmental release. In this study, specimens of the clam Ruditapes philippinarum were exposed to three main BPA analogues, namely BPAF, BPF, BPS and their mixture at an environmentally relevant concentration of 300 ng/L for 7 and 14 days. Effects on biomarkers indicative of cytotoxicity, oxidative stress and damage and neurotoxicity were evaluated. In addition, bioaccumulation of the compound tested was analysed in clam soft tissues. Results showed that BPA analogues at an environment concentration affected cellular parameters and antioxidant system causing also oxidative damage, suggesting that BPA analogues can be harmful compounds for clams

Low Level of Allergens in the Argentinean Plant Zuccagnia punctata Cav.: Screening and Quality Control of North-Western Propolis Using an LC-DAD-QTOF System

North-western Argentinean propolis (NAP), having promising bioactivity, was recently included into the National Food Code. Zuccagnia punctata Cav., a native shrub of north-western Argentina, is one of the prevalent botanical sources of NAPs, but no information on its allergenic constituents was available so far. A liquid chromatography-diode array detector -quadrupole-time of flight system (LC-DAD-QTOF) was used as a screening method for the reliable identification of sensitizing agents belonging to caffeic acid derivatives in Z. punctata and in two NAPs collected in the provinces of Catamarca and Tucum\ue1n. Caffeic acid phenethyl ester, one of the most active allergens in propolis, was never detected in either Z. punctata or NAP. Among 31 sensitizers, only geranyl caffeate was alleged in Z. punctata as <10% of its major constituent, whereas three caffeic acid derivatives with strong allergenic effect, i.e., geranyl, pentenyl, and benzyl caffeates, occurred in NAP samples (29%\u201336% of the Z. punctata major constituent), indicating other minor botanical sources. However, the high content of chalcones and flavonoids ascribed to Z. punctata significantly contributes to the antiallergenic and antioxidant character of these NAPs. This peculiar chemical profile depends on the extremophile condition in which this shrub grows and suggests other studies to characterize such raw materials for oral and topical formulation

Phthalates and heavy metals as endocrine disruptors in food: A study on pre-packed coffee products

Phthalate plasticizers and heavy metals are widely recognized to be pollutants that interfere with key developmental processes such as masculinization. We investigated the release of phthalates and heavy metals in coffee brewed from coffee packed in single-serve coffee containers made from different types of materials: metal, biodegradable and plastics. We detected with GC\u2013MS small amounts phthalates, below the tolerated daily risks levels, in all the coffees prepared from the different types of capsules. Specifically, Di (2-ethyl-hexyl)-phthalate and DiBP: Diisobuthyl-pthalate were ubiquitously present despite the high variability among the samples (respective range 0.16\u20131.87 \u3bcg/mL and 0.01\u20130.36 \u3bcg/mL). Whereas, diethyl-phthalate (range 0.20\u20130.26 \u3bcg/mL) and di-n-buthyl-phthalate (range 0.02\u20130.14 \u3bcg/mL) were detected respectively in one and three out of the four types of capsule tested. In contrast, we detected by atomic mass spectrometry on mineralized samples heavy metals lead (Pb) and nickel (Ni), in all coffee tested. PB levels (respective range 0.32\u2013211.57 \u3bcg/dose) accounted for 42\u201379%, whereas Ni levels (respective range 166.25\u20131950.26 \u3bcg/dose) accounted for >100% of the tolerable daily intake. These results add to the already present concerns related to the multiple pathways of human exposure and the ubiquitous presence of these pollutants in consumer products and their long-term effect on human health

Metallic functionalization of magnetic nanoparticles enhances the selective removal of glyphosate, AMPA, and glufosinate from surface water

