8.30 – 8.45 h Inauguració Jornada Científica

8.45 – 9.00 h

Ana Almazán Moga. Recerca translacional en càncer infantil VHIR La via hedgehog com a possible diana terapèutica en el rabdomiosarcoma

9

9.00 – 9.15 h Verónica Cánovas Hernández. Unitat de recerca biomèdica i oncologia translacional VHIR Prostate Tumor OVerexpressed 1 (PTOV1) modulates Castration Resistant Prostate Cancer (CRPC) cells resistance to docetaxel

10

9.15 – 9.30 h

Toni Jauset González. Modelització de teràpies anti-tumorals en ratolí VHIO Myc inhibition is a promising therapy in glioma and induces mitotic crisis in cancer cells

11

9.30 – 9.45 h

Noèlia Purroy i Zuriguel. Hematologia experimental VHIR L’inhibidor de la proteïna quinasa SYK, TAK659, bloqueja els senyals de supervivència, proliferació i migració procedents del microambient en cèl.lules primàries de Leucèmia Limfàtica Crònica

12

9.45 – 10.00 h

Ada Sala Hojman. Expressió gènica i càncer VHIO Molecular mechanisms involved in the regulation of glioma-initiating cells

13

10.00 – 10.15 h

Helena Plà. Direccion. i alliber. farmacològica – CIBBIM Nanomedicina Polymer-drug conjugates (PDC) based on polyglutamic acid (PGA) for the treatment of advanced colorectal cancer (CRC)

14

10.15 – 10.30 h

Paulo André Gonçalves Rodrigues. Oncologia molecular – CIBBIM Nanomedicina Loss of RHOA is important for the activation of Wnt signaling and progression of colorectal tumorigenesis

15

10.30 – 10.45 h

Anna Salas Torras. Oftalmologia VHIR In vitro studies on the anti-angiogenic effects of pigment epithelium derived factor and somatostatin

16

10.45 - 11.00 h

Cristina Sáez López. Diabetis i metabolisme VHIR El ácido oleico incrementa la producción hépatica de Sex Hormone-Binding Globulin (SHBG) en hombres

17

11.00 – 11.15 h

Irina B. Torres Rodriguez. Nefrologia VHIR Gene expression signature of tolerance and lymphocyte subsets in stable renal transplants: results of a cross-sectional study

18

11.15 – 11.30 h

Pausa descans

11.30 – 11.45 h Àstrid Brull Cañagueral. Patologia mitocondrial i neuromuscular VHIR Caracterització fenotípica d’un model murí de la malaltia de McArdle (pR50X) en fons genètic homogeni

19

11.45 – 12.00 h Mireia Campos Martorell. Malalties neurovasculars VHIR Eficàcia i seguretat de la simvastatina com a tractament en la fase aguda de la isquèmia cerebral

20

12.00 – 12.15 h Koen Galenkamp. Senyalització cel·lular i apoptosi VHIR Sensitizing neuroblastoma cell lines to FasL-induced cell death by treatment with TNFα

21

12.15 – 12.30 h Ariadna Recasens Ibabe. Malalties neurodegeneratives VHIR Cossos de Lewy de pacients amb malaltia de Parkinson inicien un procés neurodegeneratiu dependent de l’α-sinucleina en ratolins i primats no humans

22

12.30 – 12.45 h Marta Vila Pueyo. Neurologia infantil VHIR Epigenetic changes in a rat model of migraine with aura

23

12.45 – 13.00 h Suchita Panda. Fisiologia i fisiopatologia digestiva VHIR

Short-term effect of antibiotics on human gut microbiota

24

13.00 – 13.15 h Mireia Puig Asensio. Malalties infeccioses VHIR Determinants of early mortality in candidemia

25

13.15 – 13.30 h Arnald Alonso. Malalties sistemiques VHIR ASTREAM i FOCUS: Dues potents metodologies per a l’anàlisi de dades metabolòmiques obtingudes per espectrometria de masses i ressonància magnètica nuclear

26

13.30 – 13.45 h Mònica Faro Colomés. Fatiga crònica VHIR Situació laboral en els pacients amb síndrome de fatiga crónica (SFC)

27

13.45 – 14.00 h Alejandra Fernández Martín. Bioenginyeria ortopèdia i cirurgia pediàtriques VHIR Caracterización de precursores neurales en líquido amniótico de modelo ovino de MMC

28

14.00 – 14.15 h Aina Ruiz Romero. Medicina materna i fetal VHIR Maternal and fetal anti-angiogenic imbalance in congenital heart defects

29

14.15 – 14.30 h Maria Lujan Iavecchia. Farmacologia clínica VHIR Perioperative management and outcomes of patients with high or moderate thrombotic risk treated with antithrombotic therapy and hip fracture surgery

30

14.30 – 15.00 h

Pausa dinar

P1 Laura Devis. Unitat d'investigació biomèdica i oncología traslacional VHIR New actor in endometrial carcinoma invasion: alcam regularion

31

P2

Alba Gonzalez. Expressió gènica i cáncer VHIO Targeting the TGF-beta pathway as a new therapeutic aproach for glioma patients

32

P3 Luz Jubierre. Recerca translacional en càncer infantil VHIR SMARCA4/BRG1 is a novel therapeutic target against high-risk neurobla

33

P4

Lucia Lanau. Unitat de recerca biomèdica i oncologia translacional VHIR A comparative study between primary tumors, ascites and metastases reveals a new gene expression profile associated with ovarian cancer dissemination

34

P5

Daniel Massó. Modelització de teràpies anti-tumorals en ratolí VHIO Inhibiting BTK function as therapeutic strategy in pancreatic cancer

35

P6

Melania Montes. Unitat de recerca biomèdica i oncologia translacional VHIR Identification of a specific MicroRNA profile in urine samples for prostate cancer

36

P7 Tamara Sequeiros. Unitat de recerca biomèdica i oncologia translacional VHIR New biomarkers for the diagnosis and prognosis of prostate cancer from urinary extracellular vesicles

37

P8 Elena Andretta. Oncologia molecular - CIBBIM - Nanomedicina Role of EPHA3 in colorectal cancer

38

P9

Sarah Bazzocco. Oncologia molecular - CIBBIM - Nanomedicina Genes highly expressed in rapidly proliferating tumor cells as new targets for colorectal cancer treatment

39

P10 Laura Planells. Senyalització cel·lular i apoptosi VHIR MYCN repression of lifeguard is associated with increased cell migration and neuroblastoma poor prognosis

40

P11 Marianela Mego. Fisiologia i fisiopatologia digestiva VHIR

Afectación anorectal en enfermedad de chagas en médio no endèmico

41

P12 Lidia Corraliza. Diabetis i metabolisme VHIR El ratolí C57BL/KsJ-db/db és un model adequat per investigar la neurodegeneració retiniana induïda per la diabetis.

42

P13 Maria Atonieta Azancot. Nefrologia VHIR Renal biopsies from expanded criteria donors: reproducibility and predictive value on outcome

43

P14 Iolanda Riba. Malalties neurovasculars VHIR El deteriormament cognitiu lleuger es relaciona amb el mal control dels factors de risc vascular i les lesions vasculars cerebrals silents

44

P15

Jorge Urrusti. Senyalització cel·lular i apoptosi VHIR Bcl-xL is required for protection from etoposide-induced cell death by Lifeguard

45

P16 Eloísa Salvo Romero. Fisiologia i fisiopatologia digestiva VHIR Identification of mucosal eosinophil activity in the irritable bowel syndrome: modulatory role of neuromediators

46

P17 Alba Santiago Badenas. Fisiologia i fisiopatologia digestiva VHIR

Microbiome composition by pyrosequencing in the mesenteric lymph nodes of CCl4-induced cirrhotic rats

47

P18 Maddalena Peghin. Malalties infeccioses VHIR Unusual forms of invasive acute and chronic pulmonary aspergillosis in patients with solid tumors

48

16.00 – 16.15 h Pilar Meler Amella. Unitat d’Ictus HUVH Tiempo de reacción de enfermería en el código ictus

49

16.15 – 16.30 h Contxita Jiménez Gutierrez. Fatiga crònica VHIR Ensayo clínico, aleatorizado, controlado, paralelo, para evaluar la eficacia de la Acupuntura, como tratamiento en los pacientes diagnosticados del Síndrome de Fatiga Crónica

50

16.30 – 16.45 h Marisa Cebrián Batalla. Recerca en cures de salut VHIR Els professionals de salut davant el seguiment de tractaments de llarga durada en adolescents.

51

8.15 – 8.30 h María Mutuberria. Cardiologia HUVH Triple therapy (anticoagulation plus dual antiplatelet therapy) or oral anticoagulation (OAC) plus clopidogrel in patients with a mechanical prostheses undergoing coronary stenting (CS)?

53

8.30 – 8.45 h Marina García García de Acilu. CRIPS/ Servei medicina intensiva HUVH Niveles plasmáticos de ST2 como predictor de mortalidad en pacientes con síndrome de distrés respiratorio agudo

54

8.45 – 9.00 h Cristina López Sánchez. Malalties Infeccioses VHIR Liver-biopsy related infections in liver transplant recipients

55

9.00 – 9.15 h Isabel Huber Ruano. Expressió gènica i càncer VHIO TGFβ controls tumor progression through JMJD3-mediated epigenetic regulation

56

9.15 – 9.30 h Beatriz Morancho Armisen. Factors de creixement i càncer VHIO A dominant-negative N-terminal fragment of HER2 frequently expressed in breast cancers

57

9.30 – 9.45 h Bárbara Castellana Esteban. Patologia molecular VHIR Interplay of YB-1 and IL-6 in the acquisition of EMT-like characteristics

58

9.45 – 10.00 h Javier Torres Torronteras. Patologia neuromuscular i mitocondrial VHIR Corrección bioquímica del modelo animal de MNGIE mediante el uso de un AAV2/8 dirigido a hígado

59

10.00 – 10.15 h Paulina Carriba Domínguez. Senyalització cel·lular i apoptosis VHIR Amyloid beta reduces the expression of faim-l shifting the TNFa protection to aggression

60

10.15 – 10.30 h Cristina Frías García. Fisiologia i fisiopatologia digestiva VHIR Gender determines mucosal macrophage activity in the human jejunum: implications for irritable bowel syndrome

62

10.30 – 10.45 h

Adrià Curran Fabregas. Malalties Infeccioses VHIR Concentraciones plasmáticas totales y libres de Darunavir en pacientes con co-infección VIH-VHC y cirrosis hepática comparadas con pacientes mono-infectados por el VIH

63

10.45 – 11.00 h

Petra Gener. Direccionament i alliberament farmacològica -CIBBIM - Nanomedicina Paclitaxel loaded PLGA-co-PEG polymeric micelles eradicate cancer cells and effectively hamper recovery of cancer stem cells in vitro

64

10:54 – 11:15 h Pausa descans

17a CONFERÈNCIA ANUAL HUVH

29 de noviembre de 2013

11.15 h Presentació

11.30 h Presentació de resultats 2013 Dr. Joan X. Comella. Director VHIR

12.15 h Conferència Anual ‘From viral hepatitis to hepatocyte transplantation – examples for the benefits of scientific networking’ Prof. Michael Manns.

Departament de Medicina de l’Escola de Hannover.

13.15 h Lliurament Premis 7º Jornada Científica

13.45 h Cloenda

Josep Sánchez de Toledo i Josep Roma

Ana Almazán-Moga1, Josep Roma1, Carla Molist1, Pablo Velasco1, Luz Jubierre1, Aroa Soriano1, Miguel F. Segura1, José Sánchez de Toledo2, Soledad Gallego1,2 1Recerca Translacional en el Càncer Infantil, Vall d’Hebron Institut de Recerca (VHIR), Universitat Autònoma de Barcelona, Passeig Vall d’Hebron 119, Barcelona 08035, Spain; 2 Servei d’Oncologia Pediàtrica i Hematologia, Hospital Universitari Vall d’Hebron, Universitat Autònoma de Barcelona, Passeig Vall d’Hebron 119, Barcelona 08035, Spain

Objectiu: Determinar el paper oncogènic de la via Hedgehog així com el seu mecanisme d’activació en el Rabdomiosarcoma (RMS), per tal d’establir noves dianes terapèutiques pel tractament dels pacients afectats amb aquesta malaltia. Material i mètode: Es varen determinar els nivells d’activació de la via Hedgehog en línies cel•lulars i tumors de RMS mitjançant Real-time PCR, Western blot i immunohistoquímica. Per a la inhibició genètica de la via es varen generar línies cel•lulars estables que expressaven shRNA per silenciar: SHH, IHH, DHH, GLI1 i GLi2. Per a la inhibició farmacològica de la via es va utilizar Vismodegib. Es varen realitzar assajos funcionals per tal d’avaluar el efectes in vitro de la inhibició de la via Hedgehog. Resultats: La inhibició in vitro de la via Hedgehog redueix de forma significativa la proliferació i la migració cel·lular, tenint un efecte clau sobre les cèl·lules iniciadores de RMS. Les dades obtingudes de l’anàlisi dels tumors de pacients de RMS suggereixen l’activació autocrina de la via Hedgehog en el RMS, en la qual tindrien un paper clau els lligands IHH i DHH. Conclusions: Els nostres resultats in vitro situen la via Hedgehog com a diana terapèutica contra el RMS. Publicacions: AUTHORS: Roma J, Almazán-Moga A, Sánchez de Toledo J, Gallego S TITLE: Notch, Wnt, and Hedgehog Pathways in Rhabdomyosarcoma: From Single Pathways to an Integrated Network JOURNAL: Sarcoma YEAR: 2012 - ISSUE: 2012 - PAGES: 7 doi: 10.1155/2012/695603 TYPE OF PUBLICATION: R

Rosanna Paciucci Barzanti

Verónica Cánovas, Néus Marqués, Marta Sesé, Inés de Torres,Yolanda Puñal, Yolanda Fernández, Ibane Abasolo, Mercedes Marín, Santiago Ramón y Cajal, Begoña Mellado, Rosanna Paciucci.

INTRODUCTION: CRPC patients treated with Docetaxel (D) initially respond but resistance is inevitably developed. The protein PTOV1, identified in our laboratory as a novel gene and protein, is strongly expressed in metastatic carcinoma and pre-neoplastic (HG-PIN) lesions but is barely detected in benign prostate epithelium. Signaling routes that control protein synthesis are emerging as unique and critical steps in the progression of human cancers.

OBJETIVES: Study the development of drug resistance in CRPC focusing on the translatome (genome-wide pool of translated mRNAs) and identify the molecular mechanisms by which PTOV1 modulates progression of prostate cancer.

MATERIALS AND METHODS: Two chemosensitives and D-resistant (DR) CRPC cell lines were used in this study. Inducible knockdown for PTOV1 and c-Jun expression was obtained using TRIPZ vectors. Cells were studied for their invasive and tumorigenic properties in vitro and in vivo. Polysomal profiling of chemosensitives and DR cells is being performed by fractionation on sucrose gradients, RNA purification and microarrays analysis.

RESULTS AND CONCLUSIONS: PTOV1 interacts with the ribosomal protein RACK1 and modulates global protein translation and c-Jun mRNAs translation. In turn, c-Jun/AP1 activates SNAIL1 transcription and these oncoproteins promote invasion, tumor growth and metastasis in SCID-beige mice. Preliminary evidences show that DR-CRPC cells knockdown for PTOV1 are unable to survive in the presence of D, suggesting that a functional PTOV1-AP1-SNAIL1 circuit might be required for resistance to chemotherapy. Our results also provide an explanation for the few number of reports based on transcriptional profiling describing the overexpression of the c-Jun oncogene in cancer.

PUBLICATIONS: Regulation of protein translation and c-Jun expression by Prostate Tumor Overexpressed 1 (PTOV1). N. Marqués, M. Sesé, V. Cánovas, F. Valente, R. Bermudo, I. de Torres, Y. Fernández, I. Abasolo, P. L Fernández, H. Contreras, E. Castellón, T. Celià-Terrassa, R.l Méndez, S. Ramón y Cajal, T. M. Thomson and R. Paciucci. Oncogene. 2013 Mar 4. doi: 10.1038/onc.2013.51.

Laura Soucek

D. Annibali, J.R. Whitfield, E. Favuzzi, T. Jauset, E. Serrano, G. Folch, M.I. Cuartas, A. Gonzalez, L. Brown Swigart, S. Nasi, G.I. Evan, J. Seoane and L. Soucek.

