179 research outputs found

    Identification of differentially expressed genes in SHSY5Y cells exposed to okadaic acid by suppression subtractive hybridization

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    <p>Abstract</p> <p>Background</p> <p>Okadaic acid (OA), a toxin produced by several dinoflagellate species is responsible for frequent food poisonings associated to shellfish consumption. Although several studies have documented the OA effects on different processes such as cell transformation, apoptosis, DNA repair or embryogenesis, the molecular mechanistic basis for these and other effects is not completely understood and the number of controversial data on OA is increasing in the literature.</p> <p>Results</p> <p>In this study, we used suppression subtractive hybridization in SHSY5Y cells to identify genes that are differentially expressed after OA exposure for different times (3, 24 and 48 h). A total of 247 subtracted clones which shared high homology with known genes were isolated. Among these, 5 specific genes associated with cytoskeleton and neurotransmission processes (NEFM, TUBB, SEPT7, SYT4 and NPY) were selected to confirm their expression levels by real-time PCR. Significant down-regulation of these genes was obtained at the short term (3 and 24 h OA exposure), excepting for NEFM, but their expression was similar to the controls at 48 h.</p> <p>Conclusions</p> <p>From all the obtained genes, 114 genes were up-regulated and 133 were down-regulated. Based on the NCBI GenBank and Gene Ontology databases, most of these genes are involved in relevant cell functions such as metabolism, transport, translation, signal transduction and cell cycle. After quantitative PCR analysis, the observed underexpression of the selected genes could underlie the previously reported OA-induced cytoskeleton disruption, neurotransmission alterations and <it>in vivo </it>neurotoxic effects. The basal expression levels obtained at 48 h suggested that surviving cells were able to recover from OA-caused gene expression alterations.</p

    Identification of Differentially Expressed Genes in SHSY5Y Cells Exposed to Okadaic Acid by Suppression Subtractive Hybridization

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    [Abstract] Background: Okadaic acid (OA), a toxin produced by several dinoflagellate species is responsible for frequent food poisonings associated to shellfish consumption. Although several studies have documented the OA effects on different processes such as cell transformation, apoptosis, DNA repair or embryogenesis, the molecular mechanistic basis for these and other effects is not completely understood and the number of controversial data on OA is increasing in the literature. Results: In this study, we used suppression subtractive hybridization in SHSY5Y cells to identify genes that are differentially expressed after OA exposure for different times (3, 24 and 48 h). A total of 247 subtracted clones which shared high homology with known genes were isolated. Among these, 5 specific genes associated with cytoskeleton and neurotransmission processes (NEFM, TUBB, SEPT7, SYT4 and NPY) were selected to confirm their expression levels by real-time PCR. Significant down-regulation of these genes was obtained at the short term (3 and 24 h OA exposure), excepting for NEFM, but their expression was similar to the controls at 48 h. Conclusions: From all the obtained genes, 114 genes were up-regulated and 133 were down-regulated. Based on the NCBI GenBank and Gene Ontology databases, most of these genes are involved in relevant cell functions such as metabolism, transport, translation, signal transduction and cell cycle. After quantitative PCR analysis, the observed underexpression of the selected genes could underlie the previously reported OA-induced cytoskeleton disruption, neurotransmission alterations and in vivo neurotoxic effects. The basal expression levels obtained at 48 h suggested that surviving cells were able to recover from OA-caused gene expression alterations.This work was funded by a grant from the Spanish Ministry of Science and Innovation (PSI2010-15115). V. Valdiglesias was supported by a fellowship from the University of A Coruña. Authors would like to thank the Genomics Service from INIBIC (Complejo Hospitalario Universitario A Coruña) for providing their facilitie

    Cytogenetics of the razor clam "Solen marginatus" (Mollusca: Bivalvia: Solenidae)

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    [Abstract:] The razor clam Solen marginatus has a diploid chromosome number of 38. The karyotype consists of one metacentric/submetacentric, three submetacentric/metacentric, five submetacentric, one submetacentric/subtelocentric, one subtelocentric/submetacentric, six subtelocentric and two telocentric chromosome pairs. Staining with chromomycin A3 revealed bright positive bands subcentromerically in the long arms of one medium-sized subtelocentric pair, while DAPI staining showed uniform fluorescence in all chromosomes of the complement. Fluorescence in situ hybridization using an 18S-5.8S-28S rDNA probe locates these loci at the subcentromeric region of one subtelocentric pair and at the subtelomeric region of another subtelocentric pair.Xunta de Galicia; PGIDT99 MAR1030

