Differential activity of GSK-3 isoforms regulates NF-kappa B and TRAIL- or TNF alpha induced apoptosis in pancreatic cancer cells

While TRAIL is a promising anticancer agent due to its ability to selectively induce apoptosis in neoplastic cells, many tumors, including pancreatic ductal adenocarcinoma (PDA), display intrinsic resistance, highlighting the need for TRAIL-sensitizing agents. Here we report that TRAIL-induced apoptosis in PDA cell lines is enhanced by pharmacological inhibition of glycogen synthase kinase-3 (GSK-3) or by shRNA-mediated depletion of either GSK-3 alpha or GSK-3 beta. In contrast, depletion of GSK-3 beta, but not GSK-3 alpha, sensitized PDA cell lines to TNF alpha-induced cell death. Further experiments demonstrated that TNF alpha-stimulated I kappa B alpha phosphorylation and degradation as well as p65 nuclear translocation were normal in GSK-3 beta-deficient MEFs. Nonetheless, inhibition of GSK-3 beta function in MEFs or PDA cell lines impaired the expression of the NF-kappa B target genes Bcl-xL and cIAP2, but not I kappa B alpha. Significantly, the expression of Bcl-xL and cIAP2 could be reestablished by expression of GSK-3 beta targeted to the nucleus but not GSK-3 beta targeted to the cytoplasm, suggesting that GSK-3 beta regulates NF-kappa B function within the nucleus. Consistent with this notion, chromatin immunoprecipitation demonstrated that GSK-3 inhibition resulted in either decreased p65 binding to the promoter of BIR3, which encodes cIAP2, or increased p50 binding as well as recruitment of SIRT1 and HDAC3 to the promoter of BCL2L1, which encodes Bcl-xL. Importantly, depletion of Bcl-xL but not cIAP2, mimicked the sensitizing effect of GSK-3 inhibition on TRAIL-induced apoptosis, whereas Bcl-xL overexpression ameliorated the sensitization by GSK-3 inhibition. These results not only suggest that GSK-3 beta overexpression and nuclear localization contribute to TNF alpha and TRAIL resistance via anti-apoptotic NF-kappa B genes such as Bcl-xL, but also provide a rationale for further exploration of GSK-3 inhibitors combined with TRAIL for the treatment of PDA.

A Non-Parametric Dimension Test of the Term Structure

Published as an article in: Studies in Nonlinear Dynamics & Econometrics, 2004, vol. 8, issue 3, article 6.

Long memory in return structures from developed markets

[En]The present study aimed at investigating the existence of long memory properties in ten developed stock markets across the globe. When return series exhibit long memory, the series realizations are not independent over time and past returns can help predict future returns, thus violating the market efficiency hypothesis. It poses a serious challenge to the supporters of random walk behavior of the stock returns indicating a potentially predictable component in the series dynamics. We computed Hurst-Mandelbrot’s Classical R/S statistic, Lo’s statistic and semi parametric GPH statistic using spectral regression. The findings suggest existence of long memory in volatility and random walk for logarithmic return series in general for all the selected stock market indices. Findings are in line with the stylized facts of financial time series.

Respuesta a la sequía de Pinus radiata D. Don y su implicación en los procesos de tolerancia

160 p. (Bibliogr. 141-160)

Analysis of SUMOylated proteins using SUMO-traps

6 p. [+ 7 p. Supplementary Information]

Control of gene expression by modulated self-assembly

Numerous transcription factors self-assemble into different order oligomeric species in a way that is actively regulated by the cell. Until now, no general functional role has been identified for this widespread process. Here, we capture the effects of modulated self-assembly in gene expression with a novel quantitative framework. We show that this mechanism provides precision and flexibility, two seemingly antagonistic properties, to the sensing of diverse cellular signals by systems that share common elements present in transcription factors like p53, NF-kappa B, STATs, Oct and RXR. Applied to the nuclear hormone receptor RXR, this framework accurately reproduces a broad range of classical, previously unexplained, sets of gene expression data and corroborates the existence of a precise functional regime with flexible properties that can be controlled both at a genome-wide scale and at the individual promoter level.

