Ministry Of Science And Innovation Research Articles

Intron retention as a productive mechanism in human MAPT: RNA species generated by retention of intron 3

Tau is a microtubule-binding protein encoded by the MAPT gene. Tau is essential for several physiological functions and associated with pathological processes, including Alzheimer's disease (AD). Six tau isoforms are typically described in the central nervous system, but current research paints a more diverse landscape and a more nuanced balance between isoforms. Recent work has described tau isoforms generated by intron 11 and intron 12 retention. This work adds to that evidence, proving the existence of MAPT transcripts retaining intron 3. Our aim is to demonstrate the existence of mature MAPT RNA species that retain intron 3 in human brain samples and to study its correlation with Alzheimer's disease across different regions. Initial evidence of intron-3-retaining MAPT species come from in silico analysis of RNA-seq databases. We further demonstrate the existence of these mature RNA species in a human neuroepithelioma cell line and human brain samples by quantitative PCR. We also use digital droplet PCR to demonstrate the existence of RNA species that retain either intron 3, intron 12 or both introns. Intron-3-retaining species are even more prominently present that intron-12-retaining ones. We show the presence of MAPT transcripts that retain both introns 3 and 12. These intron-retaining species are diminished in brain samples of patients with Alzheimer's disease with respect to individuals without dementia. Conversely, relative abundance of intron-3- or intron-12-retaining MAPT species with respect to double-retaining species as well as their percentage of expression with respect to total MAPT are increased in patients with Alzheimer's disease, especially in hippocampal samples. Among these TIR-MAPT species, TIR3+12 double truncation allows better classification potential of Alzheimer's disease samples. Moreover, we find a significant increase in intron-3- or intron-12-retaining species and its relative abundance with respect to double-retaining MAPT species in cerebellum in contrast to frontal lateral cortex and hippocampus in individuals with no signs of dementia. Intron retention constitutes a potential mechanism to generate Tau isoforms whose mature RNA expression levels correlate with Alzheimer's pathology showing its potential as a biomarker associated to the disease. This research was funded by the Spanish Ministry of Science, Innovation and Universities: PGC2018-096177-B-I00 (J.A.); Spanish Ministry of Science and Innovation (MCIN): PID2020-113204GB-I00 (F.H.) and PID2021-123859OB-100 from MCIN/AEI/10.13039/501100011033/FEDER, UE (J.A.). It was also supported by CSIC through an intramural grant (201920E104) (J.A.) and the Centre for Networked Biomedical Research on Neurodegenerative Diseases (J.A.). The Centro de Biología Molecular Severo Ochoa (CBMSO) is a Severo Ochoa Center of Excellence (MICIN, award CEX2021-001154-S).

ENHANCING CARTILAGE TISSUE FORMATION IN GELMA/ALGINATE-TYRAMINE INTERPENETRATED NETWORKS (IPNS) WITH LOW INTENSITY PULSE ULTRASOUND STIMULATION (LIPUS)

In the native articular cartilage microenvironment, chondrocytes are constantly subjected to dynamic physical stimuli that maintains tissue homeostasis. They produce extra cellular matrix (ECM) components such as collagens (type II mainly, 50-75%), proteoglycans (10-30%) and other type of proteins1 . While collagen offers a large resistance in tension, proteoglycans are the responsible of the viscoelastic response under compression due to the negative charge they confer to the ECM allowing it to entrap a large amount of interstitial fluid. In pathologic states (e.g. osteoarthritis), this ECM is degenerated and the negative charge becomes unbalanced, losing the chondroprotective properties and resulting on an overloaded chondrocytes that further degenerate the matrix.Low-Intensity Pulsed Ultrasound Stimulation (LIPUS) has been used to generate acoustic (pressure) waves that create bubbles that collapse with cells, inducing a stimulus that can modulate cell response2. This mechanical stimulation promotes the expression of type II collagen, type X collagen, aggrecan and TGF-β, appearing as a great strategy to regenerate cartilage. However, current strategies make use of extrinsic forces to stimulate cartilage formation overlooking the physico-chemical properties of the degenerated cartilage, resulting in an excessive load-transfer to chondrocytes and the consequent hypertrophy and degeneration.Here, interpenetrated networks (IPNs) with different compositions were created using methacrylated gelatin (GelMA), to mimic the collagen, and alginate functionalized with tyramine (Alg-tyr) to mimic glycosaminoglycans and to introduce a negative charge in the model. Within the matrix chondrocytes where encapsulated and stimulated under different conditions to identify the ultrasound parameters that enhance tissue formation. Samples with and without stimulation were compared analysing the expression and deposition of collagen II, aggrecan, collagen X and TGF-β. The results suggested that the chondrogenic marker expression of the samples stimulated for 10 minutes per day for 28 days, was two times higher overall in all of the cases, which was correlated to the tissue formation detected.Acknowledgments: The authors would like to thank the Basque Government for the “Predoctoral Training Program for Non-Doctoral Research Staff 2021-2022” (Grant ref.: PRE_2021_1_0403). This work was supported by the RETOS grant PID2020-114901RA-I00 of the Ministry of Science and Innovation (MICINN).

