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New real-time-PCR method to identify single point mutations in hepatitis C virus

World Journal of Gastroenterology 2016年第43期22卷 9604-9612页

作者： Qian Chen Irene Belmonte Maria Buti Leonardo Nieto Damir Garcia-Cehic Josep Gregori Celia Perales Laura Ordeig meritxell llorens Maria Eugenia Soria Rafael Esteban Juan Ignacio Esteban Francisco Rodriguez-Frias Josep Quer Liver Unit Internal Medicine Lab. Malalties Hepàtiques Vall d’Hebron Institut Recerca-Hospital Universitari Vall d’ Hebron08035 Barcelona Spain Centro de Investigación Biomédica en Red Enfermedades Hepáticas y Digestivas Instituto de Salud Carlos Ⅲ28029 madrid Spain Liver Pathology Unit Departments of Biochemistry and Microbiology Hospital Universitari Vall d’Hebron Autonomous University of Barcelona 08035 Bareelona Spain Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas del Instituto de Salud Carlos Ⅲ 28029 Madrid Spain Universitat Autònoma de Barcelona 08193 Bareelona Spain

AIM To develop a fast, low-cost diagnostic strategy to identify single point mutations in highly variable genomes such as hepatitis C virus(HCV).METHODS In patients with HCV infection, resistance-associated amino acid substitutions within the viral quasispecies prior to therapy can confer decreased susceptibility to direct-acting antiviral agents and lead to treatment failure and virological relapse. One such naturally occurring mutation is the Q80 K substitution in the HCV-NS3 protease gene, which confers resistance to PI inhibitors, particularly simeprevir. Low-cost, highly sensitive techniques enabling routine detection of these single point mutations would be useful to identify patients at a risk of treatment failure. Light Cycler methods, based on real-time PCR with sequencespecific probe hybridization, have been implemented in most diagnostic laboratories. However, this technique cannot identify single point mutations in highly variable genetic environments, such as the HCV genome. To circumvent this problem, we developed a new method to homogenize all nucleotides present in a region except the point mutation of interest. RESULTS Using nucleotide-specific probes Q, K, and R substitutions at position 80 were clearly identified at a sensitivity of 10%(mutations present at a frequency of at least 10% were detected). The technique was successfully applied to identify the Q80 K substitution in 240 HCV G1 serum samples, with performance comparable to that of direct Sanger sequencing, the current standard procedure for this purpose. The new method was then validated in a Catalonian population of 202 HCV G1-infected individuals. Q80 K was detected in 14.6% of G1 a patients and 0% of G1 b in our setting. CONCLUSION A fast, low-cost diagnostic strategy based on real-time PCR and fluorescence resonance energy transfer probe melting curve analysis has been successfully developed to identify single point mutations in highly variable genomes such as hepatitis C virus. This

关键词： Hepatitis C virus Resistance-associated amino acid substitutions Low-cost test Single-point mutations Q80K

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A new player in the beneficial effects of exercise on the aged brain

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Signal Transduction and Targeted Therapy 2020年第1期5卷 901-902页

作者： M.llorens-Martín Department of Molecular Neuropathology Centro de Biología Molecular“Severo Ochoa”CBMSOCSIC-UAMMadridSpain Center for Networked Biomedical Research on Neurodegenerative Diseases(CIBERNED) MadridSpain Department of Molecular Biology Faculty of SciencesUniversidad Autónoma de MadridMadridSpain

In a recent article in Science,Horowitz et *** glycosylphosphatidylinositol(GPI)-degrading enzyme Gpld1 as the most critical player in the blood that can transfer the effects of exercise on adult neurogenesis and cognition to sedentary aged *** also demonstrate the importance of this liver-to-brain axis in ameliorating age-related regenerative and cognitive impairments.

