Infección por VIH-2 en España: aspectos epidemiológicos y respuesta al tratamiento con inhibidores de la integrasa

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Bioquímica. Fecha de lectura: 13-09-2019En 1985 se identificó un patrón atípico de reactividad serológica frente al VIH-1 en prostitutas senegalesas. De forma inequívoca los sueros reconocían antígenos del SIV. Un año más tarde, Clavel y colaboradores aislaron un nuevo virus de inmunodeficiencia humana, denominado VIH- 2, en africanos occidentales. En Europa, la mayoría de las personas VIH-2+ son originarias de África occidental. En España, la llegada de africanos con infección por VIH-2, llevó a la creación del grupo español de estudio de la infección por VIH-2 en 1989. Hasta diciembre de 2018, se han comunicado 373 casos en nuestro país y aproximadamente el 10% presenta coinfección por VIH-1. La infección por VIH-2 presenta peculiaridades diagnósticas, epidemiológicas y terapéuticas que deben ser consideradas para un correcto manejo de las personas con esta infección. En esta tesis hemos abordado aspectos epidemiológicos y terapéuticos de la infección por VIH-2 en España. Para empezar, analizamos la prevalencia de las personas con coinfección VIH-1+VIH-2, así como su seguimiento clínico y terapéutico. En total se identificaron 34 sujetos, un 9% de las personas VIH-2+. Se describió una mayoría de subtipos CRF02_AG y B para el VIH-1 y de A para el VIH-2. Tras 34 meses en tratamiento antirretroviral de gran actividad (TARGA) basado en inhibidores de proteasa o inhibidores de la integrasa (INI), el porcentaje de pacientes con supresión virológica para VIH-1, VIH-2 y VIH-1+VIH-2 fue del 87%, 80% y 70%, respectivamente. También se describió un aumento de +212 linfocitos T-CD4+/μL. El segundo objetivo que se persiguió en esta tesis fue la evaluación de la eficacia del TARGA que incluía INI en pacientes VIH-2+ naïve (18 sujetos) y pretratados (26 sujetos). Después de una mediana de 13 meses en tratamiento, el porcentaje de pacientes con carga viral indetectable y aumento de linfocitos T CD4+ fue del 89% y +82 células/L y del 65,4% y +126 células/L, respectivamente. En 12/15 sujetos con fracaso virológico (FV) se detectaron mutaciones asociadas a resistencia en la integrasa: 2 en naïve y 10 en pretratados. Estas fueron: N155H (5), Q18H/R (3), Y143C/G (3) y por primera vez en el VIH-2, se identificó el cambio R263K (1). Por último, se analizaron secuencias de la región de la integrasa del VIH-2 en 20 sujetos naïve para los INI y en 10 individuos con FV a raltegravir (RAL). En las secuencias de sujetos naïve, solo se identificó el cambio S138A en un paciente, mientras que los patrones de resistencia identificados en 9 pacientes con FV a RAL fueron Y143C/G (3), Q148R+G140A/S (2) y N155H+A153S/G (4), los mismos que los observados en el VIH-1. Cinco de estos sujetos fueron rescatados con dolutegravir. Aunque inicialmente se observó buena respuesta en 4 de ellos, 3 hicieron un rebrote viral a los 6 meses. Tras este segundo fracaso se identificaron nuevos cambios: Q148K, Q148Q/R y G118R

Impacts of pure shocks in the BHR71 bipolar outflow

During the formation of a star, material is ejected along powerful jets that impact the ambient material. This outflow regulates star formation by e.g. inducing turbulence and heating the surrounding gas. Understanding the associated shocks is therefore essential to the study of star formation. We present comparisons of shock models with CO, H2, and SiO observations in a 'pure' shock position in the BHR71 bipolar outflow. These comparisons provide an insight into the shock and pre-shock characteristics, and allow us to understand the energetic and chemical feedback of star formation on Galactic scales. New CO (Jup = 16, 11, 7, 6, 4, 3) observations from the shocked regions with the SOFIA and APEX telescopes are presented and combined with earlier H2 and SiO data (from the Spitzer and APEX telescopes). The integrated intensities are compared to a grid of models that were obtained from a magneto-hydrodynamical shock code which calculates the dynamical and chemical structure of these regions combined with a radiative transfer module based on the 'large velocity gradient' approximation. The CO emission leads us to update the conclusions of our previous shock analysis: pre-shock densities of 1e4 cm-3 and shock velocities around 20-25 km s-1 are still constrained, but older ages are inferred ( 4000 years). We evaluate the contribution of shocks to the excitation of CO around forming stars. The SiO observations are compatible with a scenario where less than 4% of the pre-shock SiO belongs to the grain mantles. We infer outflow parameters: a mass of 1.8x1e-2 Msun was measured in our beam, in which a momentum of 0.4 Msun km s-1 is dissipated, for an energy of 4.2x1e43erg. We analyse the energetics of the outflow species by species. Comparing our results with previous studies highlights their dependence on the method: H2 observations only are not sufficient to evaluate the mass of outflows.Comment: 14 pages, 10 figures, 4 Tables, accepted in A&

