Publications by authors named "Eva Muñoz"

58 Publications

Acute kidney injury as a risk factor for mortality in oncological patients receiving check-point inhibitors.

Nephrol Dial Transplant 2021 Feb 6. Epub 2021 Feb 6.

Nephrology Department, Vall d'Hebron University Hospital, Vall d'Hebron Institue of Research, Barcelona, Spain.

Background: Checkpoint inhibitors (CPI) have drastically improved metastatic cancer outcomes. However, immunotherapy is associated to multiple toxicities, including acute renal injury (AKI). Data about CPI related AKI are limited. Our aim was to determine risk factors for CPI related AKI, as well as its clinical characteristics and its impact on mortality in patients undergoing immunotherapy.

Methods: All patients under CPI at our center between March 2018 and May 2019, and with a follow up until April 2020, were included. Demographical, clinical data and laboratory results were collected. AKI was defined according to KDIGO guidelines. We performed a logistic regression model to identify independent risk factors for AKI and actuarial survival analysis to establish risk factors for mortality in this population.

Results: 759 patients were included, with a median age of 64 years. 59% were men and baseline median creatinine was 0.80 mg/dL. Most frequent malignance was lung cancer and 56% were receiving anti-PD1. 15.5% developed AKI during the follow-up. Age and baseline kidney function were identified as independent risk factors for AKI related ICI. At the end of follow-up, 52.3% patients had died. Type of cancer (not melanoma, lung or urogenital malignance), type of CPI (not CTLA4, PD-1, PD-L1 or their combination) and the presence of an episode of AKI were identified as risk factors for mortality.

Conclusions: 15.5% of patients under immunotherapy presented AKI. A single AKI episode was identified as an independent risk factor for mortality in these patients and age and baseline renal function were risks factors for the development of AKI.
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http://dx.doi.org/10.1093/ndt/gfab034DOI Listing
February 2021

Neuroprotection with the cannabigerol quinone derivative VCE-003.2 and its analogs CBGA-Q and CBGA-Q-Salt in Parkinson's disease using 6-hydroxydopamine-lesioned mice.

Mol Cell Neurosci 2021 01 16;110:103583. Epub 2020 Dec 16.

Instituto Universitario de Investigación en Neuroquímica, Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad Complutense, Madrid, Spain; Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CIBERNED), Madrid, Spain; Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), Madrid, Spain. Electronic address:

The quinone derivative of the non-psychotropic cannabinoid cannabigerol (CBG), so-called VCE-003.2, has been recently investigated for its neuroprotective properties in inflammatory models of Parkinson's disease (PD) in mice. Such potential derives from its activity at the peroxisome proliferator-activated receptor-γ (PPAR-γ). In the present study, we investigated the neuroprotective properties of VCE-003.2 against the parkinsonian neurotoxin 6-hydroxydopamine (6-OHDA), in comparison with two new CBG-related derivatives, the cannabigerolic acid quinone (CBGA-Q) and its sodium salt CBGA-Q-Salt, which, similarly to VCE-003.2, were found to be active at the PPAR-γ receptor, but not at the cannabinoid CB and CB receptors. First, we investigated their cytoprotective properties in vitro by analyzing cell survival in cultured SH-SY5Y cells exposed to 6-OHDA. We found an important cytoprotective effect of VCE-003.2 at a concentration of 20 μM, which was not reversed by the blockade of PPAR-γ receptors with GW9662, supporting its activity at an alternative site (non-sensitive to classic antagonists) in this receptor. We also found CBGA-Q and CBGA-Q-Salt being cytoprotective in this cell assay, but their effects were completely eliminated by GW9662, thus indicating that they are active at the canonical site in the PPAR-γ receptor. Then, we moved to in vivo testing using mice unilaterally lesioned with 6-OHDA. Our data confirmed that VCE-003.2 administered orally (20 mg/kg) preserved tyrosine hydroxylase (TH)-positive nigral neurons against 6-OHDA-induced damage, whereas it completely attenuated the astroglial (GFAP) and microglial (CD68) reactivity found in the substantia nigra of lesioned mice. Such neuroprotective effects caused an important recovery in the motor deficiencies displayed by 6-OHDA-lesioned mice in the pole test and the cylinder rearing test. We also investigated CBGA-Q, given orally (20 mg/kg) or intraperitoneally (10 mg/kg, i.p.), having similar benefits compared to VCE-003.2 against the loss of TH-positive nigral neurons, glial reactivity and motor defects caused by 6-OHDA. Lastly, the sodium salt of CBGA-Q, given orally (40 mg/kg) to 6-OHDA-lesioned mice, also showed benefits at behavioral and histopathological levels, but to a lower extent compared to the other two compounds. In contrast, when given i.p., CBGA-Q-Salt (10 mg/kg) was poorly active. We also analyzed the concentrations of dopamine and its metabolite DOPAC in the striatum of 6-OHDA-lesioned mice after the treatment with the different compounds, but recovery in the contents of both dopamine and DOPAC was only found after the treatment with VCE-003.2. In summary, our data confirmed the neuroprotective potential of VCE-003.2 in 6-OHDA-lesioned mice, which adds to its previous activity found in an inflammatory model of PD (LPS-lesioned mice). Additional phytocannabinoid derivatives, CBGA-Q and CBGA-Q-Salt, also afforded neuroprotection in 6-OHDA-lesioned mice, but their effects were lower compared to VCE-003.2, in particular in the case of CBGA-Q-Salt. In vitro studies confirmed the relevance of PPAR-γ receptors for these effects.
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http://dx.doi.org/10.1016/j.mcn.2020.103583DOI Listing
January 2021

Diagnosis of atypical mycobacterial and fungal coinfection.

