Concentrative nucleoside transporter 1 (hCNT1) promotes phenotypic changes relevant to tumor biology in a translocation independent manner

Nucleoside transporters (NTs) mediate the uptake of nucleosides and nucleobases across the plasma membrane, mostly for salvage purposes. The canonical NTs belong to two gene families, SLC29 and SLC28. The former encode equilibrative nucleoside transporter proteins (ENTs), which mediate the facilitative diffusion of natural nucleosides with broad selectivity, whereas the latter encode concentrative nucleoside transporters (CNTs), which are sodium-coupled and show high affinity for substrates with variable selectivity. These proteins are expressed in most cell types, exhibiting apparent functional redundancy. This might indicate that CNTs play specific roles in the physiology of the cell beyond nucleoside salvage. Here, we addressed this possibility using adenoviral vectors to restore tumor cell expression of hCNT1 or a polymorphic variant (hCNT1S546P) lacking nucleoside translocation ability. We found that hCNT1 restoration in pancreatic cancer cells significantly altered cell-cycle progression and phosphorylation status of key signal-transducing kinases, promoted poly-(ADP ribose) polymerase hyperactivation and cell death, and reduced tumor growth and cell migration. Importantly, the translocation-defective transporter triggered these same effects on cell physiology. These data predict a novel and totally unexpected biological role for the nucleoside transporter protein hCNT1 that appears to be independent of its role as mediator of nucleoside uptake by cells, thereby suggesting a transceptor function. Cell Death & Disease Anastasis Stephanou Receiving Editor Cell Death & Disease 19th Apr 2013 Dr Perez-Torras Av/ Diagonal 643. Edif. Prevosti, Pl -1 Barcelona 08028 Spain RE: Manuscript CDDIS-13-0136R, 'CDDIS-13-0136R' Dear Dr Perez-Torras, It is a pleasure to inform you that your manuscript has been evaluated at the editorial level and has now been officially accepted for publication in Cell Death & Disease, pending you meet the following editorial requirements: 1) the list of the abbreviations is missing please include Could you send us the revised text as word file via e-mail and we will proceed and transfer the paper onto our typesetters. Please download, print, sign, and return the Licence to Publish Form using the link below. This must be returned via FAX to ++ 39 06 7259 6977 before your manuscript can be published:

Aquaporin 3 (AQP3) participates in the cytotoxic response to nucleoside-derived drugs

Background: Nucleoside analogs used in the chemotherapy of solid tumors, such as the capecitabine catabolite50-deoxy-5-fluorouridine (50-DFUR) trigger a transcriptomic response that involves the aquaglyceroporin aquaporin 3 along with other p53-dependent genes. Here, we examined whether up-regulation of aquaporin 3 (AQP3) mRNA incancer cells treated with 50-DFUR represents a collateral transcriptomic effect of the drug, or conversely, AQP3participates in the activity of genotoxic agents. Methods: The role of AQP3 in cell volume increase, cytotoxicity and cell cycle arrest was analyzed using loss-of-function approaches. Results: 50-DFUR and gemcitabine, but not cisplatin, stimulated AQP3 expression and cell volume, which was partially and significantly blocked by knockdown of AQP3. Moreover, AQP3 siRNA significantly blocked other effects of nucleoside analogs, including G1/S cell cycle arrest, p21 and FAS up-regulation, and cell growth inhibition. Short incubations with 5-fluorouracil (5-FU) also induced AQP3 expression and increased cell volume, and the inhibition of AQP3 expression significantly blocked growth inhibition triggered by this drug. To further establish whether AQP3 induction is related to cell cycle arrest and apoptosis, cells were exposed to long incubations with escalating doses of 5-FU. AQP3 was highly up-regulated at doses associated with cell cycle arrest, whereas at doses promoting apoptosis induction of AQP3 mRNA expression was reduced. Conclusions: Based on the results, we propose that the aquaglyceroporin AQP3 is required for cytotoxic activity of 5’-DFUR and gemcitabine in the breast cancer cell line MCF7 and the colon adenocarcinoma cell line HT29, and is implicated in cell volume increase and cell cycle arrest.

