Study of metabolic changes in human erythrocytes and plasma under simulated microgravity conditions

The absence of gravity is considered as an extreme biological stressor and the human body's adaptive response to microgravity conditions triggers a series of metabolic changes that are often associated with bone density loss, skeletal muscle atrophy, nervous system dysfunction, and anaemia. To understand the biological mechanisms underlying aerospace anaemia we investigated the structural and/or metabolic changes that occur in the erythrocytes under low gravity, as well as we compared the profile of human plasma polar metabolite under normal- and micro-g conditions. Erythrocytes exposed to simulated microgravity showed morphological changes, a constant increase in reactive oxygen species (ROS), a significant reduction in total antioxidant capacity (TAC), a remarkable and constant decrease in total glutathione (GSH) concentration, and an increase of malondialdehyde (MDA) levels. In this PhD work, experiments were also performed to evaluate the lipid profile of erythrocyte membranes changes under microgravity conditions. Lipidomics results showed an upregulation of the levels of complex lipids such as phosphocholines and sphingomyelins. Alterations in the lipid composition of the membrane determine membrane rigidity and fluidity, and plays a crucial role in membrane organization, dynamics, and function. No less important is the role that lipids have in the signalling of programmed cell death. In conjunction with these results, the metabolomics analysis revealed that microgravity in human plasma induced an increase of glycolysis, Krebs cycle and fatty acids β-oxidation perturbations, and lactic acid production. These findings indicate an evident damage of circulating mitochondria causing a defective respiratory chain, oxidative environment, and incomplete β-oxidation of fatty acids with consequent accumulation of carnitines, in particular propionyl carnitine

Compositional Characteristics of Mediterranean Buffalo Milk and Whey

The main objective of this review is to summarize the compositional characteristics and the health and functional properties of Mediterranean buffalo milk and whey derived from mozzarella cheese production. Several studies have investigated the composition of buffalo milk and in particular its fat, protein, and carbohydrates contents. These characteristics may change depending on the breed, feeding regime, and rearing system of the animals involved in the study, and also with the seasons. In particular, buffalo milk showed a higher nutritional value and higher levels of proteins, vitamins, and minerals when compared to milks produced by other animal species. Additionally, buffalo milk contains beneficial compounds such as gangliosides that can provide antioxidant protection and neuronal protection, and can improve bone, heart, and gastrointestinal health in humans

An Untargeted Metabolomic Comparison of Milk Composition from Sheep Kept Under Dierent Grazing Systems

This study aimed to evaluate the effects of different feedings on main traits and polar and semi-polar metabolite profiles of ovine milk. The milk metabolome of two groups of Sarda sheep kept under different grazing systems were analyzed by gas chromatography coupled with mass spectrometry (GC-MS) and multivariate statistical analysis (MVA). The results of discriminant analysis indicated that the two groups showed a different metabolite profile, i.e., milk samples of sheep kept under Grazing System 1 (GS1) were richer in nucleosides, inositols, hippuric acid, and organic acids, while milk of sheep under Grazing System 2 (GS2) showed higher levels of phosphate. Statistical analysis of milk main traits indicates that fat content was significantly higher in GS1 samples while milk from GS2 sheep had more urea, trans-vaccenic acid, and rumenic acid. MVA studies of the associations between milk main traits and metabolite profile indicated that the latter reflects primarily the long chain fatty acid content, the somatic cell count (SCC), and lactose levels. All together, these results demonstrated that an integrated holistic approach could be applied to deepen knowledge about the effects of feeding on sheep’s milk composition

Untargeted Lipidomics of Erythrocytes under Simulated Microgravity Conditions

Lipidomics and metabolomics are nowadays widely used to provide promising insights into the pathophysiology of cellular stress disorders. Our study expands, with the use of a hyphenated ion mobility mass spectrometric platform, the understanding of the cellular processes and stress due to microgravity. By lipid profiling of human erythrocytes, we annotated complex lipids such as oxidized phosphocholines, phosphocholines bearing arachidonic in their moiety, as well as sphingomyelins and hexosyl ceramides associated with microgravity conditions. Overall, our findings give an insight into the molecular alterations and identify erythrocyte lipidomics signatures associated with microgravity conditions. If the present results are confirmed in future studies, they may help to develop suitable treatments for astronauts after return to Earth

LC-QTOF/MS Untargeted Metabolomics of Sheep Milk under Cocoa Husks Enriched Diet

The aim of this work was to evaluate, by an untargeted metabolomics approach, changes of milk metabolites induced by the replacement of soybean hulls with cocoa husks in the ewes’ diet. Animals were fed with a soybean diet integrated with 50 or 100 g/d of cacao husks. Milk samples were analyzed by an ultra high performance liquid chromatograph coupled to a time of flight mass spectrometer (UHPLC-QTOF-MS) platform. Multivariate statistical analysis showed that the time of sampling profoundly affected metabolite levels, while differences between treatments were evident at the fourth week of sampling. Cocoa husks seem to induce level changes of milk metabolites implicated in the thyroid hormone metabolism and ubiquinol-10 biosynthesis

Ion Mobility–Mass Spectrometry Approach for the Comparison of Sheep and Goat Milk Lipidomes