Int J Mycobacteriol 2020 Oct-Dec;9(4):435-437

Department of Internal Medicine, General Universitary Hospital Gregorio Marañón, Doctor Esquerdo Street, 46, Madrid, 28007, Spain.

A 74-year-old female was hospitalized for further study of chest computed tomography (CT) scan compatible with mycobacterial infection. She had no history of underlying lung disease or immune alteration. At the moment of admission, she was completely asymptomatic. Scedosporium apiospermum a filamentous fungus was isolated first in the bronchoalveolar lavage. Weeks later, Mycobacterium avium complex grows in the mycobacterial culture sample.
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http://dx.doi.org/10.4103/ijmy.ijmy_98_20DOI Listing
December 2020

Cardiac tamponade and adrenal insufficiency due to pembrolizumab: a case report.

Eur Heart J Case Rep 2018 Jun 3;2(2):yty038. Epub 2018 Apr 3.

Department of Oncology, Hospital Universitario Vall d'Hebrón, Vall d'Hebrón Institute of Oncology, Paseo Vall d'Hebrón, Barcelona, Spain.

Introduction: Patients who receive or have received anti-programmed cell-death-1 (PD-1) monoclonal antibodies can develop immune-related adverse events due to activation of the immune system.

Case Presentation: We report a case of a patient who received pembrolizumab and presented with cardiac tamponade. Despite pericardial drainage, she persisted with refractory arterial hypotension due to secondary adrenal insufficiency. After initiating corticosteroid therapy, the patient recovered successfully.

Discussion: The association of pericarditis, hypophysitis and thyroid dysfunction support the diagnosis of a life-threatening immune-related adverse event due to pembrolizumab. In case of immune-related adverse events secondary to anti-PD-1 monoclonal antibodies, corticosteroid therapy should be promptly initiated in order to avoid major complications.
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http://dx.doi.org/10.1093/ehjcr/yty038DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6177032PMC
June 2018

Thermodynamic and Kinetic Analysis of Isothermal Titration Calorimetry Experiments by Using KinITC in AFFINImeter.

Methods Mol Biol 2019 ;1964:225-239

AFFINImeter Scientific & Development Team, Software 4 Science Developments, S. L. Ed. Emprendia, Campus Vida, Santiago de Compostela, A Coruña, Spain.

Standard molecular binding isothermal titration calorimetric (ITC) experiments are designed to get thermodynamic information: changes in Gibbs energy, enthalpy, and entropy associated to the studied process. Traditionally, the kinetic information contained in the ITC raw signal has been ignored. For a usual one-step process, this corresponds to the rate constants for the association and the dissociation of the complex (k and k). The availability of highly sensitive ITC instruments with low response time, together with the development of theoretical methods and of public software for the proper analysis of the signal, cancels any reason for not retrieving this kinetic information. Here we describe how to further exploit ITC experiments of simple one-step interactions by using the software AFFINImeter.The method is exemplified using a standard reference system for thermodynamic and kinetic molecular binding analysis: the interaction of carbonic anhydrase (CA) with its inhibitor 4-carboxybenzenesulfonamide (4-CBS) at several temperatures. It is to be emphasized that old experiments initially designed and executed just for thermodynamic analysis can be readily recycled by using AFFINImeter to retrieve the previously ignored kinetic information.
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http://dx.doi.org/10.1007/978-1-4939-9179-2_16DOI Listing
August 2019

The Use of ITC and the Software AFFINImeter for the Quantification of the Anticoagulant Pentasaccharide in Low Molecular Weight Heparin.

Methods Mol Biol 2019 ;1964:215-223

AFFINImeter Scientific & Development Team, Software 4 Science Developments, S. L. Ed. Emprendia, Santiago de Compostela, 15782, A Coruña, Spain.

In this chapter, we describe an original protocol based on ITC experiments and data analysis with the software AFFINImeter to get information of heparin-AT interactions relevant for the elucidation of the anticoagulant activity of heparins. This protocol is used to confirm the presence of the bioactive pentasaccharide with anticoagulant activity in heparins and to determine the amount of this pentasaccharide in the sample. Here we have applied this protocol to the characterization of low molecular weight heparins.
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http://dx.doi.org/10.1007/978-1-4939-9179-2_15DOI Listing
August 2019

AFFINImeter: A software to analyze molecular recognition processes from experimental data.

Anal Biochem 2019 07 5;577:117-134. Epub 2019 Mar 5.

Fundación Pública Galega Centro Tecnolóxico de Supercomputación de Galicia (CESGA), Avda. de Vigo s/n, 15705, Santiago de Compostela, Spain.

The comprehension of molecular recognition phenomena demands the understanding of the energetic and kinetic processes involved. General equations valid for the thermodynamic analysis of any observable that is assessed as a function of the concentration of the involved compounds are described, together with their implementation in the AFFINImeter software. Here, a maximum of three different molecular species that can interact with each other to form an enormous variety of supramolecular complexes are considered. The corrections currently employed to take into account the effects of dilution, volume displacement, concentration errors and those due to external factors, especially in the case of ITC measurements, are included. The methods used to fit the model parameters to the experimental data, and to generate the uncertainties are described in detail. A simulation tool and the so called kinITC analysis to get kinetic information from calorimetric experiments are also presented. An example of how to take advantage of the AFFINImeter software for the global multi-temperature analysis of a system exhibiting cooperative 1:2 interactions is presented and the results are compared with data previously published. Some useful recommendations for the analysis of experiments aimed at studying molecular interactions are provided.
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http://dx.doi.org/10.1016/j.ab.2019.02.031DOI Listing
July 2019

Early evolution of BRAFV600 status in the blood of melanoma patients correlates with clinical outcome and identifies patients refractory to therapy.

Melanoma Res 2018 06;28(3):195-203

Laboratory of Oncology, Institute of Oncology Dr Rosell (IOR), University Hospital Quirón Dexeus.