Cómo resolver los conflictos familiares

Esta guía editada por la Comunidad de Madrid pretende facilitar y contribuir a mejorar el bienestar de las familias madrileñas, como forma constante de las actuaciones llevadas a cabo desde la Consejería de Familia y Asuntos Sociales. En la consecución de este objetivo, esta guía trata principalmente de facilitar a las familias madrileñas habilidades que les permitan gestionar con mayor facilidad los conflictos que se originan en el ámbito familiar. La Consejería de Familia y Asuntos Sociales de la Comunidad de la Comunidad de Madrid, consciente de la importancia de la familia como ente socializador, y sabiendo que la familia es el lugar donde se puede aprender a resolver los conflictos de manera consensuada, ha encargado la elaboración de la presente guía al Instituto Complutense de Mediación y Gestión de Conflictos (IMEDIA). En esta guía, nos vamos a encontrar con una familia que probablemente podría residir en la Comunidad de Madrid, y a la que vamos a acompañar en las diferentes situaciones de dificultad o conflicto propias cualquier convivencia familiar. Todos nos podremos sentir identificados con muchas de las situaciones que Iria y Alex van a tener que resolver a lo largo de estas páginas. Se tratarán momentos comunes y muy diversos dentro de la convivencia familiar. Así, se contemplan entre otros, conflictos en la relación de pareja, conflictos en la relación con los hijos, en la relación entre hermanos, en la relación de los hijos con los padres y abuelos

Aplicabilidad de la termografría para la inspección de los edificios rurales: Caso de una comarca española

This article is aimed a! establishing the chance of applying the infrared thermography techniquefor !he examination of rural buildings. The results from field worh are collected. The next issues were determined by thermography: Hidden materials and elements; dgerences behveen different materials; location of crach; localion of slructures; localion ofhumid areas; points ofheat losses and areas where the hot air is accumulated. It ispossible lo inspect a high number of buildings in a short time and get valuable results. The time where the inspection should be done is dependen1 on (he type of building: in (he evening for buildings with bearing walls and at daybreak for buildings with interior structures.En este articulo se pretende verijicar la posibilidad de utilización de la termografía infrarrojn como técnica de inspección del estado de los edificios rurales. Se reúnen los resultados obtenidos en varios trabajos de campo. Mediante esta técnica se puede analizar: localización de materiales y elementos ocultos, localización de diferentes materiales en fachada, presencia de grietas, localización de estructuras, localización de zonas húmedas y puntos de pérdidas de calot Esta técnica permite inspeccionar un alto número de edificios en un breve período de tiempo, proporcionando resultados válidos. Los resultados muestran que en función del tipo de edijicio las inspecciones se deben realizar en diferentes momeritos del dia: por la noche en los edificios con muros de catga y al amanecer en edificios con estructuras internas

A Proof of Principle Proteomic Study Detects Dystrophin in Human Plasma: Implications in DMD Diagnosis and Clinical Monitoring

Duchenne muscular dystrophy (DMD) is a rare neuromuscular disease caused by pathogenic variations in the DMD gene. There is a need for robust DMD biomarkers for diagnostic screening and to aid therapy monitoring. Creatine kinase, to date, is the only routinely used blood biomarker for DMD, although it lacks specificity and does not correlate with disease severity. To fill this critical gap, we present here novel data about dystrophin protein fragments detected in human plasma by a suspension bead immunoassay using two validated anti-dystrophin-specific antibodies. Using both antibodies, a reduction of the dystrophin signal is detected in a small cohort of plasma samples from DMD patients when compared to healthy controls, female carriers, and other neuromuscular diseases. We also demonstrate the detection of dystrophin protein by an antibody-independent method using targeted liquid chromatography mass spectrometry. This last assay detects three different dystrophin peptides in all healthy individuals analysed and supports our finding that dystrophin protein is detectable in plasma. The results of our proof-of-concept study encourage further studies in larger sample cohorts to investigate the value of dystrophin protein as a low invasive blood biomarker for diagnostic screening and clinical monitoring of DMD

