Anàlisi genètico-molecular d'una nova forma de distròfia muscular de maluc autosòmica dominant en un extens pedigrí

Consultable des del TDXTítol obtingut de la portada digitalitzadaS'ha abordat l'estudi genètico-molecular d'una extensa família espanyola afectada per una nova forma de distròfia muscular de maluc (Limb-girdle muscle dystrophy, LGMD) amb herència autosòmica dominant. Un cop definida la clínica de forma detallada s'han comparat les seves característiques amb les de les altres 5 formes autosòmiques dominants prèviament conegudes. Si bé ja alguns dels trets clínics eren diferencials, mitjançant l'anàlisi de lligament a 3 punts (2 marcadors-malaltia) de marcadors polimòrfics lligats a aquestes formes (regions 1q11-q21, 3p25, 5q31, 6q23 i 7q), s'han pogut excloure com a responsables de la malaltia en la família estudiada. Un cop establert que ens trobàvem davant d'una nova forma genèticament diferenciada, s'ha procedit a fer una anàlisi de lligament completa a 2 punts (marcador-malaltia), tot analitzant un ampli grup de marcadors polimòrfics dispersos pels 22 cromosomes autosòmics, comprovant manual i estadísticament la seva segregació respecte el fenotip clínic. Inicialment s'han utilitzat microsatèl·lits marcats amb fluorescència i pel fine mapping s'han utilitzat microsatèl·lits adicionals marcats radiactivament, havent-se inclós en aquesta darrera part fins a 55 d'individus del pedigrí, 27 afectes i 28 sans. Com a resultat, s'ha pogut trobar el locus de la malaltia al braç llarg del cromosoma 7, concretament a la regió 7q31-q32. Mitjançant l'estudi de marcadors adicionals i considerant la valuosa informació del 4 individus recombinants trobats en diferents parts del pedigrí, s'ha pogut establir una regió candidata d'unes 3,7 megabases, entre els marcadors D7S680 i D7S2544, dins els mapes del consorci humà i en la qual s'han identificat una sèrie de gens candidats i de expressed sequence tags (EST). El continu avenç en el projecte Genoma Humà ha suposat un gran ajut a l'hora de situar i escollir marcadors i gens dins el cromosoma 7. S'ha estudiat a fons un dels gens candidats, la Filamina C, proteïna que lliga actina. S'ha pogut excloure la Filamina C com a gen responsable de la nostra nova forma de LGMD després de no trobar-se cap mutació present en tots els individus afectes en co-segregació amb la malaltia ni cap alteració en el nivell d'expressió a nivell de mARN (àcid ribonucleic missatger). Podem concloure que ens trobem davant una nova forma d'LGMD autosòmica dominant genèticament diferenciada que caldria anomenar LGMD1F, segons la nomenclatura de l'OMIM (On-line Mendelian Inheritance in Man database).It has been performed the genetic and molecular analysis of a large Spanish family affected by a new limb-girdle muscle dystrophy form (LGMD) with an autosomal dominant inheritance trait. Once described the clinical features of the disease we compared them with the 5 other autosomal dominant LGMD previously described. Aditionally to some clinical specific and differential traits, 3-point linkage analysis (2 markers and disease) with linked markers to this forms (regions 1q11-q21, 3p25, 5q31, 6q23 and 7q) excluded them as responsible for the disease in our pedigree. Once stablished that we had a new genetically distinguished form, we performed a genomic wide 2-point linkage analysis (marker-disease), by the use of many polymorphic markers spread all over the 22 autosomal chromosomes, checking manually and statistically its segregation with respect to the clinical phenotype. We first performed the analysis with fluorescently labeled markers and then, for the fine mapping, we used aditional 32P labeled markers, including in this part up to 55 members of the pedigree, being 27 of them affected and 28 unaffected. As a result, we could find the locus of the disease in the long arm of chromosome 7, specifically to region 7q31-q32. By the use of aditional markers and analysing the 4 informative recombinant individuals found in different parts of the pedigree it was possible to stablish a candidate region that spans 3.7 megabases, between markers D7S680 and D7S2544, following the available human physical and genetic maps. We have identified some candidate genes and expressed sequence tags (EST) in this interval. The continuous improvement in the Human Genome Project has become a great help at the time of placing and choosing markers and genes in Chromosome 7. One of the candidate genes, Filamin C, an actin-binding protein, has been extensively studied. We were able to exclude Filamin C as the gene responsible for the disease in our LGMD pedigree after the analysis of the whole coding sequence and not finding any pathological mutation in all affected individuals co-segregating with the disease and after checking the expression levels at the mRNA level in affected individuals in comparison with unaffected individuals, not being significatively different. We can conclude that this is a new autosomal dominant LGMD form, genetically distinguished that we could name LGMD1F, following the OMIM nomenclature (On-line Mendelian Inheritance in Man Database)

