Tecnología molecular para optimizar el uso de biomarcadores en la clínica / Molecular technology to optimize the use of biomarkers in the clinic

Este capítulo pretende introducir lastecnologías moleculares que se han ido optimizandopara el análisis de los biomarcadores en los últimosaños y que están siendo aplicadas actualmente en lapráctica clínica.El capítulo está dividido en tres bloques, en los quese tratan los conceptos y técnicas de biología molecular relacionadas con los biomarcadores basados enDNA, RNA y proteínas. Además, se dan ejemplos de casos reales en los que se utilizan estas tecnologías moleculares en la rutina clínica. (AU)

logies that have been optimized for theanalysis of biomarkers in recent years and that are currently being applied in clinical practice.The chapter is divided into three blocks, which dealwith the concepts and techniques of molecular biology related to DNA-, RNA- and proteins-based biomarkers.In addition, real case examples in which these molecular technologies are used in clinical routine are given. (AU)

A preliminary analysis of association betweenplasma microRNA expression alteration andsymptomatology improvement in Major DepressiveDisorder (MDD) patients before andafter antidepressant treatment

Background and Objectives: Currently, there is a serious need to find practical biomarker(s) for Major Depressive Disorder (MDD) therapeutic target(s). This study aimed to investigate the association between microRNA (miRNA, miR) expression level in Peripheral Blood Mononuclear Cells (PBMCs) and symptomatology improvement in MDD patients before and after six-week antidepressant treatment. Methods: By using an Affymetrix array that covers 723 human miRNAs, 26 miRNAs were identified with significantly altered expression in PBMCs in MDD patients, of which10 miRNAs were selected for quantitative real-time Reverse Transcription Polymerase Chain Reaction (RT-PCR) study. Twenty out of all the 81 MDD patients were selected formiRNA expression levels testing and symptomatology assessments before and after sixweektreatment. Results: Compared with the control group, the expression levels of miR-26b, miR-4743, miR-4498, miR-4485 and miR-1972 of the MDD group were significantly higher(P < 0.05); the changes of expression levels of miR-4743, miR-4498, miR-4485 and miR-1972 were positively related to retardation improvement (P < 0.05), and the change of expression level of miR-26b negatively to the improvement of day and night change(P < 0.05); regression analysis result demonstrated that the alteration of miR-4485 expression accounted for 28.8% of retardation improvement (P < 0.05). Conclusions: These five miRNAs (miR-4743, miR-4498, miR-4485, miR-1972 andmiR-26b) may serve as biomarker for MDD diagnosis and therapeutic targets for MDDtreatment (AU)

Direct sample preparation methods for the detection of Plum pox virus by real-time RT-PCR

Direct systems to process plant materials allowed high-throughput testing of Plum pox virus (PPV) by real-time reverse transcription (RT)-PCR without nucleic acids purification. Crude plant extracts were diluted in buffer or spotted on membranes to be used as templates. Alternatively, immobilized PPV targets were amplified from fresh sections of plant tissues printed or squashed onto the same supports, without extract preparation. Spot real-time RT-PCR was validated as a PPV diagnostic method in samples collected during the dormancy period and showed high sensitivity (93.6%), specificity (98.0%), and post-test probability (97.9%) towards sharka disease. In an analysis of 2919 Prunus samples by spot real-time RT-PCR and DASI-ELISA 90.8% of the results coincided, demonstrating high agreement (k = 0.77 +/- 0.01) between the two techniques. These results validate the use of immobilized PPV targets and spot real-time RT-PCR as screening method for largescale analyses (AU)

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Ciclo replicativo del VIH. Dianas terapéuticas consolidadas y dianas potenciales / The HIV replication cycle. Established therapeutic targets and potential targets

La investigación realizada sobre la infección por el virus de la inmunodeficiencia humana (VIH) es un paradigma de cómo el conocimiento obtenido mediante la investigación básica se traslada al tratamiento de los pacientes. Gracias al estudio del ciclo viral y de la caracterización estructural y funcional de las proteínas del VIH disponemos en el momento actual de más de 20 fármacos para el tratamiento de los pacientes infectados. En este artículo se realiza una revisión del ciclo biológico del VIH y se destacan los pasos que representan dianas preferentes para la intervención farmacológica. En la segunda parte del trabajo se resumen las características de las principales familias que forman el esqueleto del tratamiento antirretroviral clásico y se revisan los fármacos recientemente introducidos que van dirigidos frente a nuevas dianas. Por último se realiza una aproximación a los nuevos prototipos que se encuentran en fase de desarrollo preclínico y que engrosarán en el futuro el arsenal terapéutico contra la infección por el VIH(AU)

Research carried out on human immunodeficiency virus (HIV) infection illustrates the speed in the transfer of knowledge obtained from basic research to the treatment of patients. Due to studying the viral life cycle and structure and function of HIV proteins, there are currently more than 20 drugs available to treat infected patients. In this article a review is carried out on the biological cycle of HIV, highlighting those steps that represent preferential targets for pharmacological intervention. In the second part of the article, the characteristics of the main antiretroviral families that form the basis of classic antiretroviral treatment are summarised as well as reviewing the recently introduced drugs which are directed at new targets. Finally, an assessment is made of the new prototypes that are in the pre-clinical development phase and which will strengthen the future therapeutic arsenal against HIV infection(AU)