Objectives: The aim of the project is to analyze the therapeutic impact of Myc inhibition making use of our Myc dominant negative (termed Omomyc), in both progression and maintenance of glioma and elucidate its possible mechanism of action. Materials and Methods: We made use of the GFAP-V12Ha-Ras mouse model of glioma and of a model expressing our switchable Myc inhibitor Omomyc, in order to assess Myc inhibition as a therapeutic strategy in glioma. We also used neuroprogenitor cells derived from the same mice and human glioblastoma cell lines to further study the effect of Omomyc expression in vitro and created orthotopic xenografts from patient-derived tumors to test Myc inhibition in human cells in vivo. Results: Myc inhibition by Omomyc is able to prevent tumor progression in GFAP-V12Ha-Ras mice, conferring a dramatic survival advantage to the treated group. Human glioblastoma cell lines and mouse neuroprogenitors expressing Omomyc showed a reduction in proliferation and survival. Moreover, severe mitotic catastrophe was seen in human cell lines when the inhibitor was present. Omomyc expression in patient-derived tumor samples reduced the overall number of neurospheres and significantly increased overall survival of mice orthotopically transplanted with disaggregated neurospheres. Conclusion: Myc inhibition is a potent therapeutic strategy in glioma and its effects include induction of mitotic crisis in tumor cells. We are currently further investigating the precise mechanism by which Omomyc functions and developing clinical approaches to Myc inhibition. Publications: MYC inhibition as a potential therapy in glioma inducing mitotic crisis in cancer cells. D. Annibali1,2, J.R. Whitfield3, E. Favuzzi2, T. Jauset3, E. Serrano3, G. Folch3, M.I. Cuartas3, A. Gonzalez3, L. Brown Swigart1, S. Nasi2, G.I. Evan4, J. Seoane3 and L. Soucek3 1UCSF, San Francisco, CA, USA; 2IBPM, University La Sapienza, Roma, Italy; 3Vall d’Hebron Institute of

Oncology (VHIO), Barcelona, Spain; 4University of Cambridge, Cambridge, UK.

Francesc Bosch

Marta Crespo, Júlia Carabia, Pau Abrisqueta

INTRODUCCIÓ: El microambient dels ganglis limfàtics i el moll de l’òs (MO) és fonamental per la supervivència i proliferació de les cèl.lules de Leucèmia Limfàtica Crònica (LLC). La tirosina quinasa Syk és considerada un element clau de la senyalització derivada tant del BCR com de diversos receptors de quimioquines.

OBJECTIUS: Avaluar l'efecte de la inhibició de la proteína Syk per part de TAK659 en cèl.lules d’LLC cultivades en condicions que recreen el microambient dels centres proliferatius.

MATERIAL I MÈTODES: Cultiu de cèl•lules d’LLC primàries en suspensió o en co-cultiu amb cèl.lules estromals del MO, CD40L, oligodeoxynucleotides-CpGs i anti-IgM. Anàlisi de l’efecte de TAK659 en la senyalització a través del BCR, la viabilitat, la proliferació i la quimiotaxi cap a les quimioquines derivades de les cèl.lules estromals CXCL12 i CXCL13.

RESULTATS: El tractament amb TAK659 va suprimir la fosforilació induïda per l’estimulació del BCR amb anti-IgM de Syk, AKT i ERK1/2. El co-cultiu va induir quimioresistència al tractament amb fludarabina, mentre que la DL50 de TAK659 va resultar de 39.75uM i 17.22uM per a les cèl.lules d’LLC en suspensió i en co-cultiu, respectivament. L’addició de TAK659 a la fludarabina aconseguí combatre sinèrgicament aquesta quimioresistència. El tractament amb TAK659 també va disminuir significativament la proliferació induïda pel co-cultiu, i la migració cap a CXCL12 i CXCL13.

CONCLUSIONS: Els senyals provinents del microambient reproduïts ex-vivo promouen proliferació i quimioresistència a les cèl.lules d’LLC. La TAK659 és capaç de combatre aquesta proliferació i supervivència, potenciar la citotoxicitat de la fludarabina i inhibir la quimiotaxi.

Joan Seoane

Ada Sala Hojman, Andrea Sáez Borderías, Joan Seoane Suárez

TGFβ pathway is extensively implicated in glioma. We have shown that TGFβ induces the self-renewal capacity of glioma-initiaing cells (GICs) through LIF, and that Id1 induction by TGFβ is required for GICs maintenance. However, TGFβ represses Id1 in epithelial cells through the same promoter region. Due to the relevance of Id1 in the regulation of GICs, our goal is to study how TGFβ and LIF induce Id1. Our hypothesis is that in GBM TGFβ induces Smad3 phosphorylation and LIF transcription, and LIF induces the phosphorylation of CREB. Finally, both factors (pSmad3 and pCREB) might bind to Id1 promoter and induce its transcription. Addressing the differential mechanism of Id1 induction, we reported that this antagonistic context-dependent response to TGFβ depends on the presence or absence of the transcriptional repressor ATF3. In epithelial cells, ATF3 is recruited to the Id1 promoter by an activated Smad complex. But ATF3 is not expressed in GICs, and the Smad complex can act as a transcriptional activator of the Id1 promoter. Interestingly, ATF3 and CREB, which belong to the same family, have the same binding site in Id1 promoter. It has been reported that pCREB is increased in GBM in comparison to non-tumoral cells. Thus, the dual response of Id1 to TGFβ may be controlled by a competition between ATF3 and pCREB for binding to Id1 promoter.

S. Schwarts, Jr

H.Pla1,2,3, D.Pulido1,3, N.García Aranda2, Y.Fernández2,3, F.Albericio3,4,5, I.Abasolo2,3 , M.Royo1,3 and S.Schwartz J,2,3 1 UQC-PCB,Combinatorial Chemistry Unit, Parc Científic de Barcelona, c/ Baldiri Reixac 10, 08028 Barcelona 2CIBBIM-Nanomedicine, Hospital Universitari Vall d’Hebron & Vall d’Hebron Institut de Recerca, Passeig Vall d’Hebron (19-129), Universitat Autònoma de Barcelona, 08035 Barcelona 3 CIBER-BBN, Bioingeniería, Biomateriales y Nanomedicina, Barcelona 4 IRB, Institut of Research in Biomedicine, Parc Científic de Barcelona, c/ Baldiri Reixac 10, 08028 Barcelona 5 UB, Department of Organic Chemistry, Universitat de Barcelona, Martí Franquès 1, 08028 Barcelonar.2,3

Goals: We aim to improve the efficacy of the actual treatment of advanced CRC and reduce its side effects by the use of polymeric nanoconjugates based on 5-Fluorouracil (5FU). Precisely, two PDC of PGA biodegradable carrier and 5FU will be studied, one of them with a matrix metaloprotease sensitive linker. Material and Method: Two different PDC (PGA-5FU and PGA-MMP7-5FU) were synthesized according to the linker nature between the carrier and the drug. In PGA-MMP7-5FU the linker is an MMP7-sensitive peptide (RPLALWRS), whereas in PGA-5FU the linker is non-specific. PDCs nanometric size was confirmed by DLS. Total-drug loading (TDL) of PDCs and 5FU release kinetics were measured by HPLC. PGA- 5FU cell internalization was studied by confocal microscopy and its cytotoxicity vs free 5FU was evaluated by MTT assays in different CRC cell lines. In vivo biodistribution of flourophore labeled PGA-5FU were studied in human colon cancer tumours growing in athymic nude female mice. Results: Both PDCs, PGA-5FU and PGA-MMP7-5FU, showed therapeutic activity in CRC cell lines, and obtained IC50 values were in the same order of magnitude as 5FU. Cell internalization of PGA-5FU and lisosomal colocalization was confirmed after 30 min incubation. Moreover, in vivo Fluorescence Imaging showed PGA-5FU tumour accumulation (maximal at 4 h post-administration) and compound kidney accumulation reported renal excretion route. Conclusions: The PDCs synthesized are rising candidates to increase 5FU chemotherapy treatment. Both showed therapeutic efficacy in vitro and in vivo, PGA-5FU demonstrated a promising biodistribution profile, with a fast tumour-accumulation and renal excretion. biodistribution profile, with a fast tumour-accumulation and renal excretion.

Diego Arango del Corro

Paulo Rodrigues, Irati Macaya, Sarah Bazzoco, Fernando Cartón, Elena Andretta Diego Arango.

RHOA has been found to be deregulated in distinct types of cancer but little is known of its functional relevance in colorectal cancer-CRC. Cell systems with stable downregulation of RHOA were generated and used to study the effects of the loss of RHOA in the tumorigenic capabilities of CRC cells. The “cross-talk” between RHOA and Wnt signaling pathway was studied through the analysis of the activity/expression of TCF4, cMYC and other downstream targets of Wnt. A mouse model expressing an inducible dominant negative-DN mutant of RHOA in the intestine was used to study the effects of the loss of RHOA on in vivo tumor progression. Cells in which RHOA was inactivated demonstrated significantly higher proliferation, migration, invasion, anchorage independent growth, and significant reduction in polarization and markers of enterocyte differentiation. This is consistent with the observation that cell lines where RHOA was downregulated showed significantly higher levels of Wnt activity, a hallmark of CRC. In vivo xenografts of the generated models, showed higher tumorogenic growth in the cells with abolished RHOA expression. Animals expressing a DN mutant of RHOA showed compromised intestinal brush border architecture, and increase in the number of intestinal tumors which was accompanied by a significant decrease in survival. RHOA expression was associated with shorter overall survival and disease-free survival in CRC patients, and low RHOA levels were observed in lymph node and distant metastases sites when compared to the primary tumors of CRC patients. These results indicate that RHOA is a tumor suppressor gene in CRC.

José García Arumí

Dr. Ibane Abasolo Olaortua, Dr. Marina Riera Gibernau, Carla Sanjurjo Soriano, Dr. Simo Schwartz Navarro

Objective: The aim of this study is to evaluate the anti-angiogenic and anti-proliferative properties of Pigment Epithelium-Derived Factor (PEDF) and Somatostatin (SST) in several in vitro models, as a preliminary proof-of-concept assay before testing their in vivo efficacy for the treatment of proliferative diabetic retinopathy (PDR). Methods: Human umbilical vein endothelial cells (HUVEC) were used as an endothelium model. Three different assays were performed: a cell viability MTT assay, a migration assay using the wound healing protocol, and a tubule-formation assay with a co-culture of fibroblasts and HUVEC. In all cases, cells were treated with the growth factor VEGF to assess the inhibitory activity of PEDF and SST. Results: both PEDF and SST were shown to inhibit HUVEC proliferation under VEGF treatment in a dose-dependent manner, obtaining the best results at 10 nM (30.5%) and 100 uM (49.1%) respectively. Both PEDF and SST at their highest concentrations inhibit the migratory action of VEGF by 33.9% and 27.4%. Finally, the tubule-formation assay showed the anti-angiogenic effects of PEDF and SST, reducing significantly the number of tubules in 28.8% and 50.7%, and the connections between them in 57.1% and 67.8%. Conclusions: Both PEDF and SST at 10 nM and 100 uM respectively inhibit HUVEC proliferation, migration and tubule formation under VEGF treatment, confirming its anti-angiogenic effect on endothelial cells. This in vitro data suggest that PEDF and SST could be promising tools for the treatment of PDR.

Rafael Simó Canonge

Cristina Sáez López, Federico Soriguer, Cristina Hernández, Gemma Rojo Martínez, Elehazara Rubio Martín, Rafael Simó y David Martínez Selva.

OBJETIVOS: Los niveles bajos de SHBG en plasma son un factor independiente de riesgo de padecer enfermedad cardiovascular (EC). La dieta Mediterránea está asociada con un menor riesgo de padecer EC. La finalidad de este estudio es investigar si el aumento de ácidos grasos monoinsaturados (MUFAs) asociado al consumo de aceite de oliva incrementa los niveles plasmáticos de SHBG e investigar los mecanismos moleculares asociados. MATERIALES Y MÉTODOS: Estudio observacional: 315 hombres (cohorte Pizarra) usaron aceite de oliva vs otros tipos de aceite para cocinar durante un mes. Se midieron los niveles plasmáticos de SHBG y MUFAs en los fosfolípidos. En experimentos in vitro, las células HepG2 fueron tratadas con vehículo, oleil-CoA y linoleil-CoA. Se midió la SHBG secretada al medio por ELISA y los niveles de mRNA y proteína de los factores de transcripción HNF-4α y PPARγ en las células. RESULTADOS: Los niveles circulantes de SHBG fueron significativamente mayores en los consumidores de aceite de oliva. Los niveles de SHBG se correlacionaron positivamente con MUFAs y negativamente con ácidos grasos saturados. Los estudios in vitro revelaron que oleil-CoA incrementa la producción de SHBG disminuyendo la transcripción de PPARγ en HepG2. CONCLUSIONES: Los consumidores de aceite de oliva presentan un aumento en los niveles de SHBG circulante. Nuestros resultados sugieren que el aumento en SHBG es debido a una inhibición de la transcripción de PPARγ hepático mediada por el ácido oleico. PUBLICACIONES: Artículo “Oleic Acid Increases Hepatic Sex Hormone-Binding Globulin Production in Men”, aceptado en la revista Molecular Nutrition & Food Research. Sáez-López C, Soriguer F, Hernández C, Rojo-Martínez G, Rubio-Martín E, Simó R, Selva DM Póster "Effects of Oleic and Linoleic Acids on Hepatic Sex Hormone-Binding Globulin Production". Cristina Sáez-López, Cristina Hernández, Rafael Simó y David M. Selva en el European Congress of Endocrinology (ECE 2013), Copenhague, Mayo 2013.

Daniel Serón Micas

Francesc Moreso, MD, PhD1; Irina B. Torres, MD1; Monica Martínez-Gallo, MD, PhD2; Susana Benlloch, PhD3; Carme Cantarell, MD1; Manel Perelló, MD1; José Jimeno, MD, PhD3; Ricardo Pujol-Borrell, MD, PhD2; Daniel Seron, MD, PhD1. Affiliations: Nephrology1 and Immunology2 Divisions. Hospital Universitari Vall d’Hebron. Universitat Autònoma Barcelona. Pangaea Biotech S.L3.

Background and objectives. In kidney transplants operational tolerance has been associated with up-regulation of B cell differentiation genes and an increased number of total, naive and transitional peripheral B cells. The aim is to evaluate tolerance biomarkers in different cohorts of stable renal transplants under immunosuppression. Design, setting participants and measurements. This is a cross-sectional study conducted in renal transplants. We evaluate genetic tolerance signature and lymphocyte subsets in stable transplants treated with calcineurin inhibitors (CNI) at 1 (n=15), 5 (n=14) and 10 (n=16) years, and azathioprine-treated transplants followed 30 years (n=8). Healthy volunteers (n=10) and patients with chronic rejection (n=15) served as controls. Results. We confirm that peripheral expression of IGKV1D-13 and IGKV4-1 genes by RT-PCR distinguish true tolerant (n=10) from stable transplants (n=10) provided by the International Tolerance Network. Tolerance signature was defined as the lowest expression for both genes in tolerant patients. In CNI-treated patients, genetic signature of tolerance and B cells showed a time-dependent increase not observed in azathioprine-treated patients (p<0.01). Genetic tolerance signature was observed in 0% at 1, 7% at 5 and 25% at 10-years while it was not observed in azathioprine-treated and chronic rejection patients. Fifteen out of 16 CNI-treated transplants at 10 years were revaluated 3 months apart. Nine did not show the tolerance signature in any determination, 4 in one and 2 in both determinations. Genetic signature of tolerance was associated with an increase of total, naive and transitional B cells (p<0.05). Conclusions. IGKV1D-13 and IGKV4-1 gene expression and its linked B cell populations increase during follow up in CNI-treated patients. At 10 years, 2 out of 15 CNI treated patients consistently express biomarkers associated with true tolerance. In azathioprine-treated patients these biomarkers were down-regulated

Ramon Martí Seves

Àstrid Brull Cañagueral, Tomàs Pinós Figueras, Noemí de Luna Salvà, Rosa Maria Escorihuela

La malaltia de McArdle és una miopatia autosòmica recessiva causada per mutacions al gen de la glicogen fosforilasa muscular (PYGM) generant un dèficit de l’enzim miofosforilasa. Clínicament, aquesta malaltia es caracteritza per una intolerància a l’exercici amb fatiga prematura, rampes, miàlgies, contractures, i nivells elevats de creatina quinasa en sèrum. També poden presentar episodis de rabdomiolisi i mioglobinúria i en els casos més greus, la fallada renal. Recentment, el nostre laboratori ha desenvolupat un model murí per aquesta malaltia que presenta la mutació més comú en PYGM (pR50X). Amb l’objectiu de dur a terme una caracterització fenotípica del model animal murí amb fons genètic homogeni, i avaluar les capacitats motores i cognitives, s’han realitzat tant anàlisis bioquímiques, moleculars com histològiques, així com tests motors i cognitius. Els ratolins homozigots per a la mutació pR50X no presenten activitat de la isoforma muscular de la glicogen fosforilasa ni expressió d’aquesta en múscul. Les anàlisis histològiques i bioquímiques revelen una gran acumulació de glicogen en múscul en els ratolins homozigots, cosa que no s’observa en els ratolins control. Pel que fa als tests motors i cognitius, s’observa una gran disminució de les capacitats motores en els ratolins homozigots, així com petites diferències pel que fa al seu comportament respecte als ratolins control. Aquest model reprodueix la simptomatologia clínica que s’observa en pacients, i representa una eina potent per futurs estudis de la fisiopatologia de la malaltia i altres trastorns neuromusulars, i per trobar possibles aproximacions terapèutiques. Publicacions: Knock-in mice for the R50X mutation in the PYGM gene present with McArdle disease. Nogales-Gadea G, Pinós T, Lucia A, Arenas J, Camara Y, Brull A, de Luna N, Martín MA, Garcia-Arumí E, Martí R, Andreu AL. Brain. 2012 Jul;135(Pt 7):2048-57.