    Genome sizes and karyotypes in the razor clams "Ensis arcuatus" (Jeffreys, 1865) and "E. siliqua" (Linnaeus, 1758)

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    [Abstract] The razor clams Ensis arcuatus and E. siliqua show a diploid DNA content of 3.85 ± 0.049 pg and 4.00 ± 0.050 pg, respectively. Both have a diploid chromosome number of 38 although their karyotypes show remarkable differences. The karyotype of E. arcuatus consists of 4 metacentric, 1 metacentric-submetacentric, 7 submetacentric and 7 telocentric chromosome pairs, whereas that of E. siliqua possesses 3 metacentric, 7 submetacentric and 9 telocentric pairs. In situ hybridization using an 18S-5.8S-28S rDNA probe located this ribosomal locus on one chromosome pair for both species. Results demonstrate that large differences exist between them, probably caused by chromosome rearrangements along evolution of these two species, and increase the number of studies on bivalve cytogenetics.Xunta de Galicia; PGIDT99 MAR1030

    Asociación entre morfología del pie y rendimiento en gimnasia rítmica

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    [Abstract] The objectives of the study were to determine the influence of foot morphology on performance in Rhythmic Gymnastics and to analyze the influence of years of practice. The sample consisted of 48 gymnasts who had practiced federated gymnastics and competed during the last year. The results indicate that the gymnasts have predominantly a neutral foot and with a normal footprint, presenting enough asymmetry between feet, not significant, which may be a consequence of asymmetric work and should be corrected in training. Only the range of amplitude of the talocrural joint seems to be a characteristic of foot morphology that affects technical performance and seems more an innate characteristic. The practice of rhythmic gymnastics might not be as decisive a factor as could be supposed in the morphological modifications of the footprint.[Resumen] Los objetivos del estudio fueron determinar la influencia de la morfología del pie en el rendimiento en Gimnasia Rítmica y analizar la influencia de los años de práctica. La muestra estaba compuesta por 48 gimnastas que habían practicado gimnasia federada y competido durante el último año. Los resultados indican que las gimnastas tienen predominantemente un pie neutro y con huella normal, presentando bastante asimetría entre pies, no significativa, lo que puede ser consecuencia de un trabajo asimétrico y debería ser corregido en el entrenamiento. Sólo el rango de amplitud de la articulación talocrural parece ser una característica de la morfología del pie que incida sobre el rendimiento técnico y parece más una característica innata. La práctica de la Gimnasia Rítmica podría no ser un factor tan decisivo como podía suponerse en las modificaciones morfológicas de la huella plantar

    Mitochondrial DNA (mtDNA) haplogroups J and H are differentially associated with the methylation status of articular cartilage: potential role in apoptosis and metabolic and developmental processes

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    [Abstract] Objective. To analyze the influence of mitochondrial genome variation on the DNA methylome of articular cartilage. Methods. DNA methylation profiling was performed using data deposited in the NCBI Gene Expression Omnibus database (accession no. GSE 43269). Data were obtained for 14 cartilage samples from subjects with haplogroup J and 20 cartilage samples from subjects with haplogroup H. Subsequent validation was performed in an independent subset of 7 subjects with haplogroup J and 9 with haplogroup H by RNA ‐seq. Correlated genes were validated by real‐time polymerase chain reaction in an independent cohort of 12 subjects with haplogroup J and 12 with haplogroup H. Appropriate analyses were performed using R Bioconductor and qB asePlus software, and gene ontology analysis was conducted using DAVID version 6.8. Results. DNA methylation profiling revealed 538 differentially methylated loci, while whole‐transcriptome profiling identified 2,384 differentially expressed genes, between cartilage samples from subjects with haplogroup H and those with haplogroup J. Seventeen genes showed an inverse correlation between methylation and expression. In terms of gene ontology, differences in correlations between methylation and expression were also detected between cartilage from subjects with haplogroup H and those with haplogroup J, highlighting a significantly enhanced apoptotic process in cartilage from subjects with haplogroup H (P = 0.007 for methylation and P = 0.019 for expression) and repressed apoptotic process in cartilage from subjects with haplogroup J (P = 0.021 for methylation), as well as a significant enrichment of genes related to metabolic processes (P = 1.93 × 10−4 for methylation and P = 6.79 x 10−4 for expression) and regulation of gene expression (P = 0.012 for methylation) in cartilage from subjects with haplogroup H, and to developmental processes (P = 0.015 for methylation and P = 8.25 x 10−12 for expression) in cartilage from subjects with haplogroup J. Conclusion. Mitochondrial DNA variation differentially associates with the methylation status of articular cartilage by acting on key mechanisms involved in osteoarthritis, such as apoptosis and metabolic and developmental processes.Instituto de Salud Carlos III; CIBERCB06/01/0040‐SpainInstituto de Salud Carlos III; CPII17/00026Instituto de Salud Carlos III; PI14/01254Instituto de Salud Carlos III; PI16/02124Instituto de Salud Carlos III; PI17/0021

