Role of non-coding RNAs in the transgenerational epigenetic transmission of the effects of reprotoxicants

13 p.-1 fig.Non-coding RNAs (ncRNAs) are regulatory elements of gene expression and chromatin structure. Both long and small ncRNAs can also act as inductors and targets of epigenetic programs. Epigenetic patterns can be transmitted from one cell to the daughter cell, but, importantly, also through generations. Diversity of ncRNAs is emerging with new and surprising roles. Functional interactions among ncRNAs and between specific ncRNAs and structural elements of the chromatin are drawing a complex landscape. In this scenario, epigenetic changes induced by environmental stressors, including reprotoxicants, can explain some transgenerationally-transmitted phenotypes in non-Mendelian ways. In this review, we analyze mechanisms of action of reprotoxicants upon different types of ncRNAs and epigenetic modifications causing transgenerationally transmitted characters through germ cells but affecting germ cells and reproductive systems. A functional model of epigenetic mechanisms of transgenerational transmission ncRNAs-mediated is also proposed.This work was supported by grant from MINECO (BFU2013-42164-R), Spain.We acknowledge support by the CSIC Open Access Publication Initiative through its Unit of Information Resources for Research (URICI).Peer reviewe

Endocrine disrupters, microRNAs, and primordial germ cells: a dangerous cocktail

29 p.-2 fig.Endocrine-disrupting chemicals (EDCs) are environmental pollutants that may change the homeostasis of the endocrine system, altering the differentiation of germ cells with consequences for reproduction. In mammals, germ cell differentiation begins with primordial germ cells (PGCs) during embryogenesis. Primordial germ cell development and gametogenesis are genetically regulated processes, in which the posttranscriptional gene regulation could be mediated by small noncoding RNAs (sncRNAs) such as microRNAs (miRNAs). Here, we review the deleterious effects of exposure during fetal life to EDCs mediated by deregulation of ncRNAs, and specifically miRNAs on PGC differentiation. Moreover, the environmental stress induced by exposure to some EDCs during the embryonic window of development could trigger reproductive dysfunctions transgenerationally transmitted by epigenetic mechanisms with the involvement of miRNAs expressed in germ line cells.Del Mazo lab was supported by a grant from MINECO (BFU2013-42164-R), Spain.Peer reviewe

The expression patterns of genes involved in the RNAi pathways are tissue-dependent and differ in the germ and somatic cells of mouse testis

6 páginas, 3 figuras, 2 tablas -- PAGS nros. 306-311Different RNA interference (RNAi) components participate in post-transcriptional regulation via RNA silencing. The expression pattern of the genes Drosha and Dicer and the members of the Argonaute family Ago1, Ago2, Ago3 and Ago4, all elements participating in the RNAi pathways, were investigated in mouse somatic tissues and testis using quantitative RT-PCR. Expression patterns of different testis cells and those emerging during testis development were also investigated. The differential patterns of expression seen suggest potential pleiotropic roles for certain components of the RNAi machinery. Both spermatocytes and spermatids showed a defined gene expression pattern. The strong expression of Ago4 in germ cells suggests that this protein plays a key role in germ-cell differentiation in the seminiferous epitheliumThis work was supported by grants from the MEC (BFU2004-03977/BFI) and the MSC (FIS PI071007)Peer reviewe

Gene silencing by RNAi in mouse Sertoli cells

<p>Abstract</p> <p>Background</p> <p>RNA interference (RNAi) is a valuable tool in the investigation of gene function. The purpose of this study was to examine the availability, target cell types and efficiency of RNAi in the mouse seminiferous epithelium.</p> <p>Methods</p> <p>The experimental model was based on transgenic mice expressing EGFP (enhanced green fluorescent protein). RNAi was induced by in vivo transfection of plasmid vectors encoding for short hairpin RNAs (shRNAs) targeting EGFP. shRNAs were transfected in vivo by microinjection into the seminiferous tubules via the rete testis followed by square wave electroporation. As a transfection reporter, expression of red fluorescent protein (HcRed 1) was used. Cell types, the efficiency of both transfections and RNAi were all evaluated.</p> <p>Results</p> <p>Sertoli cells were the main transfected cells. A reduction of about 40% in the level of EGFP protein was detected in cells successfully transfected both in vivo and in vitro. However, the efficiency of in vivo transfection was low.</p> <p>Conclusion</p> <p>In adult seminiferous epithelial cells, in vivo post-transcriptional gene silencing mediated by RNAi via shRNA is efficient in Sertoli cells. Similar levels of RNAi were detected both in vivo and in vitro. This also indicates that Sertoli cells have the necessary silencing machinery to repress the expression of endogenous genes via RNAi.</p

Global characterization and target identification of piRNAs and endo-siRNAs in mouse gametes and zygotes

