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The identification of traces of life beyond Earth (e.g., Mars, icy moons) is a challenging task because terrestrial chemical-based molecules may be destroyed by the harsh conditions experienced on extraterrestrial planetary surfaces. For this reason, studying the effects on biomolecules of extremophilic microorganisms through astrobiological ground-based space simulation experiments is significant to support the interpretation of the data that will be gained and collected during the ongoing and future space exploration missions. Here, the stability of the biomolecules of the cryptoendolithic black fungus Cryomyces antarcticus, grown on two Martian regolith analogues and on Antarctic sandstone, were analysed through a metabolomic approach, after its exposure to Science Verification Tests (SVTs) performed in the frame of the European Space Agency (ESA) Biology and Mars Experiment (BIOMEX) project. These tests are building a set of ground-based experiments performed before the space exposure aboard the International Space Station (ISS). The analysis aimed to investigate the effects of different mineral mixtures on fungal colonies and the stability of the biomolecules synthetised by the fungus under simulated Martian and space conditions. The identification of a specific group of molecules showing good stability after the treatments allow the creation of a molecular database that should support the analysis of future data sets that will be collected in the ongoing and next space exploration missions.

Final published version: Ljiljana Kuzmanović, Giulia Mandalà, Silvio Tundo, Roberto Ciorba, Matteo Frangella, Roberto Ruggeri, Francesco Rossini, Federica Gevi, Sara Rinalducci, Carla Ceoloni: Equipping durum wheat -Thinopyrum ponticum recombinant lines with a Thinopyrum elongatum major QTL for resistance to Fusarium diseases through a cytogenetic strategy. In: Frontiers in Plant Science Vol. 10, 2019, Article 1324, https://doi.org/10.3389/fpls.2019.01324

Cryptoendolithic microbial communities, discovered in the extremely cold, hyper-arid McMurdo Dry Valleys of Antarctica (Friedmann 1982), the most similar terrestrial environments to Mars surface (Wynn-Williams and Edwards 2000; Onofri et al., 2004), have been considered as a candidate in supporting the search of life in Mars exploration. In such harsh conditions, microorganisms grow in airspaces among mineral grains and show some adaptations, as the accumulation of protective pigments and compatible solutes, assuring their survival. In the frame of the Lichen and Fungi experiment (LIFE, P.I. Silvano Onofri; Onofri et al. 2012, 2015), small samples of these communities, were exposed to space, in the EXPOSE-E facility for 1.5 years, with an exposition to vacuum (10-7 to 10-4 Pa) (Horneck et al., 2010), galactic cosmic radiation (≤190mGy) (Berger et al., 2012), and the full spectrum of solar extraterrestrial electromagnetic radiation to which cryptoendolithic microorganisms demonstrated to survive (Scalzi et al., 2012). The search for trace of extant or extinct life is one of the main goals of the future space mission beyond Earth. The future rover missions ExoMars 2020 (ESA-Roscomos) and Mars 2020 (NASA) are exploring chemical and biological indicators of life, called biomarkers. A good biomarker must have a biogenic origin and thus must unequivocally be identified as possible trace of life. The detection of biomarkers on Mars was the aim of BIOMEX (BIOlogy and Mars Experiment) project, in which has been investigated the alteration of different biomarkers, after exposure to space and Mars-like conditions outside the International Space Station. In this contest, the aim of this work was to characterize fungal biomarkers from these exposed rock samples, with different approaches: i) Raman spectroscopy and InfraRed analyses, which has been considered excellent tools for the detection of inorganic and organic molecules, such as microbial pigments and ii) -omics approaches, as lipidomic and metabolomic techniques, performed to detect biological macromolecules and to determine their stability after space exposure. The focus of lipidomics and metabolomics has been on biomarker discovery, with the aim of identifying metabolites that are correlated with environmental exposures. The results suggest that microbial molecules can be detected through different techniques. In particular, our attention was focused on pigments, such as melanin and carotenoids that maintain their stability also after 1.5 years of space exposure. These results are of importance for the upcoming life-detection missions on Mars finalized for the search for past, extant or extinct life outside the Earth.

Embryo assessment is currently performed through the analysis of morphology and cleavage rate. Recent studies have sought to identify a correlation between quali-quantitative profiles of small molecules of metabolic interest and the outcome of embryo transfer. Approaches relying on both optical and non-optical spectroscopy have been proposed to non-invasively monitor the embryo culture media. However, the non-invasive approach only offers an indirect strategy to monitor embryos and a turn-around solution to bypass the limits of detection of these analytical techniques. In this paper we pave the way for direct metabolic assessment of embryos through the mass-spectrometry-based analysis of blastocoele fluid, which is withdrawn from the blastocoele cavity prior to cryostorage of blastocysts. We conclude that it is possible to detect most of the metabolites of potential interest right at the very heart of the blastocyst, without disrupting the workflow of a classic laboratory pipeline.

