The urinary proteomic profile of arterial stiffness in the general population

Background Although arterial stiffness is an independent predictor of cardiovascular outcomes, its physiopathology remains unclear. Purpose This study aimed to investigate the urinary proteomic profile of aortic stiffness and provide insights into pathogenetic processes of arterial stiffness by pathway analysis. Methods In 669 participants (mean age, 50.5 years; 48.9% men) randomly recruited from the Flemish population, we measured carotid-femoral pulse wave velocity (PWV) by applanation tonometry. The proteomics of urine samples was quantified by using capillary electrophoresis coupled mass spectrometry. The proteomic data were analysed by the orthogonal projections to latent structures, a supervised dimensional reduction statistic method and summarised as a urinary proteomic (UP) score. Results The mean values were 7.56±2.02 m/s for PWV and 7.59±1.95 unit for the UP score. PWV was significantly associated with the UP score before and after adjustment for the potential covariates (β coefficient: 0.81 and 0.75, respectively; p<0.001). The significant proteins in the urinary proteomic profile consisted of 43 kinds of proteins, including collagen I, II and III, fibrinogen, matrix Gla-protein, apolipoprotein A-I and A-VI. The pathways annotated by the significant proteins mainly involved in fibrosis, signal conduction, platelet activation and aggregation. Conclusions In conclusion, the urinary proteomic profile could be a new biomarker of aortic stiffness and the altered proteins may link to the underlying mechanisms and holds the potential to discover novel therapeutic targets for arterial stiffness. FUNDunding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): The Internal Funds KU Leuven (STG-18-00379) Distribution and Correlation The Enrichment Pathways

Brachyspira Species Avidity to Colonic Mucins from Pigs with and without Brachyspira hyodysenteriae Infection is Species-Specific and Varies between Strains

Brachyspira hyodysenteriae is commonly associated with swine dysentery (SD), a disease that has an economic impact in the swine industry. B. hyodysenteriae infection results in changes to the colonic mucus niche with a massive mucus induction, which substantially increases the amount of B. hyodysenteriae binding sites in the mucus. We have previously determined that a B. hyodysenteriae strain binds to colon mucins in a manner that differs between pigs and mucin types. Here, we investigated if adhesion to mucins is a trait observed across a broad set of B. hyodysenteriae strains and isolates and furthermore at a genus level ( B. innocens, B. pilosicoli, B. murdochii, B. hampsonii and B. intermedia strains). Our results show that binding to mucins appears to be specific to B. hyodysenteriae , and within this species, the binding ability to mucins varies between strains/isolates, increases to mucins from pigs with SD, and is associated to sialic acid epitopes on mucins. Infection with B. hyodysenteriae strain 8dII results in mucin glycosylation changes in the colon including a shift in sialic acid containing structures. Thus, we demonstrate through hierarchical cluster analysis and Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) models of the relative abundances of sialic acid-containing glycans, that sialic acid containing structures in the mucin O -glycome are good predictors of B. hyodysenteriae strain 8dII infection in pigs. The results emphasize the role of sialic acids in governing B. hyodysenteriae interactions with its host, which may open perspectives for therapeutic strategies.

Seminal Plasma Lipidomics Profiling to Identify Signatures of Kallmann Syndrome

BackgroundKallmann syndrome (KS) is a rare developmental disorder. Our previous metabolomics work showed substantial changes in linoleic acid and glycerophospholipid metabolism in KS. Here, we performed targeted lipidomics to further identify the differential lipid species in KS.MethodsTwenty-one patients with KS (treatment group) and twenty-two age-matched healthy controls (HC, control group) were enrolled. Seminal plasma samples and medical records were collected. Targeted lipidomics analysis of these samples was performed using ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS).ResultsLipidomics profiling of patients with KS and the HCs showed clear separation in the orthogonal projections to latent structures-discriminant analysis (OPLS-DA). There were many differential lipids identified, with the main differential lipid species being triacylglycerols (TAGs), phosphatidylcholines (PCs) and phosphatidylethanolamine (PE).ConclusionsThe lipidomics profile of patients with KS changed. It was also determined that TAGs, PCs and PE are promising biomarkers for KS diagnosis. To our knowledge, this is the first report to analyze lipidomics in men with Kallmann syndrome.