关键词： tar player critical

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Cerebrospinal fluid neurogranin as a new player in prion disease diagnosis and prognosis

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Neural Regeneration Research 2020年第5期15卷 861-862页

作者： Anna Villar-Piqué Inga Zerr Franc llorens Network Center for Biomedical Research in Neurodegenerative Diseases (CIBERNED) Institute Carlos IIIMinistry of HealthHospitalet de LlobregatSpain Bellvitge Biomedical Research Institute (IDIBELL) Hospitalet de LlobregatSpain Department of Neurology Clinical Dementia Center and National Reference Center for CJD SurveillanceUniversity Medical SchoolGotingenGermany German Center for Neurodegenerative Diseases (DZNE) GotingenGermany

Neurogranin (Ng) and its role as Alzheimer’s disease (AD) biomarker: Ng is a calmodulin-binding protein mainly expressed in cerebral structures such as the cortex,hippocampus and *** is mainly located in the dendritic processes,particularly in post-synaptic compartments,but also in the cytosolic compartment,being likely involved in the regulation of the intracellular calcium-calmodulin signaling pathway (Represa et al.,1990).In the last decade,a plethora of studies have demonstrated that cerebrospinal fluid (CSF) Ng is increased in AD patients and in individuals with an ADlike CSF profile (Kester et al.,2015a).This increase seems to be disease-specific because other neurodegenerative conditions including frontotemporal dementia,Lewy body dementia,Parkinson’s disease,progressive supranuclear palsy,multiple system atrophy or Huntington’s disease,present CSF Ng concentrations similar to controls (Wellington et al.,2016).Ng levels in CSF appear to be elevated in mild cognitive impairment (MCI)-affected individuals who progress to AD and are highly related to memory and cognitive function (Kester et al.,2015a;Tarawneh et al.,2016),which indicates that this protein may serve as an early AD biomarker with diagnostic utility in pre-dementia disease stages,and with prognostic utility to predict cognitive decline and MCI-to-AD conversion.

关键词： Represa cerebrospinal fluid multiple system

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The impact of the design of learning spaces on attention and memory from a neuroarchitectural approach:A systematic review

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Frontiers of Architectural Research 2022年第3期11卷 542-560页

作者： Mar llorens-Gamez Juan Luis Higuera-Trujillo Carla Sentieri Omarrementeria Carmen Llinares Department of Architectural Projects Universitat Politecnica de ValenciaValencia 46022Spain Institute for Research and Innovation in Bioengineering(i3B) Universitat Politecnica de ValenciaValencia 46022Spain

Enriched environments in animal models have demonstrated that exposure to an optimal stimulus improves behavior,cognition,and ***,the evidence base for the neurophysiological influence of human environment enrichment has not been extensively *** systematic review compiles indicators about the effect of built,indoor environments on the cognitive processes of memory and attention in *** work pursues two main objectives:(1)to define current knowledge and the methods that are useful and identify whether previously published studies indicate consistencies and(2)to report the approaches and strategies that can be used in evaluating cognitive processes affected by environment *** of this systematic review show that(1)form and geometry,(2)space distribution and context,(3)color and texture,(4)height,width,and enclosure,(5)transition and circulation,and(6)light,sound,and temperature have an impact on memory and/or attention,and they can be assessed *** all the advances in this field,methodological limitations and a lack of cross-validated standard protocols are ***,future research is necessary to provide a deep insight into how human cognition can be heightened by the environment to which it is exposed.

关键词： Built environment Neuroarchitecture Cognition Attention Memory Learning spaces

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Green tea catechins inhibit microglial activation which prevents the development of neurological disorders

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Neural Regeneration Research 2020年第10期15卷 1792-1798页

