Bide metabolikoen ebaluaketarako protokolo kliniko exekutagarria

Proiektu pilotu honekin medikuntzarako tresna lagungarri bat sortu da Jaiotzetiko Sortzen Diren Errore Metabolikoak (JASODEM) diagnostikatzeko. Horretarako, Gurutzeta Ospitaleko Metabolismo Laborategiko adituarekin eta UPV/EHUko Informatika Fakultateko ERABAKI taldearen arteko elkar-lana funtsezkoa izan da. Amaierako produktua Web zerbitzari batean egongo da exekutagarri. Web aplikazioaren bitartez, medikuak gaixo ezberdinei diagnostikoak egiteko aukera izango du. Horretarako aski da Praktika Klinikorako Gidaren exekuzio bat abiaraztea eta bertan pazienteari inguruan eskatzen diren datuak sartzea. Exekuzio prozesuan zehar badago sistematik irten eta ondoren lanarekin berriz jarraitzea, modu honetan froga klinikoak beharren arabera egiten direlarik, diagnostiko prozesuko kostua murriztuz. Bestalde, Praktika Klinikorako Gidan agertzen diren kontzeptuen inguruko informazioa jasotzeko funtzionalitatea eskaintzen zaio erabiltzaileari. Proiektua garatzeko aukeratu den metodologiaren jarraipen zehatza egitea eta kalitatezko dokumentazioa sortzea ezinbestekoa da. Proiektu honen garapenerako, Rational Unified Process (RUP) metodologia eta honi euskarria ematen dioten tresneria erabili da. Proiektuaren analisirako eta Jakintza Ingeniaritzarako CommonKADS metodologia erabili da.

Estrategia de internacionalización en el sector vitivinícola: análisis de casos

[ES]En este Trabajo de Fin de Grado se trata el tema de la internacionalización en el sector vitivinícola, expresando brevemente la importancia del sector agroalimentario, del cual es parte el área analizada. Será analizado mediante casos prácticos, se estudiarán dos de las bodegas más importantes de España, establecidas en dos comunidades autónomas que son productoras y distribuidoras de vino históricamente; Bodegas Eguren-Ugarte, afincada en el País Vasco y una de las marcas más conocidas a nivel nacional; Bodegas Torres, afincada en la Cataluña y una de las marcas más conocidas tanto a nivel nacional como mundial. Mediante la realización de entrevistas a responsables de comercio exterior busco por una parte afianzar mis aptitudes en cuanto a la relación con grandes empresarios, intentando conocer mejor como trabajan e intentando mejorar el cara a cara con personas influyentes. Por otro lado, pretendo identificar las características diferenciales del proceso de internacionalización del sector vitivinícola, para conocer mejor el sector y la manera en la que las empresas bodegueras realizan su expansión internacional. En tercer lugar, y debido a que el sector vitivinícola crece y se moderniza a grandes velocidades, conocer las nuevas vías de comercialización de producto. Por último, y en menor grado, debido a la falta de información, trato de valorarla participación de los entes públicos (estatales, autonómicos o europeos) dentro del proceso de internacionalización y si esta participación es influyente.

Hidrogenoaren ekoizpena hiri hondakin plastikoen pirolisi eta erreformatuaren bidez

Material plastikoen kontsumoak izugarri gora egin du azken mendean. Hori dela eta, material hauek erabiltzearen ondorioz sortutako hondakinak asko handitu dira. Europar Batasuneko herrialdeetan 250 milioi tona baino gehiago hiri - hondaki n solido ( RSU ) sortzen dira urtero, urteko %3ko hazkunt zarekin. K antitate honen %7a plastiko hondakinei dagokie, hots, 17.5 milioi tona. Itsasoko uretan ere aurki daitezke plastikoak, urtero sei milioi tona eta erdi botatzen baitira itsasora, mediterraneo itsasoa izanik munduko plastiko biltegirik handiena. Itsasoan 90 urteraino iraun dezake te eta urte hauetan zehar distantzia handiak egin ditzakete aldatu gabe. Horregatik esaten da plastikoak iraunkorrak direla. Egun, hondakin plastikoen portzentaia txiki bat bakarrik birziklatzen da eta bai biltegiratzea bai erreketa bidezko eliminazioak ingurumen arazoak dituzte . Gainera, plastiko gehienak degradaezi nak dira, urte luzez ingurugiro kalteak eraginez . Hori dela eta, material hauen balorizaziorako teknologia berrien sustapena beharrezkoa da, eskala handian eraginkorrak, ekonomikoki bideragarriak eta ingurugiroa errespetatuko dutenak. Hondakin plastikoetatik abiatuz hidrogenoa lortzeko prozesua interesgarria eta bideragarria da, hidrogenoaren kontsumoaren igoe ra kontuan hartuz. Gaur egun erregai fosilen prozesaketatik lortzen da hidrogenoa, CO 2 - a ren emisio handiak sortzen direlarik. Emisio hauen murrizketa beharrezkotzat hartu da. Hau guztiagatik, Gradu Amaierako Lan honen helburu nagusia plastikoen balorizazio a ikertzea da, konkretuki hiri - hondakin solidoetan aurkitzen diren plast ikoena . Pirolisi eta ur baporearen bidezko erreformatua erabili dira hidrogenoa lortzeko, azken hau balio handiko produktua izanik. Horretarako lehenengo etapa iturri ohantze konikoan, 500 ºC - tan, buruturiko pirolisia izan da eta bigarrenik ohantze fluidizatu batean ur baporearen bidezko erreformatua burutu da , 700 ºC - tan . Helburu nagusi hau betetzeko bestelako helburu zehatzak ezarri dira, hiri - hondakin solidoetan aurkitzen diren HDPE, PP, PS eta PET plastiko nahaste baten bideragarritasuna aztertu delarik aurrez aipatutako bi etapen bidez:  Plastiko nahastearen pirolisian sorturiko konposatu hegazkorren erreformatua era jarraian burutzea.  Zero denboran e rreakzio indizeak (konbertsioa et a etekinak) eta lortutako gasaren konposizioa determinatze a .  Erreformatuan erabilitako katalizatzailearen desaktibatzeak erreakzioa ren konbertsio eta etekina n duen eragina aztertzea.