Glyphosate (GLY) is the most used herbicide worldwide, raising concerns due to its toxicity and mobility in water. The concurrent spread of similar herbicides, i.e., glufosinate (GLUF) and aminomethylphosphonic acid (AMPA, a metabolite of GLY), also causes a serious concern to the environment. The application of magnetic nanoparticles (MNPs) gained wide attention as a promising approach for the environmental remediation of GLY. However, the fast agglomeration, low removal efficiency, and saturation of MNPs by non-target chemicals remain a challenge. Herein, we used polydopamine as a coating agent followed by functionalization with different metal ions, i.e. Ti(iv), Zr(iv), and Cu(ii), for selective removal of GLY, AMPA, and GLUF from deionised water in laboratory trials. Finally, we tested the performance of MNPs in surface waters contaminated with GLY at 0.17 & PLUSMN; 0.02 & mu;g L-1. The GLY removal efficiency (RE, %) of MNPs was optimized by using different GLY to MNP ratios and incubation times, in the presence of GLY-analogues and competitive species, i.e., phosphates. The results indicate that all metallic functionalized MNPs are more stable toward aggregation and effective in removing GLY than bare MNPs, up to 150 & mu;g L-1. The optimal ratio was 500 & mu;g(GLY) g(MNPs)(-1) (50 & mu;g GLY to 100 mg MNP), with RE > 80%. MNPs functionalized with Ti(iv) and Zr(iv) performed more efficiently than MNPs functionalized with Cu(ii), reaching an RE of 99.9% after a incubation time of 15 min. The presence of Ti(iv) and Zr(iv) in the MNPs increased the selectivity of the particles toward GLY and GLY analogues that can be removed with similar efficiency, and prevented competition with phosphates at much higher concentrations (1000 & mu;g L-1). Finally, GLY analogues can be easily re-eluted with ammonia, and the functionalized MNPs can be efficiently re-used up to four cycles. The use of metal-functionalized MNPs is a promising approach for the removal of target pollutants from contaminated water

Methiocarb metabolites are systemically distributed throughout corn plants grown from coated seeds

Funder: Università degli Studi di PadovaAbstract: Systemic insecticides such as neonicotinoids are widely used in seed coating practices for pest control in many crops, e.g., corn. Their success is due to their ability to protect the whole plant, from the roots to the upper leaves, but their use at high amounts is causing possible adverse effects on non-target animals exposed to contaminated pollen, nectar, leaves, and dust emitted during sowing. In 2018, the European Union banned some neonicotinoids and fipronil as seed coating insecticides in open fields. Consequently, the methylcarbamate methiocarb and less-toxic neonicotinoids, e.g., thiacloprid, have been authorized and largely used as alternative pesticides for corn seed coating. Here, an analytical protocol based on QuEChERS extraction/purification procedure and analysis by liquid chromatography-mass spectrometry has been optimized for the identification and the quantification of methiocarb, thiamethoxam, thiacloprid, and their metabolites in guttation drops, the xylem fluid excreted at leaf margins, and in leaves of corn plants grown from coated seeds. Although methiocarb is a non-systemic pesticide, we unexpectedly found high concentrations of its metabolites in both guttations and leaves, whereas methiocarb itself was below detection limits in most of the samples. The methiocarb main metabolite, methiocarb sulfoxide, was found at a mean concentration of 0.61 ± 1.12 µg mL−1 in guttation drops and 4.4 ± 2.1 µg g−1 in leaves. Conversely, parent compounds of neonicotinoids (thiamethoxam, thiacloprid) are systemically distributed in corn seedlings. This result raises safety concerns given that methiocarb sulfoxide is more toxic than the parent compound for some non-target species

Western Diet-Induced Metabolic Alterations Affect Circulating Markers of Liver Function before the Development of Steatosis

Since nutrition might have a significant impact on liver function, we analyzed the early effect of Western-type diet on hepatic tissue and lipid and drug metabolism in Wistar-Kyoto rats (n = 8); eight rats fed with a standard diet were used as controls. Histological analysis of liver tissue was performed, and plasma biochemical parameters were measured. Plasma concentration of six bile acids was determined by ultra-liquid chromatography-tandem mass spectrometry UHPLC-MS/MS. Hepatic gene expressions of enzymes involved in drug and lipid metabolism were assessed by means of real-time reverse transcription (qRT)-PCR. Liver of rats fed with a Western diet did not show macroscopic histological alterations, but number and diameter of lipid droplets increased, as well as DGAT1, GPAT4, SCD, FASN and SREBP2 expression. Furthermore, Western diet-fed animals showed an increase in the activation of hepatic stellate cells and macrophage number in liver tissue, as well as a significant increase in AST and bilirubin levels (p < 0.01), and in the LDL:HDL cholesterol ratio (p < 0.001). Plasma chenodeoxycholic acid concentration increased significantly, whereas cholic acid decreased (p < 0.05), and cytochrome P450 genes were generally downregulated. Significant changes in hepatic lipid and drug metabolism are early induced by the Western diet, prior to steatosis development. Such changes are associated with a peculiar alteration in circulating bile acids, which could represent an early marker of non-alcoholic fatty liver disease (NAFLD) development

