Impact of Transcriptome and Gut Microbiome on the Response of HIV-1 Infected Individuals to a Dendritic Cell-Based HIV Therapeutic Vaccine

Therapeutic vaccines based on dendritic cells offer a good approach to HIV-specific T-cell responses and partial control of the viral load after antiretroviral therapy interruption. The aim of the present study was to identify mRNA expression profiles and to assess the impact of the gut microbiome composition for predicting the viral load control after antiretroviral therapy interruption. We enrolled 29 patients to receive either placebo or a monocyte-derived dendritic cell vaccine. Patients with a decrease in their viral load of >0.5 log10 copies/mL by 12 weeks after antiretroviral therapy interruption were considered responders. In total, 66 genes were considered differentially expressed between responders and non-responders. Enrichment analysis revealed several upregulated pathways involved in the host defense response to a virus via the type I interferon signaling pathway. Regarding the gut microbiota, responders showed enriched levels of Bacteroidetes (p < 0.005) and Verrucomicrobia (p = 0.017), while non-responders were enriched with Tenericutes (p = 0.049) and Actinobacteria (p < 0.005). We also found important differences at the genus level. However, we did not discover any effect of the dendritic cell vaccine on the transcriptome or the gut microbiota. An alternative analysis did characterize that the microbiota from responders were associated with the metabolic production of short-chain fatty acids, which are key metabolites in the regulation of intestinal homeostasis. The evidence now consistently shows that short-chain fatty acid depletion occurs in HIV-infected individuals receiving antiretroviral treatment.This study was partially supported by the SPANISH AIDS Research Network (RIS) projects RD16/0025/0002 and RD16/0025/0013 integrated in the State Plan for Scientific and Technical Research and Innovation from the General Sub-Directorate for research assessment and promotion; Spanish Carlos III Institute of Health (ISCIII) co-funded by the European Regional Development Fund (FEDER); the Fondo de Investigación Sanitaria (FIS), AC16/00051 and PI18/00699; Gilead grants in Biomedical research in HIV, hepatic, and hemo-oncology from the Spanish Ministry of Economy (MINECO) (grants: SAF2015-66193-R and RTI2018-096309-B-I00); and the income found from Europe in the Regional Development (FEDER) and the CERCA Programme/Generalitat de Catalunya SGR 615 and SGR 653. Felipe García received the support of José María Segovia de Arana contracts.S

Metodologia analítica per a l’especiació de mercuri en mostres del medi aquàtic

[cat] Degut a la necessitat d’estudis sistemàtics per a l’establiment de mètodes eficaços per a la determinació d’espècies de mercuri en matrius ambientals i biològiques complexes, que assegurin la inalterabilitat de les mateixes durant la seva manipulació, aspectes que són avui dia els punts més febles en l’especiació analítica, l’objectiu de la present Tesi es basa en l’optimització de mètodes analítics per a estudiar l’especiació de mercuri en mostres del medi aquàtic mitjançant la tècnica acoblada HPLC-UV-CV-AFS. Dins d’aquest marc, la Tesi aborda uns objectius específics que es descriuen a continuació: - Optimització de totes les condicions experimentals implicades en la determinació de mercuri inorgànic i metilmercuri mitjançant CV-AFS establint, en primer lloc, les condiciones òptimes de detecció i, a continuació, estudiant la separació de les espècies mitjançant cromatografia de líquids en fase inversa; tot establint els paràmetres de qualitat per al mètode analític proposat. - Modificació i/o optimització del mètode analític per tal de determinar altres espècies de mercuri que de manera poc habitual es puguin trobar en mostres ambientals i/o biològiques. - Desenvolupament d’un mètode de preconcentració on-line per a l’especiació de mercuri en aigües. - Establiment dels efectes de pretractament en la determinació de mercuri en mostres de sediments. - Aplicació de la metodologia analítica desenvolupada a mostres ambientals i biològiques (aigües, sediments i biota) prèviament estudiant diferents mètodes d’extracció d’espècies emprant materials de referència. Els resultats obtinguts dins del tres primers objectius es mostren i es discuteixen al Capítol 3 de la present memòria; els estudis realitzats per a l’avaluació dels efectes de pretractament de mostra en sediments es descriuen al Capítol 4, i l’últim capítol descriu l’aplicació a mostres ambientals i biològiques del medi aquàtic. Als annexos I i II s’inclouen els articles científics publicats fins al moment. L’Annex I consta d’un article sobre la determinació de Hg2+ i MeHg+ en aigües contaminades mitjançant HPLC-UV-CV-AFS, mentre que a l’Annex II es troba un review sobre materials de referència certificats per a l’especiació de mercuri en matrius biològiques i ambientals.[eng] In the present thesis, analytical methodology has been developed for mercury speciation in aquatic samples. The first part of the work consists of the optimization of the method for the separation of the species by the technique HPLC-UV-CV-AFS, another part focuses on the effects of sample pretreatment for the determination of total mercury and its species in sediments, and finally the last part consists of the application of the developed methodology to environmental and biological aquatic matrices (water, sediment and biota). Obtained results in the first part of the work are shown and discussed in Chapter 3; studies about sample pretreatment in sediments are described in Chapter 4; and the last part of the work is described in Chapter 5. Appendix I includes an article about the determination of Hg2+ and MeHg+ in polluted waters by HPLC-UV-CV-AFS, while Appendix II includes a review of certified reference materials for mercury speciation in environmental and biological matrices

