Biomonitorización citogenética de cuatro poblaciones agrícolas europeas, expuestas a plaguicidas, mediante el ensayo de micronúcleos

Consultable des del TDXTítol obtingut de la portada digitalitzadaLa importancia de los plaguicidas recae tanto en los numerosos beneficios que tienen para la sociedad (protectores de plagas de las cosechas, erradicación de vectores que propagan enfermedades, etc.); como en los efectos negativos que representan para la salud (problemas reproductivos, sarcomas, linfomas, etc.). Los colectivos en contacto más directo con estos productos, como son los agricultores, tienen un mayor riesgo de sufrir los efectos adversos que se han asociado con los plaguicidas. Esta Tesis comprende una serie de trabajos sobre la evaluación del daño genotóxico de los plaguicidas en cuatro poblaciones agrícolas europeas, mediante la técnica de micronúcleos (MN) en dos tipos celulares: linfocitos de sangre periférica y células de descamación de la mucosa bucal. Paralelamente, también se determinó el índice de proliferación celular (CBPI). Las poblaciones que se han estudiado pertenecen a colectivos agrícolas con una continua exposición a plaguicidas, procedentes de Almería (España), Malopolska (Polonia), Nea Makri (Grecia) y del sudeste de Hungría. Cada población ha sido evaluada individualmente y finalmente se han agrupado y analizado los datos conjuntos, para obtener una visión general de los efectos de la exposición. Además, en la población almeriense se realizó un estudio longitudinal, para evaluar la existencia de diferencias estacionales en dos periodos de distinta aplicación de plaguicidas. Asimismo, en esta población se evaluó la posible influencia de los genotipos GSTM1 y GSTT1 sobre la susceptibilidad individual en al respuesta genotóxica a la exposición a plaguicidas. Únicamente se observó una relación entre GSTM1 y CBPI disminuyendo este último en presencia del producto del gen GSTT1 en los agricultores. A pesar de que en alguna población se observa una tendencia a un mayor número de células binucleadas con micronúcleos (BNMN), en los linfocitos de los individuos expuestos a plaguicidas, en ninguna de las poblaciones evaluadas individualmente ni en el análisis global de la población, las diferencias observadas llegan a alcanzar un nivel estadísticamente significativo. Lo mismo ocurre con las células bucales con MN (CBMN). Respecto del CBPI, en la mayoría de poblaciones se observan descensos, significativos o no, de los valores del mismo en el colectivo agrícola. Al valorar toda la población en global estas variaciones se traducen en un descenso significativo del CBPI en los individuos expuestos a plaguicidas, lo que refleja el efecto citotóxico de los mismos. En el análisis global, observamos que el hecho de pertenecer a un país u otro confiere una serie de características particulares a cada grupo, que se reflejan en una mayor frecuencia de un determinado tipo de daño genético según el sistema celular estudiado. Otro de los objetivos de la Tesis era valorar los factores higiénicos, de salud, de exposición a plaguicidas, de protección laboral, de alimentación, etc., que podían influir en las variaciones de la frecuencia de micronúcleos y en la proliferación celular. Entre los factores que han mostrado tener un efecto significativo en la variación de las frecuencias de las variables de estudio están la edad, el hecho de ser mujer, el tabaco, la exposición a rayos X por diagnóstico médico, así como factores de la dieta (consumo de vegetales crudos, pescado y aceite de oliva). Hay que destacar el incremento significativo en el porcentaje de abortos encontrados en la población agrícola de Polonia, lo que junto con el elevado número de intoxicaciones y el descenso del CBPI en este colectivo nos indica que, a pesar de no observarse un incremento significativo de daño genético detectable mediante el ensayo de MN, los plaguicidas están ejerciendo efectos biológicos adversos sobre los agricultores.The importance of pesticides can be reflected as much on the benefits that they represent for society (crops pest prevention, vector erradication); as on the negative effects they have on health (reproductive problems, sarcomas, lymphomas, etc.). These adverse effects mainly affect occupationally exposed populations, such as farmers. This PhD Thesis comprises a series of works on the evaluation of genotoxic damage caused by pesticide exposure of four agricultural European populations, using the micronucleus assay (MN). Both lymphocytes of peripheral blood and exfoliated cells of the oral mucosa were used. The cytokinesis-block proliferation index (CBPI) was also calculated to detect possible variations in the proliferative kinetics of lymphocytes (due to the pesticide exposure). Populations from Greece, Spain, Poland and Hungary, all with an intense and continuous agriculture activity, were evaluated individually. Finally, in order to obtain a general effect of the exposure to pesticides, all the data was pooled and analysed together. Furthermore, a follow-up study was carried out in the Spanish population to determine, at two different periods of time and different pesticide intensity application, if there were season-related differences in the MN frequency. In this population, to look for relationship between the genotypes and the cytogenetic response, each donor was assessed for the presence or absence of glutathione S-transferase M1 and GSTT1. Only in the exposed individuals, the CBPI appeared to be affected, decreasing in the presence of GSTT1. Although in some populations we observed a trend to a high number of BNMN (binucleated cells with micronucleus) in the lymphocytes of farmers exposed to pesticides, none of the statistical analysis showed significant differences between controls and farmers, for neither lymphocytes nor buccal cells with MN. Regarding the CBPI, decreases were observed in most of the pesticide-exposed populations. In the evaluation of all the populations, the group occupationally exposed to pesticides showed a significantly lower CBPI level, indicating some cytotoxicity due to the exposure. The global analysis showed us that the increases in genetic damage seemed to be related to different cellular systems for each country, revealing the particular characteristics of the populations evaluated. One of the aims was to assess the hygienic and health factors, exposure to pesticides, work protective measures, diet, etc., that could influence on the frequency variations of MN and cell proliferation. From the studied variables, those with a significant effect on the frequencies studied were age, the fact of being woman, tobacco, X-ray medical exposure and some diet factors (consuming raw vegetables, fish and olive oil). The agricultural Polish population showed a significant increase in the percentage of miscarriages, which together with the high number of intoxications and the CBPI decrease in this group, showed us that pesticides do have biological adverse effects on the farmers despite the lack of a significant increase of cytogenetic damage evaluated by the MN assay

