Identificación y validación de genes expresados diferencialmente en el mejillón "Mytilus galloprovincialis" a partir de genotecas normalizadas procedentes de individuos control y tratados con la biotoxina ácido ocadaico

Las floraciones algales nocivas (FANs) constituyen uno de las principales fuentes de contaminación de los océanos, produciendo altas concentraciones de biotoxinas que son acumuladas a lo largo de las cadenas tróficas. Una de éstas, el ácido okadaico (OA), es producida por dinoflagelados marinos y posteriormente acumulada en los tejidos de los organismos marinos, extendiéndose en la cadena trófica y eventualmente llegando a los consumidores humanos, donde causa el síndrome diarreico (Diarrhetic Shellfish Poisoning, DSP). En este trabajo se estudia la expresión diferencial de los genes ferritin, myticin c, hemolectin y las regiones génicas unigen1 y unigen2 en branquia y hepatopáncreas del mejillón Mytilus galloprovincialis como respuesta a la baja exposición del OA (200 cél/mL de Prorocentrum lima durante un día) por medio de la aplicación de la técnica de PCR en tiempo real. Los resultados obtenidos muestran como los genes y regiones génicas seleccionadas tienen un patrón de expresión diferente en los dos tejidos. Las regiones génicas unigen1 y unigen2 y el gen hemolectin debido a su sobreexpresión significativa podrían considerarse como posibles biomarcadores de contaminación temprana por OA en branquia. Por lo que respecta al gen myticin c, podría ser considerado como biomarcador en hepatopáncreas.Traballo fin de grao (UDC.CIE). Bioloxía. Curso 2013/201

Infección por el Virus de la Hepatitis C en la era de los antivirales de acción directa: optimización del diagnóstico, tratamiento y progresión de la enfermedad