Optimization of Enzyme Production from Streptomyces by Solid-state Fermentation of Agricultural Wastes using the Orthogonal Projections to Latent Structures-Discriminant Analysis (OPLS-DA) Method

This study used Orthogonal Projections to Latent Structures-Discriminant Analysis (OPLS-DA) to identify and optimize the combined effect of the temperature, fermentation time, substrate-type, and fermenting microorganism on solid-state fermentation (SSF) culture of different agricultural wastes, in order to evidence the interactions among variables that allow maximizing the production of each enzyme. Thus, barley and wheat straw were fermented using two Streptomyces strains (S. MDG 301 and S. MDG 147) under SSF conditions. Key degradative enzymes (xylanases, carboxylmethilcellulase (CMCase), mannanase, laccase and peroxidase) and lignin solubilisation were evaluated. Our results highlighted that the highest xylanases and CMCase levels were obtained after treatment of wheat straw for two days with the S. MDG 301 strain at 45°C. Similarly, we found that the highest mannanase level was achieved using the S. MDG 147 strain on the barley straw at 28°C. Therefore, the integrated approach used based on analytical and bio statistical methods, proved to be a valuable tool for an accurate and rapid determination of the most significant parameters for controlling the bio-transformation process in terms of the enzymatic activity and lignin solubilisation.

Chemometric Discrimination of the Geographical Origin of Three Greek Cultivars of Olive Oils by Stable Isotope Ratio Analysis

Α stable isotope ratio mass spectrometer was used for stable isotope ratio (i.e., δ13C, δ18O, and δ2H) measurements, achieving geographical discrimination using orthogonal projections to latent structures discriminant analysis. A total of 100 Greek monovarietal olive oil samples from three different olive cultivars (cv. Koroneiki, cv. Lianolia Kerkyras, and cv. Maurolia), derived from Central Greece and Peloponnese, were collected during the 2019–2020 harvest year aiming to investigate the effect of botanical and geographical origin on their discrimination through isotopic data. The selection of these samples was made from traditionally olive-growing areas in which no significant research has been done so far. Samples were discriminated mainly by olive cultivar and, partially, by geographical origin, which is congruent with other authors. Based on this model, correct recognition of 93.75% in the training samples and correct prediction of 100% in the test set were achieved. The overall correct classification of the model was 91%. The predictability based on the externally validated method of discrimination was good (Q2 (cum) = 0.681) and illustrated that δ18O and δ2H were the most important isotope markers for the discrimination of olive oil samples. The authenticity of olive oil based on the examined olive varieties can be determined using this technique.

Leaves and Spiny Burs of Castanea Sativa from an Experimental Chestnut Grove: Metabolomic Analysis and Anti-Neuroinflammatory Activity

Castanea sativa cultivation has been present in Mediterranean regions since ancient times. In order to promote a circular economy, it is of great importance to valorize chestnut groves’ by-products. In this study, leaves and spiny burs from twenty-four Castanea trees were analyzed by 1H NMR metabolomics to provide an overview of their phytochemical profile. The Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) performed on these data allowed us to distinguish ‘Marrone’ from ‘Castagna’, since the latter were generally more enriched with secondary metabolites, in particular, flavonoids (astragalin, isorhamnetin glucoside, and myricitrin) were dominant. Knowing that microglia are involved in mediating the oxidative and inflammatory response of the central nervous system, the potential anti-inflammatory effects of extracts derived from leaves and spiny burs were evaluated in a neuroinflammatory cell model: BV-2 microglia cells. The tested extracts showed cytoprotective activity (at 0.1 and 0.5 mg/mL) after inflammation induction by 5 µg/mL lipopolysaccharide (LPS). In addition, the transcriptional levels of IL-1β, TNF-α, and NF-kB expression induced by LPS were significantly decreased by cell incubation with spiny burs and leaves extracts. Taken together, the obtained results are promising and represent an important step to encourage recycling and valorization of chestnut byproducts, usually considered “waste”.

