Cytomolecular evaluation of bystander effect in human lung fibroblast cells under conditions of radiotherapeutical exposition

Durante los últimos años uno de los principales paradigmas de la radiobiología ha ido cambiando. Originalmente, se creía que sólo aquellas células irradiadas directamente podrían presentar daño en su material genético. Actualmente, la idea de que existe riesgo genotóxico para las células no directamente irradiadas ha crecido considerablemente. El propósito del presente trabajo fue mostrar la existencia de este fenómeno, conocido como efectos de la vecindad, en fibroblastos de pulmón humano, expuestos a dosis utilizadas en radioterapia fraccionada. El diseño experimental consistió en tratar células no irradiadas, con medio de cultivo proveniente de células previamente irradiadas (MCI). Se obtuvo MCI formado durante diferentes tiempos de incubación: 30, 60, 120 y 180 minutos tras irradiación. Posteriormente las células no irradiadas fueron tratadas con estos medios durante una hora. El daño en el material genético fue analizado mediante el ensayo de electroforesis de células individuales (Ensayo Cometa). Los resultados muestran considerables diferencias entre el daño genotóxico de las células tratadas con el MCI y su correspondiente control. Por otra parte, se ha observado una respuesta cíclica en los niveles de daño, en referencia al tratamiento con los distintos medios condicionados. Este hecho podría ser explicado a través de un desbalance entre factores pro y antioxidantes producidos en respuesta a la radiación. Estos hallazgos podrían contribuir a la optimización de metodologías implementadas tanto en radioterapia como en radioprotección.During the last years one of the major paradigms of radiobiology has changed. Originally, it was belived that only those cells directly hit by radiation were the ones that could damage their genetic material. At present the idea of neighboring cells being at genotoxic risk is growing considerably. The aim of this research was to show the existance of this phenomenon, usually called bystander effect, specifically in human lung fibroblasts exposed to a dose which is frecuently used in fractioned radiotherapy. The experimental design consisted of treating non-irradiated cells with the culture medium coming from cells that had been previously irradiated (ICM). We recollected ICM formed during different times of incubation: 30, 60, 120 and 180 minutes. After that, non-irradiated cells were treated with these medium during one hour in all the cases. The damage in the genetic material was analyzed by the Comet assay. The results show considerable differences in the genomic damage among the cells treated with ICM and their corresponding control. A cyclic response in such levels has also been observed. This might be due to the disbalance between the proxidant and the antioxidant factors produced in response to radiation. These findings would contribute to optimize the implemented methodology both in radiotherapy and radioprotection.Instituto de Genética Veterinari

Cytomolecular evaluation of bystander effect in human lung fibroblast cells under conditions of radiotherapeutical exposition

Durante los últimos años uno de los principales paradigmas de la radiobiología ha ido cambiando. Originalmente, se creía que sólo aquellas células irradiadas directamente podrían presentar daño en su material genético. Actualmente, la idea de que existe riesgo genotóxico para las células no directamente irradiadas ha crecido considerablemente. El propósito del presente trabajo fue mostrar la existencia de este fenómeno, conocido como efectos de la vecindad, en fibroblastos de pulmón humano, expuestos a dosis utilizadas en radioterapia fraccionada. El diseño experimental consistió en tratar células no irradiadas, con medio de cultivo proveniente de células previamente irradiadas (MCI). Se obtuvo MCI formado durante diferentes tiempos de incubación: 30, 60, 120 y 180 minutos tras irradiación. Posteriormente las células no irradiadas fueron tratadas con estos medios durante una hora. El daño en el material genético fue analizado mediante el ensayo de electroforesis de células individuales (Ensayo Cometa). Los resultados muestran considerables diferencias entre el daño genotóxico de las células tratadas con el MCI y su correspondiente control. Por otra parte, se ha observado una respuesta cíclica en los niveles de daño, en referencia al tratamiento con los distintos medios condicionados. Este hecho podría ser explicado a través de un desbalance entre factores pro y antioxidantes producidos en respuesta a la radiación. Estos hallazgos podrían contribuir a la optimización de metodologías implementadas tanto en radioterapia como en radioprotección.During the last years one of the major paradigms of radiobiology has changed. Originally, it was belived that only those cells directly hit by radiation were the ones that could damage their genetic material. At present the idea of neighboring cells being at genotoxic risk is growing considerably. The aim of this research was to show the existance of this phenomenon, usually called bystander effect, specifically in human lung fibroblasts exposed to a dose which is frecuently used in fractioned radiotherapy. The experimental design consisted of treating non-irradiated cells with the culture medium coming from cells that had been previously irradiated (ICM). We recollected ICM formed during different times of incubation: 30, 60, 120 and 180 minutes. After that, non-irradiated cells were treated with these medium during one hour in all the cases. The damage in the genetic material was analyzed by the Comet assay. The results show considerable differences in the genomic damage among the cells treated with ICM and their corresponding control. A cyclic response in such levels has also been observed. This might be due to the disbalance between the proxidant and the antioxidant factors produced in response to radiation. These findings would contribute to optimize the implemented methodology both in radiotherapy and radioprotection.Instituto de Genética Veterinari