CD69-oxLDL ligand engagement induces Programmed Cell Death 1 (PD-1) expression in human CD4 + T lymphocytes

The mechanisms that control the inflammatory–immune response play a key role in tissue remodelling in cardiovascular diseases. T cell activation receptor CD69 binds to oxidized low-density lipoprotein (oxLDL), inducing the expression of anti-inflammatory NR4A nuclear receptors and modulating inflammation in atherosclerosis. To understand the downstream T cell responses triggered by the CD69-oxLDL binding, we incubated CD69-expressing Jurkat T cells with oxLDL. RNA sequencing revealed a differential gene expression profile dependent on the presence of CD69 and the degree of LDL oxidation. CD69-oxLDL binding induced the expression of NR4A receptors (NR4A1 and NR4A3), but also of PD-1. These results were confirmed using oxLDL and a monoclonal antibody against CD69 in CD69-expressing Jurkat and primary CD4 + lymphocytes. CD69-mediated induction of PD-1 and NR4A3 was dependent on NFAT activation. Silencing NR4A3 slightly increased PD-1 levels, suggesting a potential regulation of PD-1 by this receptor. Moreover, expression of PD-1, CD69 and NR4A3 was increased in human arteries with chronic inflammation compared to healthy controls, with a strong correlation between PD-1 and CD69 mRNA expression (r = 0.655 P < 0.0001). Moreover, PD-1 was expressed in areas enriched in CD3 infiltrating T cells. Our results underscore a novel mechanism of PD-1 induction independent of TCR signalling that might contribute to the role of CD69 in the modulation of inflammation and vascular remodelling in cardiovascular diseasesOpen Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. This work was supported by grant S2017/BMD-3671-INFLAMUNE-CM from the Comunidad de Madrid, a grant from the Ramón Areces Foundation “Ciencias de la Vida y la Salud”, “La Caixa” Banking Foundation (HR17-00016) to FSM; grants PDC2021-121719-I00 and PDI-2020-120412RBI00 to FSM, and RTI2018-094727-B-100 to JMG funded by MCIN/ AEI/10.13039/501100011033 and by “ERDF A way of making Europe”; the Agència de Gestió d'Ajuts Universitaris i de Recerca (AGAUR) (2017-SGR-00333) to JMG; and a grant from the Instituto de Salud Carlos III (PI18/0919) to CR. M. Jiménez-Fernández is supported by a FPI-Severo Ochoa-CNIC (PRE2019-087941); C. Ballester-Servera is supported by a FPU fellowship (Ministerio de Universidades). Data availability The data underlying this article are available in the article and in its online Supplementary material. Declarations Conflict of interest The authors have no confict of interest to declare. Ethical approval Written consent was obtained from all participating subjects. The procedure was approved by the Ethics Committee of the Hospital de la Santa Creu i Sant Pau (Barcelona, Spain) and was conducted in accordance with the Declaration of Helsinki. Consent for publication Consent to publish has been received from all participant

Celia’s Encephalopathy (BSCL2-Gene-Related): Current Understanding

Seipin, encoded by the BSCL2 gene, is a protein that in humans is expressed mainly in the central nervous system. Uniquely, certain variants in BSCL2 can cause both generalized congenital lipodystrophy type 2, upper and/or lower motor neuron diseases, or progressive encephalopathy, with a poor prognosis during childhood. The latter, Celia’s encephalopathy, which may or may not be associated with generalized lipodystrophy, is caused by the c.985C >T variant. This cytosine to thymine transition creates a cryptic splicing zone that leads to intronization of exon 7, resulting in an aberrant form of seipin, Celia seipin. It has been proposed that the accumulation of this protein, both in the endoplasmic reticulum and in the nucleus of neurons, might be the pathogenetic mechanism of this neurodegenerative condition. In recent years, other variants in BSCL2 associated with generalized lipodystrophy and progressive epileptic encephalopathy have been reported. Interestingly, most of these variants could also lead to the loss of exon 7. In this review, we analyzed the molecular bases of Celia’s encephalopathy and its pathogenic mechanisms, the clinical features of the different variants, and a therapeutic approach in order to slow down the progression of this fatal neurological disorderThis work was supported by the Instituto de Salud Carlos III and the European Regional Development Fund, FEDER (grant numbers PI10/02873 and PI13/00314), by the Consellería de Industria, Xunta de Galicia (grant numbers 10PXIB208013PR, ED341b 2017/19 and ED431B 2020/37), and by Fundación Mutua Madrileña (Call 2015). S.S.I. was awarded a Research Fellowship, granted by the Asociación Española de Familiares y Afectados de Lipodistrofias (AELIP)S