Serial analysis of BRAF mutations in circulating-free DNA (cfDNA) could be of prognostic value in melanoma patients. We collected blood samples from 63 advanced BRAFV600E/K melanoma patients and determined BRAFV600E/K status in cfDNA using a quantitative 5'-nuclease PCR-based assay. Levels of BRAF mutation in pre-cfDNAs were associated significantly with tumour burden, progression-free survival and overall survival. Changes in BRAF status in cfDNA after initiation of treatment (early-cfDNA) had a significant correlation with outcome. In patients with persistent BRAF mutations (n=12), progression-free survival and overall survival were 3.5 months [95% confidence interval (CI): 1.6-4.6] and 5.3 months (95% CI: 3.4-8.1) compared with 16.6 months (95% CI: 8.2-22.3) and 21.9 months (95% CI: 10.2-NR) in patients with BRAF negativization (n=16), and 15.1 months (95% CI: 2.3-NR) and NR (95% CI: 5.1-NR) in patients who maintained their initial negative status (n=12) (P<0.0001). The median duration of response in patients with radiological response, but persistence of BRAFV600 in early-cfDNA (n=5) was 4 months. Our study indicates that serial BRAF testing in the blood of advanced melanoma identifies patients refractory to therapy.
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http://dx.doi.org/10.1097/CMR.0000000000000432DOI Listing
June 2018

Mitochondria sustain store-operated currents in colon cancer cells but not in normal colonic cells: reversal by non-steroidal anti-inflammatory drugs.

Oncotarget 2017 Aug 21;8(33):55332-55352. Epub 2017 Jul 21.

Institute of Molecular Biology and Genetics (IBGM), Spanish National Research Council (CSIC), Valladolid, Spain.

Tumor cells undergo a critical remodeling of intracellular Ca homeostasis that contribute to important cancer hallmarks. Store-operated Ca entry (SOCE), a Ca entry pathway modulated by mitochondria, is dramatically enhanced in colon cancer cells. In addition, most cancer cells display the Warburg effect, a metabolic switch from mitochondrial metabolism to glycolysis that provides survival advantages. Accordingly, we investigated mitochondria control of store-operated currents (SOCs) in two cell lines previously selected for representing human normal colonic cells and colon cancer cells. We found that, in normal cells, mitochondria are important for SOCs activity but they are unable to prevent current inactivation. In contrast, in colon cancer cells, mitochondria are dispensable for SOCs activation but are able to prevent the slow, Ca-dependent inactivation of SOCs. This effect is associated to increased ability of tumor cell mitochondria to take up Ca due to increased mitochondrial potential (ΔΨ) linked to the Warburg effect. Consistently with this view, selected non-steroidal anti-inflammatory drugs (NSAIDs) depolarize mitochondria, inhibit mitochondrial Ca uptake and promote SOC inactivation, leading to inhibition of both SOCE and cancer cell proliferation. Thus, mitochondria sustain store-operated currents in colon cancer cells but not in normal colonic cells and this effect is counteracted by selected NSAIDs providing a mechanism for cancer chemoprevention.
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http://dx.doi.org/10.18632/oncotarget.19430DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5589662PMC
August 2017

Adverse systemic reaction to benznidazole.

Rev Soc Bras Med Trop 2017 Jan-Feb;50(1):145-147

Department of Allergy, Hospital Univesitari i Politècnic La Fe, Valencia, España.

Benznidazole, drug of choice for Chagas disease (CD), has been associated with a high incidence of adverse reactions that can become serious, necessitating discontinuation of the drug. We describe the case of a Bolivian patient living in Spain for 9 years, who, following treatment with benznidazole for CD in indeterminate chronic phase, presented with fever, skin lesions, digestive symptoms, general malaise, and laboratory abnormalities. After the discontinuation of benznidazole and, the intake of antihistamines and systemic corticosteroids, the patient presented a complete resolution of the symptoms. Optimization of dose strategies and development of more effective, and better-tolerated drugs is advisable.
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http://dx.doi.org/10.1590/0037-8682-0071-2016DOI Listing
May 2017

Simultaneous determination of thermodynamic and kinetic parameters of aminopolycarbonate complexes of cobalt(II) and nickel(II) based on isothermal titration calorimetry data.

J Mol Recognit 2017 04 25;30(4). Epub 2016 Nov 25.

Faculty of Chemistry, University of Gdańsk, Gdańsk, Poland.

The influence of the different side chain residues on the thermodynamic and kinetic parameters for complexation reactions of the Co and Ni ions has been investigated by using the isothermal titration calorimetry (ITC) technique supported by potentiometric titration data. The study was concerned with the 2 common tripodal aminocarboxylate ligands, namely, nitrilotriacetic acid and N-(2-hydroxyethyl) iminodiacetic acid. Calorimetric measurements (ITC) were run in the 2-(N-morpholino)ethanesulfonic acid hydrate (2-(N-morpholino) ethanesulfonic acid), piperazine-N,N'-bis(2-ethanesulfonic acid), and dimethylarsenic acid buffers (0.1 mol L , pH 6) at 298.15 K. The quantification of the metal-buffer interactions and their incorporation into the ITC data analysis enabled to obtain the pH-independent and buffer-independent thermodynamic parameters (K, ΔG, ΔH, and ΔS) for the reactions under study. Furthermore, the kinITC method was applied to obtain kinetic information on complexation reactions from the ITC data. Correlations, based on kinetic and thermodynamic data, between the kinetics of formation of Co and Ni complexes and their thermodynamic stabilities are discussed.
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http://dx.doi.org/10.1002/jmr.2589DOI Listing
April 2017

Molecular Insights into the Mechanism of Calmodulin Inhibition of the EAG1 Potassium Channel.