Hazard assessment of different-sized polystyrene nanoplastics in hematopoietic human cell lines

Altres ajuts: acords transformatius de la UABThe environmental presence of micro/nanoplastics (MNPLs) is an environmental and human health concern. Such MNPLs can result from the physicochemical/biological degradation of plastic goods (secondary MNPLs) or can result from industrial production at that size, for different commercial purposes (primary MNPLs). Independently of their origin, the toxicological profile of MNPLs can be modulated by their size, as well as by the ability of cells/organisms to internalize them. To get more information on these topics we have determined the ability of three different sizes of polystyrene MNPLs (50, 200, and 500 nm) to produce different biological effects in three different human hematopoietic cell lines (Raji-B, THP-1, and TK6). Results show that none of the three sizes was able to induce toxicity (growth ability) in any of the tested cell types. Although transmission electron microscopy and confocal images showed cell internalization in all the cases, their quantification by flow cytometry demonstrated an important uptake by Raji-B and THP-1 cells, in comparison with TK6 cells. For the first ones, the uptake was negatively associated with the size. Interestingly, when the loss of mitochondrial membrane potential was determined, dose-related effects were observed for Raji-B and THP-1 cells, but not for TK6 cells. These effects were observed for the three different sizes. Finally, when oxidative stress induction was evaluated, no clear effects were observed for the different tested combinations. Our conclusion is that size, biological endpoint, and cell type are aspects modulating the toxicological profile of MNPLs

Long-term effects of polystyrene nanoplastics in human intestinal Caco-2 cells

The increasing presence of micro- and nanoplastics (MNPLs) in the environment, and their consequent accumulation in trophic niches, could pose a potential health threat to humans, especially due to their chronic ingestion. In vitro studies using human cells are considered pertinent approaches to determine potential health risks to humans. Nevertheless, most of such studies have been conducted using short exposure times and high concentrations. Since human exposure to MNPLs is supposed to be chronic, there is a lack of information regarding the potential in vitro MNPLs effects under chronic exposure conditions. To this aim, we assessed the accumulation and potential outcomes of polystyrene nanoparticles (PSNPs), as a model of MNPLs, in undifferentiated Caco-2 cells (as models of cell target in ingestion exposures) under a relevant long-term exposure scenario, consisting of eight weeks of exposure to sub-toxic PSNPs concentrations. In such exposure conditions, culture-media was changed every 2-3 days to maintain constant exposure. The different analyzed endpoints were cytotoxicity, dysregulation of stress-related genes, genotoxicity, oxidative DNA damage, and intracellular ROS levels. These are endpoints that showed to be sensitive enough in different studies. The obtained results attest that PSNPs accumulate in the cells through time, inducing changes at the ultrastructural and molecular levels. Nevertheless, minor changes in the different evaluated genotoxicity-related biomarkers were observed. This would indicate that no DNA damage or oxidative stress is observed in the human intestinal Caco-2 cells after long-term exposure to PSNPs. This is the first study dealing with the long-term effects of PSNPs on human cultured cells