Genomic and serologic characterization of enterovirus A71 brainstem encephalitis

OBJECTIVE: In 2016, Catalonia experienced a pediatric brainstem encephalitis outbreak caused by enterovirus A71 (EV-A71). Conventional testing identified EV in the periphery but rarely in CSF. Metagenomic next-generation sequencing (mNGS) and CSF pan-viral serology (VirScan) were deployed to enhance viral detection and characterization. METHODS: RNA was extracted from the CSF (n = 20), plasma (n = 9), stool (n = 15), and nasopharyngeal samples (n = 16) from 10 children with brainstem encephalitis and 10 children with meningitis or encephalitis. Pathogens were identified using mNGS. Available CSF from cases (n = 12) and pediatric other neurologic disease controls (n = 54) were analyzed with VirScan with a subset (n = 9 and n = 50) validated by ELISA. RESULTS: mNGS detected EV in all samples positive by quantitative reverse transcription polymerase chain reaction (qRT-PCR) (n = 25). In qRT-PCR-negative samples (n = 35), mNGS found virus in 23% (n = 8, 3 CSF samples). Overall, mNGS enhanced EV detection from 42% (25/60) to 57% (33/60) (p-value = 0.013). VirScan and ELISA increased detection to 92% (11/12) compared with 46% (4/12) for CSF mNGS and qRT-PCR (p-value = 0.023). Phylogenetic analysis confirmed the EV-A71 strain clustered with a neurovirulent German EV-A71. A single amino acid substitution (S241P) in the EVA71 VP1 protein was exclusive to the CNS in one subject. CONCLUSION: mNGS with VirScan significantly increased the CNS detection of EVs relative to qRT-PCR, and the latter generated an antigenic profile of the acute EV-A71 immune response. Genomic analysis confirmed the close relation of the outbreak EV-A71 and neuroinvasive German EV-A71. A S241P substitution in VP1 was found exclusively in the CSF.Grants supporting this project include the National Multiple Sclerosis Society and the American Academy of Neurology award FAN-1608-25607 (R.D.S.), Clinical Research Training Scholarship P0534134 (P.S.R.), Sandler and William K. Bowes Jr Foundations (M.R.W., J.L.D., L.M.K., H.A.S., K.C.Z.), Rachleff Family Foundation (M.R.W.), and NINDS of the NIH under award K08NS096117 (M.R.W.) and F31NS113432 (K.E.L.). This study was partially supported by a grant from the Spanish National Health Institute [grant number PI15CIII-00020] and the European Regional Development Fund (FEDER funds). UCSF Biomedical Sciences Program (I.A.H., K.E.L.), UCSF Medical Scientist Training Program (K.E.L.), and the Chan Zuckerberg Biohub (J.E.P., W.W., C.K.C., J.L.D., E.D.C.) also supported this project.S

Predicting River Macroinvertebrate Communities Distributional Shifts under Future Global Change Scenarios in the Spanish Mediterranean Area