Programa Oficial de Doutoramento en Ciencias da Saúde. 5007V01[Resumen] La infección por el Virus de la Hepatitis C (VHC) es un problema grave de salud pública a escala mundial que afecta a más de 71 millones de personas en todo el mundo. La infección por VHC se caracteriza por la acumulación progresiva de fibrosis hepática que, en estadios finales de la enfermedad, lo que conduce al desarrollo de cirrosis y carcinoma hepatocelular. La introducción de los antivirales de acción directa (AADs) ha marcado un hito en la historia del tratamiento frente al VHC, al permitir alcanzar tasas de respuesta viral sostenida (RVS) superiores al 90%, haciendo cada vez más plausible la erradicación de la infección. No obstante, en este nuevo escenario terapéutico sigue siendo necesaria una optimización tanto de los tratamientos como de su monitorización, así como la búsqueda de marcadores que permitan identificar poblaciones con mayor riesgo de progresión de la enfermedad. Además, cabe destacar que, pese a la alta eficacia de los AADs, se estima que un 80% de la infección por VHC a nivel mundial continua sin ser diagnosticada, por lo que se requiere de la implementación y optimización de programas de cribado activos que permitan identificar aquellas personas infectadas por el VHC. En este contexto los objetivos de este trabajo fueron, en primer lugar, caracterizar y evaluar el impacto de los haplogrupos europeos de ADN mitocondrial en la evolución clínica de la infección por VHC. En segundo lugar, analizar y describir la dinámica viral y su utilidad en la optimización de la duración y monitorización de los tratamientos basados en AADs. El último de los objetivos se basó en el desarrollo de un programa de cribado, con el fin de identificar posibles infecciones no diagnosticada y estudiar la seroprevalencia del VHC en la ciudad de A Coruña. Los resultados obtenidos mostraron una asociación entre la fibrosis hepática y los haplogrupos europeos de ADN mitocondrial, poniendo de manifiesto su posible utilidad como marcadores de progresión del daño hepático. En relación a la dinámica viral en los regímenes basados en AADs, se observó una caída bifásica de la carga viral del VHC, caracterizada por un rápido descenso en los primeros días de tratamiento. Además, a pesar de que la proporción de pacientes con cargas virales indetectables a semana 4 y 8 fue baja, todos alcanzaron RVS. Estos resultados sugerían la necesidad de re-evaluar las reglas de monitorización de la carga viral en el contexto de las nuevas terapias con AADs, así como la duración de las mismas. Finalmente, la realización del programa de cribado mostró que la seroprevalencia de la infección por VHC en sujetos nacidos entre 1960-1969 fue del 2,05% y que tan solo el 29,8% de la población contaba con pruebas serológicas previas. La prevalencia de infección por VHC en personas que desconocían su estado serológico fue del 0,26%. Los resultados obtenidos ha permitido una mejora en el conocimiento de la epidemiología de la infección por VHC en la ciudad de A Coruña y deberían de ser considerados a la hora de implementar futuras estrategias de cribado en busca de la erradicación de la infección por VHC en España.[Resumo] A infección polo Virus da Hepatite C (VHC) é un problema grave de saúde pública a escala mundial que afecta a máis de 71 millóns de persoas en todo o mundo. A infección por VHC caracterízase pola acumulación progresiva de fibrose hepática que, en estadios finais da enfermidade, conduce ao desenvolvemento de cirrosis e carcinoma hepatocelular. A introdución dos antivirais de acción directa (AADs) marcou un fito na historia do tratamento fronte ao VHC, ao permitir alcanzar taxas de resposta viral sostida (RVS) superiores ao 90%, facendo cada vez máis plausible a erradicación da infección. Con todo, neste novo escenario terapéutico segue sendo necesaria unha optimización tanto dos tratamentos como da súa monitoraxe, así como a procura de marcadores que permitan identificar poboacións con maior risco de progresión da enfermidade. Ademais, cabe destacar que, a pesar da alta eficacia dos AADs, estímase que un 80% da infección por VHC a nivel mundial continua sen ser diagnosticada, polo que se require da implementación e optimización de programas de cribado activos que permitan identificar aquelas persoas infectadas polo VHC. Neste contexto os obxectivos deste traballo foron, en primeiro lugar, caracterizar e avaliar o impacto dos haplogrupos europeos de ADN mitocondrial na evolución clínica da infección polo VHC. En segundo lugar, analizar e describir a dinámica viral e a súa utilidade na optimización da duración e monitoraxe dos tratamentos baseados en AADs. O último dos obxectivos baseouse no desenvolvemento dun programa de cribado, co fin de identificar posibles infeccións non diagnosticada e estudar a seroprevalencia do VHC na cidade da Coruña. Os resultados obtidos mostraron unha asociación entre a fibrose hepática e os haplogrupos europeos de ADN mitocondrial, poñendo de manifesto a súa posible utilidade como marcadores de progresión do dano hepático. En relación á dinámica viral nos réximes baseados en AADs, observouse unha caída bifásica da carga viral do VHC, caracterizada por un rápido descenso nos primeiros días de tratamento. Ademais, a pesar de que a proporción de pacientes con cargas virales indetectables a semana 4 e 8 foi baixa, todos alcanzaron RVS. Estes resultados suxiren a necesidade de re-avaliar as regras de monitoraxe da carga viral no contexto das novas terapias con AADs, así como a duración das mesmas. Finalmente, a realización do programa de cribado mostrou que a seroprevalencia da infección por VHC en suxeitos nados entre 1960-1969 foi do 2,05% e que tan só o 29,8% da poboación contaba con probas serolóxicas previas. A prevalencia de infección por VHC en persoas que descoñecían o seu estado serolóxico foi do 0,26%. Os resultados obtidos permitiron mellorar o coñecemento da epidemioloxía da infección polo VHC na cidade da Coruña e deberían de ser considerados á hora de implementar futuras estratexias de cribado en busca da erradicación da infección polo VHC en España.[Abstract] Hepatitis C Virus (HCV) infection is a major worldwide health problem with more than 71 million infected people around the world. HCV infection is characterized by progressive liver fibrosis accumulation leading to cirrhosis and hepatocellular carcinoma development in end-stages of the disease. Direct-acting antivirals (DAAs) has represented a remarkable milestone in the history of HCV treatment, since they have allowed to reach more than 90% of sustained viral response (SVR) and therefore, making more plausible the HCV global eradication. Nonetheless, in this new therapeutic scenario, it is still necessary to optimize not only the treatments and their monitoring, but also the seek for markers which allows the identification of high-risk populations. Moreover, it is important to highlight that even the high efficacy rates of DAAs, it has been estimated that approximately 80% of worldwide infected-patients are not aware of their diagnose. Thus, it is necessary to implement and optimize active screening programs in order to identify this kind of populations. In this context, the objectives of this study have been, firstly, to characterize and evaluate the impact of European mitochondrial DNA haplogroups in the outcome of HCV infection. Secondly, to analyze and describe viral dynamics and its usefulness for the optimization of treatment monitoring and duration. Finally, the last objective has been to develop a screening program that allows identifying non-diagnosed HCV infections in the city of A Coruña, and evaluating the HCV seroprevalence in this area. The results obtained showed an association between hepatic fibrosis and European haplogroups of mitochondrial DNA, thus highlighting their possible usefulness as markers for liver damage progression. According to the viral dynamics in regimens based on DAAs, a biphasic decline of HCV viral load was observed. It was characterized by a rapid decrease during first days of treatment. In addition, although the proportion of patients with undetectable viral loads at week 4 and 8 was low, all of them reached SVR. These results suggest the need of re-evaluatig viral load monitoring rules, as well as their duration in the context of new therapies with DAAs. Finally, screening program implementation showed that the seroprevalence of HCV infection in subjects born between 1960-1969 was 2.05%, and that only 29.8% of the population had been serologically tested. The prevalence of HCV infection in people who did not know their serological status was 0.26%. The results obtained have allowed an improvement in the knowledge of HCV-infection epidemiology in the city of A Coruña, and it should be considered when implementing future screening strategies in the search for the eradication of HCV infection in Spain