Recursos i Restriccions de la Xarxa en Ciències de Biomèdiques

La xarxa Internet, entesa com un consorci format per usuaris, ordinadors, elements de comunicació, coneixements i aplicacions, és una innovació tecnològica que obre la porta a noves modalitats de treball i de relació entre les persones i les organitzacions en què estan agrupades. En el cas de les ciències biomèdiques, una gran part del seu èxit rau a oferir un substrat metodològic molt adient a la necessitat intrínseca de tractar dades i informacions i comunicar-les a d"altres professionals perquè la recerca progressi o a la societat per a difondre els coneixements. La medicina es beneficia en el mateix sentit, però l"impacte és mes gran, ja que hom pot preveure un canvi radical en el paradigma de l"actuació clínica. La clàssica relació metge-pacient haurà d"incorporar nous elements amb el grau de responsabilitat corresponent. Cal acceptar el fet d"Internet amb un esperit científic alhora suficientment crític per a copsar tots els beneficis que se"n poden obtenir i totes les restriccions i limitacions que per raons tècniques o ètiques s"hagin d"acceptar.

Aquaporin 3 (AQP3) participates in the cytotoxic response to nucleoside-derived drugs

Background: Nucleoside analogs used in the chemotherapy of solid tumors, such as the capecitabine catabolite50-deoxy-5-fluorouridine (50-DFUR) trigger a transcriptomic response that involves the aquaglyceroporin aquaporin 3 along with other p53-dependent genes. Here, we examined whether up-regulation of aquaporin 3 (AQP3) mRNA incancer cells treated with 50-DFUR represents a collateral transcriptomic effect of the drug, or conversely, AQP3participates in the activity of genotoxic agents. Methods: The role of AQP3 in cell volume increase, cytotoxicity and cell cycle arrest was analyzed using loss-of-function approaches. Results: 50-DFUR and gemcitabine, but not cisplatin, stimulated AQP3 expression and cell volume, which was partially and significantly blocked by knockdown of AQP3. Moreover, AQP3 siRNA significantly blocked other effects of nucleoside analogs, including G1/S cell cycle arrest, p21 and FAS up-regulation, and cell growth inhibition. Short incubations with 5-fluorouracil (5-FU) also induced AQP3 expression and increased cell volume, and the inhibition of AQP3 expression significantly blocked growth inhibition triggered by this drug. To further establish whether AQP3 induction is related to cell cycle arrest and apoptosis, cells were exposed to long incubations with escalating doses of 5-FU. AQP3 was highly up-regulated at doses associated with cell cycle arrest, whereas at doses promoting apoptosis induction of AQP3 mRNA expression was reduced. Conclusions: Based on the results, we propose that the aquaglyceroporin AQP3 is required for cytotoxic activity of 5’-DFUR and gemcitabine in the breast cancer cell line MCF7 and the colon adenocarcinoma cell line HT29, and is implicated in cell volume increase and cell cycle arrest.

Delitos de posición y delitos con elementos de autoría meramente tipificadores: nuevas bases para una distinción necesaria.

El presente trabajo tiene por objeto dar respuesta a tres de las principales cuestiones concernientes a la teoría general de los delitos especiales. En primer lugar, se trata de conocer en qué consiste esta categoría delictiva. Esto es, abundar en el concepto de delito especial. En segunda instancia, resulta obligado saber cuál es el fundamento material de la restricción del círculo de posibles autores que caracteriza a los delitos especiales. Y, por último, es preciso preguntarse si el extraneus que participa en un delito especial debe responder penalmente, y, en caso afirmativo, si debe hacerlo con la misma pena que el intraneus o bien con una pena atenuada. Todo ello se desarrollará con especial atención al actual art. 65.3 CP, el precepto que nació con la vocación de dar respuesta, al menos en parte, a las cuestiones que han sido formuladas, y que tanta controversia viene generando en la doctrina española prácticamente desde el momento en que entró en vigor.

El abrigo del Filador (Margalef de Montsant, Tarragona) y su contextualizacion cultural y cronologica en el Nordeste Peninsular

En este articulo se pasa revista, de forma resumida, a 20 años de excavaciones en el abrigo del Filador y se dan a conocer los resultados más importantes (sedimentología, fauna, polen, industria, dataciones, etc..), obtenidos con la aplicación de nuevas técnicas de estudio desde 1979. Del mismo modo replanteamos su ubicación cronocultural dentro del marco geográfico del NE peninsular a partir de las dataciones radiocarbónicas conocidas hasta el momento. Se trata del primer trabajo de síntesis sobre el yacimiento realizado con posterioridad al estudio de J. Fortea (Fortea 1973); el Filador sigue siendo un referente obligado del Epipaleolítico de la zona en tanto que muestra la mayoría de las facies cronoculturales que definen esta fase en el NE ibérico.