Diagrama de secuencias de reducción (dsr): aproximación metodológica para el análisis de núcleos líticos y remontajes

Homenaje a Georges Laplace, realizado en Vitoria-Gasteiz el 13,14 y 15 de noviembre de 2012. Edición a cargo de Aitor Calvo, Aitor Sánchez, Maite García-Rojas y Mónica Alonso-Eguíluz.

La Tipología Analítica aplicada al estudio de materiales líticos de época histórica

Homenaje a Georges Laplace, realizado en Vitoria-Gasteiz el 13, 14 y 15 de noviembre de 2012. Edición a cargo de Aitor Calvo, Aitor Sánchez, Maite García-Rojas y Mónica Alonso-Eguíluz.

Diseño y fabricación de un conjunto de estriberas y semimanillares para moto de competición.

[ES]El objeto del presente trabajo consiste en diseñar un conjunto de estriberas y semimanillares para una moto de competición, que cumplan con una serie de requerimientos geométricos y que garanticen las mejores prestaciones posibles en cuanto a competición, entre las que destacan la seguridad y comodidad del piloto. A continuación se procederá a elegir los procesos más adecuados para la fabricación de los mismos, teniendo en cuenta requisitos económicos y de calidad, para sacar al mercado un producto lo más competitivo posible. El estudio se basa principalmente en el análisis de diferentes alternativas que se pueden adoptar para obtener el diseño de los productos y posterior fabricación. Para ello se estudiarán los objetivos y condiciones que se deben cumplir. Cabe destacar que la idea de desarrollo de este proyecto surgió dentro de otro de mayor envergadura, la competición MotoStudent. MotoStudent es una competición internacional entre universidades de todo el mundo en la que los equipos de estudiantes se enfrentan al desafío de diseñar y desarrollar un prototipo de motocicleta de competición similar a la categoría mundialista de Moto3.

Androgen-regulated metabolism and biosynthesis in prostate cancer

Metabolic changes are a well-described hallmark of cancer and are responses to changes in the activity of diverse oncogenes and tumour suppressors. For example, steroid hormone biosynthesis is intimately associated with changes in lipid metabolism and represents a therapeutic intervention point in the treatment of prostate cancer (PCa). Both prostate gland development and tumorigenesis rely on the activity of a steroid hormone receptor family member, the androgen receptor (AR). Recent studies have sought to define the biological effect of the AR on PCa by defining the whole-genome binding sites and gene networks that are regulated by the AR. These studies have provided the first systematic evidence that the AR influences metabolism and biosynthesis at key regulatory steps within pathways that have also been defined as points of influence for other oncogenes, including c-Myc, p53 and hypoxia-inducible factor 1α, in other cancers. The success of interfering with these pathways in a therapeutic setting will, however, hinge on our ability to manage the concomitant stress and survival responses induced by such treatments and to define appropriate therapeutic windows.