La producción de energías renovables en el ámbito de los puertos españoles

Los puertos en general, como eslabones de la cadena del transporte, son importantes plataformas logísticas donde se dan, y cada vez se darán más en el futuro, consumos energéticos importantes. El ámbito portuario reúne características especialmente favorables para la instalación de generadores de energías renovables, probablemente en exceso a los consumos de las propias instalaciones portuarias y de los procesos de transformación. El nuevo concepto de puerto verde, los arcos marítimos azules, los cambios de combustible en los propulsores de los buques y la paralización en muelles de los motores auxiliares generadores, abren un nuevo panorama energético en el sistema portuario, y de necesidades de nuevas instalaciones y servicios. Se hace necesario en la planificación estratégica portuaria de futuros planes y programas, considerar, junto al establecimiento de nuevas infraestructuras, la posibilidad de generación de energía limpia. Entre las energías renovables susceptibles de instalarse en el ámbito portuario tenemos la eólica, la solar y la undimotriz, al menos. La ubicación y características de cada puerto permitirán desarrollar un tipo u otro de energía, delimitando zonas en las que las características de dicha energía puede hacer rentable o no la instalación, para autoconsumo o incluso suministro a la red

Adherence to a supplemented Mediterranean diet drives changes in the gut microbiota of HIV-1-infected individuals