Proteomica e metallomica: studio del tessuto placentare in presenza di diabete gestazionale

The present research work concerns the development of new analytical methods aimed to the study of the proteome and the metallome of placental tissue and the evaluation of its possible modifications and/or alterations due to gestational diabetes. Proteomics analysis was performed by different experimental approaches based on mass spectrometry (ESI-MS and MALDI-MS), and on one- and two-dimensional electrophoresis. These analytical techniques are presently the most suitable experimental approaches to be used in omics applications thanks to their high sensitivity and specificity. Results showed the presence of protein species differently expressed in the case of gestational diabetes; in particular, it was measured an increment of the chorionic somatomammotrophin level and a decrease in the concentration of alpha, beta and gamma chains of fibrinogen and tubulointerstitial nephritis antigen-like in the case of placental tissues impaired by GDM. Metallomics analysis was carried out by ICP-MS, an analytical technique characterized by high sensitivity and robustness. The study reported an increased concentration of Se and a decrease of Cd level in pathological placental tissues compared to healthy ones. Furthermore, this approach allowed the evaluation of the effectiveness of the sampling procedure, the influence of the blood on the obtained results and showed the importance of other parameters, such as the lifestyle of the patients, in the statistical evaluation of the experimental data. In conclusion, the obtained data might be useful for the study and the explanation of the biochemical processes that characterize the disease and for the determination of new biomarkers aimed to the development of innovative diagnostic tests for gestational diabetes.Il lavoro di ricerca presentato concerne lo sviluppo e il perfezionamento di metodi analitici atti allo studio del proteoma e del metalloma del tessuto placentare e alla valutazione di eventuali sue modificazioni e/o alterazioni in presenza di diabete gestazionale. Le analisi di proteomica sono state effettuate mediante metodologie analitiche basate sulla spettrometria di massa (in particolare sono state utilizzate le tecniche di ionizzazione MALDI e ESI) e sull’elettroforesi mono e bi-dimensionale. Allo stato attuale dell’arte queste tecniche analitiche risultano essere gli approcci sperimentali più adatti a condurre indagini di questo tipo grazie all’elevata sensibilità e specificità che le caratterizza. I risultati analitici hanno evidenziato la presenza di specie proteiche diversamente espresse nel caso di diabete gestazionale; in particolare è stato notato un incremento dei livelli di somatomammotropina corionica e una diminuzione della concentrazione delle catene alfa, beta e gamma del fibrinogeno e di Tubulointerstitial nephritis antigen-like nelle placente complicate da GDM. Le analisi di metallomica sono state condotte mediante ICP-MS, a sua volta tecnica analitica caratterizzata da elevata sensibilità e robustezza. Le analisi hanno rivelato un aumento della concentrazione di Se e una diminuzione dei livelli di Cd nelle placente patologiche rispetto a quelle sane. L’approccio seguito ha permesso di valutare l’efficacia del campionamento e delle procedure pre-analitiche, l’influenza del tessuto ematico sui risultati ottenuti e ha mostrato l’importanza di altri parametri, come ad esempio lo stile di vita della gestante, nella valutazione statistica dei dati sperimentali. In conclusione, si può affermare che le informazioni ottenute possono essere utili per lo studio e la delucidazione dei processi biochimici che caratterizzano la patologia e per la determinazione di nuovi bio-marcatori che permettano lo sviluppo di test diagnostici innovativi per il diabete gestazionale.

Effects of the Fragrance Galaxolide on the Biomarker Responses of the Clam Ruditapes philippinarum

The musk fragrance Galaxolide\uae (HHCB) is widely used in personal care and household products. Its large use leads to a continuous release of the compound into aquatic environments. Although some studies on the presence of HHCB in ecosystems and biota have been conducted, limited data about its effects on organism biomarkers are available. This study aimed at investigating both cellular and biochemical effects of HHCB in the clam Ruditapes philippinarum. Mussels were exposed for 7, 14 and 21 days to 100 ng/L and 500 ng/L of HHCB in seawater, and the effects on haemocyte parameters and antioxidant enzyme activities in the gills and digestive gland were evaluated. In addition, the neurotoxic potential of HHCB and its capacity to cause oxidative damage to proteins were assessed. Overall, our results demonstrated that exposure to HHCB was able to induce changes in biomarker responses of mussels, mainly at the cellular level