Structure 2016 Oct 8;24(10):1742-1754. Epub 2016 Sep 8.

Instituto de Biologia Molecular e Celular, Rua Alfredo Allen, 208, 4200-135 Porto, Portugal; Instituto de Investigação e Inovação em Saúde, Universidade do Porto, 4200-135 Porto, Portugal. Electronic address:

The human EAG1 potassium channel belongs to the superfamily of KCNH voltage-gated potassium channels that have roles in cardiac repolarization and neuronal excitability. EAG1 is strongly inhibited by Ca/calmodulin (CaM) through a mechanism that is not understood. We determined the binding properties of CaM with each one of three previously identified binding sites (BDN, BDC1, and BDC2), analyzed binding to protein stretches that include more than one site, and determined the effect of neighboring globular domains on the binding properties. The determination of the crystal structure of CaM bound to BDC2 shows the channel fragment interacting with only the C lobe of calmodulin and adopting an unusual bent conformation. Based on this structure and on a functional and biochemical analysis of mutants, we propose a model for the mechanism of inhibition whereby the local conformational change induced by CaM binding at BDC2 lies at the basis of channel modulation.
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http://dx.doi.org/10.1016/j.str.2016.07.020DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5176025PMC
October 2016

A genome-wide perspective about the diversity and demographic history of seven Spanish goat breeds.

Genet Sel Evol 2016 07 25;48(1):52. Epub 2016 Jul 25.

Department of Animal Genetics, Center for Research in Agricultural Genomics (CSIC-IRTA-UAB-UB), Campus Universitat Autònoma de Barcelona, 08193, Bellaterra, Spain.

Background: The main goal of the current work was to infer the demographic history of seven Spanish goat breeds (Malagueña, Murciano-Granadina, Florida, Palmera, Mallorquina, Bermeya and Blanca de Rasquera) based on genome-wide diversity data generated with the Illumina Goat SNP50 BeadChip (population size, N = 176). Five additional populations from Europe (Saanen and Carpathian) and Africa (Tunisian, Djallonké and Sahel) were also included in this analysis (N = 80) for comparative purposes.

Results: Our results show that the genetic background of Spanish goats traces back mainly to European breeds although signs of North African admixture were detected in two Andalusian breeds (Malagueña and Murciano-Granadina). In general, observed and expected heterozygosities were quite similar across the seven Spanish goat breeds under analysis irrespective of their population size and conservation status. For the Mallorquina and Blanca de Rasquera breeds, which have suffered strong population declines during the past decades, we observed increased frequencies of large-sized (ROH), a finding that is consistent with recent inbreeding. In contrast, a substantial part of the genome of the Palmera goat breed comprised short ROH, which suggests a strong and ancient founder effect.

Conclusions: Admixture with African goats, genetic drift and inbreeding have had different effects across the seven Spanish goat breeds analysed in the current work. This has generated distinct patterns of genome-wide diversity that provide new clues about the demographic history of these populations.
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http://dx.doi.org/10.1186/s12711-016-0229-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4960707PMC
July 2016

Genetic profile of GNAQ-mutated blue melanocytic neoplasms reveals mutations in genes linked to genomic instability and the PI3K pathway.

Oncotarget 2016 May;7(19):28086-95

Biomedical Research in Melanoma-Animal Models and Cancer Laboratory, Oncology Program, Vall d'Hebron Research institute, VHIR-Vall d'Hebron Hospital, Barcelona-UAB 08035, Barcelona, Spain.

Melanomas arising in association with a common or cellular blue nevus (MABN) comprise a relatively rare and heterogeneous group of lethal melanomas. Although GNAQ is known to be frequently mutated in common blue nevus, cellular blue nevus (CBN) and MABN and these malignant lesions present gross chromosome alterations harboring BAP1 mutations, little is known about other mutations that contribute to the development and progression of these neoplasms. Thus, the genetic profile of these tumors is important to increase the number of intervention and treatment modalities. Here, we characterized and genetically profiled two different sections of a rare MABN and two CBNs from three different patients. All of the samples harbored a GNAQ mutation, exhibited RAS pathway activation, and harbored additional mutations in genes associated with genomic instability and epigenetic regulation (KMT2C, FANCD2, ATR, ATRX, NBN, ERCC2, SETD2, and WHSC1). In addition, all neoplasms harbored mutations that directly or indirectly affected either the regulation or activation of the PI3K pathway (PIK3CA, NF1, INPP5B and GSK3B). Our results not only help understand the genetic complexity of these blue melanocytic lesions but provide a rationale to use the combination of PI3K/MTOR and MEK1/2 inhibitors against these types of tumors.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5053711PMC
http://dx.doi.org/10.18632/oncotarget.8578DOI Listing
May 2016

Extending ITC to Kinetics with kinITC.

Methods Enzymol 2016 30;567:157-80. Epub 2015 Oct 30.

AFFINImeter Scientific & Development Team, Software 4 Science Developments, S. L. Ed. Emprendia, Campus Vida, Santiago de Compostela, A Coruña, Spain.

Isothermal titration calorimetry (ITC) has long been used for kinetic studies in chemistry, but this remained confined to enzymatic studies in the biological field. In fact, the biological community has long had the tendency of ignoring the kinetic possibilities of ITC considering it solely as a thermodynamic technique, whereas surface plasmon resonance is seen as the kinetic technique par excellence. However, the primary signal recorded by ITC is a heat power which is directly related to the kinetics of the reaction. Here, it is shown how this kinetic signal can be recovered by using kinITC, the kinetic extension of ITC. The theoretical basis of kinITC is detailed for the most common situation of a second-order reaction A+B Ω C characterized by kinetic parameters kon, koff. A simplified kinITC-ETC method based upon the determination of an "Equilibration Time Curve" (ETC) is presented. The ETC is obtained by automatic determination of the "effective end" of each injection. The method is illustrated with experimental results with a comparison to Surface Plasmon Resonance (SPR) data. kon values were obtained in a wide range, from 10(3) to 0.5×10(6) M(-1) s(-1). All procedures were implemented in the program AFFINImeter (https://www.affinimeter.com/).
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http://dx.doi.org/10.1016/bs.mie.2015.08.026DOI Listing
October 2016

Genetic evolution of nevus of Ota reveals clonal heterogeneity acquiring BAP1 and TP53 mutations.