Exploration of Chemical Diversity and Antitrypanosomal Activity of Some Red Sea-Derived Actinomycetes Using the OSMAC Approach Supported by LC-MS-Based Metabolomics and Molecular Modelling

In the present study, we investigated the actinomycetes associated with the Red Sea-derived soft coral Sarcophyton glaucum in terms of biological and chemical diversity. Three strains were cultivated and identified to be members of genera Micromonospora, Streptomyces, and Nocardiopsis; out of them, Micromonospora sp. UR17 was putatively characterized as a new species. In order to explore the chemical diversity of these actinobacteria as far as possible, they were subjected to a series of fermentation experiments under altering conditions, that is, solid and liquid fermentation along with co-fermentation with a mycolic acid-containing strain, namely Nocardia sp. UR23. Each treatment was found to affect these actinomycetes differently in terms of biological activity (i.e., antitrypanosomal activity) and chemical profiles evidenced by LC-HRES-MS-based metabolomics and multivariate analysis. Thereafter, orthogonal projections to latent structures discriminant analysis (OPLS-DA) suggested a number of metabolites to be associated with the antitrypanosomal activity of the active extracts. The subsequent in silico screenings (neural networking-based and docking-based) further supported the OPLS-DA results and prioritized desferrioxamine B (3), bafilomycin D (10), and bafilomycin A1 (11) as possible antitrypanosomal agents. Our approach in this study can be applied as a primary step in the exploration of bioactive natural products, particularly those from actinomycetes.

Διερεύνηση της μητρικής πρόσληψης θρεπτικών συστατικών και συσχέτιση της με τους μεταβολίτες του αμνιακού υγρού

Σκοπός: Η παρούσα μελέτη παρατήρησης, δεδομένης χρονικής στιγμής, είχε ως σκοπό την ανάπτυξη και τον έλεγχο εγκυρότητας ενός ερωτηματολογίου συχνότητας κατανάλωσης τροφίμων (FFQ) για τη διερεύνηση της μητρικής πρόσληψης θρεπτικών συστατικών και τη συσχέτισή της με τους μεταβολίτες του αμνιακού υγρού (ΑΥ). Μέθοδος: Για τον έλεγχο εγκυρότητας του FFQ, τα αποτελέσματα του προτεινόμενου ερωτηματολογίου συγκρίθηκαν με τον μέσο όρο δύο, μη διαδοχικών, εικοσιτετραώρων ανάκλησης (μέθοδος αναφοράς). Για τη συσχέτιση της μητρικής διατροφικής πρόσληψης με τη σύσταση του ΑΥ, καταγράφηκαν διατροφικά δεδομένα εγκύων οι οποίες επρόκειτο να υποβληθούν σε αμνιοπαρακέντηση για προγεννητικό έλεγχο, χρησιμοποιώντας το έγκυρο FFQ και κατόπιν προσωπικής συνέντευξης. Μετά το πέρας της προγραμματισμένης προγεννητικής ανάλυσης, καταψύχονταν 2-3 ml ΑΥ (-80oC). Η μητρική διατροφική πρόσληψη εκφράστηκε ως πηλίκο πρωτεΐνης / υδατάνθρακα και λίπος (p/np) και το ΑΥ αναλύθηκε με τη μέθοδο του πυρηνικού μαγνητικού συντονισμού (NMR). Η κατηγοριοποίηση των γυναικών πραγματοποιήθηκε σε ποσοστημόρια (δεκατημόρια, τεταρτημόρια, τρεις κλάσεις ίσου περιεχομένου), και σε κάθε περίπτωση συγκρίθηκαν οι ακραίες ομάδες. Εφαρμόστηκε διακριτή ανάλυση με ορθογώνια περιστροφή (Οrthogonal Projections to Latent Structures Discriminant Analysis), με στόχο την ταυτοποίηση μεταβολιτών. Αποτελέσματα: Στον έλεγχο εγκυρότητας του FFQ συμμετείχαν 179 υγιείς έγκυες, με μονήρη κύηση. Αν και οι μέσες τιμές πρόσληψης για τη πλειοψηφία των θρεπτικών συστατικών και την ενέργεια, έτειναν να υπερεκτιμούνται από το FFQ σε σύγκριση με τη μέθοδο αναφοράς, η παράμετρος d του Cohen ήταν μικρότερη του 0,3 στην πλειοψηφία τους. Τα όρια συμφωνίας μεταξύ των δύο μεθόδων, σύμφωνα με τη μέθοδο Bland-Altman, ήταν αποδεκτά. Οι στατιστικά σημαντικές θετικές συσχετίσεις κυμαίνονταν από 0,35 έως 0,77. Το ποσοστό των γυναικών που ταξινομήθηκε ορθά ήταν από 73,2 έως 92,2%, ενώ η εσφαλμένη ταξινόμηση ήταν χαμηλή σε όλες τις περιπτώσεις. Ο συντελεστής k του Cohen κυμαινόταν μεταξύ 0,31 έως 0,78 και ο ICC μεταξύ 0,49 έως 0,89. Για τη συσχέτιση της μητρικής διατροφικής πρόσληψης με τη σύσταση του ΑΥ, ελέγχθηκαν 78 υγιείς έγκυες, με μονήρη κύηση, μεταξύ της 18ης-26ης εβδομάδας κύησης. Όπως φάνηκε από τον οπτικό διακριτό διαχωρισμό των διαγραμμάτων σκέδασης των ακραίων ομάδων, η μητρική διατροφική πρόσληψη επηρεάζει τη σύσταση του ΑΥ. Το ΑΥ των γυναικών οι οποίες ανήκαν στα ακραία ανώτερα ποσοστημόρια, χαρακτηρίζονταν από αυξημένα επίπεδα γλυκόζης, φαινυλαλανίνης, ιστιδίνης, βαλίνης και αλανίνης ενώ το ΑΥ εκείνων στα ακραία χαμηλότερα ποσοστημόρια, από αυξημένα επίπεδα γαλακτικού οξέος και χολίνης. Η επίδραση της μητρικής διατροφικής πρόσληψης στη σύσταση του ΑΥ ήταν πιο διακριτή, όσο η σύγκριση γινόταν μεταξύ ομάδων που είχαν τη μεγαλύτερη διαφορά πηλίκου πρωτεΐνης / υδατάνθρακα και λίπος (p/np). Συμπεράσματα: Το FFQ του οποίου ελέγχθηκε η εγκυρότητα, μπορεί να αποτυπώσει σε ικανοποιητικό βαθμό τη μητρική διατροφική πρόσληψη, κατά το δεύτερο τρίμηνο της εγκυμοσύνης, σε επίπεδο ενέργειας και θρεπτικών συστατικών. Η μητρική διατροφική πρόσληψη επηρεάζει το μεταβολικό προφίλ του ΑΥ, κατά το δεύτερο τρίμηνο της κύησης. Τα αποτελέσματα αναδεικνύουν την αναγκαιότητα εύρεσης της ιδανικής αναλογίας μακροθρεπτικών και μικροθρεπτικών συστατικών της μητρικής διατροφικής πρόσληψης.