Joan Montaner

Lidia García-Bonilla, Mar Hernández-Guillamón, Anna Rosell

Objectius: Comprovar l'eficàcia i la seguretat de l'administració de simvastatina durant la fase aguda de la isquèmia cerebral mitjançant un model embòlic en rata. Mètodes: Aquest estudi es composa de 4 subestudis. En el primer s’estudià l’eficàcia avaluant el dèficit neurològic i el volum d'infart en administrar simvastatina 15 minuts post isquèmia. Al segon s’avaluaren les complicacions hemorràgiques associades a la isquèmia cerebral i condicionades pel tractament amb rt-PA utilitzant rates hipertenses a les quals s’administrà rt-PA en combinació o no de simvastatina. En el tercer s’estudiaren les infeccions respiratòries secundàries a la isquèmia avaluant mostres de teixit pulmonar. En el quart s’exploraren els mecanismes del tractament amb simvastatina mitjançant un estudi proteòmic a partir d’homogenats cerebrals de rata. Resultats: Es comprovà un efecte beneficiós de la simvastatina en termes de millora del dèficit neurològic i reducció del volum d'infart i no s’observaren diferències entre les rates que van rebre únicament rt-PA i les que van rebre rt-PA i simvastatina pel que fa a la incidència d'hemorràgies. El model embòlic en rata no va resultar adequat per a valorar l'efecte de la simvastatina sobre l'aparició d'infeccions secundàries. Finalment, es demostrà que l’efecte neuroprotector de la simvastatina podria explicar-se per l’atenuació de l’estrès oxidatiu i la protecció de la barrera-hemato-encefàlica després de la isquèmia. Conclusions: Els resultats confirmen tant l'eficàcia com la seguretat enfront de l'aparició de transformacions hemorràgiques de les estatines (simvastatina) en la isquèmia cerebral i suggereixen el seu ús com a medicament neuroprotector en el tractament de l'ictus. Publicacions: 1. García-Bonilla L, Campos M, Giralt D, Salat D, Chacón P, Hernández-Guillamon M, Rosell A, Montaner J.Evidence for the efficacy of statins in animal stroke models: a meta-analysis. Journal of Neurochemistry 2012; 122: 233–243 2. Combining statins with tPA treatment after experimental and human stroke: a safety study on hemorrhagic transformation. Mireia Campos, Lidia García-Bonilla, Mar Hernández-Guillamon, Verónica Barceló, Anna Morancho, Manolo Quintana, Marta

Rubiera, Anna Rosell, Joan Montaner. CNS Neuroscience and therapeutics (Accepeted August 2013. In press) 3. Brain proteomics identifies novel neuroprotective actions of simvastatin in a rat embolic stroke model. Mireia Campos, Nelida Salvador, Marta Monge, Francesc Canals, Lidia García-Bonilla, Mar Hernández-Guillamon, Pilar Chacón, Anna Rosell, Alberto Alcazar, Joan Montaner (Submitted to Journal of Proteomics in September). 4. Embolic Middle Cerebral Artery occlusion in rat is not a suitable model for the study of stroke-induced spontaneous infections. Mireia Campos, Mar Hernández-Guillamón, Anna Rosell, Mª Ángeles Montero, Javier Gomis, David Salat, Lidia García-Bonilla, Joan Montaner (Submitted to Neuroscience Letters in October 2013).

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Joan X. Comella

Koen M.O. Galenkamp, Paulina Carriba, Jorge Urresti Ibañez, Fernando Marques-Fernandez, Laura Planells-Ferrer, Joquin Lopez-Soriano, Rana S. Moubarak, Joan X. Comella

Objective: Neuroblastoma cell lines are known to lack an apoptotic response after death receptor activation, due to inhibition of their signalling or downregulation of caspase 8. In this study we aimed to sensitize neuroblastoma cell lines to death receptor-induced apoptosis by treating with death ligands TNFα and FasL. Results: SK-N-AS neuroblastoma cells double-treated for 24H with FasL and TNFα showed an increase in cell death when compared to the untreated and single-treated controls. Pretreatment for 24H with FasL or TNFα revealed that TNFα sensitizes to FasL-induced cell death. In SK-N-AS cells, TNFα induced upregulation of Fas expression and thereby increased its cell surface exposure. Inhibition of PI3K, MEK1, JNK and NF-κB indicated that the TNFα–induced upregulation of Fas is mediated through NF-κB-induced transcription of the Fas gene. All studied neuroblastoma cell lines showed NF-κB activation, as assessed by IκBα degradation, however, only the cell lines SK-N-AS, SK-N-SH and Tet21N responded with an increase in Fas expression. In these cells, the upregulation of Fas expression increased the amount of cell death induced by FasL (SK-N-AS and Tet21N) or sensitized the cells to FasL (SK-N-SH). Conclusions: In some neuroblastoma cell lines, TNFα is able to induce upregulation of Fas expression through the activation of NF-κB-mediated transcription of the Fas gene. Upregulation of Fas expression is concomitant with cell surface exposure of the death

receptor and the sensitization of neuroblastoma cell lines to FasL-induced cell death and/or increase the apoptotic-response to FasL treatment.

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Miquel Vila

A. Recasens, B. Dehay, J. Bové, I. Carballo-Carbajal, S. Dovero, A. Pérez, P.O. Fernagut, J. Blesa, A. Parent, C. Perier, I. Fariñas, J. A. Obeso, E. Bezard i M. Vila.

Objectiu: Evidències recents indiquen que les lesions neuropatològiques d’α-sinucleina agregada característiques de la malaltia de Parkinson (MP), anomenades cossos de Lewy (CL), podrien autopropagar-se a diferents regions cerebrals per un mecanisme de transmissió cèl•lula-cèl•lula, contribuint així a la progressió de la MP. Per exemple, s’ha demostrat que la injecció de fibril•les sintètiques d’α-sinucleina al cervell de ratolins donen lloc a una patologia similar a la de la MP. Tot i així, el significat fisiopatològic de l’α-sinucleina patològica humana, present als CL, és desconegut. Mètodes: Hem injectat estereotàxicament CL purificats de teixit cerebral postmortem de pacients amb MP a la substancia negra o estriat de ratolins wild-type o primats no humans (PNH). Resultats: En ratolins, una injecció nigral de CL resulta en una neurodegeneració progressiva de la via dopaminèrgica nigroestriatal que comença als terminals dopaminèrgics de l’estriat i dóna lloc a dèficits motors. A l’inici de la denervació induïda pels CL, l’α-sinucleina endògena murina adopta una conformació patològica i s’acumula al citoplasma de neurones nigrals. El procés patogènic induït pels CL requereix tan l’α-sinucleina humana present als CL com l‘expressió endògena d’α-sinucleina. En PNH, la injecció nigral o estriatal de CL causa una degeneració nigroestriatal i una acumulació d’α-sinucleina patològica en vàries regions cerebrals anatòmicament interconnectades. Conclusions: Aquests resultats revelen un efecte patogènic de l’α-sinucleina humana present als CL tant en ratolins com en PNH. Publicacions: A. Recasens, B. Dehay, J. Bové, I. Carballo-Carbajal, S. Dovero, A. Pérez, P.O. Fernagut, J. Blesa, A. Parent, C. Perier, I. Fariñas, J. A. Obeso, E. Bezard i M. Vila. Lewy Bodies from Parkinson Disease Brains Transmit alpha-Synuclein Pathology in vivo. Under 2nd revision, Annals of Neurology.

Alfons Macaya Ruiz

Marta Vila-Pueyo, Noèlia Fernández-Castillo, Bru Cormand, Patricia Pozo-Rosich, Alfons Macaya

Introduction: Migraine with aura (MA) is a subtype of migraine characterized by reversible neurological disturbances preceding headache. Epigenetic mechanisms are postulated to mediate migraine susceptibility. MA neurophysiological correlate is cortical spreading depression (CSD), a wave of neuronal depolarization and depression that is suppressed by chronic administration of the migraine preventive drugs valproate and topiramate. Objective: Using a CSD rat model, we investigated if valproate and topiramate lower the susceptibility to develop CSD by inducing changes in brain DNA methylation. Materials and methods: Adult male Sprague-Dawley rats were treated with valproate, topiramate or saline for 4 weeks. CSDs were KCl-induced for 1 hour. Cortices were removed and DNA was extracted to perform MBD-based genome-wide methylation sequencing. Results were analyzed by MEDIPS software to obtain differentially methylated regions (DMRs). Genes containing DMRs were analyzed for GO and KEGG pathways enrichment and gene networks were constructed using IPA software. Results: Both treatments, as expected, reduced CSD intensity. Both drugs induced differential methylation (DM) of genes belonging to “synapse activity” pathways including Rimbp2, Pcdhb3 and Grip1. Treatment with valproate was associated with DM of ion channel encoding-genes such as Asic2 and Kcnd2. CSD, in turn, promoted DM of genes involved in axon growth (Cdh13), modulation of pain (Mrgprx3) or neuronal proliferation (Mapk10). Conclusions: These results confirm that valproate and topiramate protect against CSD in the rat and that this is associated with changes in brain DNA methylation. Further analyses are required to determine the potential of the identified targets in determining susceptibility to human migraine.

Dr. C. Manichanh

Suchita Panda, Ismail El khader, Francesc Casellas, Josefa López Vivancos, Montserrat García Cors, Silvia Cuenca, Francisco Guarner and Chaysavanh Manichanh

Introduction and goals: From birth onwards, the human gut microbiota rapidly increases in diversity and reaches an adult-like stage at three years of age. After this age, the composition may fluctuate in response to external factors such as antibiotics. Previous studies have shown that resilience is not complete months after cessation of antibiotic intake. We aim to know about the short-term effects of antibiotic intake on the gut microbial community. Methods: We examined the load and composition of the faecal microbiota immediately after treatment in 21 patients, who received broad-spectrum antibiotics such as fluoroquinolones and beta-lactams. A faecal sample was collected from all participants before treatment and one week after for microbial load and community composition analyses by quantitative PCR and pyrosequencing of the 16S rRNA gene, respectively. Results: Fluoroquinolones and beta-lactams significantly decreased microbial diversity by 25% and reduced the core phylogenetic microbiota from 29 to 12 taxa. However, at phylum level, these antibiotics increased the Bacteroidetes/Firmicutes ratio (P = 0.0007, FDR = 0.002). At species level, our findings unexpectedly revealed that both antibiotic types increased the proportion of several unknown taxa belonging to the Bacteroides genus, a Gram-negative group of bacteria (P = 0.0003, FDR < 0.016). Counter intuitively, the average microbial load was not decreased after treatment. Rather, the beta-lactams increased it significantly by two-fold (P = 0.04). Conclusions: The unexpected increase detected in microbial load and the selection of Gram-negative over Gram-positive bacteria breaks the idea generally held about the effect of broad-spectrum antibiotics on gut microbiota. Publicacions: Panda S, et al., Short-term Effect of Antibiotics on Human Gut Microbiota. 2013; Aliment Pharmacol Ther. (UNDER REVIEW)

Benito Almirante

Mireia Puig, Benito Almirante

Background. The epidemiology of Candida bloodstream infections varies over time and between geographical areas, and should be monitored. We aimed to describe the present epidemiology in Spain and investigate risk factors for death. Methods. A prospective, multicenter, population-based surveillance program for candidemia was implemented in 5 metropolitan areas of Spain from May 2010-April 2011. Candida isolates were centralized to a reference laboratory for species identification by DNA sequencing and for susceptibility testing by EUCAST reference procedure. Prognostic factors associated with early (3-7 days) and late (8-30 days) death were analyzed using logistic regression modelling. Results. We detected 773 episodes: annual incidence 8.1 cases/100 000 inhabitants, 0.89/1000 admissions, and 1.36/10 000 patient-days. Highest incidence was found in infants < 1 year (96.4/100 000 inhabitants). C. albicans was the leading agent (45.4%), followed by C. parapsilosis (24.9%), C. glabrata (13.4%), and C. tropicalis (7.7%). Overall, 79% of Candida isolates were susceptible to fluconazole. Cumulative mortality at 7 and 30 days was 13.2% and 31.3%, respectively. Multivariate analysis showed that therapeutic measures within the first 48h may improve early mortality: 0.51, 95% confidence interval [CI] 0.27-0.95) and central venous catheter removal (OR 0.43, 95%CI 0.21-0.87). Predictors of late death included host factors (eg, patients’ comorbid status and signs of organ dysfunction), primary source (OR 1.63, 95%CI 1.03-2.61), and severe sepsis or septic shock (OR 1.77, 95%CI 1.05-3.00). Conclusions. The proportion of Candida isolates categorized as intermediate or resistant to fluconazole is remarkable. Early mortality may be improved with strict adherence to guidelines. Publications: Puig-Asensio M, Padilla B, Garnacho-Montero M, et al. Epidemiology and predictive factors for early and late mortality in Candida bloodstream infections: a population-based surveillance in Spain. Clin Microbiol Infect 2013 [epub ahead of print]. doi: 10.1111/1469-0691.12380.

Sara Marsal

Arnald Alonso, Raül Tortosa, Antonio Julià, Sara Marsal

Objectius: Desenvolupar noves eines bioinformàtiques destinades a automatitzar, integrar i millorar el workflow d’anàlisi metabolòmic sobre dades d’espectrometria de masses (EM) i de ressonància magnètica nuclear (RMN) Material i mètode: El primer mètode, AStream, permet processar dades d’EM per accelerar el procés d’identificació de metabòlits amb significat biològic dins el context de la malaltia estudiada. D’altra banda, FOCUS és un workflow d’anàlisi complet per a estudis high-throughput de RMN: des del processat del senyal, passant per la normalització i finalment per la identificació dels metabòlits associats a malalties. Demostrem el rendiment d’ambdues tècniques utilitzant dades d’estudis biomèdics per identificar i quantificar els metabòlits presents en mostres d’orina (N=60) i d’extractes de fetge (N=120). Els resultats varen ser validats per personal tècnic especialitzat en metabolòmica. Resultats: El procés d’anàlisi metabolòmic, fins ara manual i amb un gran requeriment d’hores de personal tècnic especialitzat, pot ésser realitzat en qüestió de minuts amb els nous softwares AStream i FOCUS. Sobre els dos conjunts de dades estudiats (i.e. orina/fetge) es varen obtenir, respectivament, 240/228 pics dels quals 43/63 s’associen inequívocament a un metabòlit. Aquests resultats demostren una millora molt significativa respecte a metodologies prèvies, integrant totes les etapes d’anàlisi necessàries, reduint considerablement els temps necessari per al processat i facilitant la interpretació biològica dels resultats. Conclusions: AStream i FOCUS suposen un salt qualitatiu en l’anàlisi de dades metabolòmiques, permetent a l’investigador biomèdic analitzar, per primera vegada, dades metabolòmiques de grans nombres de mostres de forma integrada, ràpida i robusta. PUBLICACIONS: FOCUS: A robust workflow for one-dimensional NMR spectral analysis. A Alonso, MA Rodriguez, M Vinaixa, R Tortosa, X Correig, A Julià, S Marsal. Enviada a: Analytical Chemistry (2013)

AStream: an R package for annotating LC/MS metabolomic data. A Alonso, A Julià, A Beltran, M Vinaixa, M Díaz, L Ibañez, X Correig, S Marsal. Bioinformatics 2011, 27 (9), 1339

José Alegre Martín

Eva Ruiz, Naia Saez, Luisa Aliste, Jesus Castro, Natalia Calvo.