Influence of previous mental state on psychological outcomes of Spanish out-of-hospital professionals during the COVID-19 pandemic

Producción CientíficaThis study aimed to describe factors relating to the psychological distress of healthcare workers (HCWs) in Spanish out-of-hospital emergency medical services (EMS), according to the previous or non-use of psychotropic drugs or psychotherapy. A multicentre, cross-sectional descriptive study was designed. The study population were all physicians, nurses, and emergency medical technicians (EMTs) working in any Spanish out-of-hospital EMS between February and April 2021. The main outcomes were the levels of stress, anxiety, depression, and self-efficacy, assessed by DASS-21 and G-SES. Differences in levels of stress, anxiety, depression, and self-efficacy, according to sex, age, previous use of psychotropic drug or psychotherapy, work experience, professional category, type of work, and modification of working conditions were measured using the Student’s t-test for independent samples, one-way ANOVA, Pearson’s correlation, or 2-factor analysis of covariance. A total of 1636 HCWs were included, of whom one in three had severe mental disorders because of the pandemic. The interaction of the previous or non-use of psychotropic drugs or psychotherapy with the rest of the factors considered did not modify the levels of stress, anxiety, depression, and self-efficacy. However, HCWs with a history of psychotropic drug or psychotherapy use had a more intense negative emotional response and lower self-efficacy, regardless of their sex, professional category, type of work, or change in the working conditions. These HCWs are considered particularly vulnerable to the development or recurrence of new disorders or other comorbidities; therefore, the implementation of monitoring and follow-up strategies should be a priority

Women in limnology: From a historical perspective to a present-day evaluation

Research in limnology is nurtured by the work of many fascinating and passionate women, who have contributed enormously to our understanding of inland waters. Female limnologists have promoted and established the bases of our knowledge about inland waters and fostered the need of protecting the values of those ecosystems. However, on numerous occasions, their contribution to the advancement of limnology has not been duly recognized. Here, we review the presence of women in limnology through the history of the discipline: from the pioneers who contributed to the origins to present day' developments. We aim at visibilizing those scientists and establish them as role models. We also analyze in a simple and illustrative way the current situation of women in limnology, the scientific barriers they must deal with, and their future prospects. Multiple aspects fostering the visibility of a scientist, such as their presence in conferences, awards, or representation in societal or editorial boards show a significant gap, with none of those aspects showing a similar visibility of women and men in limnology. This article raises awareness of the obstacles that women in limnology faced and still face, and encourages to embrace models of leadership, scientific management, and assessment of research performance far from those commonly established.info:eu-repo/semantics/publishedVersio

Fluorescent labeling of micro/nanoplastics for biological applications with a focus on "true-to-life" tracking

Altres ajuts: acords transformatius de la UABThe increased environmental presence of micro-/nanoplastics (MNPLs) and the potential health risks associated with their exposure classify them as environmental pollutants with special environmental and health concerns. Consequently, there is an urgent need to investigate the potential risks associated with secondary MNPLs. In this context, using "true-to-life" MNPLs, resulting from the laboratory degradation of plastic goods, may be a sound approach. These non-commercial secondary MNPLs must be labeled to track their presence/journeys inside cells or organisms. Because the cell internalization of MNPLs is commonly analyzed using fluorescence techniques, the use of fluorescent dyes may be a sound method to label them. Five different compounds comprising two chemical dyes (Nile Red and Rhodamine-B), one optical brightener (Opticol), and two industrial dyes (Amarillo Luminoso and iDye PolyPink) were tested to determine their potential for such applications. Using commercial standards of polystyrene nanoplastics (PSNPLs) with an average size of 170 nm, different characteristics of the selected dyes such as the absence of impact on cell viability, specificity for plastic staining, no leaching, and lack of interference with other fluorochromes were analyzed. Based on the overall data obtained in the wide battery of assays performed, iDye PolyPink exhibited the most advantages, with respect to the other compounds, and was selected to effectively label "true-to-life" MNPLs. These advantages were confirmed using a proposed protocol, and labeling titanium-doped PETNPLs (obtained from the degradation of milk PET plastic bottles), as an example of "true-to-life" secondary NPLs. These results confirmed the usefulness of iDye PolyPink for labeling MNPLs and detecting cell internalization

Novel genetic variants in differentiated thyroid cancer and assessment of the cumulative risk