Pigment Cell Melanoma Res 2016 Mar;29(2):247-53

Biomedical Research in Melanoma-Animal Models and Cancer Laboratory, Vall dHebron Research Institute-VHIR Vall d'Hebron Hospital, Autonomous University of Barcelona, Barcelona, Spain.

Melanoma presents molecular alterations based on its anatomical location and exposure to environmental factors. Due to its intrinsic genetic heterogeneity, a simple snapshot of a tumor's genetic alterations does not reflect the tumor clonal complexity or specific gene-gene cooperation. Here, we studied the genetic alterations and clonal evolution of a unique patient with a Nevus of Ota that developed into a recurring uveal-like dermal melanoma. The Nevus of Ota and ulterior lesions contained GNAQ mutations were c-KIT positive, and tumors showed an increased RAS pathway activity during progression. Whole-exome sequencing of these lesions revealed the acquisition of BAP1 and TP53 mutations during tumor evolution, thereby unmasking clonal heterogeneity and allowing the identification of cooperating genes within the same tumor. Our results highlight the importance of studying tumor genetic evolution to identify cooperating mechanisms and delineate effective therapies.
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http://dx.doi.org/10.1111/pcmr.12452DOI Listing
March 2016

SIMON: Integration of mobility and parking solutions for people with disabilities.

Stud Health Technol Inform 2015 ;217:332-6

ETRA Investigacion y Desarrollo, S.A., Spain.

Mobility and parking in urban areas are often difficult for people with disabilities. Obstacles include lack of accessible information on routes, transport alternatives and parking availability, as well as fraud in the use of the specific services intended for these citizens. The SIMON project aims to improve this situation through the integration of different ICT solutions. SIMON is enhancing the European Parking Card for disable people with contactless technologies and integrates mobile solutions to support user unique identification in existing parking areas whilst preserving privacy. SIMON will also promote better mobility solutions for mobility including information, navigation and access to restricted areas.
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April 2017

Ipilimumab for advanced melanoma: experience from the Spanish Expanded Access Program.

Melanoma Res 2014 Dec;24(6):577-83

aMedical Oncology, Hospital General Valencia bMedical Oncology, Instituto Valenciano de Oncologia, Valencia cMedical Oncology, Hospital Clinic i Provincial dMedical Oncology, Hospital Vall d´Hebron, Barcelona eMedical Oncology, Hospital 12 de Octubre fMedical Oncology, Hospital La Paz gMedical Oncology, Hospital MD Anderson Cancer Center, Madrid hMedical Oncology, Hospital ClinicoVirgen de la Victoria, Malaga iMedical Oncology, Hospital Virgen de las Nieves, Granada jMedical Oncology, Clinica Universitaria de Navarra, Pamplona, Spain.

Ipilimumab, a fully human, recombinant, monoclonal antibody to cytotoxic T-lymphocyte antigen 4 improves overall survival (OS) in previously treated and untreated metastatic melanoma. This retrospective analysis reports data gathered by a questionnaire on the demographics, outcomes, and toxicity of ipilimumab administered through an Expanded Access Program (EAP). Ipilimumab 3 mg/kg was administered intravenously every 3 weeks for four cycles to adults with metastatic melanoma. Efficacy outcomes included complete response, partial response (PR), progressive disease, stabilized disease, and OS. EAP data were collected from EAP physicians. A subgroup analysis examined efficacy in elderly patients (≥70 years) and factors predictive of survival were identified. Of 355 requests for ipilimumab, resulting in 288 treatments, completed questionnaires were received for 153 ipilimumab recipients (median age 58 years, 57.2% men). Efficacy was evaluated in 144 patients: complete response in 1.3%, PR in 9.6%, PR with previous progression 8.4%, stabilized disease in 14.5%, and progressive disease in 66.2%. The median OS was 6.5 months (199 days); 1-year survival was 32.9%. Predictive survival factors included lymphocytes over 1000/ml (P=0.0008) and lactate dehydrogenase more than 1.5×upper limit of normal (P=0.003). Cutaneous, hepatic, and gastrointestinal toxicities were mild. In 30 patients aged more than 70 years, ipilimumab efficacy and tolerability was similar to that of the overall population. In the clinical practice setting, ipilimumab is effective and well tolerated in patients with advanced melanoma, including elderly patients, when administered at the recommended dosage. Ipilimumab improves treatment options for patients who, until recently, have had little hope of an improved prognosis.
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http://dx.doi.org/10.1097/CMR.0000000000000108DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4457498PMC
December 2014

Intracellular Ca(2+) remodeling during the phenotypic journey of human coronary smooth muscle cells.

Cell Calcium 2013 Nov 8;54(5):375-85. Epub 2013 Sep 8.

Institute of Molecular Biology and Genetics (IBGM), Spanish National Research Council (CSIC), 47003 Valladolid, Spain.