    Low Vitamin D Levels and Frailty Status in Older Adults: A Systematic Review and Meta-Analysis

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    Serum vitamin D deficiency is widespread among older adults and is a potential modifiable risk factor for frailty. Moreover, frailty has been suggested as an intermediate step in the association between low levels of vitamin D and mortality. Hence, we conducted a systematic review of the literature and meta-analysis to test the possible association of low concentrations of serum 25-hydroxyvitamin D (25(OH)D), a marker of vitamin D status, with frailty in later life. We reviewed cross-sectional or longitudinal studies evaluating populations of older adults and identifying frailty by a currently validated scale. Meta-analyses were restricted to cross-sectional data from studies using Fried’s phenotype to identify frailty. Twenty-six studies were considered in the qualitative synthesis, and thirteen studies were included in the meta-analyses. Quantitative analyses showed significant differences in the comparisons of frail (standardized mean difference (SMD)—1.31, 95% confidence interval (CI) (−2.47, −0.15), p = 0.0271) and pre-frail (SMD—0.79, 95% CI (−1.58, −0.003), p = 0.0491) subjects vs. non-frail subjects. Sensitivity analyses reduced heterogeneity, resulting in a smaller but still highly significant between-groups difference. Results obtained indicate that lower 25(OH)D levels are significantly associated with increasing frailty severity. Future challenges include interventional studies testing the possible benefits of vitamin D supplementation in older adults to prevent/palliate frailty and its associated outcomes.This research was funded by Xunta de Galicia [ED431B 2019/02]; Ministerio de Educación, Cultura y Deporte [BEAGAL18/00142 to V.V, PRX19/00353 to B.L.]; and Deputación Provincial de A Coruña [to D.M.-P. and M.S.-F.]info:eu-repo/semantics/publishedVersio

    The CHROMEVALOA Database: A Resource for the Evaluation of Okadaic Acid Contamination in the Marine Environment Based on the Chromatin-Associated Transcriptome of the Mussel Mytilus galloprovincialis

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    Okadaic Acid (OA) constitutes the main active principle in Diarrhetic Shellfish Poisoning (DSP) toxins produced during Harmful Algal Blooms (HABs), representing a serious threat for human consumers of edible shellfish. Furthermore, OA conveys critical deleterious effects for marine organisms due to its genotoxic potential. Many efforts have been dedicated to OA biomonitoring during the last three decades. However, it is only now with the current availability of detailed molecular information on DNA organization and the mechanisms involved in the maintenance of genome integrity, that a new arena starts opening up for the study of OA contamination. In the present work we address the links between OA genotoxicity and chromatin by combining Next Generation Sequencing (NGS) technologies and bioinformatics. To this end, we introduce CHROMEVALOAdb, a public database containing the chromatin-associated transcriptome of the mussel Mytilus galloprovincialis (a sentinel model organism) in response to OA exposure. This resource constitutes a leap forward for the development of chromatin-based biomarkers, paving the road towards the generation of powerful and sensitive tests for the detection and evaluation of the genotoxic effects of OA in coastal areas

    Community-based screening enhances hepatitis B virus linkage to care among West African migrants in Spain