Several studies on global change over the next century predict increases in mean air temperatures of between 1°C to 5°C that would affect not only water temperature but also river flow. Climate is the predominant environmental driver of thermal and flow regimes of freshwater ecosystems, determining survival, growth, metabolism, phenology and behaviour as well as biotic interactions of aquatic fauna. Thus, these changes would also have consequences for species phenology, their distribution range, and the composition and dynamics of communities. These effects are expected to be especially severe in the Mediterranean basin due its particular climate conditions, seriously threatening Southern European ecosystems. In addition, species with restricted distributions and narrow ecological requirements, such as those living in the headwaters of rivers, will be severely affected. The study area corresponds to the Spanish Mediterranean and Balearic Islands, delimited by the Köppen climate boundary. With the application of the MEDPACS (MEDiterranean Prediction And Classification System) predictive approach, the macroinvertebrate community was predicted for current conditions and compared with three posible scenarios of watertemperature increase and its associated water flow reductions. The results indicate that the aquatic macroinvertebrate communities will undergo a drastic impact, with reductions in taxa richness for each scenario in relation to simulated current conditions, accompanied by changes in the taxa distribution pattern. Accordingly, the distribution area of most of the taxa (65.96%) inhabiting the mid-high elevations would contract and rise in altitude. Thus, families containing a great number of generalist species will move upstream to colonize new zones with lower water temperatures. By contrast, more vulnerable taxa will undergo reductions in their distribution area.This work was funded by GUADALMED-II (REN2001-3438-C07-06/HID), a project of excellence from “Junta de Andalucía” (RNM-02654/FEDER), the Spanish “Ministerio de Ciencia e Innovación” (CGL2007-61856/BOS), projects and a collaboration agreement between the “Spanish Ministerio de Medio Ambiente, Medio Rural y Marino” and the University of Granada (21.812-0062/8511)

Molecular epidemiology of an enterovirus A71 outbreak associated with severe neurological disease, Spain, 2016

Altres ajuts: We wish to thank I Bustillo, H del Pozo and P Higueras for their technical assistance. We also sincerely wish to thank all technical staff from microbiology departments and medical staff from paediatrics departments from all participating hospitals. Some of the samples are included in an ongoing project (PI15CIII-00020) which was supported by a grant by the Health Research System (AES).Introduction: Enterovirus A71 (EV-A71) is an emerging pathogen that causes a wide range of disorders including severe neurological manifestations. In the past 20 years, this virus has been associated with large outbreaks of hand, foot and mouth disease with neurological complications in the Asia-Pacific region, while in Europe mainly sporadic cases have been reported. In spring 2016, however, an EV-A71 outbreak associated with severe neurological cases was reported in Catalonia and spread further to other Spanish regions. Aim: Our objective was to investigate the epidemiology and clinical characteristics of the outbreak. Methods: We carried out a retrospective study which included 233 EV-A71-positive samples collected during 2016 from hospitalised patients. We analysed the clinical manifestations associated with EV-A71 infections and performed phylogenetic analyses of the 3'-VP1 and 3Dpol regions from all Spanish strains and a set of EV-A71 from other countries. Results: Most EV-A71 infections were reported in children (mean age: 2.6 years) and the highest incidence was between May and July 2016 (83%). Most isolates (218/233) were classified as subgenogroup C1 and 217 of them were grouped in one cluster phylogenetically related to a new recombinant variant strain associated with severe neurological diseases in Germany and France in 2015 and 2016. Moreover, we found a clear association of EV-A71-C1 infection with severe neurological disorders, brainstem encephalitis being the most commonly reported. Conclusion: An emerging recombinant variant of EV-A71-C1 was responsible for the large outbreak in 2016 in Spain that was associated with many severe neurological cases

Molecular epidemiology of an enterovirus A71 outbreak associated with severe neurological disease, Spain, 2016

IntroductionEnterovirus A71 (EV-A71) is an emerging pathogen that causes a wide range of disorders including severe neurological manifestations. In the past 20 years, this virus has been associated with large outbreaks of hand, foot and mouth disease with neurological complications in the Asia-Pacific region, while in Europe mainly sporadic cases have been reported. In spring 2016, however, an EV-A71 outbreak associated with severe neurological cases was reported in Catalonia and spread further to other Spanish regions.AimOur objective was to investigate the epidemiology and clinical characteristics of the outbreak.MethodsWe carried out a retrospective study which included 233 EV-A71-positive samples collected during 2016 from hospitalised patients. We analysed the clinical manifestations associated with EV-A71 infections and performed phylogenetic analyses of the 3'-VP1 and 3Dpol regions from all Spanish strains and a set of EV-A71 from other countries.ResultsMost EV-A71 infections were reported in children (mean age: 2.6 years) and the highest incidence was between May and July 2016 (83%). Most isolates (218/233) were classified as subgenogroup C1 and 217 of them were grouped in one cluster phylogenetically related to a new recombinant variant strain associated with severe neurological diseases in Germany and France in 2015 and 2016. Moreover, we found a clear association of EV-A71-C1 infection with severe neurological disorders, brainstem encephalitis being the most commonly reported.ConclusionAn emerging recombinant variant of EV-A71-C1 was responsible for the large outbreak in 2016 in Spain that was associated with many severe neurological cases.S