Objectius: analitzar les diferències clíniques i en la valoració de la fatiga en pacients amb SFC segons la seva situación laboral. Material i mètodes: estudi transversal amb casos consecutius sobre un registre de base poblacional. Ambit: hospital universitari. Subjectes: pacients diagnosticats de SFC Periode d’estudi: gener 2008 – març 2011. Criteris d’ inclusió: diagnòstic de SFC segons els criteris de Fukuda. Acceptació de participació mitjançant consentiment informat. Variables: Sociodemogràfiques: edat, sexe, situació laboral. Clíniques: edat d’inici, temps d’evolució i nivell de fatiga i de dolor; presència de son no reparador i de cefalea recurrent; simptomatologia cognitiva, neurològica, disfunció neurovegetativa. Qüestionaris: impacte de fatiga, intensitat de fatiga, qualitat de vida SF-36. Resultats: es van incloure 822 pacients dels quals el 25% (210) estaven actius. El percentatge i la severitat de simptomatologia cognitiva, neurològica i disfunció neurovegetativa van ser majors als pacients inactius (p<0.05). L’edat d’inici de la fatiga i del dolor va ser posterior als pacients inactius (p<0.05). La intensitat de la fatiga va ser major als pacients inactius (p<0.05). Els pacients inactius van puntuar més alt a les escales d’impacte de fatiga així com a l’escala d’intensitat de fatiga (p<0.005). Les puntuacions del qüestionari de qualitat de vida van ser menors els pacients inactius (p<0.05). Conclusions: elevat percentatge de minusvalia laboral als pacients amb SFC. Els pacients inactius presentaven més simptomatologia neurológica, cognitiva i neurovegetativa i majors puntuacions a les escales d’impacte de fatiga i menor qualitat de vida.

José Luis Peiró

1. Alejandra Fernández Martín - a 2. Mario Marotta Baleriola - a, c 3. Cesar García Fontecha - a, b 3. José Luis Peiró Ibáñez- a, b, c a: Grupo de Bioingeniería, Ortopedia y Cirugía Pediátricas, Institut de Recerca Vall d’ Hebrón, Barcelona, Spain. b: Servicio de Cirugía Hospital Materno Infantil. Hospital Universitario Vall d’ Hebrón. Barcelona, Spain. c: Center for Fetal, Cellular and Molecular Therapy. Cincinnati Children's Hospital Medical Center (CCHMC). Cincinnati (Ohio), USA.

OBJETIVOS: El mielomeningocele (MMC) es una malformación por falta de cierre del tubo neural que provoca secuelas devastadoras en la vida postnatal. La terapia celular se plantea como regenerativa de la lesión neural progresiva. En este estudio, en modelo ovino de MMC se analizaron los líquidos amnióticos (LA) de ovejas sanas y con MMC para caracterizar los distintos tipos celulares e identificar y aislar precursores neurales. MATERIALES Y MÉTODOS: En modelo ovino, la lesión de mielomeningocele se creó a día 75 de gestación, y se obtuvo muestra de LA (control). A día 95 de gestación la lesión se reparó y antes se tomó muestra de LA, representativa de MMC. Ambos LA se mantuvieron en cultivo durante quince días, y en las células obtenidas se analizaron marcadores de pluripotencialidad, precursores neurales y células de linaje neural mediante inmunocitofluorescencia. RESULTADOS: Células específicas aisladas de las muestras de MMC mostraron expresión de marcadores específicos de progenitores neurales, que por el contrario no se observaron en células de LA sano. Además, las células de MMC no mostraron expresión de marcadores normalmente negativos en progenitores neurales, corroborando su linaje neural. CONCLUSIONES: El LA de muestras de MMC contiene células con características de progenitores neurales que podrían ser utilizadas para la reparación de la médula espinal. En caso de demostrar regeneración medular mediante la utilización de precursores neurales obtenidos de LA, se generarían nuevas estrategias terapéuticas para el tratamiento del MMC.

Elisa Llurba Olivé

E. Llurba, O. Sánchez, Q. Ferrer, KH. Nicolaides, A. Ruiz, J. Sánchez-de-Toledo, G. Soro, S. Arévalo, M. Goya, B. García-García, S. Pérez-Hoyos, J. Alijotas-Reig, MC. Domínguez, E. Carreras, L. Cabero.

Animal models showed that angiogenesis is related to abnormal heart development. Our objectives were to ascertain whether a relationship exists between congenital heart defects (CHD) and angiogenic/anti-angiogenic imbalance in maternal and fetal blood. Methods and Results: Maternal and cord blood placental growth factor (PlGF), soluble fms-like tyrosine kinase-1(sFlt-1) and soluble endoglin(sEng) at 18-37 weeks’ gestation were compared in 61cases of isolated major fetal heart defects and 204normal controls. Angiogenic factor expression and markers of hypoxia were measured in heart tissue from 23fetuses with CHD and 8controls from terminated pregnancies. In the heart defect group, compared to controls, the maternal PlGF multiple of median (MoM) was lower (0.94, IQR0.85-1.03 vs. 1.03, IQR0.95-1.09; p<0.0001), sFlt1 MoM was higher (1.03, IQR0.95-1.08 vs. 0.99, IQR0.93-1.07 ; p=0.0325) and no differences were found in sEng MoM (1.03, IQR0.89-1.17 vs. 1.01, IQR0.83-1.18; p=0.3504). Fetuses with CHD had higher cord plasma sFlt-1 (324pg/mL, IQR238-585 vs. 241pg/mL, IQR192-335; p=0.0102) and sEng (6.83ng/mL, IQR5.61-8.82 vs. 4.50ng/mL, IQR2.53-7.45, p=0.0006) levels and no differences in PlGF (12.90pg/mL,IQR2.50-31.29 vs. 13.50pg/mL, IQR4.69-21.99; p=0.4798). Expression of VEGF, sFlt1, markers of chronic hypoxia and antioxidant activity were significantly higher in heart tissue from fetuses with major CHD compared to normal hearts (VEGF,1.59-fold; sFlt-1, 1.92-fold; HIF- -fold; HO-1,1.62-fold; SOD1,1.31-fold ). Conclusions: An intrinsically angiogenic impairment exists in CHD that appears to be present in both the maternal and fetal circulation and fetal heart. Our data suggest that the role of angiogenesis is essential in human heart development. Publicacions: - Maternal serum placental growth factor at 11–13 weeks’gestation and fetal cardiac defects. E. LLURBA*†, A. SYNGELAKI‡§, O. SA´ NCHEZ†, E. CARRERAS*, L. CABERO* and K. H. NICOLAIDES. Ultrasound Obstet Gynecol 2013; 42: 169–174 Published online 26 June 2013 - Maternal and fetal anti-angiogenic imbalance in congenital heart defects. *E. Llurba1,2, O. Sánchez2, Q. Ferrer3, KH. Nicolaides4,5, A. Ruiz1, J. Sánchez-de-Toledo6, G. Soro1, S. Arévalo1, M. Goya1, B. García-García1, S. Pérez-Hoyos7, J. Alijotas-Reig8, MC. Domínguez2, E. Carreras1, L. Cabero1. Acceptada per a publicació a "European heart Journal"

Antonia Agustí Escasany

Mª Luján Iavecchia, Ahmad Safiya, Antonia Agustí Escasany, Mònica Sabaté Gallego, Montse Bosch Ferrer, Alfons Biarnés Suñe, Mª Angels Camps Cervantes, Dolors Castellà, Pilar Lalueza Broto, Veronica Pons Escoll, Jordi Teixidor Serra, Maria del Mar Villar Casares

OBJECTIVE. To describe the perioperative management and outcomes of patients with high-moderate thrombotic risk treated with antithombotic therapy and admitted for hip fracture surgery. METHODS. A retrospective cohort of patients admitted from October 2011-March 2012 was analyzed. All patients were followed-up for three months after surgery. Treatment recommendations and thrombotic risk classification from the ACCP were used and information on haemorrhagic and thromboembolic events was collected. RESULTS. A total of 113 patients were admitted (median [IQR] age 86 [89-80]; 71% women) and 87% had a III-IV ASA risk category. Median hospital stay was 16 days (21-12). Ninety-eight (86.7%) patients had a high (13.3%) or moderate (73.5%) thrombotic risk. Of these, 16% were treated with oral anticoagulants and 84% with antiplatelet agents. Acenocumarol, clopidogrel and triflusal were interrupted in all cases before surgery and aspirin in 70% of cases. Acenocumarol was stopped a median of 7 [9-6] days before surgery, clopidogrel 9 [14-8], triflusal 4 [7.5-2.75] and aspirin 6 [8-3]. Acenocumarol treatment was resumed a median of 12 [26-7] days after surgery, clopidogrel 8.5 [10-6], triflusal 11 [19.5-5.5] and aspirin 6 [12-1]. In 7 cases antithrombotic therapy wasn’t resumed. Sixty-two patients had 80 bleeding events (70% major haemorrhages and 30% minors). Thirteen experienced 14 thrombotic events and 24 patients died (two due to haemorrhages and two to thromboembolic events). CONCLUSIONS. In most cases antithrombotic therapy was interrupted before surgery, even in patients treated with aspirin. Moreover, antithrombotic therapy was stopped more days before surgery and resumed later after surgery than is recommended. PUBLICATIONS: PERIOPERATIVE MANAGEMENT AND OUTCOMES OF PATIENTS WITH HIGH OR MODERATE THROMBOTIC RISK TREATED WITH ANTITHROMBOTIC THERAPY AND HIP FRACTURE SURGERY. Clinical

pharmacology congress of the Spanish Society of Clinical Pharmacology, 17 and 18 october, 2013, Cadiz. Communication poster.

Jaume Reventós

Devis L, Thomas W, Campoy I, Martinez E, Castellví J, Mancebo G, Carreras R, Cabrera S, Gil-Moreno A, Dufour S, Alameda F, Reventós J*, Colas E*

We have identified an ETS transcription factor, ETV5, associated with myometrial infiltration in endometrial carcinoma (EC). We have unveiled that ETV5 promotes EMT by direct regulation on E-Cadherin repressors, degrades the extracellular matrix through MMP2 activation and prevents apoptosis of tumor cells under oxidative stress conditions through Hep27 overexpression. A cDNA microarray comparing ETV5 overexpressing EC cell lines with control cells evidenced that most of the functions altered were related to cell adhesion, cell-cell contact and cellular junction, being the IgG superfamily one of the main families altered in ETV5 overexpressing cells. We have now confirmed that ETV5 regulates ALCAM mRNA expression by binding to its promoter region. At protein level, we observed that ETV5 increases ALCAM protein half-life but protein levels are not modified. In order to understand ALCAM’s role in adhesion, we used a micropipette dual assay for measuring the forces required to separate two adherent cells always in suspension to avoid impact of cell-matrix adhesion and signaling. Those experiments, performed with EC cells overexpressing ETV5 and ALCAM full length and/or ALCAM cytoplasmic-tail less, confirmed the necessity of ALCAM full length in adhesion formation and maturation and its relation to invasiveness. In relation to these in vitro observations, ALCAM was found to be downregulated in invasive front vs non-invasive sections of EC human tissues, and in tumors presenting an advanced stage vs tumors in initial stages. Those results were obtained by means of immunohistochemistry and immunblot. Altogether, we have characterized a new mechanism of ETV5-related invasion in EC. Publicacions: Autores: Núria Pedrola, Eva Colás, Marta Llauradó, Laura Devis, Irene Campoy, Elena Martínez, Marta Garcia, Laura Muinelo, Lorena Alonso, Miguel Abal, Francesc Alameda, Josep Castellví, Sílvia Cabrera, Antonio Gil, Xavier Matias-Guiu, Juan L. Iovanna, Jaume Reventós, Anna Ruiz

Título: NID1 and NUPR1 function downstream the ETV5 transcription factor to promote endometrial tumor progression and dissemination Referencia: Int J Cancer. 2013 (under review) Factor de Impacto: 6,198 Quartil/Decil: Q1 Autores: Colas E, Muinelo-Romay L, Alonso-Alconada L, Llaurado M, Monge M, Barbazan J, Gonzalez M, Schoumacher M, Pedrola N, Ertekin T, Devis L, Ruiz A, Castellvi J, Doll A, Gil-Moreno A, Vazquez-Levin M, Lapyckyj L, Lopez-Lopez R, Robine S, Friederich E, Castro M, Reventos J, Vignjevic D, Abal M. Título: ETV5 cooperates with LPP as a sensor of extracellular signals and promotes EMT in endometrial carcinomas. Referencia: Oncogene. 2012 Jan 23. Factor de Impacto: 6,373 Quartil/Decil: Q1/D1

Joan Seoane

Alba Gonzàlez Juncà, Fran Martínez Ricarte, Isabel Cuartas Maza, Sara Sanchez-Redondo Campos, Joan Seoane

Glioma, and its most malignant form, Glioblastoma (GBM) is one of the most lethal cancers, and despite the recent advances of the field, patients only have a median survival of 15 months. The standard therapy for GBM patients is surgical resection together with a combination of temozolomide and radiotherapy. Most GBM patients relapse shortly after treatment, which points out to the need of new therapeutic approaches to treat this disease. The TGF-beta pathway has been described to be crucial for many tumor types. Among others, TGF-beta controls glioma progression (Bruna et al, C Cell 2008) and initiation, (Penuelas et al, C Cell 2009) and hiperactivation of this signaling pathway correlates with poor prognosis in patients. For this reason, several companies have become interested in inhibiting the TGF-beta signalling and there are several clinical trials using inhibitors of the TGF-beta pathway in glioma patients. In our hospital, we are participating in a clinical trial using a small molecule inhibitor of the TGF-beta Receptor Type I (TBRI). We are using our preclinical patient derived xenograft (PDXs) mouse model in order to study the molecular mechanisms of the TGF-beta oncogenic effect in glioma as well as to study the response to the TBRI inhibitor.

In our collection of GBM-derived cultures and PDXs we are able to study the response to conventional therapies and to TGF-beta inhibitors. Our aim is to better understand the molecular mechanisms of response to TGF-beta inhibition in order find new biomarkers that could improve the therapeutical management of these GBM patients. Publicacions: Anido J, Sáez-Borderías A, Gonzàlez-Juncà A et al TGF-β Receptor Inhibitors Target the CD44(high)/Id1(high) Glioma-Initiating Cell Population in Human Glioblastoma.Cancer Cell. 2010 Dec 14;18(6):655-68.

Miguel F. Segura Guinard

Luz Jubierre, Aroa Soriano, Octavio A. Romero, Laia Paris, Rana Moubarak, Ana Almazán-Moga, Carla Molist, Josep Roma, Samuel Navarro, Rosa Noguera, Montserrat Sánchez-Céspedes, Soledad Gallego,José Sánchez de Toledo, Miguel F. Segura

GOALS: Neuroblastoma (NBL) is the most common solid tumor of infancy. High-risk categorized patients have poor prognosis and less than 40% of patients achieve long-term cure. Relapsed and metastatic tumors acquire multi-drug resistance raising the need of alternative treatments. Recently, new small molecule drugs that modulate the epigenetic landscape of tumors have been found to improve patient’s survival in adult cancers, indicating that epigenetic factors are promising therapeutic targets. Our goal is to analyze the implication of epigenetic genes in the progression and resistance to therapy of high-risk neuroblastomas. This project will lead to the identification new therapeutic targets that control the gene expression program of neuroblastoma cells. METHODS: To identify deregulated epigenetic genes, we analyzed multipe mRNA neuroblastoma data sets (GSE3960, GSE16237 and GSE16476). Epigenetic genes that showed differential expression in the high-risk NBL group with the same trend in the 3 independent data sets were selected for functional characterization. Loss of function experiments were performed by shRNA lentiviral infection of chemoresistant NBL cell lines. The effect of gene knockdown alone and/or in combination with chemotherapy was evaluated. RESULTS: We found that one member of the SWI/SNF chromatin-remodeling complex SMARCA4/BRG1, was consistently upregulated in the worst prognostic group of NBL (Stage 4 with MYCN amplified). SMARCA4 knockdown reduced the proliferation and viability of chemoresistant neuroblastoma cells. CONCLUSIONS: SMARCA4 is essential for the proliferation and viability of NBL cells and may represent a novel therapeutic target against chemoresistant NBL.