作者： Tahereh Farkhondeh Ali Mohammad Pourbagher-Shahri Milad Ashrafizadeh Silvia llorens Folgado Ali Rajabpour-Sanati Mohammad Reza Khazdair Saeed Samarghandian Cardiovascular Diseases Research Center Birjand University of Medical SciencesBirjandIran Faculty of Medicine Birjand University of Medical SciencesBirjandIran Department of Basic Science Faculty of Veterinary MedicineUniversity of TabrizTabrizIran Department of Medical Sciences Faculty of Medicine of AlbaceteCentro Regional de Investigaciones Biomedicas(CRIB)University of Castilla-La ManchaAlbaceteSpain Healthy Ageing Research Center Neyshabur University of Medical SciencesNeyshaburIran Department of Basic Medical Sciences Neyshabur University of Medical SciencesNeyshaburIran

The over-activated microglial cells induce neuroinflammation which has the main role in neurological *** over-activated microglia can disturb neuronal function by releasing inflammatory mediators leading to neuronal dysfunctions and ***,inhibition of over-activated microglia may be an effective therapeutic approach for modulating *** studies have indicated anti-neuroinflammatory effects of flavonoids such as green tea *** current research was aimed to review the effect of green tea catechins in inhibiting microglial cells,inflammatory cascades,and subsequent neurological diseases.

关键词： catechins green tea microglia neuroinflammation neurological disease

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Effect of the micro-environment on α-synuclein conversion and implication in seeded conversion assays

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Translational Neurodegeneration 2020年第1期9卷 39-54页

作者： Niccolo Candelise Matthias Schmitz Katrin Thüne Maria Cramm Alberto Rabano Saima Zafar Erik Stoops Hugo Vanderstichele Anna Villar-Pique Franc llorens Inga Zerr Department of Neurology University Medicine Goettingen and the German Center for Neurodegenerative Diseases(DZNE)Robert-Koch-Stra?e 4037075 G?ttingenGermany Biomedical Engineering and Sciences Department School of Mechanical and Manufacturing Engineering(SMME)National University of Sciences and Technology(NUST)IslamabadPakistan Department of Experimental Diagnostic and Speciality MedicineUniversity of BolognaBolognaItaly Departamento de Neuropatología y Banco de Tejidos(BT-CIEN) Fundación CIENInstituto de Salud Carlos III Centro Alzheimer Fundación Reina SofíacValderrebollo n°528031 MadridSpain ADx NeuroSciences Technologiepark 4GhentBelgium CIBERNED(Network center for biomedical research of neurodegenerative diseases) Institute Carlos IIIMadridSpain Bellvitge Biomedical Research Institute(IDIBELL) Hospitalet de LlobregatBarcelonaSpain

Background: α-Synuclein is a small soluble protein,whose physiological function in the healthy brain is poorly *** inclusions of α-synuclein,referred to as Lewy bodies(LBs),are pathological hallmarks ofαsynucleinopathies,such as Parkinson’s disease(PD)or dementia with Lewy bodies(DLB).Main body:Understanding of the molecular basis as well as the factors or conditions promoting α-synuclein misfolding and aggregation is an important step towards the comprehension of pathological mechanism ofαsynucleinopathies and for the development of efficient therapeutic *** on the conversion and aggregation mechanism of α-synuclein,novel diagnostic tests,such as protein misfolding seeded conversion assays,*** real-time quaking-induced conversion(RT-QuIC),had been *** diagnostics, α-synuclein RT-QuIC exhibits a specificity between 82 and 100%while the sensitivity varies between 70 and 100%among different *** addition,the α-synuclein RT-QuIC can be used to study the α-synuclein-seeding-characteristics of different α-synucleinopathies and to differentiate between DLB and ***:The variable diagnostic accuracy of current α-synuclein RT-QuIC occurs due to different protocols,cohorts and material etc..An impact of micro-environmental factors on the α-synuclein aggregation and conversion process and the occurrence and detection of differential misfolded α-synuclein types or strains might underpin the clinical heterogeneity of α-synucleinopathies.