Age at menopause in Latin America

OBJECTIVE: To assess the age at menopause (AM) in Latin America urban areas. DESIGN: A total of 17,150 healthy women, aged 40 to 59 years, accompanying patients to healthcare centers in 47 cities of 15 Latin American countries, were surveyed regarding their age, educational level, healthcare coverage, history of gynecological surgery, smoking habit, presence of menses, and the use of contraception or hormone therapy at menopause. The AM was calculated using logit analysis. RESULTS: The mean age of the entire sample was 49.4 ± 5.5 years. Mean educational level was 9.9 ± 4.5 years, and the use of hormone therapy and oral contraception was 22.1% and 7.9%, respectively. The median AM of women in all centers was 48.6 years, ranging from 43.8 years in Asuncion (Paraguay) to 53 years in Cartagena de Indias (Colombia). Logistic regression analysis determined that women aged 49 living in cities at 2,000 meters or more above sea level (OR = 2.0, 95% CI: 1.4-2.9, P less than 0.001) and those with lower educational level (OR = 1.9, 95% CI: 1.3-2.8, P less than 0.001) or living in countries with low gross national product (OR = 2.1, 95% CI: 1.5-2.9, P less than 0.001) were more prone to an earlier onset of menopause. CONCLUSIONS: The AM varies widely in Latin America. Lower income and related poverty conditions influence the onset of menopause. © 2006 by The North American Menopause Society

Objectively Measured Sleep Duration and Health-Related Quality of Life in Older Adults with Metabolic Syndrome: A One-Year Longitudinal Analysis of the PREDIMED-Plus Cohort

The aim of our cross-sectional and longitudinal study is to assess the relationship between daytime and night-time sleep duration and health-related quality of life (HRQoL) in adults with metabolic syndrome after a 1-year healthy lifestyle intervention. Analysis of the data from 2119 Spanish adults aged 55–75 years from the PREDIMED-Plus study was performed. Sleep duration was assessed using a wrist-worn accelerometer. HRQoL was measured using the SF-36 questionnaire. Linear regression models adjusted for socioeconomic and lifestyle factors and morbidity were developed. In cross-sectional analyses, participants with extreme night-time sleep duration categories showed lower physical component summary scores in Models 1 and 2 [β-coefficient (95% confidence interval) 9 h vs. 7–9 h: −1.1 (−2.0 to −0.3); p = 0.01]. Participants who sleep less than 7 h a night and take a nap are associated with higher mental component summary scores [β-coefficient (95% confidence interval) 6.3 (1.3 to 11.3); p = 0.01]. No differences between night-time sleep categories and 12-month changes in HRQoL were observed. In conclusion, in cross-sectional analyses, extremes in nocturnal sleep duration are related to lower physical component summary scores and napping is associated with higher mental component summary scores in older adults who sleep less than 7 h a night.The PREDIMED-Plus trial was supported by the European Research Council (Advanced Research Grant 2013–2018, 340918) to Dr Martínez-González, and the official funding agency for biomedical research of the Spanish government, Instituto de Salud Carlos III, through the Fondo de Investigación para la Salud, which is cofunded by the European Regional Development Fund (five coordinated Fondo de Investigación para la Salud projects led by Dr. Salas-Salvadó and Dr Vidal, including the following projects: PI13/00673, PI13/00492, PI13/00272, PI13/01123, PI13/00462, PI13/00233, PI13/02184, PI13/00728, PI13/01090, PI13/01056, PI14/01722, PI14/00636, PI14/00618, PI14/00696, PI14/01206, PI14/01919, PI14/00853, PI14/01374, PI14/00972, PI14/00728, PI14/01471, PI16/00473, PI16/00662, PI16/01873, PI16/01094, PI16/00501, PI16/00533, PI16/00381, PI16/00366, PI16/01522, PI16/01120, PI17/00764, PI17/01183, PI17/00855, PI17/01347, PI17/00525, PI17/01827, PI17/00532, PI17/00215, PI17/01441, PI17/00508, PI17/01732, PI17/00926, PI19/00957, PI19/00386, PI19/00309, PI19/01032, PI19/00576, PI19/00017, PI19/01226, PI19/00781, PI19/01560 and PI19/01332), the Especial Action Project titled Implementación y evaluación de una intervención intensiva sobre la actividad física, a Cohorte PREDIMED-Plus grant to Dr Salas-Salvadó, the Recercaixa grant to Dr Salas-Salvadó (2013ACUP00194), a CICYT (Consejo Interinstitucional de Ciencia y Tecnología) grant (AGL2016–75329-R), a grant from the Generalitat Valenciana (APOSTD/2019/136 to R.B.) and Generalitat de Catalunya (SGR-2019 to R.E.), grants from the Consejería de Salud de la Junta de Andalucía (PI0458/2013, PS0358/2016 and PI0137/2018), grants from the Generalitat Valenciana (PROMETEO/2017/017), a SEMERGEN (Sociedad Española de Médicos de Atención Primaria) grant, EU-COST (European Cooperation in Science and Technology) Action CA16112, a grant of support to research groups number 35/2011 from the Balearic Islands Government, grants from IDISBA (Instituto de Investigación Sanitaria Islas Baleares), funds from the European Regional Development Fund (CIBEROBN CB06/03 and CB12/03), from the European Commission (EAT2BENI-CE_H2020_SFS2016) and Universidad de León for funding the manuscript. The funding sponsors had no role in the design of the study; in the collection, analyses or interpretation of the data; in the writing of the article or in the decision to publish the results