Jaume Reventós Puigjaner

Lucia Lanau, Marina Rigau, Blanca Majem, Tatiana Altadill, Josep Castellví, Assumpció Pérez-Benavente, Jose Luís Sánchez-Iglesias, Silvia Cabrera, Josep-Maria del Campo, Antonio Gil-Moreno, Andreas Doll, Mireia Olivan, Eva Colás, Anna Ruiz, Marta Llauradó and Jaume Reventós

Aim: Epithelial ovarian cancer (OC) is the most lethal gynecological malignancy and the fifth cause of cancer deaths in women in the Western-world. Largely asymptomatic, OC is frequently (75-80%) detected at late stage. Five-year survival rate for women with advanced stage disease is less than 20%, while the cure rate is 90% when OC is diagnosed at early stages. Epithelial OC metastasizes by direct extension from the ovary seeding the cells into the peritoneal cavity and to neighboring organs. Our aim is to study the OC dissemination comparing human paired ovarian primary tumors, ascites and metastases, in order to identify tumor progression biomarkers that could lead to the development of new specific therapies against metastatic OC. Methods: Fresh ovarian primary tumor, ascites and peritoneal metastases from patients suffering advanced serous OC were collected from surgery room and processed for further analysis. Formalin-fixed, paraffin-embedded (FFPE) tissues were collected from Pathology Department for immunohistochemical analysis. Discovery phase: We determined the global gene expression profile by microarray analysis from 5-paired fresh samples. Validation phase: We analyzed mRNA (RTqPCR) and protein (Western-blot), from 7 unpaired fresh samples. We further evaluated protein expression by immunohistochemistry from 10-paired tumor and metastasis FFPE-samples. Results: INHBA and FABP4 were significantly over-expressed in metastases versus primary tumors at both RNA and protein level. Conclusions: The present study highlights the role of novel candidates in OC dissemination that might be used as tumor biomarkers, to clinically monitor the progression of the disease, or as target therapies, to block OC dissemination.

Laura Soucek

Daniel Massó-Vallés, Erika Serrano, Nicole M. Sodir, Roderik Kortlever, Toni Jauset, Marie-Eve Beaulieu and Laura Soucek

Objectives: Assess the efficacy of Ibrutinib, an inhibitor of Bruton’s tyrosine kinase, in a mouse model of pancreatic ductal carcinoma. Materials and methods: Ibrutinib (PCI-32765) is a novel inhibitor of Bruton's tyrosine kinase (BTK) that blocks mast cell degranulation and is currently in clinical trials as a therapy for B cell malignancies that include chronic lymphocytic leukemia, multiple myeloma and B cell non-Hodgkin's lymphoma. We assessed the effect of Ibrutinib on mice bearing pancreatic ductal carcinomas (PDAC) and compared their overall survival with non-treated mice. We also compared overall survival of mice treated with the standard of care for PDAC, Gemcitabine, with mice treated with a combination of Ibrutinib and Gemcitabine. Results: Here, we show that systemic treatment of PDAC-bearing mice with Ibrutinib induces shrinkage of the tumor stroma by reducing inflammatory cell infiltration and collagen deposition, and seems to confer a survival advantage. Combinatorial therapy of Ibrutinib with standard of care significantly increased survival when compared to standard of care alone. Conclusions: Our data reinforce the notion that mast cell function is required for maintenance of the tumor stroma. We also demonstrate that the combination of standard of care chemotherapy with the BTK inhibitor Ibrutinib may be useful in treating PDAC.

Andreas Doll

Melània Montes, Mireia Olivan, Marina Rigau, Tamara Sequeiros, Eva Colás, Juan Morote, Jaume Reventós, and Andreas Doll

Background and aims: Prostate cancer (PCa) is the most common malignant neoplasia and the second leading cause of cancer death in men in developed countries. Although prostate specific antigen (PSA) has increased its detection during the last years, is not specific for PCa; therefore, it is necessary to search for more accurate diagnostic biomarkers. The aim of this study was to evaluate if microRNAs known to be deregulated in blood or tissue for PCa could also be detected in urine sediment, and characterize “in vitro” the most promising of them. Methods: Total RNA was isolated from 40 post-massage urine samples, which will undergo a prostatic biopsy as part of their routine medical care, due to the suspicion of PCa. Firstly, the expression level of 20 microRNAs was analyzed in PCa by RT-qPCR. Thereafter, there was a screening for more than 700 microRNAs by low density TaqMan arrays. Results: Data showed that several microRNAs were significantly deregulated in the urine of cancer patients compared with the benign controls (p-value<0.05). Subsequently, became functional analysis for 9 of these deregulated microRNAs in 3 PCa cell lines, were found 3 microRNAs that significantly affected proliferation, viability and/or migration. Conclusion: Results suggest that changes in the microRNA profiles can be detected in urine samples and could be good candidates as biomarkers to improve PCa diagnosis. Publications: AUTHORS: Melania Montes, Mireia Olivan, Tamara Sequeiros, Marta Garcia, Marina Rigau, Juan Morote, Jaume Reventós, and Andreas Doll TITLE: MicroRNA profiles as predictors of significant prostate cancer TYPE OF PRESENTATION: Poster CONGRESS: 20th Meeting of the EAU Section of Urological Research MEETING PLACE: Strasbourg. YEAR: 25-27 October 2012 AUTHORS: Melania Montes, Mireia Olivan, Tamara Sequeiros, Marta Garcia, Marina Rigau, Juan Morote, Jaume Reventós, and Andreas Doll TITLE: MicroRNA expression profiles in urine as diagnostic biomarkers for prostate cancer TYPE OF PRESENTATION: Poster

CONGRESS: SCB 1st Congres, Global Questions on Advanced Biology MEETING PLACE: Barcelona, Spain. YEAR: 9-12 July 2012

Andreas Doll

Sequeiros T, Rigau M, Campoy I, Montes M, de Torres I, Morote J, Reventós J, Oliván M and Doll A

Fast and reliable diagnosis of prostate cancer (PCa) is highly desirable but, due to the low sensitivity and high failure rate of current diagnostic methods, advanced disease is often encountered. Because of the location of the prostate in the body, its secreted products, including extracellular vesicles (EVs), can be detected in urine. The aim of this project is to identify new protein biomarkers from urinary EVs to improve PCa detection. Firstly, we established the ultracentrifugation based-method for isolation of EVs from urine samples. The obtained vesicles were characterized by electronic microscopy and Western blot, and quantified by Nanoparticle Tracking Analysis. Next, we carried out a discovery phase by label-free LC-MS/MS. We selected 24 samples: 8 benign samples, 8 low risk PCa samples (Gleason=7(3+4)) and 8 high risk PCa samples (Gleason>7). Twenty µg of protein were digested following the method called Filter-Aided Sample Preparation. Peptides were analyzed using an Orbitrap Velos mass spectrometer. Analysis of the data was carried out in two parallel ways: ion intensities and spectral counting. We have identified several differentially expressed proteinsbetween the 3 study groups. The next step will be the validation of these protein candidate markers, by targeted techniques such as Selected Reaction Monitoring in a new set of samples. In the future, this could improve current PCa detection, distinguishing aggressive from clinically insignificant PCa and other benign conditions and, therefore, avoiding PCa related over-diagnosis and over-treatment.

Diego Arango

Sarah Bazzocco, Paulo Rodrigues

Altered expression of EPH receptors has been shown to be an important step in the tumorigenic process in many types of human cancer. In a genome wide analysis EPHA3 was reported as the sixth most frequently mutated in human colorectal cancer. Many EPHA3 mutations identified impaired kinase activity or ephrin ligand binding and/or decreased the level of receptor cell surface localization, suggesting that EPHA3 has ephrin and kinase dependent tumor suppressing activities. To investigate the role of EPHA3 as tumor suppressor gene in colorectal tumorigenesis, we used inducible isogenic in vitro systems, xenografts and Epha3 Knockout (KO) mice models. We selected DLD1 and LS174T colon cancer cell lines which show low endogenous levels of EPHA3 and high EphrinA5 ligand expression to conditionally overexpress EPHA3 under doxycycline control. We found that the reintroduction of EPHA3 in DLD1 and LS174T did not alter their motility, the growth on solid substrate, soft agar or in a xenograft model. Moreover the ectopic expression of EPHA3 does not regulate invasive migration in colon cancer cells on a Boyden chamber assay. Also an Epha3 Knockout mouse model was used to show that Epha3 inactivation does not efficiently initiate tumorigenesis in the intestinal tract. In addition, when intestinal tumorigenesis is initiated either genetically (heterozygosis Apc mutations) or pharmacologically (azoxymethane), no differences were observed between EphA3+/+ and EphA3-/- mice in animal survival, tumor multiplicity size or histology. Our data indicate that the loss of EPHA3 is not an important event in the oncogenic colorectal cancer.

Diego Arango

Hafid Alazzouzi

A major hallmark of cancer is the deregulation of cell division, and rapid proliferation is often associated with poor patient prognosis. Here we investigated the molecular mechanisms responsible for rapid cancer cell growth. We characterized the proliferation rates of 54 colorectal cancer cell lines, and found that the doubling time ranges from 21h-69h (>3.2-fold). We then used mRNA microarray analysis of 31 of these cell lines to investigate the molecular determinants of growth differences. We found a significant enrichment in the number of genes with expression differences between rapid/slow proliferating lines that participate in different biological processes like mRNA processing (p=5.45x10e-18), translation (p=1.53x10e-15), and cell cycle (p=2.69x10e-8). We then hypothesized that genes showing high levels of expression in rapidly proliferating tumor cells could be good therapeutic targets. In good agreement, thymidylate synthetase (TS) was among the genes whose expression was best correlated with proliferation rates. Importantly, TS is the direct target of 5-fluorouracil, the gold standard for colorectal cancer treatment for >50 years. Other genes with significant expression/proliferation associations were protoporphyrinogen oxidase (PPOX) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). As extensively documented for TS, using specific chemical inhibitors we demonstrated that inactivation of PPOX and GAPDH efficiently inhibits the growth of colon cancer cells in vitro and in vivo using a xenograft model. In conclusion, we identified PPOX as a novel candidate chemotherapeutic target and using the specific inhibitor of PPOX acifluorfen, we confirmed that targeting of PPOX in vitro and in vivo significantly interferes with tumor growth. Publications: Sarah Bazzocco, Paulo Rodrigues, Miriam Garrido, Hafid Alazzouzi, Elena Andretta, Fernando Cartón, Irati Macaya, John M. Mariadason, Simo Schwartz Jr, Diego Arango. Genes highly expressed in rapidly proliferating tumor cells as new targets for colorectal cancer treatment - Manuscript in preparation

Joan X. Comella Carnicé

Planells-Ferrer, Rana S. Moubarak, Aroa Soriano, Derek M. Murphy, Joan Carles Ferreres, Francesc Borràs, Soledad Gallego, Raymond L. Stallings, Miguel F. Segura, Joan X. Comella

Objective: MYCN-amplified neuroblastoma represents the most aggressive group of neuroblastoma, generally chemoresistant and highly metastatic, resulting with overall survival below 5%. These properties have been linked to defects in the apoptotic machinery, either by silencing components of the extrinsic apoptotic pathway (e.g. caspase-8) or by overexpression of antiapoptotic regulators (e.g. Bcl-2, Mcl-1 or FLIP). Very little is known on the implication of death receptors and their antagonists in neuroblastoma prognosis. In this work, we aimed to evaluate the role of death receptor antagonists in the prognostic and oncogenic properties of neuroblastoma. Experimental design: The expression levels of death receptor antagonists were analyzed in multiple neuroblastoma data sets. We then correlate mRNA expression and clinical variables such as stage and overall survival. The function of the most differentially expressed death receptor antagonist was analyzed in neuroblastoma cell lines in vitro and in vivo. Results: We report the implication of Lifeguard (LFG/FAIM2/NMP35) in neuroblastoma prognosis. Intriguingly, although LFG has been initially characterized as an antiapoptotic protein, LFG was downregulated in the most aggressive and undifferentiated tumors. LFG is associated with neuroblastoma differentiation and its downregulation reduces cell adhesion, increases sphere growth and enhances migration, thus conferring a higher metastatic potential on NBL cells. Furthermore, LFG expression was found to be directly repressed by MYCN at transcriptional level. Conclusion: Our data, which support a new functional role for a hitherto undiscovered MYCN target, provide a new link between MYCN overexpression and increased NBL metastatic properties. Publications: MYCN repression of Lifeguard/FAIM2 is associated with increased cell migration and neuroblastoma poor prognosis (Submitted to Clinical Cancer Research)

Fernando Azpiroz

Marianela Mego, Maria Moris, Fernando Salvador(*), Adrian Sanchez(*), Israel Molina(*), Anna Accarino, Juan R. Malagelada, Fernando Azpiroz. (*)Servicio de Enfermedades Infecciosas. Hospital Universitario Valle Hebrón. Barcelona.

OBJETIVOS: Determinar la prevalencia de afectación anorectal en pacientes diagnosticados de Chagas no tratados. MATERIAL Y METODOS: Se estudiaron de forma prospectiva un año los pacientes afectos de enfermedad de Chagas de diagnóstico reciente antes de iniciar tratamiento. Los síntomas digestivos se recogieron mediante un cuestionario estructurado y se realizó una manometria anorectal. RESULTADOS: Se incluyeron 77 pacientes (54 mujeres y 23 hombres) con enfermedad de Chagas diagnosticada mediante dos técnicas serológicas. 47 pacientes no referían síntomas y de estos 21% mostraban alteraciones en la manometria anorectal (7 un defecto expulsivo, 2 insuficiencia esfinteriana; solo uno presentaba ausencia del reflejo recto-anal inhibitorio); los restantes 37 pacientes (79%) tenían una manometría anorectal normal. Treinta pacientes (39%) tenían síntomas (18 estreñimiento crónico y 11 de SII-E y 1 dolor abdominal funcional); el 73% de estos pacientes sintomáticos presentaban alteraciones en la manometría anorectal (17 defecto expulsivo, 1 hipertonía del esfínter y 4 contracción paradójica, pero ninguno alteración del reflejo recto-anal inhibitorio) y la proporción de alteraciones manométricas fue significativamente superior que en los pacientes asintomáticos (p<0.0001). CONCLUSION: Los pacientes con enfermedad de Chagas presentan una gran proporción de síntomas de estreñimiento y defecto obstructivo, pero baja incidencia de alteración del reflejo recto-anal inhibitorio.

