Emanuela Salviati

Background/objectives: This study aimed to characterize the fatty acid (FA) profile of breast milk (BM) at 7 days (T7) and 1 month postpartum (T30) using gas chromatography–mass spectrometry (GC-MS) and to evaluate associations between maternal diet during pregnancy and BM FA composition. Methods: A prospective observational cohort study was conducted from March 2022 to October 2023, involving mothers grouped by gestational age at delivery (32 weeks, 32–36.6 weeks, and >37 weeks). Results: BM lipid profiles were generally similar across gestational groups, with notable differences at T7 in saturated fatty acids (SFAs), myristic acid, monounsaturated fatty acids (MUFAs), erucic acid, nervonic acid, and some FA ratios. At T30, differences persisted in SFAs, MUFAs, myristic acid, and MUFA/SFA. At T7, red meat intake was positively correlated with stearic acid; white meat intake was negatively associated with multiple FAs (including ω-3) but positively with linoleic. Cheese correlated positively with caprylic acid; milk negatively with pentadecylic acid; and dried fruit positively with MUFA. At T30, fish consumption was prevalently positively related to DHA, EPA, and Omega-3, while red meat was positively associated with arachidic acid and margaric acid and negatively with di-homo-gamma linolenic acid. White meat showed a predominantly negative correlation with DHA, EPA and MUFA. Milk intake showed both positive (i.e., caproic acid) and multiple negative FA associations. Cheese was positively associated with caprylic acid, while dried fruit intake was positively linked to oleic acid and MUFA. Conclusions: Despite stable total lipid content, gestational age influenced specific FA profiles. These shifts may reflect adaptive responses to neonatal metabolic and neurodevelopmental needs. Understanding such mechanisms could guide tailored nutritional strategies, especially for preterm infants.

SAMP mice develop progressive Crohn’s disease (CD)-like ileitis without spontaneous colitis that worsens over time without chemical, genetic, or immunological manipulation. Even growing in an identical vivarium and fed with the same diet, SAMP mice reveal a distinct fecal microbiome, metabolome, and lipidome profile compared to AKR mice, their non-inflamed parental control strain. Differences are already present in 5-week-old mice, with a tendency to increase in 15-week-old mice. SAMP and AKR mice metabolome and lipidome profiles were substantially different, belonging to two clusters in line with the progression of intestinal disease. Similarly, the 16S analysis confirmed differences between 15-week-old AKR and SAMP mice. The protective role of dietary polyphenols has been documented in inflammatory bowel diseases (IBD); thus, we supplemented the chow diet with an anthocyanin-rich extract (RED) to evaluate disease reduction in SAMP mice and changes in fecal microbiota/metabolome. Our data reveal that 10-week supplementation with anthocyanin-rich extract ameliorated disease severity in SAMP mice despite limited fecal microbiota/metabolome differences.

Endocannabinoids (eCBs) are endogenous lipid messengers that primarily bind cannabinoid receptors CB1/CB2 and together with the enzymes that regulate their biosynthesis and degradation define the endocannabinoid system. The eCB signaling system plays a key role in the central nervous system, resulting altered in most neurological disorders. The analysis of eCBs is challenging for their low concentration in biospecimens, and this is exacerbated in Mass Spectrometry Imaging (MSI) where low sensitivity and tissue dependent ion suppression obscure their spatial visualization. In this work we address this limitation by the application of laser-induced post-ionization (MALDI-2). Herein we demonstrate that MALDI-2 boosts the detection of 2-arachidonylglycerol (2-AG) and N-acylethanolamines (AEA, PEA, OEA) with respect to MALDI, and that eCBs can be visualized in brain at endogenous concentration only by MALDI-2-MSI. Both root-mean-square (RMS) and deuterated internal standards (I.S.) normalization were evaluated, with I.S. normalization providing improved pixel to pixel variation and more uniform distribution in specific brain regions, especially for 2-AG and PEA. Furthermore, high lateral resolution up to 5 µm pixel size was evaluated, resulting in the detection of all eCBs and confirming the MALDI-2 potential even reducing the ablated tissue amount. Lastly the method was applied as proof of concept in a mouse model of mild traumatic brain injury demonstrating the ability to reveal valuable biological insights for neuropharmacology.