Objectively Measured Sleep Duration and Health-Related Quality of Life in Older Adults with Metabolic Syndrome: A One-Year Longitudinal Analysis of the PREDIMED-Plus Cohort

[EN] The aim of our cross-sectional and longitudinal study is to assess the relationship between daytime and night-time sleep duration and health-related quality of life (HRQoL) in adults with metabolic syndrome after a 1-year healthy lifestyle intervention. Analysis of the data from 2119 Spanish adults aged 55–75 years from the PREDIMED-Plus study was performed. Sleep duration was assessed using a wrist-worn accelerometer. HRQoL was measured using the SF-36 questionnaire. Linear regression models adjusted for socioeconomic and lifestyle factors and morbidity were developed. In cross-sectional analyses, participants with extreme night-time sleep duration categories showed lower physical component summary scores in Models 1 and 2 [β-coefficient (95% confidence interval) 9 h vs. 7–9 h: −1.1 (−2.0 to −0.3); p = 0.01]. Participants who sleep less than 7 h a night and take a nap are associated with higher mental component summary scores [β-coefficient (95% confidence interval) 6.3 (1.3 to 11.3); p = 0.01]. No differences between night-time sleep categories and 12-month changes in HRQoL were observed. In conclusion, in cross-sectional analyses, extremes in nocturnal sleep duration are related to lower physical component summary scores and napping is associated with higher mental component summary scores in older adults who sleep less than 7 h a night.S

Choice of the initial antiretroviral treatment for HIV-positive individuals in the era of integrase inhibitors

BACKGROUND: We aimed to describe the most frequently prescribed initial antiretroviral therapy (ART) regimens in recent years in HIV-positive persons in the Cohort of the Spanish HIV/AIDS Research Network (CoRIS) and to investigate factors associated with the choice of each regimen. METHODS: We analyzed initial ART regimens prescribed in adults participating in CoRIS from 2014 to 2017. Only regimens prescribed in >5% of patients were considered. We used multivariable multinomial regression to estimate Relative Risk Ratios (RRRs) for the association between sociodemographic and clinical characteristics and the choice of the initial regimen. RESULTS: Among 2874 participants, abacavir(ABC)/lamivudine(3TC)/dolutegavir(DTG) was the most frequently prescribed regimen (32.1%), followed by tenofovir disoproxil fumarate (TDF)/emtricitabine (FTC)/elvitegravir(EVG)/cobicistat(COBI) (14.9%), TDF/FTC/rilpivirine (RPV) (14.0%), tenofovir alafenamide (TAF)/FTC/EVG/COBI (13.7%), TDF/FTC+DTG (10.0%), TDF/FTC+darunavir/ritonavir or darunavir/cobicistat (bDRV) (9.8%) and TDF/FTC+raltegravir (RAL) (5.6%). Compared with ABC/3TC/DTG, starting TDF/FTC/RPV was less likely in patients with CD4100.000 copies/mL. TDF/FTC+DTG was more frequent in those with CD4100.000 copies/mL. TDF/FTC+RAL and TDF/FTC+bDRV were also more frequent among patients with CD4<200 cells//muL and with transmission categories other than men who have sex with men. Compared with ABC/3TC/DTG, the prescription of other initial ART regimens decreased from 2014-2015 to 2016-2017 with the exception of TDF/FTC+DTG. Differences in the choice of the initial ART regimen were observed by hospitals' location. CONCLUSIONS: The choice of initial ART regimens is consistent with Spanish guidelines' recommendations, but is also clearly influenced by physician's perception based on patient's clinical and sociodemographic variables and by the prescribing hospital location