44 p.-1 tab.-10 fig.A set of small RNAs known as rasRNAs (repeat-associated small RNAs) have been related to the down-regulation of Transposable Elements (TEs) to safeguard genome integrity. Two key members of the rasRNAs group are piRNAs and endo-siRNAs. We have performed a comparative analysis of piRNAs and endo-siRNAs present in mouse oocytes, spermatozoa and zygotes, identified by deep sequencing and bioinformatic analysis. The detection of piRNAs and endo-siRNAs in the spermatozoa and revealed also in zygotes, hints to their potential delivery to oocytes during fertilization. However, a comparative assessment of the three cell types indicates that both piRNAs and endo-siRNAs are mainly maternally inherited. Finally, we have assessed the role of the different rasRNA molecules in connection with amplification processes by way of the "ping-pong cycle". Our results suggest that the ping-pong cycle can act on other rasRNAs, such as tRNA- and rRNA-derived fragments, thus not only being restricted to TEs during gametogenesis. © 2014 Elsevier B.V.This work was funded by grants from The European Chemical Industry Council Long-range Research Initiative (CEFIC-LRi), from the MEDDTL (11-MRESPNRPE-9-CVS-072), France, from the CSIC (PIE 201020E016), Spain.Peer reviewe

Especificidad de los alteradores endocrinos en la expresión génica durante el desarrollo

3 p.Los estudios de genes y los efectos de los alteradores endocrinos sobre la expresión génica llevados a cabo en el laboratorio de J. del Mazo han sido llevados a cabo en proyectos financiados parcialmente por: EC (QLK4-CT-2002-02403), CEFIC-LRI; MEDDTL (11-MRES-PNRPE-9-CVS -072) Francia, y CSIC (PIE 201020E016), España.Peer reviewe

Experimenta. 60 experimentos con materiales sencillos

ReviewRese&ntilde;

Numerical simulation of the micro-extrusion process of printable biomaterials

This work aims to gain a better understanding of how the rheological properties of printable materials affect their processability, as well as the quality of the final product, which at the end can lead to reducing time and costs of the process and increase product development. As the first step, the proper rheological non-Newtonian models are extracted from experimental studies. Later, three-dimensional numerical simulation of extrusion process is performed in the context of Direct Numerical Simulation (DNS) of governing equations, where the whole physics of fluid motion is taken into account. A finite-volume fractional step approach is used to solve the Navier-Stocks equations on collocated arbitrary meshes. Geometrical volume-of-fluid (GVOF) interface capturing approach is used to resolve the topological changes of the moving interface. The governing equations are solved using High-Performance Computing (HPC) parallel approaches. Besides the contribution of this work to the advancement of numerical techniques applied to multiphase complex flows, obtained results will shed light on the nature of non-Newtonian extrusion process with vast applications in the 3D printer industrial sectors.This work was developed in the context of a research project (BASE3D 001-P-001646) co-financed by the European Union Regional Development Fund within the framework of the ERDF Operational Program of Catalonia 2014-2020 with a grant of 50% of total cost eligible.Postprint (published version

Advanced monitoring of rail breakage in double-track railway lines by means of PCA techniques

This work describes a classifier designed to identify rail breakages in double-track railway lines, completing the electronic equipment carried out by authors. The main objective of this proposal is to guarantee the integrity of tracks before the railway traffic starts working. In addition, it facilitates maintenance tasks providing information about possible breakages. The detection of breakages is based on the analysis of eight currents provided by the electronic equipment, one per rail, at the ends of the section (emitting and receiving nodes). The imbalance that occurs among the value of these currents implies that there is at least a breakage in the track section under analysis. This analysis is conducted according to three phases. The first one identifies whether there is a breakage, and, in that case, the damaged track is identified. The second phase provides information about which rail is broken (internal, external or both of them) in the previously identified track. Finally, if there is only one breakage, the third phase estimates its most likely zone along the track section. This situation is considered as a classification problem, and solved by means of the Principal Component Analysis technique. This means that a significant number of measurements is required for every breakage pattern (types of breakages) to be considered. Due to the difficulty of having real data, the proposal has been validated using an 8km-long double-track hardware simulator specially designed by the authors, with specific localizations for breakages

Los cathedraticos de mathematicas de la Universidad de Salamanca, el jubilado y actual D. Diego de Torres y D. Isidoro Ortiz impugnados en carta respuesta a un amigo suyo por Fr. Miguel de Jesus Maria y Hualde, religioso de la vida activa del Orden de Nra. Sra. del Carmen sobre el mas exacto concierto de la cuenta con que deben intercalarse los bisiestos y sobre la averiguacion del dia y año cierto de la Pasion y Muerte de nuestro redentor Jesu-Christo

Copia digital. Valladolid : Junta de Castilla y León. Consejería de Cultura y Turismo, 2011En pag.2 consta 1766Sign.: []2, A-K