Vascular smooth muscle cells undergo phenotypic switches after damage which may contribute to proliferative disorders of the vessel wall. This process has been related to remodeling of Ca(2+) channels. We have tested the ability of cultured human coronary artery smooth muscle cells (hCASMCs) to return from a proliferative to a quiescent behavior and the contribution of intracellular Ca(2+) remodeling to the process. We found that cultured, early passage hCASMCs showed a high proliferation rate, sustained increases in cytosolic [Ca(2+)] in response to angiotensin II, residual voltage-operated Ca(2+) entry, increased Stim1 and enhanced store-operated currents. Non-steroidal anti-inflammatory drugs inhibited store-operated Ca(2+) entry and abolished cell proliferation in a mitochondria-dependent manner. After a few passages, hCASMCs turned to a quiescent phenotype characterized by lack of proliferation, oscillatory Ca(2+) response to angiotensin II, increased Ca(2+) store content, enhanced voltage-operated Ca(2+) entry and Cav1.2 expression, and decreases in Stim1, store-operated current and store-operated Ca(2+) entry. We conclude that proliferating hCASMCs return to quiescence and this switch is associated to a remodeling of Ca(2+) channels and their control by subcellular organelles, thus providing a window of opportunity for targeting phenotype-specific Ca(2+) channels involved in proliferation.
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http://dx.doi.org/10.1016/j.ceca.2013.08.006DOI Listing
November 2013

Thermodynamic signature of substrates and substrate analogs binding to human blood group B galactosyltransferase from isothermal titration calorimetry experiments.

Biopolymers 2013 Oct;99(10):784-95

Center of Structural and Cell Biology in Medicine, Institute of Chemistry, University of Luebeck, Ratzeburger Allee 160, 23562, Luebeck, Germany.

It has been observed earlier that human blood group B galactosyltransferase (GTB) hydrolyzes its donor substrate UDP-Galactose (UDP-Gal) in the absence of acceptor substrate, and that this reaction is promoted by the presence of an acceptor substrate analog, α-L-Fuc-(1,2)-β-D-3-deoxy-Gal-O-octyl (3DD). This acceleration of enzymatic hydrolysis of UDP-Gal was traced back to an increased affinity of GTB toward the donor substrate in the presence of 3DD. Herein, we present new thermodynamic data from isothermal titration calorimetry (ITC) on the binding of donor and acceptor substrates and analogs to GTB. ITC data are supplemented by surface plasmon resonance and STD-NMR titration experiments. These new data validate mutual allosteric control of binding of donor and acceptor substrates to GTB. It is of note that ITC experiments reveal significant differences in enthalpic and entropic contributions to binding of the natural donor substrate UDP-Gal, when compared with its analog UDP-Glucose (UDP-Glc). This may reflect different degrees of ordering of an internal loop (amino acids 176-194) and the C-terminus (amino acids 346-354), which close the binding pocket on binding of UDP-Gal or UDP-Glc. As both ligands have rather similar dissociation constants KD and almost identical modes of binding this finding is unexpected. Another surprising finding is that an acceptor analog, α-L-Fuc-(1,2)-β-D-3-amino-3-deoxy-Gal-O-octyl (3AD) as well as the constituent monosaccharide β-D-3-amino-3-deoxy-Gal-O-octyl (3AM) effectively inhibit enzymatic hydrolysis of UDP-Gal. This is unexpected, too, because in analogy to the effects of 3DD one would have predicted acceleration of enzymatic hydrolysis of UDP-Gal. It is difficult to explain these observations based on structural data alone. Therefore, our results highlight that there is an urgent need of experimental studies into the dynamic properties of GTB.
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http://dx.doi.org/10.1002/bip.22297DOI Listing
October 2013

Real-time evaluation of binding mechanisms in multivalent interactions: a surface plasmon resonance kinetic approach.

J Am Chem Soc 2013 Apr 12;135(16):5966-9. Epub 2013 Apr 12.

Department of Organic Chemistry and Center for Research in Biological Chemistry and Molecular Materials, University of Santiago de Compostela, Jenaro de la Fuente s/n, 15782 Santiago de Compostela, Spain.

Multivalency is a key, ubiquitous phenomenon in nature characterized by a complex combination of binding mechanisms, with special relevance in carbohydrate-lectin recognition. Herein we introduce an original surface plasmon resonance kinetic approach to analyze multivalent interactions that has been validated with dendrimers as monodisperse multivalent analytes binding to lectin clusters. The method, based on the analysis of early association and late dissociation phases of the sensorgrams provides robust information of the glycoconjugate binding efficiency and real-time structural data of the binding events under the complex scenario of the glyco-cluster effect. Notably, it reveals the dynamic nature of the interaction and offers experimental evidence on the contribution of binding mechanisms.
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http://dx.doi.org/10.1021/ja400951gDOI Listing
April 2013

Multiresidue method for pesticide residue analysis in food of animal and plant origin based on GC or LC and MS or MS/MS.

J AOAC Int 2012 Nov-Dec;95(6):1777-96

Servei de Química, Laboratori de l'Agència de Salut Pública de Barcelona, Avinguda Drassanes 13, 08001 Barcelona, Spain.

A multiresidue method based on GC or LC and MS or MS/MS for the determination of 204 pesticides in diverse food matrixes of animal and plant origin is described. The method can include different stages of cleanup according to the chemical characteristics of each sample. Samples were extracted using accelerated solvent extraction. Those with a high fat content or that contained chlorophyll required further purification by gel permeation chromatography and/or SPE (ENVI-Carb). The methodology developed here was fully validated; the LOQs for the 204 pesticides are presented. The LOQ values lie between 0.01 to 0.02 mg/kg. However, in some cases, mainly in baby food, they were as low as 0.003 mg/kg, thereby meeting European Union requirements on maximum residue levels for pesticides, as outlined in European regulation 396/2005 and the Commission Directive 2003/13/EC. The procedure has been accredited for a wide scope of pesticides and matrixes by the Spanish Accreditation Body (ENAC) following ISO/IEC 17025:2005, as outlined in ENAC technical note NT-19.
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http://dx.doi.org/10.5740/jaoacint.11-036DOI Listing
April 2013

The power of hyphenated chromatography/time-of-flight mass spectrometry in public health laboratories.