pTINCR microprotein promotes epithelial differentiation and suppresses tumor growth through CDC42 SUMOylation and activation

Cancer; Mechanisms of diseaseCàncer; Mecanismes de la malaltiaCáncer; Mecanismos de la enfermedadThe human transcriptome contains thousands of small open reading frames (sORFs) that encode microproteins whose functions remain largely unexplored. Here, we show that TINCR lncRNA encodes pTINCR, an evolutionary conserved ubiquitin-like protein (UBL) expressed in many epithelia and upregulated upon differentiation and under cellular stress. By gain- and loss-of-function studies, we demonstrate that pTINCR is a key inducer of epithelial differentiation in vitro and in vivo. Interestingly, low expression of TINCR associates with worse prognosis in several epithelial cancers, and pTINCR overexpression reduces malignancy in patient-derived xenografts. At the molecular level, pTINCR binds to SUMO through its SUMO interacting motif (SIM) and to CDC42, a Rho-GTPase critical for actin cytoskeleton remodeling and epithelial differentiation. Moreover, pTINCR increases CDC42 SUMOylation and promotes its activation, triggering a pro-differentiation cascade. Our findings suggest that the microproteome is a source of new regulators of cell identity relevant for cancer.Work in the Abad lab is supported by VHIO, Fero Foundation, La Caixa Foundation, Asociación Española Contra el Cancer (AECC), La Mutua Foundation and by grants from the Spanish Ministry of Science and Innovation (SAF2015-69413-R; RTI2018-102046-B-I00). M.A. was recipient of a Ramón y Cajal contract from the Spanish Ministry of Science and Innovation (RYC-2013-14747). O.B. is recipient of a FPI-AGAUR fellowship from Generalitat de Catalunya. We also acknowledge funding from grant PGC2018-094091-B-I00 from the Spanish Government

pTINCR microprotein promotes epithelial differentiation and suppresses tumor growth through CDC42 SUMOylation and activation

The human transcriptome contains thousands of small open reading frames (sORFs) that encode microproteins whose functions remain largely unexplored. Here, we show that TINCR lncRNA encodes pTINCR, an evolutionary conserved ubiquitin-like protein (UBL) expressed in many epithelia and upregulated upon differentiation and under cellular stress. By gain- and loss-of-function studies, we demonstrate that pTINCR is a key inducer of epithelial differentiation in vitro and in vivo. Interestingly, low expression of TINCR associates with worse prognosis in several epithelial cancers, and pTINCR overexpression reduces malignancy in patient-derived xenografts. At the molecular level, pTINCR binds to SUMO through its SUMO interacting motif (SIM) and to CDC42, a Rho-GTPase critical for actin cytoskeleton remodeling and epithelial differentiation. Moreover, pTINCR increases CDC42 SUMOylation and promotes its activation, triggering a pro-differentiation cascade. Our findings suggest that the microproteome is a source of new regulators of cell identity relevant for cancer.Acknowledgements: The authors thank VHIO Proteomics, Molecular Oncology and Genomics Core Facilities for technical assistance. We are grateful to Manuel Serrano for providing several reagents, advice and critical discussion on the manuscript. We also thank Alonso García and Raquel Pérez for their help in processing and analyzing digital images, Gemma Serra and Sandra Peiró for their assistance with subcellular fractionation and immunoprecipitation experiments, Sara Arce and Joaquín Mateo for providing several reagents during the development of critical experiments of this manuscript, and Juan Angel Recio for his help with the cSCC cohort. We are immensely grateful to all the members of the Abad lab for generating the know-how for the identification of novel sORFs, for the critical reading on the manuscript and in general for their constant support to this project. Work in the Abad lab is supported by VHIO, Fero Foundation, La Caixa Foundation, Asociación Española Contra el Cancer (AECC), La Mutua Foundation and by grants from the Spanish Ministry of Science and Innovation (SAF2015-69413-R; RTI2018-102046-B-I00). M.A. was recipient of a Ramón y Cajal contract from the Spanish Ministry of Science and Innovation (RYC-2013-14747). O.B. is recipient of a FPIAGAUR fellowship from Generalitat de Catalunya. We also acknowledge funding from grant PGC2018-094091-B-I00 from the Spanish Government