N-linked glycosylation supports cross-talk between receptor tyrosine kinases and androgen receptor

Prostate cancer is the second most common cause of cancer-associated deaths in men and signalling via a transcription factor called androgen receptor (AR) is an important driver of the disease. Androgen treatment is known to affect the expression and activity of other oncogenes including receptor tyrosine kinases (RTKs). In this study we report that AR-positive prostate cancer cell-lines express 50% higher levels of enzymes in the hexosamine biosynthesis pathway (HBP) than AR-negative prostate cell-lines. HBP produces hexosamines that are used by endoplasmic reticulum and golgi enzymes to glycosylate proteins targeted to plasma-membrane and secretion. Inhibition of O-linked glycosylation by ST045849 or N-linked glycosylation with tunicamycin decreased cell viability by 20%. In addition, tunicamycin inhibited the androgen-induced expression of AR target genes KLK3 and CaMKK2 by 50%. RTKs have been shown to enhance AR activity and we used an antibody array to identify changes in the phosphorylation status of RTKs in response to androgen stimulation. Hormone treatment increased the activity of Insulin like Growth Factor 1-Receptor (IGF-1R) ten-fold and this was associated with a concomitant increase in the N-linked glycosylation of the receptor, analyzed by lectin enrichment experiments. Glycosylation is known to be important for the processing and stability of RTKs. Inhibition of N-linked glycosylation resulted in accumulation of IGF-1R pro-receptor with altered mobility as shown by immunoprecipitation. Confocal imaging revealed that androgen induced plasma-membrane localization of IGF-1R was blocked by tunicamycin. In conclusion we have established that the glycosylation of IGF-1R is necessary for the full activation of the receptor in response to androgen treatment and that perturbing this process can break the feedback loop between AR and IGF-1R activation in prostate cells. Achieving similar results selectively in a clinical setting will be an important challenge in the future.

O-GlcNAc transferase integrates metabolic pathways to regulate the stability of c-MYC in human prostate cancer cells

Metabolic disruptions that occur widely in cancers offer an attractive focus for generalized treatment strategies. The hexosamine biosynthetic pathway (HBP) senses metabolic status and produces an essential substrate for O-linked β-N-acetylglucosamine transferase (OGT), which glycosylates and thereby modulates the function of its target proteins. Here, we report that the HBP is activated in prostate cancer cells and that OGT is a central regulator of c-Myc stability in this setting. HBP genes were overexpressed in human prostate cancers and androgen regulated in cultured human cancer cell lines. Immunohistochemical analysis of human specimens (n = 1987) established that OGT is upregulated at the protein level and that its expression correlates with high Gleason score, pT and pN stages, and biochemical recurrence. RNA interference-mediated siliencing or pharmacologic inhibition of OGT was sufficient to decrease prostate cancer cell growth. Microarray profiling showed that the principal effects of OGT inhibition in prostate cancer cells were related to cell-cycle progression and DNA replication. In particular, c-MYC was identified as a candidate upstream regulator of OGT target genes and OGT inhibition elicited a dose-dependent decrease in the levels of c-MYC protein but not c-MYC mRNA in cell lines. Supporting this relationship, expression of c-MYC and OGT was tightly correlated in human prostate cancer samples (n = 1306). Our findings identify HBP as a modulator of prostate cancer growth and c-MYC as a key target of OGT function in prostate cancer cells.

Molecular subtyping of primary prostate cancer reveals specific and shared target genes of different ETS rearrangements

This work aimed to evaluate whether ETS transcription factors frequently involved in rearrangements in prostate carcinomas (PCa), namely ERG and ETV1, regulate specific or shared target genes. We performed differential expression analysis on nine normal prostate tissues and 50 PCa enriched for different ETS rearrangements using exon-level expression microarrays, followed by in vitro validation using cell line models. We found specific deregulation of 57 genes in ERG-positive PCa and 15 genes in ETV1-positive PCa, whereas deregulation of 27 genes was shared in both tumor subtypes. We further showed that the expression of seven tumor-associated ERG target genes (PLA1A, CACNA1D, ATP8A2, HLA-DMB, PDE3B, TDRD1, and TMBIM1) and two tumor-associated ETV1 target genes (FKBP10 and GLYATL2) was significantly affected by specific ETS silencing in VCaP and LNCaP cell line models, respectively, whereas the expression of three candidate ERG and ETV1 shared targets (GRPR, KCNH8, and TMEM45B) was significantly affected by silencing of either ETS. Interestingly, we demonstrate that the expression of TDRD1, the topmost overexpressed gene of our list of ERG-specific candidate targets, is inversely correlated with the methylation levels of a CpG island found at -66 bp of the transcription start site in PCa and that TDRD1 expression is regulated by direct binding of ERG to the CpG island in VCaP cells. We conclude that ETS transcription factors regulate specific and shared target genes and that TDRD1, FKBP10, and GRPR are promising therapeutic targets and can serve as diagnostic markers for molecular subtypes of PCa harboring specific fusion gene rearrangements.