J Agric Food Chem 2012 May 18;60(21):5311-23. Epub 2012 May 18.

Research Institute for Pesticides and Water, University Jaume I, E-12071 Castellón, Spain.

Laboratories devoted to the public health field have to face the analysis of a large number of organic contaminants/residues in many different types of samples. Analytical techniques applied in this field are normally focused on quantification of a limited number of analytes. At present, most of these techniques are based on gas chromatography (GC) or liquid chromatography (LC) coupled to tandem mass spectrometry (MS/MS). Using these techniques only analyte-specific information is acquired, and many other compounds that might be present in the samples would be ignored. In this paper, we explore the potential of time-of-flight (TOF) MS hyphenated to GC or LC to provide additional information, highly useful in this field. Thus, all positives reported by standard reference targeted LC-MS/MS methods were unequivocally confirmed by LC-QTOF MS. Only 61% of positives reported by targeted GC-MS/MS could be confirmed by GC-TOF MS, which was due to its lower sensitivity as nonconfirmations corresponded to analytes that were present at very low concentrations. In addition, the use of TOF MS allowed searching for additional compounds in large-scope screening methodologies. In this way, different contaminants/residues not included in either LC or GC tandem MS analyses were detected. This was the case of the insecticide thiacloprid, the plant growth regulator paclobutrazol, the fungicide prochloraz, or the UV filter benzophenone, among others. Finally, elucidation of unknowns was another of the possibilities offered by TOF MS thanks to the accurate-mass full-acquisition data available when using this technique.
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http://dx.doi.org/10.1021/jf300796dDOI Listing
May 2012

Direct surface plasmon resonance immunosensor for in situ detection of benzoylecgonine, the major cocaine metabolite.

Biosens Bioelectron 2011 Jul 6;26(11):4423-8. Epub 2011 May 6.

Department of Organic Chemistry and Centre for Research in Biological Chemistry and Molecular Materials (CIQUS), University of Santiago de Compostela, Av de las Ciencias, 15782 Santiago de Compostela, Spain.

In this paper the development of the first direct surface plasmon resonance (SPR) immunoassay for the detection of benzoylecgonine (BZE) is described. Immunosensor chips consisting of a high affinity monoclonal anti-BZE-antibody (anti-BZE-Ab) immobilized at high density to a sensor chip were prepared. First, BZE detection in Hepes buffer was achieved by direct, real time monitoring of the binding between BZE in solution and the surface bound antibody. The detection protocol was based on calibration curves obtained from reaction rate data and end point data analysis of sensorgrams registered after injection of a series of known BZE concentrations over the chips. Moreover, immunosensor accuracy, reproducibility, stability and robustness were tested to demonstrate their good performance as reusable devices. The immunosensor was used for BZE detection in oral fluid (OF) showing that, within 180 s, our immunoassay detects BZE concentrations as low as 4 μg/L in filtered OF-buffer (1:4) samples. This value is remarkably lower than current cut off levels established by the Substance Abuse and Mental Health Services Administration. These results manifest the potential use of this direct SPR immunoassay for the in situ sensitive detection of recent cocaine abuse, of utility in roadside drug OF testing. Moreover, it exemplifies the high potential of direct SPR immunoassays for the rapid, sensitive detection of small molecules in contrast with the more established indirect methods.
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http://dx.doi.org/10.1016/j.bios.2011.04.056DOI Listing
July 2011

Irvalec inserts into the plasma membrane causing rapid loss of integrity and necrotic cell death in tumor cells.

PLoS One 2011 Apr 27;6(4):e19042. Epub 2011 Apr 27.

Departamento de Biología Celular, PharmaMar S.A., Colmenar Viejo, Madrid, Spain.

Irvalec is a marine-derived antitumor agent currently undergoing phase II clinical trials. In vitro, Irvalec induces a rapid loss of membrane integrity in tumor cells, accompanied of a significant Ca(2+) influx, perturbations of membrane conductivity, severe swelling and the formation of giant membranous vesicles. All these effects are not observed in Irvalec-resistant cells, or are significantly delayed by pretreating the cells with Zn(2+). Using fluorescent derivatives of Irvalec it was demonstrated that the compound rapidly interacts with the plasma membrane of tumor cells promoting lipid bilayer restructuration. Also, FRET experiments demonstrated that Irvalec molecules localize in the cell membrane close enough to each other as to suggest that the compound could self-organize, forming supramolecular structures that likely trigger cell death by necrosis through the disruption of membrane integrity.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0019042PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3083409PMC
April 2011

Nonsteroidal anti-inflammatory drugs inhibit vascular smooth muscle cell proliferation by enabling the Ca2+-dependent inactivation of calcium release-activated calcium/orai channels normally prevented by mitochondria.

J Biol Chem 2011 May 14;286(18):16186-96. Epub 2011 Mar 14.

Institute of Molecular Biology and Genetics, University of Valladolid and Spanish Research Council, Valladolid, Spain.