Dairy Intake in 2 American Adult Cohorts Associates with Novel and Known Targeted and Nontargeted Circulating Metabolites

ABSTRACT Background The role of dairy in health can be elucidated by investigating circulating metabolites associated with intake. Objectives We sought to identify metabolites associated with quantity and type of dairy intake in the Framingham Heart Study Offspring and Third Generation (Gen3) cohorts. Methods Dairy intake (total dairy, milk, cheese, yogurt, and cream/butter) was analyzed in relation to targeted (Offspring, n = 2205, 55.1 ± 9.8 y, 52% female, 217 signals; Gen3, n = 866, 40.5 ± 8.8 y, 54.9% female, 79 signals) and nontargeted metabolites (Gen3, ∼7031 signals) in a 2-step analysis including orthogonal projections to latent structures with discriminant analysis (OPLS-DA) in discovery subsets to identify metabolites distinguishing between high and low intake; and linear regression in confirmation subsets to assess putative associations, subsequently tested in the total samples. Previously reported associations were also investigated. Results OPLS-DA in the Offspring targeted discovery subset resulted in a variable importance in projection (VIP) >1 of 65, 60, 58, 66, and 60 metabolites for total dairy, milk, cream/butter, cheese, and yogurt, respectively, of which 5, 3, 1, 6, and 4 metabolites, respectively, remained after confirmation. In the Gen3 targeted discovery subset, OPLS-DA resulted in a VIP >1 of 17, 15, 13, 7, and 6 metabolites for total dairy, milk, cream/butter, cheese, and yogurt, respectively. In the Gen3 nontargeted discovery subset, OPLS-DA resulted in a VIP >2 of 203, 503, 78, 186, and 206 metabolites, respectively. Combining targeted and nontargeted results in Gen3, significant associations of 7 (6 unannotated), 2, 12 (11 unannotated), 0, and 61 (all unannotated) metabolites, respectively, remained. Candidate identities of unannotated signals included fatty acids and food flavorings. Results supported relations previously reported for C14:0 sphingomyelin, and marginal associations for deoxycholates. Conclusions Dairy in 2 American adult cohorts associated with numerous circulating metabolites. Reports about diet-metabolite relations and confirmation of previous findings might be limited by specificity of dietary intake and breadth of measured metabolites.