pTINCR microprotein promotes epithelial differentiation and suppresses tumor growth through CDC42 SUMOylation and activation

The human transcriptome contains thousands of small open reading frames (sORFs) that encode microproteins whose functions remain largely unexplored. Here, we show that TINCR lncRNA encodes pTINCR, an evolutionary conserved ubiquitin-like protein (UBL) expressed in many epithelia and upregulated upon differentiation and under cellular stress. By gain- and loss-of-function studies, we demonstrate that pTINCR is a key inducer of epithelial differentiation in vitro and in vivo. Interestingly, low expression of TINCR associates with worse prognosis in several epithelial cancers, and pTINCR overexpression reduces malignancy in patient-derived xenografts. At the molecular level, pTINCR binds to SUMO through its SUMO interacting motif (SIM) and to CDC42, a Rho-GTPase critical for actin cytoskeleton remodeling and epithelial differentiation. Moreover, pTINCR increases CDC42 SUMOylation and promotes its activation, triggering a pro-differentiation cascade. Our findings suggest that the microproteome is a source of new regulators of cell identity relevant for cancer

Differential requirements for Tousled-like kinases 1 and 2 in mammalian development

The regulation of chromatin structure is critical for a wide range of essential cellular processes. The Tousled-like kinases, TLK1 and TLK2, regulate ASF1, a histone H3/H4 chaperone, and likely other substrates, and their activity has been implicated in transcription, DNA replication, DNA repair, RNA interference, cell cycle progression, viral latency, chromosome segregation and mitosis. However, little is known about the functions of TLK activity in vivo or the relative functions of the highly similar TLK1 and TLK2 in any cell type. To begin to address this, we have generated Tlk1- and Tlk2-deficient mice. We found that while TLK1 was dispensable for murine viability, TLK2 loss led to late embryonic lethality because of placental failure. TLK2 was required for normal trophoblast differentiation and the phosphorylation of ASF1 was reduced in placentas lacking TLK2. Conditional bypass of the placental phenotype allowed the generation of apparently healthy Tlk2-deficient mice, while only the depletion of both TLK1 and TLK2 led to extensive genomic instability, indicating that both activities contribute to genome maintenance. Our data identifies a specific role for TLK2 in placental function during mammalian development and suggests that TLK1 and TLK2 have largely redundant roles in genome maintenance

Els efectes dels descansos actius sobre l’atenció escolar a quart de primària

[cat] Els descansos actius és una metodologia basada en la realització d‟activitat física a dins l‟aula ordinària en un període de 10 minuts amb l‟objectiu d‟interrompre el temps sedentari, aquesta metodologia està integrada a dins l‟aula i ajuda a repassar o aprendre continguts del currículum en qualsevol àrea curricular. També pretenc fer front a l‟obesitat, el sobrepès i intentar millorar l‟activitat física dels nostres alumnes. Els descansos actius han demostrat millores a nivell físic com a nivell cognitiu i el duré a la pràctica a dins una aula de quart de Primària ja que és un curs que li fa falta treballar l‟atenció selectiva. El meu objectiu del Treball Final de Grau analitzar l‟impacte sobre l‟atenció selectiva a través del projecte de descansos actius, on he pogut observar un lleu milloria en els resultats. La metodologia emprada del Treball Final de Grau és activa i dinàmica. I els resultats, amb el poc temps que he duit a pràctica el projecte, s‟observen resultats esperançadors. Com a conclusió, els descansos actius és una bona metodologia per treballar l‟atenció selectiva.[eng] Active breaks is a methodology based on totally physical response activities in the classroom, where we take 10 minutes physical activity breaks to get children moving without sacrificing time dedicated to academic learning at any curricular area. Also, I mean to deal with obesity and overweight as well as improving physical activity among our students. Through active breaks, students‟ fitness and knowledges have improved. Therefore, I will take it into practice in the fourth level class as they are a group which needs to work on selective attention. The main goal of my Degree Final Project is to analyze the active breaks‟ impact on selective attention. I have noticed some improvement. This methodology is active and dynamic. The results have, so far, been positive and hopeful taking in mind that I have had few time to put the methodology into practice. In conclusion, active breaks are a good methodology to work with selective attention in the classroom