Análisis histológico e inmunohistoquímico de los subtipos de enfermedad liquenoide oral

176 p. : il.

Bihotz-biriketako berpizte masajeak EKG-an eta bular-inpedantzian eragindako interferentziaren azterketa

[EU]Proiektu honetan bihotz-biriketako berpizte masajearen bular-sakadek elektrokardiograman eta bular-inpedantziaren seinaleetan eragindako interferentziaren azterketa egiten da. Helburu nagusia bi interferentzia hauen arteko erlazioa aztertzea da, horretarako tresna garatuz. Erlazio hau definitzeak interferentziaren eragina txikitzeko modua aurkitzen lagunduko luke, eta honek berpizteko aukerak handituko lituzke. Proiektua gauzatzeko ospitalez kanpoko geldialdien erregistro multzo batetik abiatuta datu-base propioa garatu da ezarritako irizpide batzuk jarraituz. Datu-base berri hau 37 pazienteren 237 mozketak osatzen dute, 10 segundotako luzera minimoarekin non pazienteek asistolia bitarteko kanpoko bular masajea jasotzen duten. Bestalde, interferentzia ezaugarritzeko interfaze grafiko bat garatu da, elektrokardiograma eta bular-inpedantziaren seinaleak denboran eta maiztasunean erakutsi eta hauen parametro esanguratsuak automatikoki zein eskuz ateratzeko aukera ematen duena. Parametroak seinaleen sakada bakoitzeko maximo eta minimoak, beraien kokapenak eta oinarrizko maiztasuna, bere harmonikoak eta hauen anplitudeak dira. Tresna hau erabiliz aipatutako datu-baseko episodioen prozesaketa egin da. Bukatzeko, lortutako emaitzak tratatzeko bigarren interfaze grafiko bat garatu da, non emaitzen banaketa estatistikoa eta hauen arteko erlazio lineala aztertzen diren. Proiektuaren ekarpen nagusia, beraz, bihotz-biriketako berpizte masajeak eragindako interferentzia aztertzeko tresna ahaltsuaren garapena da, jatorri desberdineko bestelako berpizte episodioak aztertzeko ere balio duena.

Diseño e implementación de una herramienta para la integración de datos de QoS en Internet publicados por el regulador español

[ES]Este proyecto tiene como objetivo el diseño e implementación de una herramienta para la integración de los datos de calidad de servicio (QoS) en Internet publicados por el regulador español. Se trata de una herramienta que pretende, por una parte, unificar los diferentes formatos en que se publican los datos de QoS y, por otra, facilitar la conservación de los datos favoreciendo la obtención de históricos, datos estadísticos e informes. En la página del regulador sólo se puede acceder a los datos de los 5 últimos trimestres y los datos anteriormente publicados no permanecen accesibles si no que son sustituidos por los más recientes por lo que, desde el punto de vista del usuario final, estos datos se pierden. La herramienta propuesta en este trabajo soluciona este problema además de unificar formatos y facilitar el acceso a los datos de interés. Para el diseño del sistema se han usado las últimas tecnologías en desarrollo de aplicaciones web con lo que la potencia y posibilidad de futuras ampliaciones son elevadas.

The promoter of cell growth- and RNA protection-associated SND1 gene is activated by endoplasmic reticulum stress in human hepatoma cells

Background: Staphyloccocal nuclease domain-containing protein 1 (SND1) is involved in the regulation of gene expression and RNA protection. While numerous studies have established that SND1 protein expression is modulated by cellular stresses associated with tumor growth, hypoxia, inflammation, heat- shock and oxidative conditions, little is known about the factors responsible for SND1 expression. Here, we have approached this question by analyzing the transcriptional response of human SND1 gene to pharmacological endoplasmic reticulum (ER) stress in liver cancer cells. Results: We provide first evidence that SND1 promoter activity is increased in human liver cancer cells upon exposure to thapsigargin or tunicamycin or by ectopic expression of ATF6, a crucial transcription factor in the unfolded protein response triggered by ER stress. Deletion analysis of the 5'-flanking region of SND1 promoter identified maximal activation in fragment (-934, +221), which contains most of the predicted ER stress response elements in proximal promoter. Quantitative real- time PCR revealed a near 3 fold increase in SND1 mRNA expression by either of the stress- inducers; whereas SND1 protein was maximally upregulated (3.4-fold) in cells exposed to tunicamycin, a protein glycosylation inhibitor. Conclusion: Promoter activity of the cell growth- and RNA-protection associated SND1 gene is up-regulated by ER stress in human hepatoma cells.

