New insights in the paternal genetic landscape of Southwestern Europe: dissection of haplogroup R1b-M269 and forensic applications

325 p.El cromosoma Y determina el sexo masculino y posee una naturaleza haploide que escapa a la recombinación, lo que hace que sólo esté presente en individuos varones y que se trasmita prácticamente sin cambios de padres a hijos, estableciendo linajes paternos. El estudio de los linajes paternos, mediante polimorfismos de un solo nucleótido (single nucleotide polimorphism, SNP) en el cromosoma Y (Y-SNP), permite la reconstrucción de la historia evolutiva de los linajes paternos de la especie humana. Los marcadores genéticos del cromosoma Y pueden ser de dos tipos, los anteriormente mencionados Y-SNPs, y los microsatélites Y-STRs (short tandem repeat, STR). Su estudio permite conocer cuáles son los linajes paternos característicos o presentes en cada población humana, permitiendo así diferenciar entre unas poblaciones y otras. Por ello, en los últimos años su estudio es relevante dentro del área de la Genética Forense y otras áreas afines como la genética de poblaciones, genealogía genética o la genética evolutiva.El estudio de los Y-SNPs ha revelado que las agrupaciones de linajes paternos, también llamadas haplogrupos, se distribuyen en áreas geográficas concretas a lo largo del mundo, tanto continental comoregionalmente. En el caso del oeste de Europa el haplogrupo más común es R1b-M269. La actual composición genética de Europa ha sido objeto de múltiples controversias centradas alrededor del origen de M269, ya que las estimaciones de edad obtenidas a partir de los distintos estudios genéticos realizados por distintos autores situaron el origen de este linaje paterno tanto durante el periodo Paleolítico, como en tiempos más recientes, en el Neolítico.El objetivo principal de este estudio se centra en la reconstrucción del escenario evolutivo más probable del principal linaje paterno europeo M269 en la Península Ibérica y el suroeste de Europa a través de la disección en sus subhaplogrupos, lo que permitirá caracterizar de manera detallada la distribución de los linajes paternos presentes en la Península Ibérica e inferir el papel de esta región en la historia evolutiva de Europa.Los resultados obtenidos han permitido caracterizar el paisaje genético paterno del suroeste de Europa, revelando, por un lado, que el origen más probable de M269 sea el Este de Europa y, por otro lado, que uno de los sublinajes principales de M269, R1b-S116, presenta un patrón de distribución distinto al propuesto anteriormente por otros autores. Además, el paragrupo de S116, S116*, fue prácticamente resuelto por la presencia del sublinaje R1b-DF27, que ha resultado ser casi específico de la Península Ibérica, haciendo de él un marcador de potencial interés forense para la determinación de la ancestralidad biogeográfica paterna. Las estimaciones de edad de DF27 indican que se originó hace 4.000-4.200 años, durante la transición entre el Neolítico y la Edad de Bronce, siendo su lugar más probable de origen la región noreste de la Península Ibérica. Por otro lado, fruto de este estudio también se han desarrollado dos paneles multiplex de Y-SNPs e Y-STRs para su uso en Genética Forense y de poblaciones.En conclusión, el presente trabajo de tesis doctoral ha proporcionado, por un lado, nuevas pistas sobre la historia evolutiva de acervo genético europeo actual y, por otro lado, dos nuevas herramientas de uso forense para el análisis multiplex de Y-SNPs e Y-STRs

Los aceites minerales pueden ser una alternativa al uso de acaricidas para el control de araña roja