Synthesis of a new stable and water-soluble tris(4-hydroxysulfonyltetrachlorophenyl)methyl radical with selective oxidative capacity

A new stable organic free radical of the PTM (perchlorotriphenylmethyl) series very soluble in water is reported. This free radical is sensitive to electron transfer processes, and the selectivity of these reactions in the presence of ascorbic acid, pyrogallol, and catechol as reducing species is described. The electron paramagnetic resonance spectrum and the electrochemical behavior are also presented.Financial support for this research from the MCI (Spain) through project AGL2009-12374-C03-03/ALI is gratefully acknowledged. J.A.M. gratefully acknowledges the Spanish Foreign Office Department for a predoctoral grant. We also thank the EPR service of the Advanced Research Institute of Catalunya (CSIC) (Spain) for recording the EPR spectra and the scientific-technical services of the University of Barcelona for recording the mass spectra.Peer reviewe

Preparation and characterization of persistent maltose-conjugated triphenylmethyl radicals

The condensation reaction of d-maltose to free radicals of the series of tris(2,4,6-trichlorophenyl)methyl (TTM) and tris(perchlorophenyl)methyl (PTM) has been described for the first time. The new persistent radicals 1 and 2 are very stable and have been characterized by EPR. Their cyclic voltammograms show a quasi-reversible process in the cathode, being reduced to the corresponding anions, with redox potentials a little lower than those of TTM and PTM, respectively. Their oxidant activity is in close relation with their reduction potentials. Therefore, while 2 is reduced by ascorbic acid, 1 remains unaltered.Financial support for this research from the MCI (Spain) through projects AGL2009-12374-C03-03/ALI and CTQ2009-13797 is gratefully acknowledged. J.A.M. gratefully acknowledges the Spanish Foreign Office Department for a predoctoral grant (AECI).Peer Reviewe

Punicalagin and catechins contain polyphenolic substructures that influence cell viability and can be monitored by radical chemosensors sensitive to electron transfer

Plant polyphenols may be free radical scavengers or generators, depending on their nature and concentration. This dual effect, mediated by electron transfer reactions, may contribute to their influence on cell viability. This study used two stable radicals (tris(2,3,5,6-tetrachloro-4-nitrophenyl)methyl (TNPTM) and tris(2,4,6-trichloro-3,5-dinitrophenyl)methyl (HNTTM)) sensitive only to electron transfer reduction reactions to monitor the redox properties of polyphenols (punicalagin and catechins) that contain phenolic hydroxyls with different reducing capacities. The use of the two radicals reveals that punicalagin's substructures consisting of gallate esters linked together by carbon-carbon (C-C) bonds are more reactive than simple gallates and less reactive than the pyrogallol moiety of green tea catechins. The most reactive hydroxyls, detected by TNPTM, are present in the compounds that affect HT-29 cell viability the most. TNPTM reacts with C-C-linked gallates and pyrogallol and provides a convenient way to detect potentially beneficial polyphenols from natural sources. © 2012 American Chemical Society.This work was supported by the Spanish Ministry of Education and Science (research grants AGL2006-12210-C03-02/ALI; SAF2008-00164; AGL2009-12374-C03-03/ALI), the Instituto de Salud Carlos III and European Regional Development Fund (ISCIII-RTICC, RD06/0020/0046), the Generalitat de Catalunya (2009SGR1308, 2009CTP 00026, and Icrea Academia award 2010 granted to M.C.), and the European Commission (Etherpaths Project KBBE-Grant Agreement 222639).Peer Reviewe