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Selenium is a well-known essential element with important roles in human reproductive health mainly due to its antioxidant character. This study aimed to investigate the potential role of selenoproteins on gut microbiota and male reproductive health. A new assay for the absolute quantification of selenoproteins in testicular tissue based on two dimensional chromatography with inductively coupled plasma mass spectrometry was performed for the first time. The gut microbiota profile was obtained by 16S rRNA gene sequencing. Numerous associations were found between testicular selenoproteins and gut microbiota (e.g. Mucispirillum, related with sperm activity and testosterone, was associated with glutathione peroxidase (GPx) and selenoalbumin (SeAlb), while Escherichia/Shigella, related to sex hormones, correlated with GPx, selenoprotein P (SelP) and SeAlb). The effects of Se-supplementation on testicular selenoproteins only occur in conventional mice, suggesting a potential selenoproteins-microbiota interplay that underlies testicular function. The selenoproteins GPx and SelP have been quantified for the first time in the testicles, and the novel identification of SeAlb, a protein with nonspecifically incorporated Se, is also reported. These findings demonstrate the significant impact of Se-supplementation on gut microbiota and male reproductive health. In addition, the analytical methodology applied here in selenoprotein quantification in testicular tissue opens new possibilities to evaluate their role in gut microbiota and reproductive health axis. ; This article was funded by the projects PGC 2018- 096608-B-C21 from the Spanish Ministry of Science and innovation (MCI) (Generación del Conocimiento. MCI/AEI/FEDER, EU "Una manera de hacer Europa") and UHU-1256905 from the FEDER Andalusian Operative Program 2014-2020 (Ministry of Economy, Knowledge, Business and Universities, Regional Government of Andalusia, Spain). S.R.A. thanks the Spanish Ministry of Economy and Competitiveness for a PhD scholarship (BES-2016-076364). The authors are grateful to FEDER (European Community) for financial support, Grant UNHU13-1E-1611. The authors would like to acknowledge the support from The Ramón Areces Foundation (ref. CIVP19A5918). ; Peer reviewed

Metals interactions ; Arsenic ; Cadmium ; Metallomics ; Metabolomics ; Mus musculus ; Mass spectrometry ; Arsenic and cadmium are toxic metals of environmental significance with harmful effects on man. To study the toxicological and biochemical effects of arsenic/cadmium in mammals a combined metallomic and metabolomic approach has been developed, complemented with the measurement of biochemical parameters in blood and histopathological evaluation of liver injury in mice Mus musculus under exposure to both xenobiotics. Size-exclusion chromatography (SEC) was combined with affinity chromatography (AF) and ICP-MS detection using species unspecific isotopic dilution analysis (SUID) to characterize the biological effects of As/Cd on selenium containing proteins in the bloodstream of exposed mice. On the other hand, both direct infusion mass spectrometry (DIMS) and gas chromatography–mass spectrometry (GC–MS) provided information about changes in metabolites caused by metals. The results show that As/Cd exposure produces interactions in the distribution of both toxics between organs and plasma of mice and antagonistic interactions with selenium containing proteins in the bloodstream. Interplay with essential metabolic pathways, such as energy metabolism and breakdown of membrane phospholipids were observed, which are more pronounced under As/Cd exposure. In addition, heavy metal and metalloid causes differential liver injury, manifested by steatosis (non-alcoholic fatty liver disease, NAFLD) and infiltration of blood cells into the space of Disse. Biological significance This work presents new contributions in the study of arsenic/cadmium interactions in mice Mus musculus under controlled exposure. With the combination of metallomic and metabolomic approaches the traffic of As and Cd from liver to kidney by means of blood was observed and excretion of As (as arsenic metabolites) or Cd (as MTCd) is inhibited with the simultaneous administration of As/Cd, and these toxic elements have important influence in the levels of seleno-proteins in the plasma. In addition, the metabolomic approach reveals inhibition of different metabolic cycles such as tricarboxylic acid and phospholipid degradation that causes membrane damage and apoptosis that is histopathologically confirmed. This article is part of a Special Issue entitled: Environmental and structural proteomics. ; The authors thank the projects CTM2012-38720-G03-01 (Ministerio de Economia y Competitividad-Spain), P08-FQM-03554 and P09-FQM-04659 (Consejeria de Innovacion, Andalusian government). Miguel Angel Garcia Sevillano thanks the Ministerio de Educacion for a predoctoral scholarship.

Speciation of seleno-metabolites and selenoproteins in Se-enriched Chlorella sorokiniana microalgae, in vitro and in vivo extracts has been performed by HPLC-ICP-MS. Selenium bioaccesibility in Se-enriched Chlorela sorokiniana evaluated by in vitro gastrointestinal digestion of the selenized microalga was 81% (79% as SeMet). Mice supplemeted with Se-enriched C. sorokiniana presented increased Se concentration in kidney, reflecting a potencial mechanism of excretion by urine. In addition, selenium bioavailavility, which was in the range of 3–15% depending on the diet was 1.13-fold higher in mice fed with Se-enriched microalgae against the basal diet, but decreased with higher amounts of selenium. Regarding bioactivity (∑Se as selenoproteins in serum/total Se in serum = 76–85% depending on the diet), selenium and selenoproteins (SeP) increased in serum from animals fed high supplemental Se levels. These results showed that Se-enriched algae can be considered as an alternative selenized food for humans, due to their high Se bioavailability. ; This work has been supported by the projects CTM2015-67902-C2-1-P and PG2018-096608-B-C21 from the Spanish Ministry of Economy and Competitiveness and P12-FQM-0442 and P09-FQM-04659 from the Regional Ministry of Economy, Innovation, Science and Employment (Andalusian Government, Spain). Finally, authors are grateful to FEDER (European Community) for financial support, grants number UNHU13-1E-1611 and UNHU15-CE-3140.