Rafael Simó Canonge

Dra. Patricia Bogdanov, Andrea R. Carvalho, Dra. Cristina Hernández, Dr. Rafael Simó

Objectiu: Caracteritzar el procés seqüencial que té lloc en la neurodegeneració retiniana al ratolí db/db. Materials i mètodes: s’utilitzaren 72 ratolins C57BL/KsJ-db/db (Harlan Lab). Es van dividir en 12 diabètics i 12 no diabètics (C57BL/KsJ-db/+) per cada edat estudiada (8, 16 i 24 setmanes). Les retines es van avaluar en termes morfològics (activació glial i apoptosi) i funcionals (electroretinograma). A més es va realitzar un estudi intervencionista per baixar els nivells de glucosa en sang utilitzant un agonista de GLP-1 (liraglutide). Per aquest estudi es van utilitzar 16 ratolins db/db de 8 setmanes d’edat que van rebre diàriament injeccions subcutànies de liraglutide (300uL/50g pes) o vehicle, durant 15 dies. Resultats: Les retines diabètiques mostren major grau d’apoptosi a la GCL i activació glial en comparació amb les no-diabètiques. Aquestes diferències són més marcades a 16 i 24 setmanes. Les anormalitats funcionals són presents des de les 16 setmanes. Trobem una reducció significativa de la apoptosi i de la activació glial després del tractament amb liraglutide als ratolins db/db i simultàniament d’una atenuació de les anormalitats de l’electroretinograma. Conclusions: La neurodegeneració a les retines dels ratolins db/db és un fet primerenc i progressiu que reprodueix les anormalitats funcionals i morfològiques que tenen lloc als pacients diabètics. El db/db és un valuós model per testar nous fàrmacs neuroprotectors. Publicacions: Bogdanov P, Corraliza L, Carvalho AR, et al. Morphological and functional characterization of retinal neurodegeneration in db/db mice. Diabetologia 2012; 55(Suppl):A1117. Hernández C, García-Ramírez M, Colomé N, et al. Identification of new pathogenic candidates for diabetic macular edema using fluorescence-based difference gel electrophoresis analysis. Diabetes Metab Res Rev. 2013 Sep;29(6):499-506. Hernández C, García-Ramírez M, Corraliza L,et al. Topical administration of somatostatin prevents retinal neurodegeneration in experimental diabetes. Diabetes. 2013 Jul;62(7):2569-78. Garcia-Ramírez M, Villarroel M, Corraliza L, et al. Measuring permeability in human retinal epithelial cells (ARPE-19): implications for the study of diabetic retinopathy.Methods Mol Biol. 2011;763:179-94. Villarroel M, Garcia-Ramírez M, Corraliza L, et al. Fenofibric acid prevents retinal pigment epithelium disruption induced by interleukin-1β by suppressing AMP-activated protein kinase (AMPK) activation.Diabetologia. 2011 Jun;54(6):1543-53

Daniel Serón Micas

Francesc Moreso, Maite Salcedo, Carme Cantarell, Manel Perello, Irina Torres, Angeles Montero, Enric Trilla, Joana Sellares, Joan Morote

Reproducibility and the predictive value on outcome are the main criteria to evaluate the utility of histological scores. We analyze the reproducibility of donor biopsies assessment by different On-call pathologists and the retrospective evaluation by a single renal pathologist blinded to clinical outcomes. We also evaluate the predictive value on graft outcome of both evaluations. Material and methods: From 2009 to 2011 a biopsy was performed in donors with any of the following: ≥55 years, hypertension, diabetes, creatinine >1.5 mg/dL or stroke. Glomerulosclerosis (0=none, 1=1-10%, 2=11-20%, 3=>20%) as well as interstitial fibrosis, tubular atrophy, intimal thickening and arteriolar hyalinosis evaluated according to the Banff criteria were added to obtain a chronic score. Biopsies were classified as mild (≤3), intermediate (4-5) or advanced (6-7) damage and unacceptable (≥8) for transplantation. Results: A total of 127 out of 182 kidneys were biopsied. Weighted kappa value between both readings was 0.41 (95%CI: 0.28-0.54). Evaluation of biopsies by the renal pathologist was independently associated with estimated 12-month GFR (52±15 for mild, 46±16 for moderate and 37±12 ml/min for advanced; p=0.004) and with a composite outcome variable including death censored graft survival and time to reach a e-GFR < 30 ml/min/1.73m2 (p=0.023). There was no association between readings of On-call pathologists and outcome. Conclusion: The lack of association between histological scores obtained by the On-call pathologists and graft outcome suggests that a specific training on renal pathology is recommended to optimize the use of kidneys retrieved from expanded criteria donors. Related publications: Azancot MA, Moreso FJ, Salcedo MT, Carme Cantarell, Manel Perello, Irina Torres, Angeles Montero, Enric Trilla, Joana Sellares, Joan Morote, Seron D. "Renal biopsies from expanded criteria donors: reproducibility and predictive value on outcome". Kidney International (in press).

Joan Montaner Villalonga

Cristina Nafría Pérez, Xavier Mundet Tudurí, Antonio López Rueda, Inés Fernández Cortiñas, Carmen J.Jarca, J.L. Tovar, F. Pujadas, Joan Montaner Villalonga i Pilar Delgado Martínez. Investigadors del projecte ISSYS.

INTRODUCCIÓ: Determinar el deteriorament cognitiu lleuger (DCL) en la cohort de l'estudi ISSYS, estudiar els seus determinants i subtipus. METODOLOGIA: Estudi observacional, prospectiu en 1037 hipertensos sense antecedents d'ictus o demència. En la visita basal, es van recollir dades demogràfiques i clíniques (factors de risc vascular, REGICOR). Es van evaluar de lesions vasculars silents en RM cerebral (infarts, microsagnats, leucoaraiosi) i es va realitzar monitorització ambulatòria de la pressió arterial de 24 hores. També estudi cognitiu amb l'escala de cribatge DRS-2. Posteriorment, es va realizar una re-evalució cognitiva neurològica i neuropsicològica complerta en aquells que tenien una puntuació ajustada per edat i escolaritat pitjor de l’esperada en la DRS-2. RESULTATS: Es van re-evaluar cognitivament a 134 participants, dels quals el 22.6% presentaven DCL (2/3 DCL amnèsic, 1/3 no amnèsic) i el 38.3% envelliment normal (EN). Comparat amb l'EN els DCL tenien major puntuació en el REGICOR (p=0.012), més infarts silents (p=0.011) i microsagnats cerebrals (p=0.065) i alta pressió de pols (PP, p=0.002). En l'anàlisi multivariant els infarts (OR; IC95% 4.775;1.029,22.168) i la PP (1.071; 1.014-1.132) són predictors independents del DCL. CONCLUSIONS: Els factors de risc vascular no clàssics (rigidesa arterial mesurada per PP) i les lesions vasculars cerebrals silents (infarts) aporten informació complementària pel diagnòstic del DCL i s'han de considerar en poblacions d'alt risc. PUBLICACIONS: - BMC Neurol. 2013 Oct 2;13(1):130. Investigating silent strokes in hypertensives: a magnetic resonance imaging study (ISSYS): rationale and protocol design. Riba-Llena I, Jarca CI, Mundet X, Tovar JL, Orfila F, López-Rueda A, Nafría C, Fernández JL, Castañé X, Domingo M, Alvarez-Sabín J, Fernández-Cortiñas I, Maisterra O, Montaner J, Delgado P. - Stroke. 2013 Apr;44(4):e42. doi: 10.1161/STROKEAHA.112.681817. Epub 2013 Jan 17. Letter by Riba-Llena et al regarding article, "Not listened or not reported rather than silent stroke".

Riba-Llena I, Montaner J, Delgado P; ISSYS Project. - J Neurol Sci. 2012 Nov 15;322(1-2):79-81. doi: 10.1016/j.jns.2012.06.015. Epub 2012 Jul 24. - Cognitive assessment protocol design in the ISSYS (Investigating Silent Strokes in hYpertensives: a magnetic resonance imaging Study). Riba I, Jarca CI, Mundet X, Tovar JL, Orfila F, Nafría C, Raga A, Girona A, Fernández-Lara P, Castañé X, Alvarez Sabin J, Fernández Cortiñas I, Maisterra O, Montaner J, Delgado P.

Joan X. Comella

Jorge Urresti, Stephanie Reix, Laura Planells, Koen M.O. Galenkamp, Joaquin López-Soriano, Rana S. Moubarak, Joan X. Comella

OBJECTIVE: Lifeguard (LFG) is a membrane protein that antagonizes activation of Fas receptor that leads to cell death. In this study we aim to establish if this protein is also able to rescue from other apoptotic stimuli and study its interactions with other anti-apoptotic proteins and its relevance to its death protection effects. RESULTS: Treatment of PC-12 cells with 10µM etoposide showed protection from death in LFG and Bcl-xL overexpressing cells compared to empty plasmid transfected cells. Bcl-xL down-regulation showed sensitization to etoposide- induced cell death compared to cells infected with Scrambled sequence. LFG down-regulation followed by Bcl-xL overexpression showed protection from etoposide-induced cell death compared to empty plasmid transfected and Scrambled infected control, while Bcl-xL down-regulation followed by LFG overexpression fails to do so. CONCLUSIONS: LFG is able to rescue cells from etoposide-induced cell death. Moreover, LFG interacts with Bcl-xL, an antiapoptotic member of Bcl-2 family proteins, and this protection is dependent on endogenous levels of Bcl-xL. When LFG is overexpressed, cells became resistant to etoposide-induced cell death, but this protection is lost when Bcl-xL is down-regulated in LFG overexpressing cells. Taken together, these results may indicate a cross-talk between extrinsic and intrinsic apoptotic pathways, with LFG being a regulator of apoptotic cell death.

Javier Santos

Salvo-Romero E, Frías C, Lobo B, González-Castro A, Pigrau M, Alonso C, Sevillano-Aguilera C, Maria D Castillo, Maria Morís, Azpiroz F, Santos J, Vicario M.

Aim: To determine the implication of eosinophils in the pathogenesis of diarrhea-predominant irritable bowel syndrome (IBS-D) and to evaluate the immunomodulatory role of substance P (SP) and acetylcholine on eosinophil (Eo) function. Methods: Healthy(H) subjects (n=12) and diarrhea-prone IBS(IBS-D) Rome III (n=17) were included. Jejunal biopsies were obtained by Watson's capsule. Mucosal Eos counts, ultrastructure, activation and secretory activity were evaluated by immunohistochemistry, transmission electron microscopy or gene expression. The Eo cell line AML14.3D10 was stimulated with SP (10-6M) or Carbachol (CCh, 10-4M), and gene and protein expression were assessed by RT-PCR and immunofluorescence. Results: The number of mucosal MBP+ cells was similar in both groups, but degranulation, increased SNAP-23, and decreased eotaxin, EDN and ECP gene expression were identified in IBS-D (P<0.05). The Eo cell line expressed NKR1 and responded to SP by upregulating ECP, downregulating NFKB and MBP, without changing EDN, EPO, IL-8 and CCL21 expression (P<0.05). Moreover, Eos expressed M1 and M2 muscarinic and NA7 nicotinic receptors, but did not respond to CCh stimulation. Secretory activity was confirmed by localization of SNAP-23 on the cell membrane only in SP-stimulated cells. Conclusions: In IBS-D, mucosal eosinophils display secretory activity with low pro-inflammatory profile, similar to that observed after in vitro SP stimulation of Eos. These results suggest a non-inflammatory role of mucosal eosinophils in the intestinal mucosa of these patients. Publications: ACTIVACIÓN DE EOSINÓFILOS EN PACIENTES CON SÍNDROME DEL INTESTINO IRRITABLE: FUENTE DE CRF EN LA MUCOSA INTESTINAL E. Salvo-Romero, C. Martínez, B. Lobo, C. Frías, A.M. González-Castro, M. Pigrau, C. Alonso, C. Sevillano-Aguilera, F. Azpiroz, J. Santos, M. Vicario. Rev.Esp.Enferm.Dig. 2013; 105 (supl.I) 27-122

Chaysavanh Manichanh

Silvia Cuenca, Elisabet Sanchez, Alba Santiago, Ismail El khader, Suchita Panda, Silvia Vidal, Juan Camilo Nieto, Cándido Juárez, Francesc Sancho, Francisco Guarner, German Soriano, Carlos Guarner, Chaysavanh Manichanh

Objective The cross-talk between the gut microbiota and the immune system, essential to maintain homeostasis, takes place at the intestinal lymphoid tissues such as the mesenteric lymph nodes (MLNs). Here we investigated the presence of bacterial DNA in MLNs of control and cirrhotic rats and its relationship with inflammatory response. Material and methods DNA from mesenteric lymph nodes (MLNs) of ci rrhotic rats with ascites, induced by carbon tetrachloride (CCl4), was extracted, analyzed and compared to control rats using qPCR and pyrosequencing of the 16S rRNA gene. Cytokines were measured in blood samples by ELISA. Results Unexpectedly, sequence analysis revealed a high microbial diversity in the MLNs of both controls and cirrhotic rats with Proteobacteria as one of the most dominant phylum. CCl4-induced liver injury was not associated with a change in bacterial load, but was linked to a decrease in microbial diversity (p < 0.05) and to a microbial community alteration in the MLNs. A high proportion of Bifidobacterium animalis was also positively correlated with an elevated IL-10 expression (p = 0.002, FDR = 0.03, r = 0.94). Conclusions For the first time, the high microbial diversity observed in MLNs both in controls and CCl4-induced cirrhotic rats provides evidence that bacterial translocation is more than a mere dichotomical phenomenon. Rats with cirrhosis presented a lower bacterial diversity and different microbiome composition in MLNs in comparison with control rats. Publications Cuenca S. et al, Microbiome composition by pyrosequencing in the mesenteric lymph nodes of CCl4-induced cirrhotic rats; Journal of Innate Immunity, 2013 (Under review)

Maddalena Peghin

M.Peghin, I. Ruiz-Camps, C.Garcia-Vidal, J.Andreu, C.Cervera, A.Moreno, C.Gudiol, M.Martin, J.Puig de la Bellacasa, J.Ayats, E.Felip, J.Carratalà, A.Pahissa

Objectives Aspergillus spp. can cause acute invasive disease in severely immunocompromised patients. However there are few cases reported in patients with solid tumors complicated with acute and chronic invasive pulmonary aspergillosis (IPA). Methods A retrospective observational cohort study of all consecutive cases in patients with primary lung cancer or secondary lung metastases complicated with IPA, in three referral centers in Barcelona, from January 2008 to December 2011.We included cases of proven or probable, acute and chronic IPA. Results During the study period 16 episodes of probable and 2 of proven IPA were diagnosed. Thirteen cases were chronic and 5 acute IPA. Thirteen episodes occurred in men, mean age 60 years (SD +/- 9.7). Eleven patients had primary lung cancer and 7 secondary lung metastases. The most common underlying oncological disease was non-small cell lung cancer (9 patients). Clinical stages were III (6) or IV (12). Chronic lung disease was a co-existing illness in 8 persons, but none had previous Aspergillus spp. colonization. The median time between cancer diagnosis and Aspergillus spp. detection was 199.5 days (IQR 49-740). One episode occurred during neutropenia. Within 30 days prior to the diagnosis of IPA, 9 patients were receiving chemotherapy and 4 systemic steroids therapy. Five patients had received previous chest radiotherapy. The most common clinical presentations were fever (7), cough (4), hemoptysis (4), dyspnoea (4), chest pain (4) and distress syndrome (2). Aspergillus was detected in 7 bronchoalveolar lavages, 6 sputum samples, 3 tracheal aspirates and 1 percutaneous lung puncture and one percutaneous biopsy. A. fumigatus (11) was the most frequently isolated, followed by A. terreus (3), A. flavus (3) and A.niger (1). Ten patients had concomitant bacterial isolation and 1 patient viral infection (influenza A H1N1). At the time of Aspergillus spp. isolation the most common radiological findings on thoracic computed tomography (TCT made in 17 out of 18 patients) were consolidations (n=15, 88.2%), that in 8 patients were cavitated and in 4 associated with nodules, and bilateral alveolar infiltrates (n=2, 11.7%). Since Aspergillus spp. detection, TCT controls were performed in 13 out of 17 cases over the time and showed that 9(69.2%) patients (5 with PD, 3 with PR) had new cavity formation or expansion of one or more existing cavities (in 5 out of 9 cases associated with progressive abscess formation), 2 improved and 2 cured (after surgery).

All but one patient were treated with azoles (15 voriconazole, 2 itraconazole) and two underwent surgical resection. Thirteen persons died after Aspergillus spp. detection (average time 18 weeks) with 3 IPA caused and 8 related death.

Carlos Molina Cateriano

Estela Sanjuan Menendez, Montserrat Rodrigo Llana, Alba Thais Aparicio Reyes, Josep Maria Solorzano Fabrega, Susana Fernandez Garcia

INTRODUCCIÓN: El tiempo de reacción de enfermería es fundamental para ofrecer al paciente un tratamiento eficaz en fase aguda OBJETIVO: Conocer el tiempo de reacción de enfermería en el Código ictus e identificar los motivos de retraso en el servicio de Urgencias MATERIAL Y MÉTODOS: Estudio prospectivo de 50 pacientes con sospecha de ictus en el área de Urgencias, entre Abril y Julio de 2013. Se recogieron datos demográficos, tiempos de actuación, registro de exploraciones clínicas y tratamiento inmediato. Definimos retraso en salida del box,cuando el tiempo es mayor a 15 minutos RESULTADOS: El 50% de los pacientes fueron varones con una edad media de 68,4 ±15 años. A su llegada al box, se realizaron: electrocardiograma (62%); toma de tensión arterial (98%); temperatura (76%); glucemia capilar (98%); canalización de via periférica (90%) y analítica (82%). Se administró insulina subcutánea en 7 pacientes (14%), anti-hipertensivos en 4 pacientes (8%) y fármaco fibrinolítico tPA (Alteplasa) en 14 pacientes (28%). El tiempo medio de actuación de enfermería dentro del box fue de 18±11 minutos. En los casos que se administró tPA, el tiempo medio fue significativamente menor (9±5 minutos frente a 21±11 minutos, p=0,001). En 32 pacientes (64%) se produjo un retraso en la salida del box, siendo las causas más importantes: la realización de eco-Doppler(52%) y la canalización de vía (8%). CONCLUSIONES: El tiempo de reacción de enfermería en el paciente con ictus agudo es menor en los casos de administración de tPA, siendo la canalización de la vía uno de los principales retrasos.