Diabetes impairs osteogenic differentiation of bone marrow mesenchymal stem cells

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Spanish Ministry of Science and Innovation (MICINN) Diabetes impairs osteogenic differentiation of bone marrow mesenchymal stem cells Background Diabetes is a severe metabolic condition that induces micro- and macrovascular disease, including atherosclerotic vascular disease, cutaneous lesions and bone necrosis. Whether it affects stem cell function is a matter of investigation. When the self-renewing capacity of mesenchymal stem cells (MSC) is compromised, their regenerative potential for cell therapy is impaired. Purpose We aimed to investigate the effects of diabetes on bone marrow- mesenchymal stem cells (BM-MSC) transcriptomics to evidence their potential for differentiation. Methods Bone marrow was extracted from femurs of Zucker rats, and BM-MSC were isolated by culturing adherent cells of extracted bone marrow in DMEM low glucose/ 20% FBS/ 1% P/S under hypoxic conditions (1% oxygen). After one week, MSC were characterized by flow cytometry. Gene expression arrays were used to analyse the expression of BM-MSC of Zucker rats with diabetes (ZD) compared to those of lean-normoglycemic controls (ZC) with the same genetic background (n=6). Results were analysed in silico to identify significantly modified signalling pathways. In addition, BM-MSC differentiation into endothelial, adipogenic and osteogenic cells was investigated in vitro. Results 95-99% of cells isolated from BM expressed CD90, a marker of MSC. Expression data analyses revealed a down-regulation of transcription factors related to the osteogenic pathway (Abcb1a, Adipoq, Fut1, Il1b, Kdr, Nes, Rhoa, Runx2, Tbx5, Tert and Tgfb3), and up-regulation of adipogenic- and angiogenic-related genes such as Bmp7, Casp3, Cd44, Csf2, Erbb2, Gdf7, Il10, Mcam, Nt5e, Pparg, Sox2, Vegfa and Vwf, in BM-MSC from animals with diabetes compared to lean controls. Concomitantly, BM-MSC from rats with diabetes showed impaired osteogenic differentiation in vitro compared to lean controls. No significant differences in adipogenesis or endothelial differentiation were observed between BM-MSC of rats with diabetes and lean controls. Conclusions Diabetes down-regulates several factors involved in the osteogenic differentiation pathway resulting in impaired osteogenesis. The imbalance in gene expression and MSC differentiation may partially explain the impaired regenerative capacity in diabetes.

Effect of Combining Different Photoperiod Regimes on the Sexual Maturation of Male Sea Bass (Dicentrarchus labrax L.) During the First Two Years of Life

The authors wish to thank C. Marin and X. Simo for their technical assistance in rearing and sampling the fish. The work of the authors in this article was supported by UE projects Q5RS-2002-01801 (PUBERTIMING) and Grants AGL2006-04672 (PUBERCONTROL) and CSD2007-00002 (AQUAGENOMICS) from the Spanish Ministry of Science and Innovation (MICINN) and PROMETEO/2010/003 (Generalitat Valenciana). RR was supported by a FPI fellowship from the MICINN (Spain).

Diastereoselective Synthesis of Pyranoquinolines on Zirconium‐Containing UiO‐66 Metal‐Organic Frameworks

The Zr terephthalate MOFs UiO‐66 and UiO‐66‐NH2 have been found to be highly diastereoselective catalysts for the synthesis of a pyrano[3,2‐c]quinoline through an inverse electron‐demand aza‐Diels–Alder [4+2] cycloaddition of an aryl imine (formed in situ from aniline and benzaldehyde) and 3,4‐dihydro‐2H‐pyran in one pot, affording the corresponding trans isomer in diastereomeric excesses of 90–95 %. The solids are stable under the reaction conditions and can be reused at least three times without significant loss of activity or diastereoselectivity.

Does cue exposure treatment improve outcomes in smoking cessation treatment? A controlled trial