Lycium barbarum, commonly recognized as goji berry or wolfberry, is highly appreciated not only for its organoleptic and nutritional properties but also as an important source of bioactive compounds such as polysaccharides, carotenoids, phenolics, and various other non-nutritive compounds. These constituents give it a multitude of health benefits, including antioxidant, anti-inflammatory, and anticancer properties. However, the precise biochemical mechanisms responsible for its anticancer effects remain unclear, and the comprehensive composition of goji berry extracts is often insufficiently explored. This study aimed to investigate the biochemical pathways modulated in breast cancer cells by an ethanolic extract of Lycium barbarum fruit (LBE). Following metabolomic profiling using UHPLC-HRMS/MS, we assessed the antitumoral properties of LBE on different breast cancer cell lines. This investigation revealed that LBE exhibited cytotoxic effects, inducing a pro-oxidant effect that triggered pyroptosis activation through endoplasmic reticulum (ER) stress and subsequent activation of the P-IRE1α/XBP1/NLRP3 axis in MCF-7 cells. In addition, LBE did not display cytotoxicity toward healthy human cells but demonstrated antioxidant properties by neutralizing ROS generated by doxorubicin. These findings underscore the potential of LBE as a highly promising natural extract in cancer therapy.

Among gynecological cancers, endometrial cancer is the most common in developed countries. Extracellular vesicles (EVs) are cell-derived membrane-surrounded vesicles that contain proteins involved in immune response and apoptosis. A deep proteomic approach can help to identify dysregulated extracellular matrix (ECM) proteins in EVs correlated to key pathways for tumor development. In this study, we used a proteomics approach correlating the two acquisitions—data-dependent acquisition (DDA) and data-independent acquisition (DIA)—on EVs from the conditioned medium of four cell lines identifying 428 ECM proteins. After protein quantification and statistical analysis, we found significant changes in the abundance (p < 0.05) of 67 proteins. Our bioinformatic analysis identified 26 pathways associated with the ECM. Western blotting analysis on 13 patients with type 1 and type 2 EC and 13 endometrial samples confirmed an altered abundance of MMP2. Our proteomics analysis identified the dysregulated ECM proteins involved in cancer growth. Our data can open the path to other studies for understanding the interaction among cancer cells and the rearrangement of the ECM.

Endocrine disruptors (EDCs) are chemicals that interfere with the endocrine system. EDC exposure may contribute to the development of obesity, type 2 diabetes, and cardiovascular diseases by impacting the composition of an infant’s gut microbiota during the first 1000 days of life. To explore the relationship between maternal urinary levels of Bisphenol-A and phthalates (UHPLC-MS/MS), and the composition of the infant gut microbiota (16S rDNA) at age 12 months (T₃) and, retrospectively, at birth (T₀), 1 month (T₁), and 6 months (T₂), stool samples from 20 infants breastfed at least once a day were analyzed. Metataxonomic bacteria relative abundances were correlated with EDC values. Based on median Bisphenol-A levels, infants were assigned to the over-exposed group (O, n = 8) and the low-exposed group (B, n = 12). The B-group exhibited higher gut colonization of the Ruminococcus torques group genus and the O-group showed higher abundances of Erysipelatoclostridium and Bifidobacterium breve. Additionally, infants were stratified as high-risk (HR, n = 12) or low-risk (LR, n = 8) exposure to phthalates, based on the presence of at least three phthalates with concentrations exceeding the cohort median values; no differences were observed in gut microbiota composition. A retrospective analysis of gut microbiota (T₀–T₂) revealed a disparity in β-diversity between the O-group and the B-group. Considering T₀–T₃, the Linear Discriminant Effect Size indicated differences in certain microbes between the O-group vs. the B-group and the HR-group vs. the LR-group. Our findings support the potential role of microbial communities as biomarkers for high EDC exposure levels. Nevertheless, further investigations are required to deeply investigate this issue.