Abnormal vascular smooth muscle cell (VSMC) proliferation contributes to occlusive and proliferative disorders of the vessel wall. Salicylate and other nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit VSMC proliferation by an unknown mechanism unrelated to anti-inflammatory activity. In search for this mechanism, we have studied the effects of salicylate and other NSAIDs on subcellular Ca(2+) homeostasis and Ca(2+)-dependent cell proliferation in rat aortic A10 cells, a model of neointimal VSMCs. We found that A10 cells displayed both store-operated Ca(2+) entry (SOCE) and voltage-operated Ca(2+) entry (VOCE), the former being more important quantitatively than the latter. Inhibition of SOCE by specific Ca(2+) released-activated Ca(2+) (CRAC/Orai) channels antagonists prevented A10 cell proliferation. Salicylate and other NSAIDs, including ibuprofen, indomethacin, and sulindac, inhibited SOCE and thereby Ca(2+)-dependent, A10 cell proliferation. SOCE, but not VOCE, induced mitochondrial Ca(2+) uptake in A10 cells, and mitochondrial depolarization prevented SOCE, thus suggesting that mitochondrial Ca(2+) uptake controls SOCE (but not VOCE) in A10 cells. NSAIDs depolarized mitochondria and prevented mitochondrial Ca(2+) uptake, suggesting that they favor the Ca(2+)-dependent inactivation of CRAC/Orai channels. NSAIDs also inhibited SOCE in rat basophilic leukemia cells where mitochondrial control of CRAC/Orai is well established. NSAIDs accelerate slow inactivation of CRAC currents in rat basophilic leukemia cells under weak Ca(2+) buffering conditions but not in strong Ca(2+) buffer, thus excluding that NSAIDs inhibit SOCE directly. Taken together, our results indicate that NSAIDs inhibit VSMC proliferation by facilitating the Ca(2+)-dependent inactivation of CRAC/Orai channels which normally is prevented by mitochondria clearing of entering Ca(2+).
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http://dx.doi.org/10.1074/jbc.M110.198952DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3091227PMC
May 2011

Computed tomography (CT) predicts accurately the pathologic tumour size in stage I non-small-cell lung cancer (NSCLC).

Clin Transl Oncol 2010 Dec;12(12):829-35

Medical Oncology Service, Vall d'Hebron University Hospital, Barcelona, Spain.

Introduction: In stage I non-small-cell lung cancer (NSCLC) tumour size has been the most consistent determinant of survival. The choice of therapy option is based on accurate definition of the stage. The aim of our study is to correlate tumour size by computed tomography scan (CT) with pathologic size and to determine possible prognostic factors in surgically resected pathologic stage IA and IB NSCLC patients.

Methods: Retrospective review of CT scans and medical history data from 89 pathologic stage I NSCLC patients. Clinical prognostic factors analysed were age, gender, smoking status, pulmonary function, performance status (PS), surgical procedure, histopathology, vessel invasion, pleural infi ltration, tumour size and number of lymph nodes resected. According to the new TNM classification for lung cancer, tumour size was divided into five groups (I: <2 cm, II: 2-3 cm, III: 3-5 cm, IV: 5-7 cm and V: >7 cm).

Results: After a median surveillance of 55.2 months, 42 patients relapsed and 55 had died. The 5-year progressionfree survival was 55.7% and 5-year overall survival (OS) 49.9% (median 58.97 months). None of the clinical parameters analysed were predictors of OS. Significant correlation was found between tumour size in CT scan and pathologic stage (Pearson 0.75).

Conclusions: In our analysis with 89 surgically resected stage IA and IB NSCLC patients we found a good correlation between clinical and pathologic tumour size by CT scan. The prognoses factors analysed had no significant impact on survival.
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http://dx.doi.org/10.1007/s12094-010-0605-6DOI Listing
December 2010

Crystallographic structure of porcine adenovirus type 4 fiber head and galectin domains.

J Virol 2010 Oct 4;84(20):10558-68. Epub 2010 Aug 4.

Departamento de Bioquímica y Biología Molecular, Facultad de Farmacia, Universidad de Santiago de Compostela, E-15782 Santiago de Compostela, Spain.

Adenovirus isolate NADC-1, a strain of porcine adenovirus type 4, has a fiber containing an N-terminal virus attachment region, shaft and head domains, and a C-terminal galectin domain connected to the head by an RGD-containing sequence. The crystal structure of the head domain is similar to previously solved adenovirus fiber head domains, but specific residues for binding the coxsackievirus and adenovirus receptor (CAR), CD46, or sialic acid are not conserved. The structure of the galectin domain reveals an interaction interface between its two carbohydrate recognition domains, locating both sugar binding sites face to face. Sequence evidence suggests other tandem-repeat galectins have the same arrangement. We show that the galectin domain binds carbohydrates containing lactose and N-acetyl-lactosamine units, and we present structures of the galectin domain with lactose, N-acetyl-lactosamine, 3-aminopropyl-lacto-N-neotetraose, and 2-aminoethyl-tri(N-acetyl-lactosamine), confirming the domain as a bona fide galectin domain.
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http://dx.doi.org/10.1128/JVI.00997-10DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2950603PMC
October 2010

Binding of an acceptor substrate analog enhances the enzymatic activity of human blood group B galactosyltransferase.

Glycobiology 2010 Jun 12;20(6):718-23. Epub 2010 Feb 12.

Institute of Chemistry, University of Luebeck, Ratzeburger Allee 160, 23538 Luebeck Germany.

The hydrolysis of the donor substrate uridine diphosphate galactose (UDP-Gal) by human blood group B galactosyltransferase (GTB) has been followed by nuclear magnetic resonance in the presence and in the absence of an acceptor substrate analog. It is observed that the presence of the acceptor substrate analog promotes hydrolysis of UDP-Gal. Subsequent analysis of the kinetics of the enzymatic hydrolysis suggests that this effect is due to an increased affinity of GTB for UDP-Gal in the presence of the acceptor analog. Isothermal titration calorimetry experiments substantiate this conclusion. As hydrolysis may be understood as a glycosyl transfer reaction where water serves as universal acceptor, we suggest that in general the binding of acceptor substrates to retaining glycosyltransferases modulates the rate of glycosyl transfer. In fact, this may point to a general mechanism used by retaining glycosyltransferases to discriminate acceptor substrates under physiological conditions.
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http://dx.doi.org/10.1093/glycob/cwq019DOI Listing
June 2010