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Despite decades of research and handling of semen for use in artificial insemination (AI) and other assisted reproductive technologies, 5-10% of selected boar sires are still considered sub-fertile, escaping current assessment methods for sperm quality and resilience to preservation. As end-product, the ejaculate (emitted spermatozoa sequentially exposed to the composite seminal plasma, the SP) ought to define the homeostasis of the testes, the epididymis, and the accessory sexual glands. Yet, linking findings in the ejaculate to sperm production biology and fertility is suboptimal. The present essay critically reviews how the ejaculate of a fertile boar can help us to diagnose both reproductive health and resilience to semen handling, focusing on methods -available and under development- to identify suitable biomarkers for cryotolerance and fertility. Bulk SP, semen proteins and microRNAs (miRNAs) have, albeit linked to sperm function and fertility after AI, failed to enhance reproductive outcomes at commercial level, perhaps for just being components of a complex functional pathway. Hence, focus is now on the interaction sperm-SP, comparing in vivo with ex vivo, and regarding nano-sized lipid bilayer seminal extracellular vesicles (sEVs) as priority. sEVs transport fragile molecules (lipids, proteins, nucleic acids) which, shielded from degradation, mediate cell-to-cell communication with spermatozoa and the female internal genital tract. Such interaction modulates essential reproductive processes, from sperm homeostasis to immunological female tolerance. sEVs can be harvested, characterized, stored, and manipulated, e.g. can be used for andrological diagnosis, selection of breeders, and alternatively be used as additives to improve cryosurvival and fertility.

BackgroundMating induces large changes in the female genital tract, warranting female homeostasis and immune preparation for pregnancy, including the preservation of crucial oxidative status among its pathways. Being highly susceptible to oxidative stress, sperm survival and preserved function depend on the seminal plasma, a protection that is removed during sperm handling but also after mating when spermatozoa enter the oviduct. Therefore, it is pertinent to consider that the female sperm reservoir takes up this protection, providing a suitable environment for sperm viability. These aspects have not been explored despite the increasing strategies in modulating the female status through diet control and nutritional supplementation. AimsTo test the hypothesis that mating modifies the expression of crucial oxidative-reductive transcripts across the entire pig female genital tract (cervix to infundibulum) and, particularly in the sperm reservoir at the utero-tubal junction, before ovulation, a period dominated by estrogen stimulation of ovarian as well as of seminal origin. MethodsThe differential expression of estrogen (ER) and progesterone (PR) receptors and of 59 oxidative-reductive transcripts were studied using a species-specific microarray platform, in specific segments of the peri-ovulatory sow reproductive tract in response to mating. ResultsMating induced changes along the entire tract, with a conspicuous downregulation of both ER and PR and an upregulation of superoxide dismutase 1 (SOD1), glutaredoxin (GLRX3), and peroxiredoxin 1 and 3 (PRDX1, PRDX3), among other NADH Dehydrogenase Ubiquinone Flavoproteins, in the distal uterus segment. These changes perhaps helped prevent oxidative stress in the area adjacent to the sperm reservoir at the utero-tubal junction. Concomitantly, there were a downregulation of catalase (CAT) and NADH dehydrogenase (ubiquinone) oxidoreductases 1 beta subcomplex, subunit 1 (NDUFB1) in the utero-tubal junction alongside an overall