Alveolar nitric oxide and its role in pediatric asthma control assessment

Resumen Background: Nitric oxide can be measured at multiple flow rates to determine proximal (maximum airway nitric oxide flux; Jaw(NO)) and distal inflammation (alveolar nitric oxide concentration; CA(NO)). The main aim was to study the association among symptoms, lung function, proximal (maximum airway nitric oxide flux) and distal (alveolar nitric oxide concentration) airway inflammation in asthmatic children treated and not treated with inhaled glucocorticoids. Methods: A cross-sectional study with prospective data collection was carried out in a consecutive sample of girls and boys aged between 6 and 16 years with a medical diagnosis of asthma. Maximum airway nitric oxide flux and alveolar nitric oxide concentration were calculated according to the two-compartment model. In asthmatic patients, the asthma control questionnaire (CAN) was completed and forced spirometry was performed. In controls, differences between the sexes in alveolar nitric oxide concentration and maximum airway nitric oxide flux and their correlation with height were studied. The correlation among the fraction of exhaled NO at 50 ml/s (FENO50), CA(NO), Jaw(NO), forced expiratory volume in 1 second (FEV1) and the CAN questionnaire was measured and the degree of agreement regarding asthma control assessment was studied using Cohen's kappa. Results: We studied 162 children; 49 healthy (group 1), 23 asthmatic participants without treatment (group 2) and 80 asthmatic patients treated with inhaled corticosteroids (group 3). CA(NO) (ppb) was 2.2 (0.1-4.5), 3 (0.2-9.2) and 2.45 (0.1-24), respectively. Jaw(NO) (pl/s) was 516 (98.3-1470), 2356.67 (120-6110) and 1426 (156-11805), respectively. There was a strong association (r = 0.97) between FENO50 and Jaw(NO) and the degree of agreement was very good in group 2 and was good in group 3. There was no agreement or only slight agreement between the measures used to monitor asthma control (FEV1, CAN questionnaire, CA(NO) and Jaw(NO)). Conclusions: The results for CA(NO) and Jaw(NO) in controls were similar to those found in other reports. There was no agreement or only slight agreement among the three measure instruments analyzed to assess asthma control. In our sample, no additional information was provided by CA(NO) and Jaw(NO).

Cytokine pathway disruption in a mouse model of schizophrenia induced by Munc18-1a overexpression in the brain

Background: An accumulating body of evidence points to the significance of neuroinflammation and immunogenetics in schizophrenia, and an imbalance of cytokines in the central nervous system (CNS) has been suggested to be associated with the disorder. Munc18-overexpressing mice (Munc18-OE) have provided a model for the study of the alterations that may underlie the symptoms of subjects with schizophrenia. The aim of the present study was to elucidate the involvement of neuroinflammation and cytokine imbalance in this model. Methods: Cytokines were evaluated in the cortex and the striatum of Munc18-OE and wild-type (WT) mice by enzyme-linked immunosorbent assay (ELISA). Protein levels of specific microglia and macrophage, astrocytic and neuroinflammation markers were quantified by western blot in the cortex and the striatum of Munc18-OE and WT mice. Results: Each cytokine evaluated (Interferon-gamma (IFN-gamma), Tumor Necrosis Factor-alpha (TNF-alpha), Interleukin-2 (IL-2) and CCL2 chemokine) was present at higher levels in the striatum of Munc18-OE mice than WT. Cortical TNF-alpha and IL-2 levels were significantly lower in Munc18-OE mice than WT mice. The microglia and macrophage marker CD11b was lower in the cortexes of Munc18-OE mice than WT, but no differences were observed in the striatum. Glial Fibrillary Acidic Protein (GFAP) and Nuclear Factor-kappaB (NF-kappa B)p65 levels were not different between the groups. Interleukin-1beta (IL-1 beta) and IL-6 levels were beneath detection limits. Conclusions: The disrupted levels of cytokines detected in the brain of Munc18-OE mice was found to be similar to clinical reports and endorses study of this type for analysis of this aspect of the disorder. The lower CD11b expression in the cortex but not in the striatum of the Munc18-OE mice may reflect differences in physiological activity. The cytokine expression pattern observed in Munc18-OE mice is similar to a previously published model of schizophrenia caused by maternal immune activation. Together, these data suggest a possible role for an immune imbalance in this disorder.