A genome-wide association study (GWAS) performed on a high-incidence Italian population followed by replications on low-incidence cohorts suggested a strong association of differentiated thyroid cancer (DTC) with single nucleotide polymorphisms (SNPs) at 9q22.33, 2q35, 20q11.22-q12 and 14q24.3. Moreover, six additional susceptibility loci were associated with the disease only among Italians. The present study had two aims, first to identify loci involved in DTC risk and then to assess the cumulative effect of the SNPs identified so far in the Italian population. The combined analysis of the previous GWAS and the present Italian study provided evidence of association with rs7935113 (GALNTL4, OR = 1.36, 95%CI 1.20-1.53, p-value = 7.41 × 10) and rs1203952 (FOXA2, OR = 1.29, 95%CI 1.16-1.44, p-value = 4.42 × 10). Experimental ENCODE and eQTL data suggested that both SNPs may influence the closest genes expression through a differential recruitment of transcription factors. The assessment of the cumulative risk of eleven SNPs showed that DTC risk increases with an increasing number of risk alleles (p-trend = 3.13 × 10 â '47). Nonetheless, only a small fraction (about 4% on the disease liability scale) of DTC is explained by these SNPs. These data are consistent with a polygenic model of DTC predisposition and highlight the importance of association studies in the discovery of the disease hereditability

Influence of the Cumulative Incidence of COVID-19 Cases on the Mental Health of the Spanish Out-of-Hospital Professionals

This study aimed to analyze the psychological affectation of health professionals (HPs) of Spanish Emergency Medical Services (EMSs) according to the cumulative incidence (CI) of COVID19 cases in the regions in which they worked. A cross-sectional descriptive study was designed, including all HPs working in any EMS of the Spanish geography between 1 February 2021 and 30 April 2021. Their level of stress, anxiety and depression (DASS-21) and the perception of self-efficacy (GSES) were the study’s main results. A 2-factor analysis of covariance was used to determine if the CI regions of COVID-19 cases determined the psychological impact on each of the studied variables. A total of 1710 HPs were included. A third presented psychological impairment classified as severe. The interaction of CI regions with the studied variables did not influence their levels of stress, anxiety, depression or self-efficacy. Women, younger HPs or those with less EMS work experience, emergency medical technicians (EMT), workers who had to modify their working conditions or those who lived with minors or dependents suffered a greater impact from the COVID-19 pandemic in certain regions. These HPs have shown high levels of stress, anxiety, depression and medium levels of self-efficacy, with similar data in the different geographical areas. Psychological support is essential to mitigate their suffering and teach them to react to adverse events.This research was funded by Fundación ASISA and Sociedad Española de Urgencias y Emergencias (SEMES)

Spread of a SARS-CoV-2 variant through Europe in the summer of 2020

[EN] Following its emergence in late 2019, the spread of SARS-CoV-21,2 has been tracked by phylogenetic analysis of viral genome sequences in unprecedented detail3,4,5. Although the virus spread globally in early 2020 before borders closed, intercontinental travel has since been greatly reduced. However, travel within Europe resumed in the summer of 2020. Here we report on a SARS-CoV-2 variant, 20E (EU1), that was identified in Spain in early summer 2020 and subsequently spread across Europe. We find no evidence that this variant has increased transmissibility, but instead demonstrate how rising incidence in Spain, resumption of travel, and lack of effective screening and containment may explain the variant’s success. Despite travel restrictions, we estimate that 20E (EU1) was introduced hundreds of times to European countries by summertime travellers, which is likely to have undermined local efforts to minimize infection with SARS-CoV-2. Our results illustrate how a variant can rapidly become dominant even in the absence of a substantial transmission advantage in favourable epidemiological settings. Genomic surveillance is critical for understanding how travel can affect transmission of SARS-CoV-2, and thus for informing future containment strategies as travel resumes.S

Assessing Copy Number of MON810 Integrations in Commercial Seed Maize Varieties by 5' Event-specific Real-time PCR Validated Method Coupled to 2-deltadelta CT Analysis

The objective of the present study was to assess the applicability of the MON 8105' event-specific method validated by the Community Reference Laboratory for Genetically Modified Food and Feed(CRL-GMFF) that is commonly used for quantitative purposes. This 5' event-specific /hmg-taxon gene real-time PCR protocol coupled to 2 analysis was the chosen approach to determine the MON 810 insert copy number per haploidgenome across 26 genetically modified (GM) commercial maize varieties. Variety DK513 containing one copy integration per haploid genome was used as calibrator in each assay. Complementary data from end-point real-time PCRs that targeted specifically the MON 810 insert were also analysed. Global results assessed and guaranteed the genetic intactness of the transgenic integration per haploid genome for 24 out of the 26 commercial varieties studied,which showed no significant differences between 2 values respect to the calibrator value. Conversely,two varieties showed no transgenic insert in their genomes. This validated analytical method was suitable for MON 810 detection and quantification purposes.JRC.I.3-Molecular Biology and Genomic