Los aceites minerales pueden ser una alternativa muy interesante a los acaricidas para el control de la araña roja, Tetranychus urticae Koch, una plaga que actualmente está produciendo importantes daños en el cultivo de clementinas. Sin embargo, la información sobre la dosificación de estos productos y su eficacia sobre este fitófago es prácticamente inexistente. En este trabajo se evalúa en laboratorio la eficacia de cuatro productos comerciales, a cinco concentraciones diferentes, sobre huevos, protoninfas y adultos de T. urticae. Así mismo, se estudia cómo se depositan estos productos, con el fin de determinar una posible relación entre los parámetros que describen la deposición (recubrimiento, tamaño y densidad de los impactos) y la eficacia acaricida. El producto que produjo mayor área media de las deposiciones fue también el más eficaz para controlar a la araña roja en todos los estadios y con una menor concentración de producto comercial, alcanzando niveles de eficacia de 90-100 %. Los demás aceites consiguieron eficacias elevadas a partir de concentraciones al 1,5 -2,0 %. También se observó que al aumentar la concentración de producto comercial en el caldo, se incrementó el recubrimiento y el área media de los impactos. Los aceites minerales han mostrado un gran potencial para controlar la araña roja en condiciones de laboratorio. El siguiente paso será determinar su eficacia en condiciones de campoMineral oils could be an alternative to acaricides for the control of the red spider mite, Tetranychus urticae, which causes major damage to clementine crops. However, the information on the appropriate dosage of these products and their effectiveness on this pest is practically non-existent. The effectiveness of four commercial mineral oil products (containing 81.49-85% oil) at 5 different concentrations (0.5 -2.0%) was evaluated in a laboratory study using eggs, protonymphs and adults of T. urticae. The study also investigated the deposition of the products to determine whether there was a possible relationship between deposition parameters (coating, size and density) and acaricidal efficacy. The product with the highest average deposition area was also the most effective in mite control at all stages of development, and reached 90-100% efficacy at a low concentration of product. The other 3 oils were also highly efficient at concentrations of 1.5-2%. Increasing concentration of the commercial product increased both coating and average area of impact. Mineral oils have proved to have a huge potential for the control or the red spider mite under laboratory conditions. The next step will be evaluating its efficacy under field conditions

Early development of anti-natalizumab antibodies in MS patients

The purpose of this study is to monitor the development of anti-natalizumab antibodies to evaluate their first appearance in multiple sclerosis patients, since their presence has been associated with a reduction in the efficacy of the treatment and an increase of adverse events. A total of 134 multiple sclerosis patients were included in the trial. Anti-natalizumab antibodies were monthly detected by ELISA up to the first year of treatment and subsequently, a determination was made at 18 months. 15.7% of the patients were positive, being 7.5% transiently positive and 8.2% persistently positive. The first appearance of anti-natalizumab antibodies occurred after the first month of treatment onset in 72% of positive patients; 18% did so after the second month, and 9.7% after the third month. Antibodies were never detected for the first time after the fourth infusion. The development of anti-natalizumab antibodies occurs very early after treatment onset. This observation should be considered when standardizing the follow up of patients treated with this drug in order to minimize the risks and optimize the treatment

Soluble Receptor Isoform of IFN-Beta (sIFNAR2) in Multiple Sclerosis Patients and Their Association With the Clinical Response to IFN-Beta Treatment

Alternative splicing; Soluble receptors; IFNAR; Interferon beta; Multiple sclerosisEmpalmament alternatiu; Receptors solubles; IFNAR; Interferó beta; Esclerosi múltipleSplicing alternativo; Receptores solubles; IFNAR; Interferón beta; Esclerosis múltiplePurpose: Interferon beta receptor 2 subunit (IFNAR2) can be produced as a transmembrane protein, but also as a soluble form (sIFNAR2) generated by alternative splicing or proteolytic cleavage, which has both agonist and antagonist activities for IFN-β. However, its role regarding the clinical response to IFN-β for relapsing-remitting multiple sclerosis (RRMS) is unknown. We aim to evaluate the in vitro short-term effects and after 6 and 12 months of IFN-β therapy on sIFNAR2 production and their association with the clinical response in MS patients. Methods: Ninety-four RRMS patients were included and evaluated at baseline, 6 and 12 months from treatment onset. A subset of 41 patients were classified as responders and non-responders to IFN-β therapy. sIFNAR2 serum levels were measured by ELISA. mRNA expression for IFNAR1, IFNAR2 splice variants, MxA and proteases were assessed by RT-PCR. The short-term effect was evaluated in PBMC from RRMS patients after IFN-β stimulation in vitro. Results: Protein and mRNA levels of sIFNAR2 increased after IFN-β treatment. According to the clinical response, only non-responders increased sIFNAR2 significantly at both protein and mRNA levels. sIFNAR2 gene expression correlated with the transmembrane isoform expression and was 2.3-fold higher. While MxA gene expression increased significantly after treatment, IFNAR1 and IFNAR2 only slightly increased. After short-term IFN-β in vitro induction of PBMC, 6/7 patients increased the sIFNAR2 expression. Conclusions: IFN-β administration induces the production of sIFNAR2 in RRMS and higher levels might be associated to the reduction of therapeutic response. Thus, levels of sIFNAR2 could be monitored to optimize an effective response to IFN-β therapy.This research was funded by grants from the Instituto de Salud Carlos III and co-funded by European Regional Development Fund (ERDF), Technological Development Project in health DTS/1800045 to BO-M. BO-M holds a contract from Red Andaluza de Investigacion Clínica y Traslacional en Neurología (Neuro-reca) (RIC-0111-2019). PA-G is supported by Promoción de Empleo Joven e Implantación de la Garantía Juvenil 2018 (PEJ2018-002719-A). JR-B is supported by grants from Red Temática de Investigación Cooperativa, Red Española de Esclerosis Multiple REEM (RD16/0015/0010). LL holds a Nicolás Monardes research contract (RC-002-2019) from the Andalusian Ministry of Health and Family. IB-M holds a pFIS contract (FI19/00139) from the Spanish Science and Innovation Ministry