PUBLICACIONES: - Bridging Intravenous–Intra-Arterial Rescue Strategy Increases Recanalization and the Likelihood of a Good Outcome in Nonresponder Intravenous Tissue Plasminogen Activator-Treated Patients A Case–Control Study. Rubiera M, Ribo M, Pagola J, Coscojuela P, Rodriguez-Luna D, Maisterra O, Ibarra B, Piñeiro S, Meler P, Romero FJ, Alvarez-Sabin J, Molina CA. Stroke 2011; 42: 993-997. - Bringing Forward Reperfusion with Oxygenated Blood Perfusion beyond Arterial Occlusion during Endovascular Procedures in Patients with Acute Ischemic Stroke. Ribo M, Rubiera M, Pagola J, Rodriguez-Luna D, Meler P, Flores A, Alvarez-Sabin J, Molina CA. Am J Neuroradiol 2010; 31: 1899-1902. - Predictors of Tissue-Type Plasminogen Activator Nonresponders According to Location of Vessel Occlusion.Mendonça N, Rodriguez-Luna D, Rubiera M, Boned-Riera S, Ribo M, Pagola J, Piñeiro S, Meler P, Alvarez-Sabin J, Montaner J, Molina CA.Stroke 2012; 43: 417-421. - Serum Low-Density Lipoprotein Cholesterol Level Predicts Hematoma Growth and Clinical Outcome After Acute Intracerebral Hemorrhage. Rodriguez-Luna D, Rubiera M, Ribo M, Coscojuela P, Pagola J, Piñeiro S, Ibarra B, Meler P, Maisterra O, Romero F, Alvarez-Sabin J, Molina CA. -Impact of blood pressure changes and course on hematoma growth in acute intracerebral hemorrhage.Rodriguez-Luna D, Piñeiro S, Rubiera M, Ribo M, Coscojuela P, Pagola J, Flores A, Muchada M, Ibarra B, Meler P, Sanjuan E, Hernandez-Guillamon M, Alvarez-Sabin J, Montaner J, Molina CA. Eur J Neurol. 2013 Sep;20(9):1277-83. doi: 10.1111/ene.12180. Epub 2013 May 5. -Difficult catheter access to the occluded vessel during endovascular treatment of acute ischemic stroke is associated with worse clinical outcome. Ribo M, Flores A, Rubiera M, Pagola J, Mendonca N, Rodriguez-Luna D, Piñeiro S, Meler P, Alvarez-Sabin J, Molina CA. J Neurointerv Surg. 2013 May;5 Suppl 1:i70-3. doi: 10.1136/neurintsurg-2012-010438. Epub 2012 Oct 31.

Contxita Jiménez Gutierrez

Dr. José Alegre Martín, Dra. Naia Sáez Francas, Dra. Eva Ruiz Ruiz, Luisa Aliste Sánchez, Cristina Domingo Gómez, Mª del Mar Caparros Granados, Josep Mª Charles Vallcanera

ANTECEDENTES: El Síndrome de Fatiga Crónica presenta muchas perturbaciones multidimensionales que afectan de forma holística a las personas que padecen esta enfermedad y que el tratamiento actual de la fatiga, dolor, ansiedad-depresión y alteraciones del sueño, presentes en dicha entidad clínica, es insatisfactorio. HIPÓTESIS: Es la acupuntura una técnica eficaz para mejorar la sintomatología de los pacientes con Síndrome de Fatiga Crónica? La hipótesis de partida de este ensayo consiste en contrastar que la acupuntura resulta más útil que placebo. OBJETIVO GENERAL. Evaluar la eficacia de la acupuntura verdadera (AV) en comparación con placebo-acupuntura o acupuntura simulada (AS) en la mejoría de la sintomatología de los pacientes diagnosticados del SFC. OBJETIVOS ESPECÍFICOS: - Evaluar si el tratamiento con AV, en comparación a la AS, se asocia a una reducción de la fatiga, medido mediante la escala FIS. - Comprobar la reducción del dolor tras el tratamiento medido mediante la escala McGill. - Constatar la disminución en el uso de analgésicos, tras el tratamiento con acupuntura mediante el recuento de estos fármacos. - Evaluar la reducción de la ansiedad y depresión tras el tratamiento medida mediante la escala HAD. - Evaluar la mejoría en la calidad del sueño tras el tratamiento medido mediante la escala Pittsburgh. - Comprobar la mejora de la calidad de vida relacionada con la salud tras el tratamiento medida mediante la escala SF-36. METODOLOGÍA: Diseño: Ensayo clínico piloto aleatorizado, de grupos paralelos, controlado con acupuntura placebo-simulada, unicéntrico, prospectivo, con paciente y evaluador ciego. Ensayo con asignación aleatoria 1:1. Después de firmar el consentimiento informado, se incluirán 60 pacientes diagnosticados del SFC, 30 en el grupo de intervención de acupuntura verdadera y 30 en el grupo control de acupuntura simulada, aplicándose un plan de tratamiento individualizado y personalizado, según la valoración energética de cada paciente. Duración del tratamiento: 4 meses. PUBLICACIONES -Publicación de este protocolo de estudio clínico en la UAB, septiembre 2013, dentro del máster de investigación clínica aplicada a ciencias de la salud ICACS, lectura y presentación en el tribunal del master ICACS de la UAB obteniendo la cualificación de: Excelente. -NEWS & VIEWS: Antioxidants & Redox Signaling. Factor de impacto: 8,45 Could mitochondrial dysfunction be a differentiating marker between Chronic Fatigue Syndrome and Fibromyalgia? Jesús Castro-Marrero1*, Mario D. Cordero2*, Naia Sáez-Francas3, Conxita Jimenez- Gutierrez1, Francisco J. Aguilar-Montilla4, Luisa Aliste1, José Alegre-Martin1 1 CFS Unit. Institut de Recerca Vall d’Hebron. Hospital Universitari Vall d’Hebron, Universitat Autònoma de Barcelona, 08035, Barcelona, Spain. 2 Departamento de Citología e Histología Normal y Patológica, Facultad de Medicina, Universidad de Sevilla, 41009, Sevilla, Spain. 3 Departamento de Psiquiatría y Medicina Legal, Centro de Investigación Biomèdica en

Red de Salud Mental (CIBERSAM), Hospital Universitari Vall d’Hebron, Universitat Autònoma de Barcelona, 08035, Barcelona, Spain. 4 División de Neurociencias, Universidad Pablo de Olavide de Sevilla, Carretera de Utrera Km. 1, 41013, Sevilla, Spain. * These authors contributed equally to this work. Running Title: Mitochondrial dysfunction in CFS and Fibromyalgia. doi:10.1089/ars.2013.5346. -Próxima presentación de casos clínicos derivados de este estudio, en el congreso de Enfermería en anestesia que se celebrará en Barcelona el 8 de Noviembre.

MªLuisa Cebrián Batalla

Pili Gil, Verónica Fernández, Sandra Lamarca, Paloma Conde, MªÁngeles Aceituno, Anna Callés, Carmen Conde, Sara Farrero, Raquel Rodrígiez, Raquel Milán, Mar López, estefanía Cámara.

Estudi descriptiu realitzat a l' Àrea Infantil de l'Hospital Universitari Vall d' Hebron . Els subjectes d'estudi van ser els professionals sanitaris ( infermeres , metges i psicòlegs ) que participen en els circuits d'atenció i seguiment als adolescents trasplantats ( hepàtics , reals , cardíacs i pulmonars ) , amb malaltia oncohematològica , diabetis , fibrosi quística i VIH . La recollida de dades es va realitzar mitjançant qüestionari autocomplimentat elaborat específicament per a aquest estudi. Es va obtenir la participació de 105 professionals ( 70% ) , dels quals el 52% ( 55 ) havia rebut formació en educació sanitària . Un 80 % ( 84 ) eren infermeres . Van manifestar disposar d'instruments per mesurar adherència terapèutica el 28% ( 29 ) . El 56 % ( 59 ) ha considerat que el compliment del tractament dels adolescents era bo i el 29 % ( 31 ) regular . Un 43 % ( 45 ) ha assenyalat que l'adherència no s'aborda bé. Els professionals no disposen d'un temps planificat per realitzar educació sanitària relacionada amb el tractament (el 79 % busca temps durant la jornada laboral ) . Els recursos que es posen a disposició dels pacients i els seus cuidadors per gestionar els seus tractaments són : fullets ( 58% ) , full de recomanacions a l'alta ( 77% ) , telèfon per fer consultes ( 63,8% ) i associacions de pacients ( 47,6% ) .El 39 % dels participants van aportar suggeriments de millora .

Antonia Sambola Ayala

Albert Alonso, Gerard Oristrell, Alba Santos, Bruno García Del Blanco, Pilar Tornos and David García-Dorado

Background: Triple therapy (TT: anticoagulation plus dual antiplatelet therapy) is recommended for patients with mechanical prosthetic valves submitted to coronary stenting (CS). This treatment carries a high risk of bleeding. Aim: to assess the benefits and risks of oral anticoagulation (OAC) and clopidogrel compared with TT in patients with mechanical prosthetic valves submitted to CS. Methods: we conducted a prospective study from 2008 to 2011 in patients with mechanical prostheses submitted to CS. We compared the incidence of thromboembolic and bleeding events in patients with TT vs OAC and clopidogrel. Follow-up was 1-year. Results: We included 72 patients (67±12.5 years), 42 (38.3%) receiving TT and 30 (41.7%) OAC plus Clopidogrel (p=0.26). Baseline clinical characteristics were similar in both groups: hypertension (71.4% vs 75%;p=0.48), diabetes mellitus (40.5% vs 28.6%;p=0.22), renal failure (10.3% vs 18.5%;p=0.27) and previous stroke (40% vs 60%;p=0.35). The overall incidence of adverse events was similar in both groups (31% vs 36.7%;p=0.39), without differences in thromboembolism (2.4% vs 0%;p=0.58) and bleeding events (21.4% vs 20%;p=0.56), overall mortality (11.9% vs 6.7%;p=0.37) and cardiovascular mortality (9.5% vs 6.7%;p=0.55). However, the incidence of ischemic events associated to target vessel failure (TVF) was significantly higher in patients receiving OAC plus Clopidogrel (2.4% vs 20%;p=0.01). A multivariate analysis showed as an independent factor of TVF the treatment with OAC plus Clopidogrel (OR 9.97, 95% CI: 1.05-94.7;p=0.04). Conclusions: Patients with mechanical prostheses submitted to CS and treated with OAC plus Clopidogrel alone, without aspirin, have a higher incidence of ischemic events due to TVF.

Jordi Rello Condomines

Marina García de Acilu, Oriol Roca, Laura Ruano, Joan R Masclans, Jordi Rello

Objetivos: examinar el valor de la medición de Interleukina 33 y su recetor ST2 en pacientes con síndrome de distrés respiratorio agudo. Material y métodos: se trata de un estudio observacional, prospectivo y unicéntrico, que incluye pacientes con SDRA que requirieron ingreso en UCI. Se realizaron determinaciones de concentración plasmática de IL-33 y ST2 los días 1 y 3 desde la instauración del SDRA. Las diferencias en las concentraciones de IL-33 y ST2 se estudiaron mediante el test de ANOVA para medidas repetidas y los valores discriminantes mediantes el área de la curva AUROC. Las curvas de supervivencia fueron comparadas empleando el test de log-rank. Para identificar las variables de ingreso asociadas a mortalidad en UCI se emplearon modelos multivariantes de regresión de Cox. Resultados: 17 pacientes con SDRA (12 [70.6%] varones) con una edad media de 61 años (rango intercuartílico [RI] 48-70) fueron incluídos. La media de APACHE II fue 24 (19-29) y el origen del SDRA fue intrapulmonar en 12 de ellos (70.6%). A día 1, 12 (70.6%) pacientes estaban conectados a ventilación mecánica y 5 (29.4%) recibían oxigenoterapia de alto flujo mediante cánulas nasales. La mortalidad en UCI fue 41.2%. La concentración media de ST-2 el día 1 (4934 [RI 3672-7547]pg/ml vs 1007 [RI 677-1987]pg/ml) y el día 3 (5720 [RI 3932-7404]pg/ml vs 823 [RI 597-1547]pg/ml) fue superior en los pacientes que no sobrevivieron (p<0.001). No se encontraron diferencias en los niveles plasmáticos de IL-33. La concentración de ST-2 el día 1 identificó con precisión los pacientes que fallecieron (AUROC 0.96 [95%IC 0.86-1.0];p<0.01), superando el APACHE y SOFA. Una concentración de ST-2 el día 1 ≥ 3672pg/ml identificó a los pacientes que fallecieron con una sensibilidad del 86% y una especificidad del 100%. La curva de supervivencia de Kaplan-Meier mostró una supervivencia mayor en los pacientes con concentraciones de ST-2 <3672 el día 1 (p<0.001). En el análisis multivariante, la única variable asociada con la mortalidad en UCI fue la concentración de ST-2 ≥3672 el día 1 (HR 14.7 [95%CI 1.7-131.1]). Conclusión: en este studio la concentración de ST-2 resultó ser un potente predictor de mortalidad en UCI en pacientes con SDRA.

Albert Pahissa

C. López MD, J. Gavaldà PhD, I. Bilbao PhD, L. Castells PhD, A. Gelabert MD, H. Allende PhD, A. Pahissa PhD, O. Len PhD.

Resum: Data from published studies regarding risk factors for liver-biopsy related infectious complications in liver transplant recipients are inconsistent. We carried out a retrospective cohort study analyzing consecutive liver biopsies in orthotopic liver transplant patients in a tertiary hospital (2001-2011), including 669 liver biopsies (557 percutaneous and 92 transjugular) in 286 liver transplant recipients. There were 25 complications in 24 patients (overall incidence 3.7%), 14 of which were infectious complications: 8 sepsis, 1 cholangitis, 3 fever, and 2 peritonitis. The causal microorganisms, isolated in 10 of the 14 cases, were mainly Pseudomonas aeruginosa (4 patients) and Enterobacteriaceae (4 patients). All complications occurred in biopsies performed in patients hospitalized for more than 48 hours (381 biopsies in 201 patients); hence, only this group was included in the risk factor analysis. The only variable statistically associated with development of infectious complications after liver biopsy was plasma albumin less than 2.4 mg/dL, which conferred an estimated 3.78-fold higher risk (95% CI 1.36-10.53, p=0.016). In conclusion, based on our experience, infectious complications secondary to liver biopsy in liver transplant recipients are related to hospitalization at the time of biopsy, particularly in the presence of low albumin values.

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Joan Seoane

I. Huber-Ruano, A. Cascante, C.Raventós, A. Arias, J. Seoane

TGFβ is a cytokine that has been widely related to tumor progression. Indeed, several TGFβ inhibitors are currently under clinical development for both primary and metastatic disease. Moreover, during last years, increasing evidence has supported the relationship between epigenetic modifications and tumor development. In order to determine the effects of TGFβ activity on the epigenetic landscape, we performed microarrays and looked for epigenetic-related genes that were being affected by this cytokine. We identified that KDM6B, which encodes for the H3K27 demethylase JMJD3, is specifically up-regulated through the Smad pathway upon TGFβ treatment. By analyzing microarray data from cells expressing shJMJD3 and performing ChIP experiments, we have shown that JMJD3 regulates H3K27 methylation at specific promoters leading to changes in the expression of a group of genes that have been previously linked to tumor growth, angiogenesis or invasion. Moreover, pharmacological inhibition of JMJD3, either with IOX-1 or with the recently developed drug GSK-J4, leads to the repression of a similar cohort of genes. By using a matrigel-coated transwell system, we have seen that JMJD3 may play a role in invasion. Strikingly, although JMJD3 does not affect proliferation in vitro, in vivo orthotopic experiments in mice using A549 cells expressing an inducible shJMJD3 showed that JMJD3 down-regulation leads to a decrease in tumor size and to fewer metastases.