Performance of HZSM-5 agglomerated in a mesoporous matrix in the fuel production from ethylene at atmospheric pressure

Ethylene oligomerization into liquid fuels at slightly over atmospheric pressure has been studied due to its interest to valorize online the excess of ethylene in sustainable olefins production processes and to intensify the production of fuel from refinery secondary streams. Runs were carried out in a fixed-bed reactor under the following conditions: 1.5 bar; 275–375 °C; space time, 2.7–16.2 gcatalyst h molC−1; time on stream, 5 h. The catalyst was prepared by agglomerating a HZSM-5 zeolite (SiO2/Al2O3 of 30) in a mesoporous matrix (γ-Al2O3/α-Al2O3). The hierarchical porous structure of the catalyst enables to reach a pseudo-steady state with a remarkable remnant activity after an initial deactivation period of 2–3 h. Temperature shows a relevant effect on ethylene conversion and product distribution, where a C5+ liquid fuel yield of 55% above 325 °C and 10.6 gcatalyst h molC−1 is obtained. At 325 °C, gasoline yield is 60%, with high olefin content (49%), which decreases at higher temperature, due to an increase in aromatic and paraffin concentration. Soft and hard coke analysis reveal the role of the matrix to attenuate deactivation. Moreover, above 325 °C the cracking of hard coke precursors deposited in the zeolite micropores prevails respect to their formation.This work has been carried out with the financial support of the Ministry of Economy and Competitiveness of the Spanish Government (Project PID2022-140584OB-I00); the Basque Government (Project IT1645-22); and the European Regional Development Funds (ERDF) and the European Commission (HORIZON H2020-MSCA RISE 2018. Contract No. 823745). Z. Tabernilla is grateful for the PhD grant from the Department of Education, University and Research of the Basque Government (PRE_2022_2_0136). The authors thank for technical and human support provided by SGIker (UPV/EHU)