The word „omics‟, is derived from the Greek suffix, „ome‟ (meaning all, every). In the mid 1990‟s, rapid evolution and growing knowledge concerning genomes led to a quick development of the full range of „omics‟ approaches (genomics, transcriptomics, proteomics, metabolomics). This PhD thesis focuses on the application of new “omics” sciences to characterized samples of biomedical interest such as pharmaceutical products. The first section is focused on the characterization of important pharmaceutical products used by hemophiliac patients, trough proteomics approaches. The first objective of this study was to compare the heterogeneity and the high purity of three rFVIII commercially available rFVIII concentrates before and after thrombin digestion. The second object was to extend the same study to plasma derived factor VIII . The second part of my PhD project was designed mainly to the development and optimization of a new HPLC-MS method for metabolomics. In the last part of my research I used this analytical approach to investigate metabolites in blastocoels fluid that is a routine waste product prior to pre-implantation blastocyst vitrification. The aim of this study was directed to identify a correlation between quali-quantitative profiles of small molecules of metabolic interest and the outcome of embryo transfer through a simple HPLC-MS assay. The last part of my research was dedicated to studing metabolism in red blood cell during storage. Proteome and metabolomic approaches, are widely used for studies complex biological systems and can be used for applications in biological areas. La parola omica deriva dal suffisso greco “ome” che significa: tutto, intero, completo. In particolar modo per quanto riguarda l’area biologica lo scopo delle scienze omiche è quello di avere una visione globale per comprendere la vita e la sua organizzazione. L’evoluzione rapida e la crescita di conoscenza riguardante il genoma hanno portato ad una rapida crescita nello sviluppo di nuove scienze (genomica, trascrittomica, proteomica, metabolomica) La mia tesi di dottorato riguarda le applicazioni delle nuove scienze omiche per caratterizzare campioni di interesse biomedico come ad esempio prodotti farmaceutici. Il primo obiettivo dello studio è stato quello di comparare e caratterizzare fattore VIII ricombinante prodotto da tre ditte farmaceutiche differenti. Questi prodotti attaualmente utilizzati per curare l’emofilia sono stati studiati prima e dopo attivazione attraverso il taglio effettuato dalla trombina. Il secondo obiettivo della mia tesi è stato quello di estendere lo studio proteomico non solo ai fattori VIII ricombinanti ma anche a quelli plasmaderivati. La seconda parte del mio progetto è stata dedicata allo sviluppo e all’ottimizzazione di un nuovo metodo basato sull’utilizzo di HPLC e spettrometro di massa per lo studio della metabolomica. Nell’ultima parte della mia tesi ho utilizzato questo tipo di approcio per studiare la variazione metabolica all’interno del fluido del blastocele. Lo scopo di questo lavoro è stato quello di individuare marker metabolici che possano definire la qualità delle blastocisti da impiantare durante il processo della FIVET. Infine ho utilizzato questo metodo per mettere in evidenza la variazione metabolica nei globuli rossi durante la loro conservazione in sacca. Dottorato di ricerca in Genetica e biologia cellulare

Background. Deficiency or dysfunction of coagulation factor VIII (FVIII) is the underlying cause of haemophilia A. Haemophilic patients are at present treated with plasma-derived FVIII (pdFVIII) or recombinant FVIII (rFVIII) in order to correct their clotting deficiency. pdFVIII concentrates are exclusively produced from human plasma upon pooling from multiple donors. It is not know whether the presence of excess of other plasma proteins, in addition to von Willebrand factor, could stimulate untoward immune responses in the recipient. Thus, information regarding the presence of contaminants in commercial products is of concern. Materials and Methods. Two commercially available pdFVIII concentrates were characterized through SDS-PAGE and mass spectrometry Emoclot® and Beriate® . Results. The components of two pdFVIII products considered in this study were well identified by mass spectrometry analysis, in both cases we found abundant components coming from blood plasma, and some other contaminants. Only in Beriate® we also found truncated form of pdFVIII. Conclusion. The two pdFVIII examined showed the presence of vWF, Fibrinogen in excess, and other substances that could be considered as contaminants or impurities. L'articolo è disponibile sul sito dell'editore http://www.bloodtransfusion.it

Demographic and clinical characteristics of the autistic sample (N = 30, unless otherwise specified). Typically developing controls were tightly sex- and age-matched, with M:F = 22–8, age 5.03 ± 0.32 years, and no clinical evidence of ASD-related DSM-IV diagnoses or intellectual disability. (DOCX 21 kb)

Supplementary methods and references. (DOCX 15 kb)

Detailed and summary statistics for metabolites displayed in Figs. 4 and 5. (DOCX 17 kb)

Supplementary methods and references. (DOCX 15 kb)