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    Hepatitis B virus; Community approach; Screening programsVirus de l'hepatitis B; Abordatge comunitari; Programes de deteccióVirus de la hepatitis B; Abordaje comunitario; Programas de detecciónBackground: Chronic infection with HBV is responsible for >50% of all hepatocellular cancer cases globally and disproportionately affects sub-Saharan African (sSA) countries. Migration from these countries to Europe has increased substantially in recent years, posing unique challenges to health systems. The aim of this study was to carry out a community-based intervention to increase HBV screening, vaccination, and linkage to care among sSA migrants in Catalonia, Spain. Methods:This was a prospective cohort study. Participants ≥18 years were offered community-based HBV screening between 20/11/20 and 21/01/22. Rapid HBV testing and blood sample collection utilizing plasma separation cards were carried out and linkage to care was offered to all participants. HBV vaccination and post-test counseling were performed at a second visit in the community. The main outcome was the odds of those with current HBV infection being successfully linked to hepatology. Rates of completing the care cascade of this model were analyzed. Results: In the present study, 444 people undergo screening, with 50.6% of participants showing evidence of past or current HBV infection, including an HBsAg prevalence of 9.2%. Migrants with current HBV infection exhibit 5.2 times higher odds of successful linkage to care compared to those in need of post-test counseling or vaccination. The study achieves a successful linkage to care rate of 72% for all participants, with specialist appointments arranged within 15.5 days.Conclusions:This community-based HBV screening program provides evidence of a successful model for identifying and providing care, including vaccination, to west African migrants at high risk of HBV infection who may otherwise not engage in care.Antecedents: La infecció crònica pel VHB és responsable del >50% de tots els casos de càncer hepatocel·lular a nivell mundial i afecta desproporcionadament els països de l'Àfrica subsahariana (SAS). La migració d'aquests països a Europa ha augmentat substancialment en els últims anys, plantejant reptes únics per als sistemes de salut. L'objectiu d'aquest estudi va ser dur a terme una intervenció basada en la comunitat per augmentar la detecció del VHB, la vacunació i la vinculació amb l'atenció entre els migrants de SSA a Catalunya, Espanya. Mètodes: Es tractava d'un estudi de cohort prospectiu. Als participants ≥18 anys se'ls va oferir un cribratge comunitari del VHB entre el 20/11/20 i el 21/01/22. Es van dur a terme proves ràpides de VHB i recollida de mostres de sang mitjançant targetes de separació de plasma i es va oferir vinculació a l'atenció a tots els participants. La vacunació contra el VHB i l'assessorament post-test es van realitzar en una segona visita a la comunitat. El resultat principal van ser les probabilitats que les persones amb infecció actual pel VHB estiguin vinculades amb èxit a l'hepatologia. Es van analitzar les taxes de completar la cascada assistencial d'aquest model. Resultats: En el present estudi, 444 persones se sotmeten a cribratge, amb el 50.6% dels participants que mostren evidència d'infecció passada o actual pel VHB, inclosa una prevalença de VHB del 9.2%. Els migrants amb infecció actual pel VHB presenten 5,2 vegades més probabilitats d'èxit en l'atenció en comparació amb aquells que necessiten assessorament o vacunació post-prova. L'estudi aconsegueix una taxa de vinculació reeixida a l'atenció del 72% per a tots els participants, amb cites amb especialistes concertades en un termini de 15,5 dies. Conclusions: Aquest programa de cribratge del VHB basat en la comunitat proporciona proves d'un model reeixit per identificar i proporcionar atenció, inclosa la vacunació, als migrants de l'Àfrica occidental amb alt risc d'infecció pel VHB que d'altra manera podrien no dedicar-se a l'atenció.C.A.P., J.V.L. and Lv.S. acknowledge support to ISGlobal from the Spanish Ministry of Science, Innovation and Universities through the “Centro de Excelencia Severo Ochoa 2019-2023” Program (CEX2018-000806-S), and from the Government of Catalonia through the “CERCA Program”. C.A.P. acknowledges support from the Secretaria d’Universitats i Recerca de la Generalitat de Catalunya and the European Social Fund as an AGAUR-funded PhD fellow. E.M. thanks the CERCA Program/Generalitat de Catalunya for their support to the Germans Trias i Pujol Research Institute (IGTP). This study was carried out by ISGlobal with competitive funding through the Gilead Sciences global HBV-CARE program (IN-ES-988–5799)
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