Low-pressure oligomerization of diluted ethylene on a HZSM-5 zeolite catalyst

The oligomerization at low pressure of diluted ethylene coming from secondary streams is an attractive route for hydrocarbon production, by means of a low cost and energy efficient process. To evaluate the viability of this process, the effect of dilution with N2 or syngas on the low-pressure oligomerization of ethylene (1.5 bar) was studied on a catalyst prepared with a HZSM-5 zeolite agglomerated in a mesoporous matrix of α- and γ-Al2O3, aiming to produce C5+ hydrocarbons (gasoline). The experiments were performed in a fixed bed reactor at 325 °C and a space time of 10.6 gcatalyst h molC−1. For an ethylene partial pressure of 0.33 bar, conversion surpassed 80 % and high C5+ hydrocarbon yield was obtained: >40 % with N2 as diluent; and, >30 % with syngas. The greater effect of syngas dilution on suppressing the formation of aromatics is explained by the role of H2 in decreasing the extent of dehydrocyclization reactions. The dilution of ethylene limits the extent of the reaction stages, but it also attenuates the stages for coke formation, by facilitating the diffusion of soft coke in the mesoporous matrix and contributing to decrease the deposition of hard coke in the zeolite micropores. Consequently, a pseudo-steady state of the catalyst is reached with a notable remnant activity for the formation of higher hydrocarbons.This work has been carried out with the financial support of the Ministry of Science, Innovation and Universities of the Spanish Government (PID2022-140584OB-I00); the Basque Government (Project IT1645-22); and the European Regional Development Funds (ERDF) and the European Commission (HORIZON H2020-MSCA RISE 2018. Contract No. 823745). Z. Tabernilla is grateful for the PhD grant from the Department of Education, University and Research of the Basque Government (PRE2023_2_0005)

Gasoline production by oligomerization of 1-butene at low pressure: Kinetic model and reactor simulation

A five lump kinetic model (C4=, C8=, C12=, C1-C4 and C5-C12) has been established for 1-butene (C4=) oligomerization at low pressure (1.5 bar) and 275 °C on an HZSM-5 zeolite catalyst agglomerated in a γ-Al2O3 mesoporous matrix. The kinetic parameters have been calculated based on the results obtained in a packed-bed reactor for different concentrations of 1-butene in the feed (with N2 as diluent) and different values of space time (0.5–9.5 gcatalyst h molC-1). The model quantifies the evolution with time on stream of the concentrations of the lumps, considering the selective deactivation of the catalyst for the oligomerization, cracking and pool of secondary reactions. Furthermore, it allows to calculate the remnant activity of the catalyst in the pseudo-steady state (>5h on stream) for the different reaction groups, as well as the corresponding product distribution. The kinetic model has been used in the simulation of the packed-bed reactor to study the effect of space time and 1-butene partial pressure variables. At the conditions range studied experimentally, 39.4 % gasoline yield (C5-C12 fraction, mainly composed of olefins) and a 1-butene conversion of 45 % are achieved in the pseudo-steady state of the catalyst, at a space time of 9.5 gcatalyst h molC-1 and a partial pressure of 1-butene of 0.8 bar. Additionally, for values exceeding those studied experimentally, the model predicts at the pseudo-steady state a gasoline yield of 60 % at a space time of 30 gcatalyst h molC-1 and a partial pressure of 1-butene of ∼ 0.5 bar.This work has been carried out with the financial support of the Ministry of Science, Innovation and Universities of the Spanish Government (Project PID2022/140584OB-I00); the Basque Government (Project IT1645-22); and the European Regional Development Funds (ERDF) and the European Commission (HORIZON H2020-MSCA RISE 2018. Contract No. 823745). Zuria Tabernilla is grateful for the financial support of the grant of the Basque Government (PRE2023_2_0005)