Spinosad bait treatments as alternative to malathion to control the Mediterranean fruit fly Ceratitis capitata (Diptera : Tephritidae) in the Mediterranean Basin

Current Ceratitis capitata (Wiedemann) control in Spain is based primarily on applications of organophosphate insecticides, especially malathion, mixed with protein baits. In Spain, one of the current research lines is focused on finding more environmentally friendly insecticides. One such alternative is the insecticide spinosad. We compared two formulations and various concentrations of spinosad bait treatments (10% Spintor Cebo(R)) and 0.05, 0.1, and 0.15% Spintor 480 SC(R)+0.5% Nu-lure) under laboratory conditions and found all treatments to be effective in causing high mortality of C. capitata adults within 6 days of exposure to 0-, 3-, or 6-day-old residues. Field trials demonstrated that 5% Spintor Cebo(R), 0.1% Spintor 480 SC(R)+0.5% Nu-lure, and 0.5% Malafin 50(R)+0.5% Nu-lure reduced C. capitata adults similarly and protected fruit from C capitata damage to a similar extent. Both spinosad treatments will be promising as a replacement for malathion in bait sprays. (C) Pesticide Science Society of Japan

TRAIL and TRAIL receptors splice variants during long-term interferon β treatment of patients with multiple sclerosis: evaluation as biomarkers for therapeutic response

Objective We aimed to assess the effects of interferon β (IFNβ) treatment on the expression of the splice variants of the Tumour necrosis factor-Related Apoptosis Inducing Ligand (TRAIL) and its receptors in different cell subpopulations (CD14+, CD4+ and CD8+) from patients with multiple sclerosis (MS), and to determine whether this expression discriminated responders from non-responders to IFNβ therapy. Methods We examined mRNA expression of the TRAIL and TRAIL receptors variants in patients with MS, at baseline and after one year of IFNβ therapy, according to responsiveness to this drug. Results Long-term therapy with IFNβ increased the expression of TRAIL-α in T cell subsets exclusively from responders and decreased the expression of the isoform 2 of TRAILR-2 in monocytes from responders as well as non-responders. Lower expression of TRAIL-α, and higher expression of TRAIL-β in monocytes and T cells, was found before the onset of IFNβ therapy in patients who will subsequently become responders. Baseline expression of TRAILR-1 was also significantly higher in monocytes and CD4+ T cells from responders. Conclusions The present study shows that long-term IFNβ treatment has a direct influence on TRAIL-α and TRAILR-2 isoform 2 expression. Besides, receiver operating characteristic analysis revealed that the baseline expression of TRAIL-α in monocytes and T cells, and that of TRAILR-1 in monocytes and CD4+ T cells, showed a predictive value of the clinical response to IFNβ therapy, pointing to a role of TRAIL system in the mechanism of action of IFNβ in MS that will need further investigation

Activation of the JAK-STAT Signaling Pathway after In Vitro Stimulation with IFN beta in Multiple Sclerosis Patients According to the Therapeutic Response to IFN beta

Interferon beta (IFNß) is a common treatment used for multiple sclerosis (MS) which acts through the activation of the JAK-STAT pathway. However, this therapy is not always effective and currently there are no reliable biomarkers to predict therapeutic response. We postulate that the heterogeneity in the response to IFNß therapy could be related to differential activation patterns of the JAK-STAT signaling pathway. Our aim was to evaluate the basal levels and the short term activation of this pathway after IFNß stimulation in untreated and IFNß treated patients, as well as according to therapeutic response. Therefore, cell surface levels of IFNAR subunits (IFNAR1 and IFNAR2) and the activated forms of STAT1 and STAT2 were assessed in peripheral blood mononuclear cells from MS patients by flow cytometry. Basal levels of each of the markers strongly correlated with the expression of the others in untreated patients, but many of these correlations lost significance in treated patients and after short term activation with IFNß. Patients who had undergone IFNß treatment showed higher basal levels of IFNAR1 and pSTAT1, but a reduced response to in vitro exposure to IFNß. Conversely, untreated patients, with lower basal levels, showed a greater ability of short term activation of this pathway. Monocytes from responder patients had lower IFNAR1 levels (p = 0.039) and higher IFNAR2 levels (p = 0.035) than non-responders just after IFNß stimulation. A cluster analysis showed that levels of IFNAR1, IFNAR2 and pSTAT1-2 in monocytes grouped 13 out of 19 responder patients with a similar expression pattern, showing an association of this pattern with the phenotype of good response to IFNß (p = 0.013)