Aims: Cue exposure techniques (CET) consist of controlled and repeated exposure to cues that provoke craving in order to extinguish the urge to smoke. Previous studies showed that cigarette craving can be reduced through CET. However, very few studies have explored the efficacy of CET for smoking cessation. This randomized clinical trial explored the effectiveness of a CET procedure when added to a Cognitive Behavioral Treatment (CBT) for smoking cessation among treatment-seeking smokers. Methods: The sample was made up of 102 treatment-seeking smokers with current DSM-IV nicotine dependence. Participants were randomly assigned to oneof two treatment conditions: CBTor CBT+CET. The CBT intervention involved group-based sessions during 6 weeks. In addition to CBT, participants in CBT+CET condition received 5 individual sessions of CET through virtual reality. Results: Of the patients in the CBT+CET group, 78% attended all treatment sessions compared to 73.1% in the CBT group (p = .56). At the end of the treatment, 56% of the patients in the CBT+CET condition achieved abstinence, versus 51.9% in the CBT condition (p = .68). At one month follow-up, 38% in the CBT+CET condition were abstinent compared to 38.5% in the CBT condition (p = .87). Lastly, 24% of patients treatedwith CBT+ CETwere abstinent at sixmonth follow-up compared to 25% who received CBT alone (p = .90). Conclusions: Results showed that neither abstinence levels nor treatment retention rates were better in the CET+CBT compared to CBTalone. TheCETproceduredoesnot appear to improveoutcomes in a smoking cessation program. Results from this randomized clinical trial suggest that CET is not an effective strategy for smoking cessation. Financial support: This work was supported by the Spanish Ministry of Science and Innovation (MICIIN) (Ref. PSI200805938/PSIC) and the predoctoral grant from the Spanish Ministry of Economy and Competitiveness (Ref. BES-2012-053988).

Epoxy resin curing reaction studied by proton multiple-quantum NMR

This study was supported by the Spanish Ministry of Science and Innovation (MICINN) under project MAT2011/23455 and MAT2013/48107-C3. M.M.G. and A.G.J. thank the Consejo Superior de Investigaciones Cientificas (CSIC) for their fellowships JAE-Pre and Jae-Pre-CP, respectively. J.L.V. thanks the Spanish Ministry of Science and Innovation (MICINN) for his Ramon y Cajal contract

Development of magnetic fabric in sedimentary rocks: insights from early compactional structures

The authors acknowledge the La Rioja Government for the permission to develop investigations within the Rioja palaeontological heritage. We also thank Dario Bilardello and an anonymous reviewer for all their constructive suggestions that improved substantially this paper. Besides, we acknowledge the use of Servicio de Preparacion de Rocas y Materiales Duros (Servicio General de Apoyo a la Investigacion-SAI, Universidad de Zaragoza) and Amelia Blasco for the calcimetries performing. This study has been financed by the research project CGL2009-08969 of the MICINN (Spanish Ministry of Science and Innovation). CGL also acknowledges the MICINN for the research grant BES-2010-037509.

Evolutionary paths among different red galaxy types at 0.3 < z < 1.5 and the build-up of massive E-S0's

AbstractSome recent observations seem to disagree with hierarchical theories of galaxy formation on the role of major mergers in a late build-up of massive early-type galaxies. We re-address this question by analysing the morphology, structural distortion level, and star formation enhancement of a sample of massive galaxies (M* > 5 × 1010M⊙) lying on the Red Sequence and its surroundings at 0.3 < z < 1.5. We have used an initial sample of ~1800 sources with Ks < 20.5 mag over an area ~155 arcmin2 on the Groth Strip, combining data from the Rainbow Extragalactic Database and the GOYA Survey. Red galaxy classes that can be directly associated to intermediate stages of major mergers and to their final products have been defined. For the first time we report observationally the existence of a dominant evolutionary path among massive red galaxies at 0.6 < z < 1.5, consisting in the conversion of irregular disks into irregular spheroids, and of these ones into regular spheroids. This result points to: 1) the massive red regular galaxies at low redshifts derive from the irregular ones populating the Red Sequence and its neighbourhood at earlier epochs up to z ~ 1.5; 2) the progenitors of the bulk of present-day massive red regular galaxies have been blue disks that have migrated to the Red Sequence majoritarily through major mergers at 0.6 < z < 1.2 (these mergers thus starting at z ~ 1.5); 3) the formation of E-S0's that end up with M* > 1011M⊙ at z = 0 through gas-rich major mergers has frozen since z ~ 0.6. Our results support that major mergers have played the dominant role in the definitive build-up of present-day E-S0's with M* > 1011M⊙ at 0.6 < z < 1.2, in good agreement with the hierarchical scenario proposed in the Eliche-Moral et al. (2010a) model (see also Eliche-Moral et al. 2010b). This study is published in Prieto et al. (2012).Supported by the Spanish Ministry of Science and Innovation (MICINN) under projects AYA2009-10368, AYA2006-12955, AYA2010-21887-C04-04, and AYA2009-11137, by the Madrid Regional Government through the AstroMadrid Project (CAM S2009/ESP-1496), and by the Spanish MICINN under the Consolider-Ingenio 2010 Program grant CSD2006-00070: “First Science with the GTC” (http://www.iac.es/consolider-ingenio-gtc/). S. D. H. & G.

Vitamin A effects on vertebral bone tissue homeostasis in gilthead sea bream (Sparus aurata) juveniles

This work was funded by Ministry of Science and Innovation (MICIIN) of the Spanish government (projects AGL2005-02478, AGL2008-03897-C04-01 and AGL2008-03897-C04-04).