Joaquin Arribas

Morancho B, Parra-Palau JL, Ibrahim YH, Bernadó Morales C, Peg V, Bech-Serra JJ, Pandiella A, Canals F, Baselga J, Rubio I, Arribas J.

The transmembrane tyrosine kinase HER2 (ErbB2, neu) is a prototypical biomarker for breast cancers and a therapeutic target. Although anti-HER2 therapies are remarkably effective, HER2-positive tumors are heterogeneous and some subtypes do not respond or develop resistance to these therapies. Here we show that H2NTF, a novel N-terminal fragment of HER2, is expressed at variable levels in 60% of the breast cancer samples analyzed. Despite this high frequency, none of the cell lines analyzed express the fragment at detectable levels, while HER2-derived patient-derived xenografts (PDX) present H2NTF expression. H2NTF was isolated and the C-termini of the fragment was identified by mass spectrometry, being located at the kinase domain. Characterization of H2NTF shows that it is devoid of the tyrosine kinase domain but it readily interacts with full-length HER2 and other HER receptors. As a consequence, H2NTF acts as a dominant negative, attenuating the signaling triggered by full-length HER receptors. Expression of H2NTF results in resistance to the treatment with low concentrations of trastuzumab in vitro. However, cells expressing H2NTF and non-expressing cells have similar sensitivity to trastuzumab in vivo, likely because H2NTF/trastuzumab complexes trigger antibody-dependent cell-mediated cytotoxicity. Publications: Morancho B, Parra-Palau J, Ibrahim YH, Bernado Morales C, Peg V, Bech-Serra JJ, Pandiella A, Canals F, Rubio I, Baselga J, Arribas J. A dominant negative N-terminal fragment of HER2 frequently expressed in breast cancers. Oncogene. 2013 Mar 14;32(11):1452-9.

Santiago Ramón y Cajal

Bàrbara Castellana and Santiago Ramón y Cajal. Clinical Part: Bàrbara Castellana, Miren Aizpurua, Irene Sansano, Vicente Peg and Santiago Ramon y Cajal.

An important step towards tumour progression and metastasis is the activation of the epithelial-mesenchymal transition (EMT) program. This complex process confers cancer cells traits such as enhanced motility, invasiveness and resistance to therapy. EMT can be triggered by the convergence of intrinsic factors and signals from the tumour microenvironment, such as aberrant expression of cytokines. Recently, the multifunctional oncoprotein YB-1 has been shown to play an important role in malignant tumours facilitating many of the “hallmarks of cancer” proposed by Hanahan and Weinberg. In this project we focused on the alliance between YB-1 and the cytokine IL-6, which has been implicated in tumour-cell metastasis and is a well known EMT inducer. Overexpression or direct addition of IL-6 up-regulates and activates YB-1 in the epithelial breast cancer cell line MCF7, leading to an upregulation of mesenchymal markers (Snail1, N-cadherin), increased cell migration and invasion, as well as morphological changes associated with an EMT-like phenotype. Conversely, in the mesenchymal-like breast cancer cell line MDA-MB-231, RNAi-mediated downregulation of YB-1 resulted in decreased IL-6 expression and an increase in epithelial-like cell characteristics. Our results describe a novel interplay whereby IL-6 regulates YB-1 and vice versa creating a positive feed-forward loop driving EMT during breast cancer progression. In the future we aim to delineate the exact signalling pathways linking IL-6 and YB-1, and how they converge to orchestrate important processes such as growth, invasion and metastasis. The understanding of this interplay will set the stage for new combinatorial treatments of aggressive breast cancers.

Ramon Martí Seves

Javier Torres, Carlo Viscomi, Raquel Cabrera, Yolanda Cámara, Ivano Di Meo, Jordi Barquinero, Michio Hirano, Massimo Zeviani, Ramon Martí

El MNGIE es una enfermedad mitocondrial causada por mutaciones en el gen TYMP, que codifica la enzima timidina fosforilasa. El déficit en el funcionamiento de esta enzima causa la acumulación sistémica de sus sustratos, la timidina y la desoxiuridina, que interfiere en la replicación y mantenimiento del ADN mitocondrial, produciendo alteraciones en la función mitocondrial. Los pacientes de MNGIE tienen una esperanza de vida de 37 años sin tratamiento, y la única terapia efectiva existente es el trasplante alogénico de células madre hematopoyéticas, aunque tiene una mortalidad y morbilidad asociadas muy elevadas. En este trabajo hemos estudiado el uso de la terapia génica mediante el uso un vector adenoasociado (AAV) que contiene una copia funcional de TYMP bajo el control de un promotor específico de hígado. La inyección intravenosa del vector a una dosis de 2•1011 gc/kg en el modelo murino de MNGIE consiguió reducir de forma estable la concentración sistémica de nucleósidos hasta valores normales en el 50 % de los animales. Dosis superiores permitieron la reducción hasta niveles normales o incluso por debajo en prácticamente todos los animales. Adicionalmente, se observó que en los animales tratados se recuperaban los niveles de dCTP mitocondrial, cuyo déficit es la causa directa de la depleción del ADN mitocondrial. Estos resultados demuestran que el uso de un AAV dirigido a la expresión específica de TYMP en hígado es una potencial alternativa terapéutica viable y segura.

Joan X. Comella

Paulina Carriba, Sebastian Jimenez, Joaquin Lopez Soriano, Ramón Trullás, Javier Vitorica and Joan X. Comella.

The main objective of the project was analyzed the role of FAIM-L in Alzheimer's disease (AD). AD, such as other neurodegenerative diseases, evolves with neuronal death. This neuronal death is due mainly by apoptosis. Apoptosis is activated by two key mechanisms: the intrinsic and the extrinsic pathways. The extrinsic pathway is initiated by the activation of Death Receptors (DRs) such as Tumor Necrosis Factor (TNF) Receptor. DR signalling is highly regulated by several proteins; one of them is FAIM-L. For the study, we analyzed by quantitative-PCR and WB post-mortem samples of human patients with the disease. We also use transgenic mice for the immunohistological analysis and for the biochemical studies (including WB, Hoechst counts, PCR) we use neuronal mice primary cultures. Biochemical results in neuronal primary cultures indicate that FAIM-L is necessary for the protection mediated by the TNFa release by glial cells, and its deletion abrogates this protection. More interesting, treatment with Amyloid beta (synthetic and from transgenic mice) reduces FAIM-L levels. In human samples we detect a significant reduction in FAIM-L mRNA and protein; also observed in transgenic mice model in entorhinal cortex and the hippocampus CA1. The temporal pattern of FAIM-L reduction matches with an increase in neuronal death in these areas. This draws the following picture, when the levels of FAIM-L are reduced by Amyloid beta, there is a shift from the protective response of the glial cells to a deleterious, accelerating the progression of the neurodegeneration.

Directly related to the work: Moubarak RS, Planells-Ferrer L, Urresti J, Reix S, Segura MF, Carriba P, Marqués-Fernàndez F, Sole C, Llecha-Cano N, Lopez-Soriano J, Sanchis D, Yuste VJ, Comella JX. FAIM-L is an IAP-Binding protein that inhibits XIAP ubiquitinylation and protects from Fas-induced apoptosis. Submitted to Journal of Neuroscience. Related to AD:

Lichtenstein M*, Carriba P*, Baltrons M.A, Woljciak-Stothard B, Peterson J.R, Garcia A, Galea E. Secretase-Independent and RhoGTPase/PAK/ERK-Dependent regulation of Cytoskeleton Dynamics in Astrocytes by NSAIDs and Derivatives. * Equal author contribution. Journal of Alzheimer’s disease. PubMed ID: 20930267 5-year impact factor: 4,814 Lichtenstein M, Carriba P, Masgrau R, Pujol A, Galea E. Staging anti-inflammatory therapy in Alzheimer’s disease. Front Aging Neurosci. 2010;2:142. PMID: 21152343 It is a new journal and has not yet determined the impact factor. Carriba P, Pardo L, Parra-Damas A, Lichtenstein MP, Saura CA, Pujol A, Masgrau R, Galea E. ATP and noradrenaline activate CREB in astrocytes via noncanonical Ca(2+) and cyclic AMP independent pathways. Glia 2012 PMID: 22593004. Impact factor 4,83 Not related to AD Agnati LF, Ferré S, Genedani S, Leo G, GuidolinD, Filaferro M, Carriba P, Casadó V, Lluis C, Franco R, Woods AS, Fuxe K. Allosteric Modulation of Dopamine D2 Receptors by Homocysteine. Journal of Proteome Research 5(11) 3077-83 (2006). PubMed ID: 17081059 5-year impact factor: 5,359 Paper cited 20 times. Carriba P, Ortiz O, Patkar K, Justinova Z, Stroik J, Themann A, Müller C, Woods AS, Hope BT, Ciruela F, Casadó V, Canela EI, Lluis C, Goldberg SR, Moratalla R, Franco R, Ferré S. Striatal Adenosine A2A and Cannabinoid CB1 Receptors Form Functional Heteromeric Complexes that Mediate the Motor Effects of Cannabinoids. Neuropsychopharmacology 32(11):2249-59 (2007). PubMed ID: 17356572 5-year impact factor: 6,813 Marcellino D, Carriba P, Filip M, Borgkvist A, Frankowska M, Bellido I, Tanganelli S, Müller CE, Fisone G, Lluis C, Agnati LF, Franco R, Fuxe K. Antagonistic Cannabinoid CB1/Dopamine D2 Receptor Interactions in Striatal CB1/D2 Heteromers. A Combined Neurochemical and Behavioural Analysis. Neuropharmacology 54(5):815-23 (2008). PubMed ID: 18262573 5-year impact factor: 3,824 Carriba P, Navarro G, Ciruela F, Ferré S, Casadó V, Agnati L, Cortés A, Mallol J, Fuxe K, Canela EI, Lluis C, Franco R. Detection of Heteromerization of More than Two Proteins by Sequential BRET-FRET. Nature Methods 5(8):727-33 (2008). PubMed ID: 18587404 5-year impact factor: 16,907 Carriba P, Navarro G, Ciruela F, Ferré S, Casadó V, Cortés A, Mallol J, Canela EI, Lluis C, Franco R. Detection of Heteromerization of More than Two Proteins by Sequential BRET-FRET. Nature Protocols. Protocols Network 10.1038/nprot.2008.126 5-year impact factor: 6,368 Navarro G, Carriba P, Gandía J, Ciruela F, Casadó V, Cortés A, Mallol J, Canela EI, Lluis C, Franco R. Detection of Heteromers Formed by Cannabinoid CB1, Dopamine D2, and Adenosine A2A G-Protein-Coupled Receptors by Combining Biomolecular Fluorescence Complementation and Bioluminescent Energy Transfer. The Scientific World Journal 8:1088-97 (2008). PubMed ID: 18956124 Impact factor (2008): 1,836

Javier Santos

C. Frías, M. Vicario, M. Pigrau, B. Lobo, E. Salvo-Romero , A. González-Castro, C. Alonso, B. Rodiño-Janeiro, M. D. Castillo, F. Azpiroz , J. Santos

AIMS&METHODS: We aimed at identifying macrophage contribution to IBS. Twenty newly diagnosed diarrheapredominant IBS (IBS-D, female F n=14) fulfilling Rome III criteria and age-matched healthy (H) participants (H, F n=9) were recruited. One mucosal biopsy was obtained from the jejunum, using the Watson’s capsule. Eosinophils (H&E), mast cells (CD117), T lymphocytes (CD3) and macrophages (CD68) were identified by immunohistochemistry. Mucosal RNA was isolated, submitted to microarray analysis and subsequent biological functions and pathways identification, associated with differential gene expression. RESULTS: Eosinophils, CD117+, CD68+ and CD3+ infiltrate was similar between H and IBS-D groups. However, in females, the number of CD68+ was higher in IBS-D compared to H (IBS-D F: 29.0±1.5 n=14; H F: 20.9±1.6 n=7; P =0.004). Considering just the H group, females displayed less CD68+ than males (H F: 20.9±1.6 n=7; H M: 27.2±2.4 n=7; P=0.051). Differential gene expression revealed the immunological response as the most altered function in IBS-D, with significant recruitment and activation of monocytes (P<0.0001), induction and activation of macrophages (P<0.0001), antigen-presenting and cytotoxic T cells(P<0.0001). CONCLUSION: Increased immune activity, specifically macrophage response, features the jejunal mucosa of IBS-D patients. The differences in the number of macrophages in women comparing to men evidence that further studies are needed to clarify gender-dependence in phagocytic activity and whether this mechanism explains increased prevalence of IBS in women. PUBLICATIONS: R EV E SP E NFERM D IG 2013; 105 (Supl. I): 27-122

Esteve Ribera

Adrian Curran1, Ramon Marti2, Mercè Perez1, Manel Crespo1, Maria Jesús Melià2, Josep Guiu2, Joaquin Burgos1, Vicenç Falcó1 and Esteve Ribera1. 1. Hospital Universitari Vall d’Hebron, Barcelona, Spain. 2. Institut de Recerca Vall d’Hebron, Barcelona, Spain.

Objectives: Ritonavir-boosted Darunavir(DRV/r) is mainly metabolized in the liver. Hepatic cirrhosis can impair liver function, potentially modifying DRV/r pharmacokinetics (PK). Unbound concentrations may be more reliable in these patients, but there is scarce data. Methods: HIV-HCV coinfected patients with compensated cirrhosis and HIV monoinfected patients as controls were included. DRV/r was given 800/100 mg once-daily. A complete steady-state12-hours PK study was performed. Drug concentrations were determined by mass spectrometry. AUC and CL/F were assessed by non-compartmental model and linear/log trapezoidal rule. Descriptive values are expressed as medians (IQR) and geometric means (GM). Geometric mean ratios (GMR) were used to compare concentrations between groups and Spearman test for correlations. Results: Thirty patients (20 cirrhosis, 10 controls) were included. Baseline characteristics for cirrhotic patients: 75% men, age 49 (43-51) years, BMI 24 (20-28) kg/m2, CD4 343 (215-677) cel/mm3. HCV genotypes were: 1 (75%), 3 (20%) and unknown (5%). Five patients had had prior clinical decompensation, median MELD was 9 (8-12) and patient’s worse Child-Pugh was C in 3 cases and A in 17. Median elastography was 20 (14-26) kPa. For controls: 40% men, age 35 (31-51) years, BMI 25 (23-27) kg/m2, CD4 414 (289-894) cel/mm3 and median elastography 5.3 (4.2-6.4) kPa. Darunavir total and unbound PK parameters are described in table 1. There were no differences in PK parameters depending on Child-Pugh, gender or concomitant medications. Conclusions: In HIV-HCV patients with compensated cirrhosis, DRV/r total and unbound concentrations are similar to those observed in non-cirrhotic patients, and dose adjustments are not necessary.

Simo Schwartz Jr

Gener P., Minara R., Sarkani V., Sofia A., Arranja A., Rafael D., Gaspar R.S., Gouveia L., Videira M., Abasalo I., Schwartz Jr S.

In many tumors, resistance to therapy and metastatic spread seem to be sustained by the presence of cancer stem cells (CSC). These cells retain the capacity of repopulating the tumor, while being insensitive to conventional anticancer therapies, antimitotic agents or radiation. Drug delivery by nanoparticles may well circumvent the CSC resistance machinery by avoiding multi drug resistance channels and by increasing the concentration of the drug inside the cells. However, to precisely evaluate the therapeutic effect of nanoparticles on CSC, cells with CSC properties need to be properly identified and characterized. Thus, we have developed an original, stable and cost effective in vitro model in which we can stably tag CSC, based on fluorescent reporter gene tdTomato, expressed under the control of a CSC specific promoter. We have separated and tracked CSC in breast (MCF7, MDA-MB-231) and colon (HCT116) cancer cell lines. To ensure specific CSC enrichment, isolated cells were tested for pluripotency, the ability to self-renew and for expression of specific stem cell markers. Using this system, we have investigated the prospect of biodegradable PEGylated poly(lactic-co- glycolic acid (PLGA-co-PEG) polymeric micelles loaded with paclitaxel (PTX). Eager to further improve nanoparticles uptake and to ameliorate selective drug delivery to CSC, a second generation of CD44 targeted micelles has been tested as well. CSC showed significant ability to recover after naked PTX treatment, but when we employed PTX encapsulated in CD44-PLGA-co-PEG micelles, the ability of CSC to recover was completely abolish in MDA-MB-231 cells line and significantly decrease in MCF7 cells.