Antiviral, Immunomodulatory and Antiproliferative Activities of Recombinant Soluble IFNAR2 without IFN-beta Mediation

Soluble receptors of cytokines are able to modify cytokine activities and therefore the immune system, and some have intrinsic biological activities without mediation from their cytokines. The soluble interferon beta (IFN-ss) receptor is generated through alternative splicing of IFNAR2 and has both agonist and antagonist properties for IFN-ss, but its role is unknown. We previously demonstrated that a recombinant human soluble IFN-ss receptor showed intrinsic therapeutic efficacy in a mouse model of multiple sclerosis. Here we evaluate the potential biological activities of recombinant sIFNAR2 without the mediation of IFN-ss in human cells. Recombinant sIFNAR2 down-regulated the production of IL-17 and IFN-? and reduced the cell proliferation rate. Moreover, it showed a strong antiviral activity, fully protecting the cell monolayer after being infected by the virus. Specific inhibitors completely abrogated the antiviral activity of IFN-ss, but not that of the recombinant sIFNAR2, and there was no activation of the JAK-STAT signaling pathway. Consequently, r-sIFNAR2 exerts immunomodulatory, antiproliferative and antiviral activities without IFN-ss mediation, and could be a promising treatment against viral infections and immune-mediated diseases

Antiviral, Immunomodulatory and Antiproliferative Activities of Recombinant Soluble IFNAR2 without IFN-ß Mediation

Soluble receptors of cytokines are able to modify cytokine activities and therefore the immune system, and some have intrinsic biological activities without mediation from their cytokines. The soluble interferon beta (IFN-ß) receptor is generated through alternative splicing of IFNAR2 and has both agonist and antagonist properties for IFN-ß, but its role is unknown. We previously demonstrated that a recombinant human soluble IFN-ß receptor showed intrinsic therapeutic efficacy in a mouse model of multiple sclerosis. Here we evaluate the potential biological activities of recombinant sIFNAR2 without the mediation of IFN-ß in human cells. Recombinant sIFNAR2 down-regulated the production of IL-17 and IFN-ɣ and reduced the cell proliferation rate. Moreover, it showed a strong antiviral activity, fully protecting the cell monolayer after being infected by the virus. Specific inhibitors completely abrogated the antiviral activity of IFN-ß, but not that of the recombinant sIFNAR2, and there was no activation of the JAK-STAT signaling pathway. Consequently, r-sIFNAR2 exerts immunomodulatory, antiproliferative and antiviral activities without IFN-ß mediation, and could be a promising treatment against viral infections and immune-mediated diseases

Decreased soluble IFN-β receptor (sIFNAR2) in multiple sclerosis patients: A potential serum diagnostic biomarker

Background: The soluble isoform of the interferon-β (IFN-β) receptor (sIFNAR2) could modulate the activity of both endogenous and systemically administered IFN-β. Previously, we described lower serum sIFNAR2 levels in untreated multiple sclerosis (MS) than in healthy controls (HCs). Objective: To assess sIFNAR2 levels in a new cohort of MS patients and HCs, as well as in patients with clinically isolated syndrome (CIS) and with other inflammatory neurological disorders (OIND) and to assess its ability as a diagnostic biomarker. Methods: The cross-sectional study included 148 MS (84 treatment naive and 64 treated), 87 CIS, 42 OIND, and 96 HCs. Longitudinal study included 94 MS pretreatment and after 1 year of therapy with IFN-β, glatiramer acetate (GA), or natalizumab. sIFNAR2 serum levels were measured by a quantitative ELISA developed and validated in our laboratory. Results: Naive MS and CIS patients showed significantly lower sIFNAR2 levels than HCs and OIND patients. The sensitivity and specificity to discriminate between MS and OIND, for a sIFNAR2 cutoff value of 122.02 ng/mL, were 70.1%, and 79.4%, respectively. sIFNAR2 increased significantly in IFN-β-treated patients during the first year of therapy in contrast to GA- and natalizumab-treated patients who showed non-significant changes. Conclusion: The results suggest that sIFNAR2 could be a potential diagnostic biomarker for MS