Immune-inflammatory response is regulated among other cells by macrophages through the secretion of various soluble molecules, as cytokines, chemokines and lipid mediators. Lipid mediators, also termed oxylipins, are generated from both n-3 and n-6 polyunsaturated fatty acids (PUFAs) through enzymatic (e.g., prostanoids, epoxy, and hydroxy fatty acids) and non-enzymatic (e.g., isoprostanes) oxidation reactions. PUFAs act as precursors of pro-inflammatory, anti-inflammatory, and specialized pro-resolving lipid mediators (SPMs) through enzymatic oxidation reactions. Pro-inflammatory mediators released at the very beginning of inflammation foster the appearance of its classic signs, whereas the switch to lipoxygenase-derived anti-inflammatory and SPMs leads to a natural resolution of inflammation. PUFAs can also undergo free radical-mediated oxidation, thus generating isoprostanes, currently known as gold standard biomarkers of oxidative stress. The analysis of oxylipins may furnish a clear picture of the oxidant and inflammatory cascade, thus revealing key information about the pathogenesis and pathophysiology of many diseases. In this scenario, we have developed a powerful in-house MS-based platform that couples the new micro-extraction by packed sorbent (MEPS) technique with ultra-high performances liquid chromatography tandem mass spectrometry for the detection of 60 oxylipins in a single run. In the recent years, we have been working on MS-based targeted metabolomics for the determination of oxylipins in different research area, such as cardiovascular, neurological, and neurodegenerative diseases, the evaluation of metabolic adaptation to extreme physical exercise or altitude induced-hypoxia, the study of fetal-to-newborn transition and of host-pathogen response upon viral infection, as in the case of the COVID-19 pandemic crisis.
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Cardiovascular diseases (CVDs) are a group of disorders of the heart and blood vessels that belong to non-communicable diseases. They include coronary artery disease, thromboembolism, myocardial infarction, cardiac arrhythmia, and stroke, among others. While molecular pathways involved in the initiation and evolution of CVDs have been extensively explored and partially established, imaging studies and routine laboratory tests lack diagnostic efficacy in 50% of CV events. Nowadays, clinical use of peptides is rapidly increasing since most of them play a crucial role in maintaining tissue homeostasis. They are smaller than proteins, hence they may rapidly enter the bloodstream (i.e., due to passive diffusion), unlike proteins that may need to be actively secreted. Furthermore, their plasma concentration can potentially reflect an altered vascular biology. Simultaneous analysis of a panel of peptides may represent a potential approach to monitor the complex and multifactorial nature of CVDs.
The aim of this project is to develop and validate the first reliable analytical protocol based on targeted top-down peptidomics and UHPLC-ESI-MS/MS analysis for the determination of an array of cardiovasculary active peptides (i.e., Endothelins, Urotensins, Cystatin C, Fibrinopeptides, Guanylins) in plasma samples. In collaboration with the Department of Clinical and Experimental Medicine of the University of Pisa, a pilot study will be conducted aimed at finding possible correlations between the target peptides and clinical variables. PERICARD would represent an innovative tool to better identify “vulnerable” individuals, diagnose disease conditions promptly, and effectively prognosticate and treat patients with the disease.
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Body odours have a great importance in chemical communication. It is widely known that breastfeeding women emit olfactive signals that elicit a response of newborns influencing their behaviour. This project aims to develop reliable and non-invasive analytical protocols for the chemical characterisation of body odour emitted from the breast area during the whole period of pregnancy. We have chosen to test a passive sampling technique called Thin Film Microextraction (TFME). This technique involves carbon mesh sheet impregnated with one or more sorptive phases capable of passively collecting a wide range of volatile and semi-volatile analytes due to high porosity and use of a combination of sorbent materials. Instrumental analysis will be mainly performed by thermal desorption coupled with a comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry (GC×GC-TOFMS), one of the most powerful separation and identification tools for non-targeted analysis and suspect screening of VOCs.
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In recent years, exhaled breath analysis has become increasingly attractive for its potential use as an easy, painless and non-invasive tool to monitor physiological and pathological conditions as well as exposure to environmental contaminants. A main advantage of this approach is that it can be used on people of all ages and conditions (e.g. newborns, infants and mechanically ventilated patients) reducing the problems associated with blood sampling (e.g. acceptance from patients, risk of infections, production of potentially infected waste, and need for trained personnel working in dedicated environment). Several studies highlighted the potential of breath analysis as an innovative diagnostic tool since the chemical composition of breath is related to the occurrence of diseases, suggesting the diagnostic utility of breath biomarkers, often complementary or even alternative to those of blood and urine. For example, acetone is potentially useful for monitoring patients suffering from diabetes and heart failure, hydrocarbons and aldehydes for monitoring abnormal lipid peroxidation. Analytical protocols based on thermal desorption coupled to gas-chromatography and mass spectrometry will be normally employed in several pilot studies, conducted in collaboration with medical partners, to monitor the chemical composition of breath sample collected from patients suffering from diabetes, heart failure and asthma.
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The project aims to analyze non-conventional biological fluids such as sweat, saliva, and exhaled breath using advanced mass spectrometry techniques (GCxGC-qTOF for untargeted analysis, GC-QqQ for targeted analysis, and PTR-TOF for real-time monitoring). The goal is to identify volatile and semi-volatile biomarkers to improve early diagnosis and disease monitoring, as well as to study chemical communication between humans, for example in mother-newborn bonding and emotional expression, providing non-invasive tools for clinical and social applications.
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Environmental pollution associated to plastic debris is gaining increasing relevance not only as a threat to ecosystems but also for its possible harmful effects on biota and human health. Plastic debris exposed to environmental weathering are prone to degradation mainly due to photo-oxidative, thermal, and hydrolytic processes. Such degradation processes lead to progressive polymer fragmentation into microplastics as well as the release of toxic volatile- and semi-volatile compounds resulting from the chemical degradation induced in particular by heat and UV-light. As example, degradation of polyolefins generates compounds belonging to the families of lactones, esters, ketones, and carboxylic acids. Most of these compounds play a key role in atmospheric chemistry and represent also a source of toxic leachate with negative impact on biota and on human health. Analytical protocols based on thermal desorption coupled to gas-chromatography and mass spectrometry will be normally employed to monitor degradation products. In this scenario, micro-extraction tools (e.g. needle trap micro extraction and stir bar sorptive extraction) will be tested too, with the aim to reduce the amount of plastic debris required for the chemical characterization. Developed methods will be employed to monitor the degradation products emitted from reference materials that will be subjected to artificial aging processes simulating the real environmental conditions.
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