Caracterización epidemiológica, clínica y virológica de los nuevos diagnósticos de infección por VHC en Galicia (2014-2015)

Abstrac

Characterization of chronic HCV infection in Northwest Spain: impact of the treatment strategic plan of the Spanish National Health Service on HCV cure

[Abstract] The aim of the study was to characterize HCV infection in Northwest Spain and assess the impact of the Spanish Strategic Plan to cure HCV infection. Overall, 387 patients were included (60.9% HIV/HCV coinfected and 28.2% cirrhotic). Of these, 72.9% of patients that were recognized as priority for HCV treatment according to the Spanish Strategic Plan (≥F2, transplant or extrahepatic manifestations), initiated treatment during 2015. Globally, SVR12 was achieved in 96.5% of patients. The implementation of the Spanish Strategic Plan has been critical to advance in HCV cure, but 27.1% of priority patients still remain awaiting HCV treatment initiation.Instituto de Salud Carlos III; CPII14/00014Instituto de Salud Carlos III; PI10/02166Instituto de Salud Carlos III; PI13/02266Instituto de Salud Carlos III; CM15/00233Instituto de Salud Carlos III; FI14/0055

Any impact of blips and low-level viraemia episodes among HIV-infected patients with sustained virological suppression on ART?

[Abstract] Objectives. The objective of this study was to evaluate the prevalence of blips and risk of virological failure (VF) among HIV-infected patients with sustained virological suppression (HIV-RNA <50 copies/mL) on ART. Methods. Newly diagnosed (2004–13) HIV-infected patients with sustained virological suppression on ART (minimum follow-up of 3 months) were identified. Risk of VF was evaluated according to different plasma HIV-RNA quantification values based on the limits of quantification/detection of current commercial assays (20 copies/mL). Kaplan–Meier and Cox proportional hazards models were used to compare the cumulative incidence of VF. Results. A total of 565 newly diagnosed HIV-infected patients were identified: 453 started ART and 354 achieved virological suppression. Prevalence of blips (isolated HIV-RNA ranging from 50 to 200 copies/mL) and VF (HIV-RNA ≥50 copies/mL) was 22.7% and 8.8%, respectively (mean follow-up of 42 months). Multivariate analysis identified differences between HIV-RNA values as an independent predictor of VF (P = 0.008); risk of VF was higher for patients with blips [HR 2.500 (95% CI 0.524–11.926)] and for those with at least three consecutive detected, but not quantified, HIV-RNA determinations (HIV-RNA 200 copies/mL [33.7% at 24 and 60 months versus <5% for other HIV-RNA values; HR 6.943 (0.728–66.261), P = 0.092]. Conclusions. Blips are frequent (22.7%) among HIV-infected patients with sustained virological suppression on ART. HIV patients with blips and at least three consecutive detected, but not quantified, HIV-RNA determinations (<20 copies/mL) had a higher risk of VF. These findings highlight the relevance of maintaining HIV-RNA levels below the limits of quantification of current assays (<20 copies/mL).Instituto de Salud Carlos III; CPII14/00014Instituto de Salud Carlos III; PI10/02166Instituto de Salud Carlos III; PI13/0226

Prevalence of drug resistance mutations among ART-naive and -experienced HIV-infected patients in Sierra Leone