Nature of the Epidemic Threshold for the Susceptible-Infected-Susceptible Dynamics in Networks

We develop an analytical approach to the susceptible-infected-susceptible epidemic model that allows us to unravel the true origin of the absence of an epidemic threshold in heterogeneous networks. We find that a delicate balance between the number of high degree nodes in the network and the topological distance between them dictates the existence or absence of such a threshold. In particular, small-world random networks with a degree distribution decaying slower than an exponential have a vanishing epidemic threshold in the thermodynamic limit.

El fill de mare diabètica: un experiment de la Natura

Del fill de mare diabètica (FMD) s´ha dit que es tracta d'un autèntic 'experiment de la natura' per tractar-se d'un ésser que al llarg dels seus creixement i desenvolupament intrauterins es relaciona amb un medi ambient diferent al d'una gestació normal. Enfront d'aquesta anomalia, l'embrió primer i el fetus més tard no resten passius, sinó que generen una resposta adaptativa al medi estrany arribant a un altre nivell d'homeostasi però definitivament compensadora de la nova situació. Ara bé, en néixer i 'normalitzar-se' l'ambient, es fa palesa la malaltia...

Cas clínic: esferocitosi hereditaria de diagnóstic tardà

Nena de 10 anys d'edat que procedeix d'un deficient nivell sòcio-cultural, de la qual es desconeixen antecedents paterns. Els materns són anodins. La malaltia actual s'inicia deus dies abans de l'ingrés per un procés intercorrent amb febre i vòmits de tres dies de durada, seguit d'astènia progressiva, mareigs, vòmits i inestabilitat a la marxa. Consulta el nostre servei d'Urgències per agreujament de la simptomatologia i certa desconexió amb l'ambient....

Molecular dynamics simulation of the spherical electrical double layer of a soft nanoparticle. Effect of the surface and counterion valence

Molecular dynamics simulations were performed to study the ion and water distribution around a spherical charged nanoparticle. A soft nanoparticle model was designed using a set of hydrophobic interaction sites distributed in six concentric spherical layers. In order to simulate the effect of charged functionalyzed groups on the nanoparticle surface, a set of charged sites were distributed in the outer layer. Four charged nanoparticle models, from a surface charge value of −0.035 Cm−2 to − 0.28 Cm−2, were studied in NaCl and CaCl2 salt solutions at 1 M and 0.1 M concentrations to evaluate the effect of the surface charge, counterion valence, and concentration of added salt. We obtain that Na + and Ca2 + ions enter inside the soft nanoparticle. Monovalent ions are more accumulated inside the nanoparticle surface, whereas divalent ions are more accumulated just in the plane of the nanoparticle surface sites. The increasing of the the salt concentration has little effect on the internalization of counterions, but significantly reduces the number of water molecules that enter inside the nanoparticle. The manner of distributing the surface charge in the nanoparticle (uniformly over all surface sites or discretely over a limited set of randomly selected sites) considerably affects the distribution of counterions in the proximities of the nanoparticle surface.

Alternatively spliced proline-rich cassettes link WNK1 to aldosterone action.