关键词： RT-QuIC α-Synucleinopathies Protein misfolding Protein strains

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Glycoprotein biomarkers for the detection of pancreatic ductal adenocarcinoma

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World Journal of Gastroenterology 2018年第24期24卷 2537-2554页

作者： Esther Llop Pedro E Guerrero AdriàDuran Sílvia Barrabés Anna Massaguer María JoséFerri Maite Albiol-Quer Rafael de llorens Rosa Peracaula Department of Biology Biochemistry and Molecular Biology UnitUniversity of GironaGirona 17003Spain Biomedical Research Institute of Girona(IdIBGi).Parc Hospitalari Martíi Julià-Edifici M2 Salt 17190Spain Clinic Laboratory University Hospital Dr Josep TruetaGirona 17007Spain Department of Surgery Hepato-biliary and Pancreatic Surgery UnitUniversity Hospital Dr Josep TruetaGirona 17007Spain

Pancreatic cancer(Pa C) shows a clear tendency to increase in the next years and therefore represents an important health and social challenge. Currently, there is an important need to find biomarkers for PaC early detection because the existing ones are not useful for that purpose. Recent studies have indicated that there is a large window of time for PaC early detection, which opens the possibility to find early biomarkers that could greatly improve the dismal prognosis of this tumor. The present manuscript reviews the state of the art of the existing PaC biomarkers. It focuses on the anomalous glycosylation process and its role in PaC. Glycan structures of glycoconjugates such as glycoproteins are modified in tumors and these modifications can be detected in biological fluids of the cancer patients. Several studies have found serum glycoproteins with altered glycan chains in PaC patients, but they have not shown enough specificity for PaC. To find more specific cancer glycoproteins we propose to analyze the glycan moieties of a battery of glycoproteins that have been reported to increase in PaC tissues and that can also be found in serum. The combination of these new candidate glycoproteins with their aberrant glycosylation together with the existing biomarkers could result in a panel, which would expect to give better results as a new tool for early diagnosis of PaC and to monitor the disease.

关键词： Pancreatic cancer Aberrant glycosylation Glycoproteins Biomarkers Diagnosis

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Influence of obstetric factors on the development of postpartum antithyroid antibodies

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Open Journal of Obstetrics and Gynecology 2012年第1期2卷 41-48页

作者： José Morales Roselló Beatriz Tirado Pelaez Nuria Peralta llorens Clínica Morales Valencia Spain

Aim: To correlate obstetric data with the appearance of antithyroid antibodies. Methods: A 6 months follow up was performed on 135 healthy women with assessment of TSH, T3, T4 and antithyroid antibodies (anti thyroglobulin and anti peroxidase). Correlation of diverse obstetrical parameters with the appearance of antithyroid antibodies at 2 and 6 months postpartum was determined. Results: Only two parameters were significant during the complete follow up: the newborn weight, which correlated with both antibodies (anti-thyroglobulin and anti-peroxidase) positivity and the maternal height, which exclusively correlated with anti-thyroglobulin positivity. Conclusions: Correlation of maternal height and newborn weight with positive autoantibodies allows to consider a future clinical screening test for this disorder.

关键词： Antithyroid Antibodies Maternal Screening Microchimerism Postpartum Thyroiditis

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Ecto-GPR37:a potential biomarker for Parkinson’s disease

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Translational Neurodegeneration 2021年第1期10卷 77-90页

作者： Xavier Morato Paula Garcia-Esparcia Josep Argerich Franc llorens Inga Zerr Wojciech Paslawski Eva Borras Eduard Sabido Ulla EPetaja-Repo Victor Fernandez-Duenas Isidro Ferrer Per Svenningsson Francisco Ciruela Pharm acology Unit Department of Pathology and Experimental TherapeuticsFaculty of Medicine and Health SciencesInstitute of NeurosciencesUniversity of BarcelonaUHospitalet de LlobregatSpain Neuroscience Program Bellvitge Biomedical Research InstituteIDIBELLL'Hospitalet de LlobregatSpain Section of Neurology Department of Clinical NeuroscienceDepartm ent of NeuroscienceKarolinska InstituteStockholmSweden Neuropathology Unit Departm ent of Pathology and Experimental TherapeuticsFaculty of Medicine and Health SciencesInstitute of NeurosciencesUniversity of BarcelonaL'Hospitalet de LlobregatSpain CIBERNED Centro de Investigacion Biomedica en Red de Enfermedades NeurodegenerativasInstituto CarlosⅢMadridSpain Department of Neurology Clinical Dementia Center and National Reference Center for CID SurveillanceUniversity Medical SchoolGottingenGermany German Center for Neurodegenerative Diseases GottingenGermany Proteomics Unit Center for Genomic RegulationBarcelonaSpain Proteomics Unit Universitat Pompeu FabraBarcelonaSpain Research Unit of Biomedicine Medical Research Center OuluUniversity of OuluOuluFinland