Objective: The health effects of a supplemented Mediterranean diet (SMD) with extra-virgin olive oil (EVOO) and nuts are well documented in non-HIV-infected individuals. We hypothesised that the benefits of an SMD could be mediated by changes in the gut microbiota, even in those with an altered intestinal microbiota such as people living with HIV. Design: Individuals living with HIV (n = 102) were randomised to receive an SMD with 50 g/day of EVOO and 30 g/day of walnuts (SMD group) or continue with their regular diet (control group) for 12 weeks. Methods: Adherence to the Mediterranean diet was assessed using the validated 14-item MD-Adherence-Screener (MEDAS) from the PREDIMED study. A sub-study classifying the participants according to their MEDAS scores was performed. Results: The lipid profile was improved in the SMD group vs. that in the control group (delta-total cholesterol and delta-B-lipoprotein). The immune activation (CD4+HLADR+CD38+ and CD8+HLADR+CD38+ cells) and IFN-γ-producing T-cells significantly decreased at week 12 compared to the baseline in the SMD group but not in the control group. The gut microbiota in those from the high-adherence group presented significantly high diversity and richness at the end of the intervention. Succinivibrio and Bifidobacterium abundances were influenced by the adherence to the MD and significantly correlated with Treg cells. Conclusion: The Mediterranean diet improved metabolic parameters, immune activation, Treg function, and the gut microbiota composition in HIV-1-infected individuals. Further, Mediterranean diet increased the Bifidobacterium abundances after the intervention, and it was associated to a beneficial profileThis study was partially supported by the SPANISH AIDS Research Network (RIS) grants RD16/0025/0002 and RD16/0025/0013 integrated in the State Plan for Scientific and Technical Research and Innovation from the General Sub-Directorate for Research Assessment and Promotion, Spanish Carlos III Institute of Health (ISCIII) co-funded by the European Regional Development Fund (ERDF), 6ª Announcement of Gilead Grants in Biomedical Research in HIV, Hepatic, and Hemo-Oncology, the Spanish Ministry of Economy (MINECO) (grant: SAF2015-66193-R), and income from Europe via the Regional Development (FEDER) and CERCA Programme/Generalitat de Catalunya SGR 615 and SGR 653. F.G. has received support from José María Segovia de Arana contracts. N.R. is a Miguel Servet II investigator from the ISCIII [grant CPII19/00025]. M.N.M. is funded by the grant IND2018/BMD-9651.Peer Reviewed"Article signat per 18 autors/es: Roque Pastor-Ibáñez, Juan Blanco-Heredia, Florencia Etcheverry, Sonsoles Sánchez-Palomino, Francisco Díez-Fuertes, Rosa Casas, María Ángeles Navarrete-Muñoz, Sara Castro-Barquero, Constanza Lucero, Irene Fernández, Lorna Leal, José Miguel Benito, Marc Noguera-Julian, Roger Paredes, Norma Rallón, Ramón Estruch, David Torrents and Felipe García"Postprint (published version

Mercury (II) and methylmercury determination in waters by liquid chromatography hyphenated to cold vapour atomic fluorescence spectrometry after online short-column preconcentration

This paper reports a method developed for the simultaneous determination of methylmercury (MeHg+) and mercury(II) (Hg2+) species in water by liquid chromatography coupled to online UV irradiation and cold vapour atomic fluorescence spectrometry (LC-UV-CV-AFS) after online short-column preconcentration. This work focused on systematic studies of several variables to establish the maximum species recoveries, preconcentration factors and good reproducibility. The optimum results obtained were the following: 0.07 mmol L 1 2-mercaptoethanol as a complexing agent, precolumn conditioning with the mobile phase: a mixture of 80% of methanol (MeOH) and 20% of the following buffer: 0.0015 mol L 1 ammonium pyrrolidine dithiocarbamate (APDC) and 0.01 mol L 1 ammonium acetate (NH4CH3COO) at pH 5.5, 2 cm precolumn length and 2 mL min 1 sample flow. This method was applied to three water samples with different mineralisation contents. Various tests, based on spikes, were performed on each sample. A breakthrough volume of 4 mL was found. The recovery values of 72 3% and 81 5% for MeHg+ and Hg2+, respectively, were obtained regardless of the matrix composition, and the PF values were 30 and 32 for MeHg+ and Hg2+, respectively. The accuracy of the preconcentration method was verified by analysing a certified reference material. The detection limits (LDs) obtained were 15 ng L 1 for MeHg+ and 2 ng L 1 for Hg2+. The quantification limits (LQs) were 50 ng L 1 for both species. The established analytical online preconcentration method is suitable for the quantification of mercury species in a wide range of environmental waters

Mercury (II) and methylmercury determination in waters by liquid chromatography hyphenated to cold vapour atomic fluorescence spectrometry after online short-column preconcentration