The thiazide-sensitive NaCl cotransporter (NCC) is important for renal salt handling and blood-pressure homeostasis. The canonical NCC-activating pathway consists of With-No-Lysine (WNK) kinases and their downstream effector kinases SPAK and OSR1, which phosphorylate NCC directly. The upstream mechanisms that connect physiological stimuli to this system remain obscure. Here, we have shown that aldosterone activates SPAK/OSR1 via WNK1. We identified 2 alternatively spliced exons embedded within a proline-rich region of WNK1 that contain PY motifs, which bind the E3 ubiquitin ligase NEDD4-2. PY motif-containing WNK1 isoforms were expressed in human kidney, and these isoforms were efficiently degraded by the ubiquitin proteasome system, an effect reversed by the aldosterone-induced kinase SGK1. In gene-edited cells, WNK1 deficiency negated regulatory effects of NEDD4-2 and SGK1 on NCC, suggesting that WNK1 mediates aldosterone-dependent activity of the WNK/SPAK/OSR1 pathway. Aldosterone infusion increased proline-rich WNK1 isoform abundance in WT mice but did not alter WNK1 abundance in hypertensive Nedd4-2 KO mice, which exhibit high baseline WNK1 and SPAK/OSR1 activity toward NCC. Conversely, hypotensive Sgk1 KO mice exhibited low WNK1 expression and activity. Together, our findings indicate that the proline-rich exons are modular cassettes that convert WNK1 into a NEDD4-2 substrate, thereby linking aldosterone and other NEDD4-2-suppressing antinatriuretic hormones to NCC phosphorylation status.

Missense mutations in TENM4, a regulator of axon guidance and central myelination, cause essential tremor.

Essential tremor (ET) is a common movement disorder with an estimated prevalence of 5% of the population aged over 65 years. In spite of intensive efforts, the genetic architecture of ET remains unknown. We used a combination of whole-exome sequencing and targeted resequencing in three ET families. In vitro and in vivo experiments in oligodendrocyte precursor cells and zebrafish were performed to test our findings. Whole-exome sequencing revealed a missense mutation in TENM4 segregating in an autosomal-dominant fashion in an ET family. Subsequent targeted resequencing of TENM4 led to the discovery of two novel missense mutations. Not only did these two mutations segregate with ET in two additional families, but we also observed significant over transmission of pathogenic TENM4 alleles across the three families. Consistent with a dominant mode of inheritance, in vitro analysis in oligodendrocyte precursor cells showed that mutant proteins mislocalize. Finally, expression of human mRNA harboring any of three patient mutations in zebrafish embryos induced defects in axon guidance, confirming a dominant-negative mode of action for these mutations. Our genetic and functional data, which is corroborated by the existence of a Tenm4 knockout mouse displaying an ET phenotype, implicates TENM4 in ET. Together with previous studies of TENM4 in model organisms, our studies intimate that processes regulating myelination in the central nervous system and axon guidance might be significant contributors to the genetic burden of this disorder.

Foreign-Medical-School Graduates object to the VISA Qualifiying Examination

To the editor; The Visa Qualifying Examination is a two-day test composed of approximately 950 multiple-choice questions conerneing the basic and clinical sciences....

Naturaleza y regimen jurídico del impuesto sobre los sueldos y salarios de los funcionarios de las Coumnidades Europeas

La prevalencia del interés comunitario para salvaguardar la igualdad entre los funcionarios de la Comunidad y la independencia de la Administración comunitaria, movieron al establecimiento de un impuesto comunitario en los Protocolos sobre Privilegios e Inmunidades anejos a los Tratados de Roma, privando a la exención de impuestos nacionales del carácter de privilegio, para configurarla como directa consecuencia de la creación de un impuesto comunitario. La realización del presente estudio nos lleva a plantearnos, en primer lugar, el problema de la naturaleza del recurso objeto del mismo.

Les relacions de suport i el component emocional com a aspectes clau per al desenvolupament del procés resilient i el benestar dels infants al sistema de protecció

La present comunicació mostra el potencial que pot tenir l’establiment de certes relacionsque ofereixen suport i recolzament emocional als infants i joves tutelats, en la promociódel benestar emocional i del procés resilient. La recerca es contextualitza al sistema de proteccióa la infància, centrant-nos en adolescents que a causa de la seva situació de desproteccióhan hagut d’abandonar el seu nucli familiar i entrar al sistema de protecció a la infància.L’afrontament d’aquesta situació afegit a l’entorn advers del qual provenen, comporta que elsadolescents hagin de lidiar amb unes necessitats específiques de l’àrea emocional i relacional.Certs tipus de relacions interpersonals poden ajudar i mediar en aquest impacte emocional,promocionant un desenvolupament del procés resilient, així com millorant el seu benestarpersonal i social. Els resultats de la recerca que es presenta, permeten identificar elements característicsd’aquest tipus de relacions i els beneficis que els adolescents perceben, essent elsaspectes emocionals els que sustenten aquestes relacions...