Drug resistance mutations in HIV-2 patients failing raltegravir and influence on dolutegravir response

Producción CientíficaBackground: A broader extent of amino acid substitutions in the integrase of HIV-2 compared with HIV-1 might enable greater cross-resistance between raltegravir and dolutegravir in HIV-2 infection. Few studies have examined the virological response to dolutegravir in HIV-2 patients that failed raltegravir. Methods: All patients recorded in the HIV-2 Spanish cohort were examined. The integrase coding region was sequenced in viraemic patients. Changes associated with resistance to raltegravir and dolutegravir in HIV-1 were recorded. Results: From 319 HIV-2-infected patients recorded in the HIV-2 Spanish cohort, 53 integrase sequences from 30 individuals were obtained (20 raltegravir naive and 10 raltegravir experienced). Only one secondary mutation (E138A) was found in one of the 20 raltegravir-naive HIV-2 patients. For raltegravir-experienced individuals, the resistance mutation profile in 9 of 10 viraemic patients was as follows: N155H + A153G/S (four); Y143G + A153S (two); Q148R + G140A/S (two); and Y143C + Q91R (one). Of note, all patients with Y143G and N155H developed a rare non-polymorphic mutation at codon 153. Rescue therapy with dolutegravir was given to 5 of these 10 patients. After >6 months on dolutegravir therapy, three patients with baseline N155H experienced viral rebound. In two of them N155H was replaced by Q148K/R and in another by G118R. Conclusions: A wide repertoire of resistance mutations in the integrase gene occur in HIV-2-infected patients failing on raltegravir. Although dolutegravir may allow successful rescue in most HIV-2 raltegravir failures, we report and characterize three cases of dolutegravir resistance in HIV-2 patients, emerging variants Q148K and Q148R and a novel change G118R.Fondo de Investigación Sanitaria-Fondos FEDER (FIS, PI13/01574; ICI14/0274, CES12/003, FI14/00264, CD14/00243

The Use of Quinacrine in Nitroimidazole-Resistant Giardia Duodenalis: An Old Drug for an Emerging Problem

Background: There is little evidence in the management of refractory giardiasis after treatment with nitroimidazoles. This study estimates the proportion of persistent giardiasis in 3 hospitals in Barcelona, describes risk factors and genotype associated and evaluates the efficacy rate of quinacrine in those with persistent giardiasis. METHODS: A clinical prospective observational study was conducted in patients with giardiasis treated with nitroimidazoles. Those with persistent giardiasis were provided quinacrine. Molecular characterization of Giardia isolates was performed by PCR amplification of a fragment of tpi and bg genes. RESULTS: Seventy-seven patients were recruited and treated with nitroimidazoles and in 14/71 of patients followed-up(20%), Giardia persisted. Refractory giardiasis was associated with malaise(p:0.007) and anorexia(p:0.019), with previous giardiasis(p:0.034) and with previous antibiotic(p:0.02) or antiparasitic(p:0.037). Quinacrine had an effectiveness rate of 100% in refractory giardiasis(n=13,95%CI 75-100). Molecular characterization showed that 17(25%) Giardia isolates belonged to assemblage A, 31(43%) to assemblage B. In refractory giardiasis, assemblage B and A were found as responsible in 6 and 4 cases respectively. CONCLUSIONS: Almost 20% of patients presented persistent giardiasis after nitroimidazole being both assemblage A and B involved. Short course of quinacrine was effective in treating refractory cases and further controlled studies should evaluate its efficacy and safety

Vigilância da qualidade microbiológica de água de consumo humano e de água destinada ao abastecimento público da província de Valência durante o período 2002-2010