Seminal plasma contains many morphologically hetero-geneous extracellular vesicles (sEVs). These are sequentially released by cells of the testis, epididymis, and accessory sex glands and involved in male and fe-male reproductive processes. This study aimed to define in depth sEV subsets isolated by ultrafiltration and size exclusion chromatography, decode their proteomic profiles using liquid chromatography-tandem mass spectrometry, and quantify identified proteins using sequential window acquisition of all theoretical mass spectra. The sEV subsets were defined as large (L-EVs) or small (S-EVs) by their protein concentration, morphology, size distribution, and EV-specific protein markers and purity. Liquid chromatography-tandem mass spectrometry identified a total of 1034 proteins, 737 of them quantified by SWATH in S-EVs, L-EVs, and non-EVs-enriched samples (18-20 size exclusion chromatography-eluted fractions). The differential expression analysis revealed 197 differentially abundant proteins between both EV subsets, S-EVs and L-EVs, and 37 and 199 between S-EVs and L-EVs versus non-EVs-enriched samples, respectively. The gene ontology enrichment analysis of differentially abundant proteins suggested, based on the type of protein detected, that S-EVs could be mainly released through an apocrine blebbing pathway and be involved in modulating the im-mune environment of the female reproductive tract as well as during sperm-oocyte interaction. In contrast, L-EVs could be released by fusion of multivesicular bodies with the plasma membrane becoming involved in sperm physiological processes, such as capacitation and avoidance of oxidative stress. In conclusion, this study provides a procedure capable of isolating subsets of EVs from pig seminal plasma with a high degree of purity and shows differences in the proteomic profile between EV subsets, indicating different sources and biological functions for the sEVs., Funding Agencies|Spanish Ministry of Science and Innovation(MCIN/AEI) [PID2019- 105713GB-I00, RED2018-102411-T, 891382]; European Union [PROMETEO/2020/071]; Generalitat Valenciana [2019-00288]; FORMAS, Stockholm; Madrid, Spain; [PID2020-113493RB-I00]

La marca de motocicletes Montesa, ha estat un referent dels vehicles de dues rodes en l’àmbit nacional i campiona indiscutible del mundial de trial durant molts anys, remarcant l’última època amb en Toni Bou. Durant la infantesa de molts nens, la Montesa Cota 25, ha estat a la carta dels reis i, tanmateix, la Cota 247 dels més grans, amb la il·lusió que tant els grans com els petits pilotessin la ‘mateixa moto’. L’objecte d’aquest treball de final de grau es centra se fer un estudi de la reconstrucció d’una Montesa, una de les marques catalanes més importants de motos. Per donar forma al treball es parlarà de la importància de Montesa al món, es farà una descripció de totes les parts que impliquen la restauració d’una ‘Montesa Cota 25 A’ i finalment es realitzarà un estudi mediambiental junt amb l’explicació de la normativa que han de complir les motos de trial clàssiques per poder circular. Un cop argumentats tots aquests punts es pretén mostrar la importància d’aquests vehicles per a la nostra història i concloure si l’impacte ambiental és tan gran com molta gent pensa. Per desenvolupar el projecte, primer tornarem als orígens d’aquesta marca de motos, per entendre la seva història, i fins on va arribar. Després explicarem el procés de restauració d’una moto de trial clàssica, i també en farem un estudi ambiental i de normatives com per exemple, tot el que comporta passar la ITV, o tenir un vehicle d’aquests com a històric. Finalment, esperem poder extreure les conclusions adients per considerar si val la pena mantenir aquests vehicles com relíquies o són objectes del passat que no val la pena guardar. Per fer-ho es presentarà una enquesta diverses persones d’èpoques diferents per veure què en pensen d’aquests vehicles i què han suposat per a ells al llarg de les seves vides.

Seminal plasma contains numerous extracellular vesicles (sEVs). Since sEVs are apparently involved in male (in)fertility, this systematic review focused on studies specifically investigating such relationship. Embase, PubMed, and Scopus databases were searched up to 31 December 2022, primarily identifying a total of 1440 articles. After processing for screening and eligibility, 305 studies were selected as they focused on sEVs, and 42 of them were considered eligible because they included the word fertility or a related word such as infertility, subfertility, fertilization, and recurrent pregnancy loss in the title, objective(s), and/or keywords. Only nine of them met the inclusion criteria, namely (a) conducting experiments aimed at associating sEVs with fertility concerns and (b) isolating and adequately characterizing sEVs. Six studies were conducted on humans, two on laboratory animals, and one on livestock. The studies highlighted some sEV molecules, specifically proteins and small non-coding RNAs, that showed differences between fertile and subfertile or infertile males. The content of sEVs was also related to sperm fertilizing capacity, embryo development, and implantation. Bioinformatic analysis revealed that several of the highlighted sEV fertility-related proteins would be cross-linked to each other and involved in biological pathways related to (i) EV release and loading and (ii) plasma membrane organization., Funding Agencies|Spanish Ministry of Science and Innovation - MCIN/AEI/10.13039/501100011033, Madrid, Spain [PID2020-113493RB-I00]; Madrid, Spain; Research Council FORMAS, Stockholm, Sweden [891382]; Ministry of Universities (Spain); European Union-Next Generation EU Funds; European Union; [2019-00288]