Korner errutinen azterketa goi errendimenduko emakumezkoen futbolean

[EUS] Lan honen abiapuntua, goi errendimenduko emakumezkoen futbol partida batean ematen diren korner errutina ezberdinak aztertu eta horietatik eraginkorrenak diren ekintza eta jokaldiak identifikatzea izan da. Ikerlanean, hiru denboralditan zehar (2012-2013; 2013-2014; 2014-2015), Espainiako emakumezkoen futboleko lehenengo mailako, hau da Superligako taldeek 51 partidutan burututako korner jaurtiketak aztertu dira. Horretarako erabili den metodologia, bideo analisi eta analisi notazionala izan dira. Bideoan jasotako informazioa, aldagai guztiak kontutan hartzen dituen taula berezi batzuetan bildu dira. Horietan, jokatutako partidua, partiduko unea (0’-15’; 15’-30’; 30’-45’; 45’-60’; 60’-75’; 75’-90’), kornerra jaurtitzeko aldea (eskuin edo ezkerra), jaurtiketa gauzatutako hanka (eskuina edo ezkerra), jaurtiketa gauzatzeko modua (luze edo motza), jaurtiketaren helmuga puntua (lehenengo zutoina, erdira edo bigarren zutoina) eta azkenik, jaurtiketaren eraginkortasuna mailakatu da (gola, errematea atera, errematea atetik kanpora, aurkariak urrundu, aurkariak kanpora bota edo jaurtiketa kanpora) aldagaiak bereizi dira. Tauletan jasotako datu guztiak Microsoft Excel eta IBM SPSS statistic 23 programan bildu dira ekintzen deskribapena (bataz besteko eta desbideratze estandarra), frekuentzia (kopuru eta portzentajeak) eta aldagaiek eraginkortasunarekin duten erlazioa ateratzeko. Emaitzek adierazten dutenez, bataz beste 7,88 korner jaurtiketa burutu dira 56 partidutan 2,97ko desbideratze tipikoaz, horietatik bi kasutan bakarrik lortu da jokaldia golean amaitzea eta jaurtiketen %22,89an errematatzea. Gainera, korner jaurtiketen bidez, talde erasotzaileek gol aukera gutxi sortzen dituztela ikusi da. Baita ohiko errutinak luzez jaurtitzea, lehenengo zutoin aldera eta eskuin hankaz direla ere. Emaitzak ikusita ondoriozta daiteke, korner jaurtiketak erasoko egoeran gehiago landu beharko liratekeela eraginkortasun handiagoa lortu eta gol aukera gehiago sortzeko. Hitz gakoak: futbola, emakumeak, goi errendimendua, partidu analisia, kornerrak, analisi notazionala.

Expression and Localization of Opioid Receptors in Male Germ Cells and the Implication for Mouse Spermatogenesis

The presence of endogenous opioid peptides in different testicular cell types has been extensively characterized and provides evidence for the participation of the opioid system in the regulation of testicular function. However, the exact role of the opioid system during the spermatogenesis has remained controversial since the presence of the mu-, delta-and kappa-opioid receptors in spermatogenic cells was yet to be demonstrated. Through a combination of quantitative real-time PCR, immunofluorescence, immunohistochemistry and flow cytometry approaches, we report for the first time the presence of active mu-, deltaand kappa-opioid receptors in mouse male germ cells. They show an exposition time-dependent response to opioid agonist, hence suggesting their active involvement in spermatogenesis. Our results contribute to understanding the role of the opioid receptors in the spermatogenesis and could help to develop new strategies to employ the opioid system as a biochemical tool for the diagnosis and treatment of male infertility.