Objective:a-Synuclein has been studied as a potential biomarker for Parkinson's disease(PD)with no concluding ***,there is an urgent need to find out reliable specific biomarkers for ***37 is an orphan G protein-coupled receptor that toxically accumulates in autosomal recessive juvenile ***,we investigated whether GPR37 is upregulated in sporadic PD,and thus a suitable potential biomarker for ***:GPR37 protein density and mRNA expression in postmortem substantia nigra(SN)from PD patients were analysed by immunoblot and RT-qPCR,*** presence of peptides from the N-terminus-cleaved domain of GPR37(***-GPR37)in human cerebrospinal fluid(CSF)was determined by liquid chromatography-mass spectrometric *** engineered in-house nanoluciferase-based immunoassay was used to quantify ecto-GPR37 in CSF samples from neurological control(NC)subjects,PD patients and Alzheimer's disease(AD)***:GPR37 protein density and mRNA expression were significantly augmented in sporadic *** amounts of ecto-GPR37 peptides in the CSF samples from PD patients were identified by mass spectrometry and quantified by the in-house ELISA ***,the CSF total a-synuclein level in PD patients did not differ from that in NC ***,the cortical GPR37 mRNA expression and CSF ecto-GPR37 levels in AD patients were also ***:GPR37 expression is increased in SN of sporadic PD *** ecto-GPR37 peptides are significantly increased in the CSF of PD patients,but not in AD *** results open perspectives and encourage further clinical studies to confirm the validity and utility of ecto-GPR37 as a potential PD biomarker.

关键词： GPR37 Pael-R Parkinson's disease Cerebrospinal fluid Biomarker Alzheimer's disease a-5ynuclein Orphan receptor

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Retail Brand Building Through Promotional Communication： A Study That Focuses on Category Killers Promotional Flyers

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Chinese Business Review 2011年第3期10卷 217-225页

作者： Roberto Manzano Anton Magdalena Ferran Aranaz Maria Puelles Gallo Miguel llorens Marin Universidad Complutense de Madrid Madrid Spain

Strong competition in retail distribution and the development of new purchasing options for the consumer generate a growing need to strengthen the retailers＇ brand image using all the communication elements, with promotional flyers being one of the most important ways to do this. Promotional flyers make-up 5.1% of the money spent on communication and rank third place in terms of importance for retailers. The aim of this investigation work is to focus on analysing the utility of a promotional tool, such as a flyer, to fulfil a strategic objective, like creating a retail brand image, within a modern dynamic sector that is growing fast, such as Category Killers. Two sources of information have been used to carry out the research. On the One hand, a survey conducted on those responsible for the definition and execution of the promotional plan, and on the other hand, the accumulation and analysis of all the flyers issued into the market thought-out a year by those retailers which belong to this format, in order to conduct a study of seven defined image indicators. This information has been largely analyzed using multidimensional scaling method and chi-square test. As a result, we have managed to gather the differential elements in flyers execution between those retailers that prize the communications of image attributes and those retailers which concentrate on communicating price. The first group makes greater use of promotional theme related flyers, and shows greater differentiation in both product assortment and the depth of the range being communicated.

关键词： retail image integrated communication store promotional flyers

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