Introduction : Pig seminal plasma (SP) is rich in active forms of all three isoforms (1-3) of transforming growth factor beta (TGF-beta), a chemokine modulatory of the immune environment in the female genital tract once semen is delivered during mating or artificial insemination (AI). The present study aimed to examine how TGF-beta s are secreted by the epithelium of the male reproductive tract and how they are transported in semen, emphasizing the interplay with seminal extracellular vesicles (sEVs). Methods : Source of TGF-beta s was examined by immunohistochemistry in testis, epididymis, and accessory sex glands, by immunocytochemistry in ejaculated spermatozoa, and by Luminex xMAP((R)) technology in SP and sEVs retrieved from healthy, fertile male pigs used as breeders in AI programs. Results : All three TGF-beta isoforms were expressed in all reproductive tissues explored and would be released into ductal lumen either in soluble form or associated with sEVs. Ejaculated spermatozoa expressed all three TGF-beta isoforms, both inside and outside, probably the outer one associated with membrane-bound sEVs. The results confirmed that pig SP contains all three TGF-beta isoforms and demonstrated that a substantial portion of them is associated with sEVs. Discussion : Seminal EVs would be involved in the cellular secretion of the active forms of seminal TGF-beta isoforms and in their safe transport from the male to the female reproductive tract., Funding Agencies|Spanish Ministry of Science and Innovation - MCIN/AEI , Madrid, Spain [PID2020-113493RB-I00]; European Union [891382]; FORMAS, Stockholm [2019-00288]

L’objectiu principal d’aquest projecte, és l’estudi d’una proposta de restauració d’un vehicle històric, concretament el bus Biblioteca, tenint en compte tot el conjunt de processos i la normativa d’aquests alhora de realitzar la restauració d’aquest vehicle històric. S’estudiarà una possible restauració per al Bus Biblioteca. S’escolliran els millors processos de restauració seguint tota una sèrie de criteris. A continuació s'exposa més detalladament la proposta de restauració més adient per a l’autobús, amb tot un seguit d’imatges que ajuden a entendre els diferents detalls de cada procés que s’ha emprat. Abans de realitzar tots els exhaustius processos que cal dur a terme en una restauració d’un vehicle històric, cal fer un estudi previ del vehicle en general (l’estat actual, número de peces originals, estat del motor, nivell de deteriorament del engranatges/peces...etc). Però abans de començar amb aquest l’estudi previ del nostre vehicle, cal conèixer una mica més a fons la seva història. Per desenvolupar aquest projecte s’ha utilitzat el Bus Biblioteca com a model de vehicle històric, ja que compleix tots els requisits per a ser-ho. Aquest vehicle ha estat proporcionat per l’empresa TMB (Transport Metropolità de Barcelona). L’autobús Biblioteca es troba al pàrquing de TMB Cotxeres Triangle Ferroviari, al polígon industrial de Montsolís. Amb l'ajuda del meu tutor hem pogut gestionar alguna quedada per anar-lo a veure i poder realitzar un estudi d’aquest, perquè en un futur poder escollir els processos més adients per a que la restauració sigui tot un èxit., The main objective of this project is to study one proposal for the restoration of an historic vehicle, specifically the Library Bus, taking into account all the processes and regulations involved in the restoration of this historic vehicle. A possible restoration for the Library Bus will be studied, choosing the best restoration processes based on a set of criteria. The most suitable restoration proposal for the bus will be presented in more detail, accompanied by a series of images that help to understand the different details of each process that has been used. Before carrying out all the exhaustive processes that are necessary in the restoration of an historical vehicle, a preliminary study of the vehicle as a whole (current condition, number of original parts, state of the engine, level of the deterioration of gears/parts etc..) must be conducted. Before starting with this preliminary study of our vehicle, it is necessary to know its history in more detail. To develop this project, the Library Bus has been used as a model of an historical vehicle, because it meets all the requirements to be considered an historical vehicle. This vehicle has been provided by the TMB company (Metropolitan Transport of Barcelona). The Library Bus is located in the TMB Cotxeres Triangle Ferroviari parking lot, in the Montosís industrial estate. With the help of my tutor, we have been able to arrange some meetings to see the vehicle and conduct a study of it, so that in the future we can choose the most suitable processes to ensure that the restoration is a success.