This study focuses on assessing the microbiological quality of drinking water and of water intended for drinking in the province of Valencia (Spain) between 2002 and 2010. Variation was found regarding space, time and source requirements in samples that did not meet the standards of quality established by Royal Decree 140/2003 for the following: total coliforms (TC); faecal coliforms (FC); Escherichia coli (EC); aerobic bacteria at 22 ºC (AB 22 ºC); faecal streptococci (FS); enterococci (EN); sulphite-reducing clostridia (SC); and Clostridium perfringens (CP). The samples were stratified by those meeting the standards (“Compliance”) and those that did not (“Non-Compliance”), as well as by their relationship with the degree of chlorination.A total of 10057 water samples were examined from various sources: springs; surface waters; denitrifying plant waters; wells; and distribution networks. They were grouped into each of the 17 districts of the province of Valencia.The total number of samples that failed to meet the standards of quality, for each criterion, were as follows: 34.0 % for TC; 16.0 % FS-EN; 13.0 % for FC-EC; 5.6 % for SC-CP; and 15.5 % for AB 22 ºC. Regarding spatial variation in samples, the highest percentages of samples in the “Non-Compliance” group were found in the interior part of the province. For time variation, the highest percentages of “Non-Compliance” were for the years: 2002 - 2004, 2008 and 2009. Regarding source variation, origin of the samples with “Non- compliance” was highest for surface waters, followed by springs and wells.For all samples studied, 39.8 % were within the “Non-Compliance” group, of which 18.3 % came from sources that supply the population (distribution networks).Of the samples within the “Compliance” group, 61 % were chlorinated, which confirms that chlorine is a powerful disinfectant and that chlorination is an effective water disinfection treatment.El presente trabajo tiene como objetivo la evaluación de la calidad microbiológica de las muestras de agua analizadas en la provincia de Valencia durante el período 2002-2010. Se observó la variación espacial, temporal y por origen de las muestras que no cumplían los requisitos especificados en el RD 140/2003 para los coliformes totales (CT), coliformes fecales (CF), Escherichia coli (EC), aerobios a 22 ºC (AB 22 ºC), estreptococos fecales (EF), enterococos (EN), Clostridium sulfito reductores (CS), y Clostridium perfringens (CP), estratificando las muestras en aquellas que cumplían la normativa “Conformes” y las que no “No Conformes”, y su relación con el grado de cloración.Se estudian un total de 10057 muestras de agua procedentes de fuentes que no manan de la red, aguas superficiales, aguas provenientes de plantas desnitrificadoras, pozos y redes de distribución. Se han agrupado en las 17 comarcas de la provincia de Valencia.Del total de las muestras, no cumplían los requisitos de calidad para CT el 34,0 %, para EF-EN el 16,0 %, para AB 22 ºC el 15.5 % para CF-EC el 13,0 % y para CS-CP el 5,6 %. Los porcentajes más elevados de muestras “No Conformes” se observaron situados en la zona interior, y en los años 2002-2004, 2008 y 2009. Respecto a la distribución por origen, se observaron en aguas superficiales, seguidas de fuentes y pozos.Del total de muestras estudiadas, el 39,8 % eran “No Conformes”, y de estas un 18,3 % procedían de abastecimientos que proveen a la población (redes de distribución).El 61,0 % de las muestras “Conformes” estaban cloradas, lo que demuestra que el cloro sigue siendo un tratamiento efectivo de desinfección.Este trabalho tem como objetivo avaliar a qualidade microbiológica de amostras de água analisadas na província de Valência durante o período de 2002 a 2010. Observou-se a variação espacial, temporal e da origem das amostras que não cumprem os requisitos estabelecidos no RD 140/2003, para coliformes totais (CT), coliformes fecais (CF), Escherichia coli (EC), germes aeróbios a 22 ºC (AB 22 ºC), estreptococos fecais (EF), enterococos (EN), Clostridium sulfito (CS) e Clostridium perfringens (CP), estratificando as amostras que cumpriam os requisitos em “conformes” e as que não cumpriam em “não conformes” e a relação com o nível de cloro. Foi estudado um total de 10057 amostras de água de diversas origens: fontes, águas superficiais, centrais de desnitrificação, poços e rede de distribuição. Foram agrupadas nos 17 municípios de Valência.Do total das amostras, não cumpriam os requisitos de qualidade para CT (34 %), EF e EN (16 %), AB 22 ºC (15,5 %), CF e EC (13 %) e CS e CP (5,6 %). Quanto à variação espacial registaram-se as maiores percentagens de amostras “não conformes” na zona interior e nos anos 2002 a 2004, 2008 e 2009. No que diz respeito à distribuição por origens foram observadas maiores percentagens de amostras “não conformes” nas águas de superfície, seguido de nascentes e poços.Do total de amostras analisadas, 39,8 % estavam “não conforme”, sendo que 18,3 % destas eram da rede que abastece a população (rede de distribuição).61 % das amostras “conformes” estão cloradas, o que significa que o cloro ainda é um tratamento eficaz de desinfeção