In this work, we report an analytical procedure to investigate the lipid compositions of sheep and goat milk. This approach is based on an ion mobility-high-resolution mass spectrometric method to facilitate the identification of complex lipid species and their regiochemistry. A common triacylglycerol profile was observed for sheep and goat milk samples, while a higher abundance of medium-chain fatty acids was observed at the sn-2 position for sheep milk. Furthermore, differences can be also observed in the levels of saturated and unsaturated fatty acids at the sn-2 position. In terms of lipid classes, goat milk showed higher levels of triacylglycerols, phosphatidylinositols and ether-linked phosphatidylethanolamines, while sheep milk showed higher levels of free fatty acids, lysophosphatidylethanolamines, lysophosphocholines and non-hydroxy fatty acid-dihydrosphingosine ceramides when compared with goat milk

Antimicrobial Activity of Grapefruit Seed Extract on Edible Mushrooms Contaminations: Efficacy in Preventing Pseudomonas spp. in Pleurotus eryngii

Pleurotus eryngii is an edible mushroom that suffers significant losses due to fungal contamination and bacteriosis. The Pseudomonadaceae family represents one of the most frequent etiologic agents. Grapefruit seed extract (GSE) is a plant extract that contains different bioactive components, such as naringin, and exhibits a strong antibacterial and antioxidant activity. Over the last decade, GSE use as an alternative to chemical treatments in the food sector has been tested. However, to our knowledge, its application on mushroom crops has never been investigated. This study focuses on evaluating GSE efficacy in preventing P. eryngii yellowing. GSE antibiotic activity, inhibitory and bactericidal concentrations, and antibiofilm activity against several microorganisms were tested with the Kirby–Bauer disk diffusion assay, the broth microdilution susceptibility test, and the Crystal violet assay, respectively. In vitro, the extract exhibited antimicrobial and antibiofilm activity against Staphylococcus aureus 6538 and MRSA (wild type), Escherichia coli ATCC 8739, and Pseudomonas spp. (Pseudomonas aeruginosa 9027, P. fluorescens (wild type)). GSE application in vivo, in pre- and post-sprouting stages, effectively prevented bacterial infections and subsequent degradation in the mushroom crops: none of the P. eryngii treated manifested bacteriosis. Our findings support the use of GSE as an eco-friendly and sustainable alternative to chemical treatments for protecting P. eryngii crops from bacterial contamination, consequently ensuring food safety and preventing financial losses due to spoilage. Furthermore, GSE’s potential health benefits due to its content in naringin and other bioactive components present new possibilities for its use as a nutraceutical in food fortification and supplementation

Investigation of Seasonal Variation in Fatty Acid and Mineral Concentrations of Pecorino Romano PDO Cheese: Imputation of Missing Values for Enhanced Classification and Metabolic Profile Reconstruction

Seasonal variation in fatty acids and minerals concentrations was investigated through the analysis of Pecorino Romano cheese samples collected in January, April, and June. A fraction of samples contained missing values in their fatty acid profiles. Probabilistic principal component analysis, coupled with Linear Discriminant Analysis, was employed to classify cheese samples on a production season basis while accounting for missing data and quantifying the missing fatty acid concentrations for the samples in which they were absent. The levels of rumenic acid, vaccenic acid, and omega-3 compounds were positively correlated with the spring season, while the length of the saturated fatty acids increased throughout the production seasons. Concerning the classification performances, the optimal number of principal components (i.e., 5) achieved an accuracy in cross-validation equal to 98%. Then, when the model was tasked with imputing the lacking fatty acid concentration values, the optimal number of principal components resulted in an R2 value in cross-validation of 99.53%

Surface Characterization of AZ31 Alloy after Long-Term Immersion in Simulated Body Fluid

The aim of the research campaign was to simulate in vitro the typical conditions for the corrosion in biofluid of a femoral bone implant manufactured with AZ31 alloy. The samples were immersed in biofluid (alpha-MEM) for time intervals of up to 56 days. For each immersion time, the chemical compositions and morphologies of the samples were studied with SEM, EDX, XRD, Raman spectroscopy, and XPS. The weight losses of the samples caused by corrosion were also measured. The results highlighted the formation of calcium phosphate crystals on the surface of the samples. This type of coating is well-known for its excellent corrosion resistance and for its ability to accelerate tissue regeneration. The deceleration of the corrosion process, observed after 28 days of immersion in biofluid, confirms the anti-corrosive effect of the coating that was spontaneously formed during the immersion tests

Lipidomics of sheep and goat Milk-based infant formulae during in vitro dynamic digestion

Lipid hydrolysis process during IF digestion, particularly the characterization of the lipidome and the resulting lipid breakdown products, has not been thoroughly investigated. This study aimed to compare the lipid hydrolysis profiles during the in vitro dynamic digestion of IFs made from whole sheep and goat milk. Using a lipidomics platform and multivariate statistical analysis, we observed changes in complex lipid levels during digestion. In the gastric compartment, we noted a progressive hydrolysis of triacylglycerols, phosphatidylcholines, and sphingomyelins. Conversely, lipolysis breakdown products like monoacylglycerols (e.g., MG(16:0), MG (18:0)), diacylglycerols, lysophosphatidylcholines (LPC 16:0, LPC 18:1, LPC 18:2), and free fatty acids increased in the intestinal compartment. The lipolysis trends were similar for both types of infant formulas, with long-chain fatty acid triglycerides (C > 46) exhibiting lower digestibility compared to medium-chain fatty acid triglycerides. Overall, these results indicate that sheep milk can be used as an ingredient in the manufacturing of IF