This paper reports a method developed for the simultaneous determination of methylmercury (MeHg+) and mercury(II) (Hg2+) species in water by liquid chromatography coupled to online UV irradiation and cold vapour atomic fluorescence spectrometry (LC-UV-CV-AFS) after online short-column preconcentration. This work focused on systematic studies of several variables to establish the maximum species recoveries, preconcentration factors and good reproducibility. The optimum results obtained were the following: 0.07 mmol L 1 2-mercaptoethanol as a complexing agent, precolumn conditioning with the mobile phase: a mixture of 80% of methanol (MeOH) and 20% of the following buffer: 0.0015 mol L 1 ammonium pyrrolidine dithiocarbamate (APDC) and 0.01 mol L 1 ammonium acetate (NH4CH3COO) at pH 5.5, 2 cm precolumn length and 2 mL min 1 sample flow. This method was applied to three water samples with different mineralisation contents. Various tests, based on spikes, were performed on each sample. A breakthrough volume of 4 mL was found. The recovery values of 72 3% and 81 5% for MeHg+ and Hg2+, respectively, were obtained regardless of the matrix composition, and the PF values were 30 and 32 for MeHg+ and Hg2+, respectively. The accuracy of the preconcentration method was verified by analysing a certified reference material. The detection limits (LDs) obtained were 15 ng L 1 for MeHg+ and 2 ng L 1 for Hg2+. The quantification limits (LQs) were 50 ng L 1 for both species. The established analytical online preconcentration method is suitable for the quantification of mercury species in a wide range of environmental waters

Method development for the simultaneous determination of methylmercury and inorganic mercury in seafood

This paper reports the method development for the simultaneous determination of methylmercury MeHgþ) and inorganic mercury (iHg) species in seafood samples. The study focused on the extraction and quantification of MeHgþ (the most toxic species) by liquid chromatography coupled to on-line UV irradiation and cold vapour atomic fluorescence spectroscopy (LC-UV-CV-AFS), using HCl 4 mol/L as the extractant agent. Accuracy of the method has been verified by analysing three certified reference materials and different spiked samples. The values found for total Hg and MeHgþ for the CRMs did not differ significantly from certified values at a 95% confidence level, and recoveries between 85% and 97% for MeHgþ, based on spikes, were achieved. The detection limits (LODs) obtained were 0.001 mg Hg/kg for total mercury, 0.0003 mg Hg/kg for MeHgþ and 0.0004 mg Hg/kg for iHg. The quantification limits (LOQs) established were 0.003 mg Hg/kg for total mercury, 0.0010 mg Hg/kg for MeHgþ and 0.0012 mg Hg/kg for iHg. Precision for each mercury species was established, being 12% in terms of RSD in all cases. Finally, the developed method was applied to 24 seafood samples from different origins and total mercury contents. The concentrations for Total Hg, MeHg and iHg ranged from 0.07 to 2.33, 0.003-2.23 and 0.006-0.085 mg Hg/kg, respectively. The established analytical method allows to obtain results for mercury speciation in less than 1 one hour including both, sample pretreatment and measuring step

Adherence to a Supplemented Mediterranean Diet Drives Changes in the Gut Microbiota of HIV-1-Infected Individuals

Objective: The health effects of a supplemented Mediterranean diet (SMD) with extra-virgin olive oil (EVOO) and nuts are well documented in non-HIV-infected individuals. We hypothesised that the benefits of an SMD could be mediated by changes in the gut microbiota, even in those with an altered intestinal microbiota such as people living with HIV. Design: Individuals living with HIV (n = 102) were randomised to receive an SMD with 50 g/day of EVOO and 30 g/day of walnuts (SMD group) or continue with their regular diet (control group) for 12 weeks. Methods: Adherence to the Mediterranean diet was assessed using the validated 14-item MD-Adherence-Screener (MEDAS) from the PREDIMED study. A sub-study classifying the participants according to their MEDAS scores was performed. Results: The lipid profile was improved in the SMD group vs. that in the control group (delta-total cholesterol and delta-B-lipoprotein). The immune activation (CD4+HLADR+CD38+ and CD8+HLADR+CD38+ cells) and IFN-γ-producing T-cells significantly decreased at week 12 compared to the baseline in the SMD group but not in the control group. The gut microbiota in those from the high-adherence group presented significantly high diversity and richness at the end of the intervention. Succinivibrio and Bifidobacterium abundances were influenced by the adherence to the MD and significantly correlated with Treg cells. Conclusion: The Mediterranean diet improved metabolic parameters, immune activation, Treg function, and the gut microbiota composition in HIV-1-infected individuals. Further, Mediterranean diet increased the Bifidobacterium abundances after the intervention, and it was associated to a beneficial profile