Objectives: The aim of this study was to assess the prevalence of HIV drug resistance (HIVDR) in HIV-infected ART-naive and -experienced patients in Sierra Leone. Patients and methods: We conducted a cross-sectional study of HIV-positive adults aged 18 years at Connaught Hospital in Freetown, Sierra Leone in November 2017. Sequencing was performed in the reverse transcriptase, protease and integrase regions, and interpreted using the Stanford HIVDR database andWHO 2009mutation list. Results: Two hundred and fifteen HIV-infected patients were included (64 ART naive and 151 ART experienced). The majority (66%) were female, the median age was 36 years and the median ART exposure was 48months. The majority (83%) were infected with HIV-1 subtype CRF02_AG. In the ART-naive group, the pretreatment drug resistance (PDR) prevalence was 36.7% (14.2% to NRTIs and 22.4% to NNRTIs). The most prevalent PDR mutations were K103N (14.3%), M184V (8.2%) and Y181C (4.1%). In the ART-experienced group, 64.4% harboured resistance-associated mutations (RAMs) and the overall prevalence of RAMs to NRTIs and NNRTIs was 85.2% (52/61) and 96.7% (59/61), respectively. The most prevalent RAMs were K103N (40.7%), M184V (28.8%), D67N (15.3%) and T215I/F/Y (15.3%). Based on the genotypic susceptibility score estimates, 22.4% of ART-naive patients and 56% of ART-experienced patients were not susceptible to first-line ART used in Sierra Leone. Conclusions: A high prevalence of circulating NRTI- and NNRTI-resistant variants was observed in ART-naive and -experienced HIV-1-infected patients in Sierra Leone. This necessitates the implementation of HIVDR surveillance programmes to inform national ART guidelines for the treatment and monitoring of HIV-infected patients in Sierra Leone.Xunta Galicia-Fondo Social Europeo | Ref. IN606A-2016/023Case Western Reserve University | Ref. NIH NIAID T32 AI07024Instituto de Salud Carlos III and Fondo Europeo de Desarrollo Regional-FEDER | Ref. RD16/0025/002

Plasma mitochondrial DNA levels are inversely associated with HIV-RNA levels and directly with CD4 counts: potential role as a biomarker of HIV replication

[Abstract] Objectives. To evaluate plasma mitochondrial DNA (mtDNA) levels among HIV-infected patients and its potential role as a biomarker of residual viral replication. Methods. HIV-infected patients on follow-up at a reference hospital in north-west Spain were selected. DNA was isolated from plasma samples and mtDNA levels were assessed using a quantitative real-time PCR assay. HIV-RNA levels and CD4+ cell counts were evaluated in the same blood samples used for plasma mtDNA quantification. Epidemiological and clinical variables were included for the analysis. Results. A total of 235 HIV-infected patients were included. Mean plasma mtDNA levels were 217 ± 656 copies/μL for naive (31.9%) and 364 ± 939 copies/μL for HIV-infected patients receiving ART and with suppressed viraemia (P = 0.043). Among the latter, mean plasma mtDNA levels were 149 ± 440 copies/μL for those with low-level viraemia (LLV; HIV-RNA 20–200 copies/mL), 265 ± 723 copies/μL for those with detected-not-quantified (DNQ) viraemia (HIV-RNA <20 copies/mL) and 644 ± 1310 copies/μL for those with not-detected (ND) viraemia. Of note, a linear trend (P = 0.006) was observed among virologically suppressed (LLV, DNQ and ND) patients. ND patients had higher mtDNA levels compared with LLV patients (P = 0.057). Moreover, mtDNA levels were inversely associated with HIV-RNA levels (Spearman’s rho −0.191, P = 0.003) and directly associated with CD4+ counts (Spearman’s rho 0.131, P = 0.046). Conclusions. Increased plasma mtDNA levels are associated with lower HIV-RNA levels and higher CD4+ cell counts. Among ART-suppressed patients, mtDNA levels were significantly higher in those with complete virological suppression (ND) than in those with LLV. These data suggest that plasma mtDNA levels might serve as a biomarker of residual HIV replication.Instituto de Salud Carlos III; CPII14/00014Instituto de Salud Carlos III; PI10/02166Instituto de Salud Carlos III; PI13/02266Instituto de Salud Carlos III; FI14/00557Instituto de Salud Carlos III; CM15/00233Instituto de Salud Carlos III; PI16/02159Instituto de Salud Carlos III; MV16/02159Instituto de Salud Carlos III; PTA2013-8277-