Chromatin binding by the androgen receptor in prostate cancer

Alterations in transcriptional programs are fundamental to the development of cancers. The androgen receptor is central to the normal development of the prostate gland and to the development of prostate cancer. To a large extent this is believed to be due to the control of gene expression through the interaction of the androgen receptor with chromatin and subsequently with coregulators and the transcriptional machinery. Unbiased genome-wide studies have recently uncovered the recruitment sites that are gene-distal and intragenic rather than associated with proximal promoter regions. Whilst expression profiles from AR-positive primary prostate tumours and cell lines can directly relate to the AR cistrome in prostate cancer cells, this distribution raises significant challenges in making direct mechanistic connections. Furthermore, extrapolating from datasets assembled in one model to other model systems or clinical samples poses challenges if we are to use the AR-directed transcriptome to guide the development of novel biomarkers or treatment decisions. This review will provide an overview of the androgen receptor before addressing the challenges and opportunities created by whole-genome studies of the interplay between the androgen receptor and chromatin.

Studying N-linked glycosylation of receptor tyrosine kinases

Metabolic alterations have been identified as a frequent event in cancer. This is often associated with increased flux through glycolysis, and also a secondary pathway to glycolysis, hexosamine biosynthetic pathway (HBP). HBP provides substrate for N-linked glycosylation, which occurs in the endoplasmic reticulum and the Golgi apparatus. N-linked glycosylation supports protein folding and correct sorting of proteins to plasma membrane and secretion. This process generates complex glycoforms, which can be recognized by other proteins and glycosylation of receptor tyrosine kinases (RTK) can also regulate their plasma-membrane retention time. Of special interest for experimental biologists, plants produce proteins, termed lectins, which bind with high specificity to glyco-conjugates. For the purposes of molecular biology, plant lectins can be conjugated to different moieties, such as agarose beads, which enable precipitation of specifically glycosylated proteins. In this chapter, we describe in detail how to perform pull-down experiments with commercially available lectins to identify changes in the glycosylation of RTKs.

Specific Noncompetitive Immunoassay for HT-2 Mycotoxin Detection

Here we demonstrate a novel homogeneous one-step immunoassay, utilizing a pair of recombinant antibody antigen-binding fragments (Fab), that is specific for HT-2 toxin and has a positive readout. Advantages over the conventional competitive immunoassay formats such as enzyme-linked immunosorbent assay (ELISA) are the specificity, speed, and simplicity of the assay. Recombinant antibody HT2-10 Fab recognizing both HT-2 and T-2 toxins was developed from a phage display antibody library containing 6 × 10(7) different antibody clones. Specificity of the immunoassay was introduced by an anti-immune complex (IC) antibody binding the primary antibody-HT-2 toxin complex. When the noncompetitive immune complex assay was compared to the traditional competitive assay, an over 10-fold improvement in sensitivity was observed. Although the HT2-10 antibody has 100% cross-reactivity for HT-2 and T-2 toxins, the immune complex assay is highly specific for HT-2 alone. The assay performance with real samples was evaluated using naturally contaminated wheat reference material. The half-maximal effective concentration (EC50) value of the time-resolved fluorescence resonance energy transfer (TR-FRET) assay was 9.6 ng/mL, and the limit of detection (LOD) was 0.38 ng/mL (19 μg/kg). The labeled antibodies can be predried to the assay vials, e.g., microtiter plate wells, and readout is ready in 10 min after the sample application.

Inhibition of O-GlcNAc transferase activity reprograms prostate cancer cell metabolism

Metabolic networks are highly connected and complex, but a single enzyme, O-GlcNAc transferase (OGT) can sense the availability of metabolites and also modify target proteins. We show that inhibition of OGT activity inhibits the proliferation of prostate cancer cells, leads to sustained loss of c-MYC and suppresses the expression of CDK1, elevated expression of which predicts prostate cancer recurrence (p=0.00179). Metabolic profiling revealed decreased glucose consumption and lactate production after OGT inhibition. This decreased glycolytic activity specifically sensitized prostate cancer cells, but not cells representing normal prostate epithelium, to inhibitors of oxidative phosphorylation (rotenone and metformin). Intra-cellular alanine was depleted upon OGT inhibitor treatment. OGT inhibitor increased the expression and activity of alanine aminotransferase (GPT2), an enzyme that can be targeted with a clinically approved drug, cycloserine. Simultaneous inhibition of OGT and GPT2 inhibited cell viability and growth rate, and additionally activated a cell death response. These combinatorial effects were predominantly seen in prostate cancer cells, but not in a cell-line derived from normal prostate epithelium. Combinatorial treatments were confirmed with two inhibitors against both OGT and GPT2. Taken together, here we report the reprogramming of energy metabolism upon inhibition of OGT activity, and identify synergistically lethal combinations that are prostate cancer cell specific.