Histone H3E73Q and H4E53A mutations cause recombinogenic DNA damage

The stability and function of eukaryotic genomes is closely linked to histones and to chromatin structure. The state of the chromatin not only affects the probability of DNA to undergo damage but also DNA repair. DNA damage can result in genetic alterations and subsequent development of cancer and other genetic diseases. Here, we identified two mutations in conserved residues of histone H3 and histone H4 (H3E73Q and H4E53A) that increase recombinogenic DNA damage. Our results suggest that the accumulation of DNA damage in these histone mutants is largely independent on transcription and might arise as a consequence of problems occurring during DNA replication. This study uncovers the relevance of H3E73 and H4E53 residues in the protection of genome integrity.European Research Council ERC2014 AdG669898 TARLOOPMinisterio de Economía y Competitividad BFU2016-75058-

Functional polymorphisms in the CYP3A4, CYP3A5, and CYP21A2 genes in the risk for hypertension in pregnancy

An intronic single nucleotide polymorphism (SNP) in the CYP3A5 gene (CYP3A5*3; SNP rs776746) affects RNA splicing and enzymatic activity. The CYP3A5*3 frequency increased with distance from the equator and natural selection has been proposed to explain the worldwide distribution of this allele. CYP3A activity has been related with the risk for hypertension in pregnancy, a major cause of morbidity and mortality among women, and CYP3A5*3 could reduce the risk for this disease in populations from regions with high sodium and water availability. The CYP3A5 genotype was related with blood pressure in the general population, but the effect on the risk for hypertension in pregnancy has not been evaluated. We compared the allele and genotype frequencies of three functional SNPs in the CYP3A5 (rs776746), CYP3A4 (rs2740574), and CYP21A2 (rs6471) genes between pregnant women who developed hypertension (n=250) or who remained normotensive (control group, n=250). In addition, we sequenced the full CYP3A5 coding sequence in 40 women from the two groups to determine whether some gene variants could explain the risk for hypertensive pregnancies in our population. Allele and genotype frequencies did not differ between hypertensive and normotensive women for the three CYP variants. We did not find CYP3A5 nucleotide changes that could explain a higher risk for hypertension in pregnancy. Our data suggests that the variation in CYP3A5, CYP3A4, and CYP21A2 did not contribute to the risk for hypertension in pregnancy in our populationThe author´s work is supported by a grant from the Spanish Fondo de Investigaciones Sanitarias/Fondos FEDER European Union (ETS PI08/90008)

Sexual and gender-based violence in migration context: Needs, gaps and good practices

The first goal of the present work was to identity needs and gaps in interventions for migrant and refugee victims of sexual and gender-based violence (SGBV) in Spain. The second goal was to develop a guide for itineraries, resources and good practices to address the gaps detected. To produce data pertaining to the first purpose, we used a qualitative approach and focus groups (FGs) with institutional operators and representatives of the Third Sector. In total, six FGs were conducted with 35 key informants. The results of the study showed that existing services and resources are, in general, sufficient to cover the needs of the target group. However, some issues, such as excessive bureaucracy, re-victimization, difficulties regarding legal regularization, cultural barriers and the scant presence of cultural mediators were revealed. Regarding the second goal, a guide was created on the legal framework, itineraries and resources for migrant and refugee victims of SGBV, to ensure good practices and incorporate cultural mediation as an essential element to guarantee optimal use of services.El primer objetivo del presente trabajo fue identificar las necesidades y lagunas en la intervención con migrantes y refugiadas víctimas de violencia sexual y de género (VSG) en España. El segundo objetivo fue desarrollar una guía sobre los itinerarios, recursos y buenas prácticas que permitan solventar las brechas detectadas. Para la producción de datos del primer propósito nos servimos del enfoque cualitativo y la herramienta de los grupos focales con representantes institucionales y operadores del Tercer Sector. Se organizaron seis grupos focales con 35 informantes clave. Los resultados mostraron que los recursos existentes dan cobertura en gran medida a las necesidades del grupo destinatario. Sin embargo, se señalaron algunas limitaciones como la burocracia excesiva, la revictimización, las dificultades en los procesos de regularización, las barreras culturales o la escasa presencia de mediadores/as culturales. Para la consecución del segundo objetivo se partió de un análisis documental y de los datos de los grupos focales. El resultado fue una guía que aúna el marco legal, los itinerarios y los recursos destinados a personas migrantes y refugiadas víctimas de VSG; proporcionando un espacio clave para las buenas prácticas y la incorporación de la mediación cultural para garantizar el mejor aprovechamiento de los servicios

Uncovering the neural control of laryngeal activity and subglottic pressure in anaesthetized rats: insights from mesencephalic regions.

To assess the possible interactions between the dorsolateral Periaqueductal Gray matter (dlPAG) and the laryngeal motor neurons of the nucleus Ambiguus (nA), we have examined the pattern of double staining c-Fos/FoxP2 protein immunoreactivity (c-Fos-ir/Fox-P2-ir) and Tyrosine Hydroxylase (TH) throughout the rostro-caudal extent of nA in spontaneously breathing anaesthetised male Sprague–Dawley rats during dlPAG electrical stimulation. Activation of the dlPAG elicited a selective increase in c-Fos-ir with an ipsilateral predominance in the somatas of the loose (p<0.05) and compact formation (p<0.01) within the nA and confirm the expression of FoxP2 bilaterally in all the domains within the nA. A second group of experiments was made to examine the importance of the dlPAG in modulating the laryngeal response evoked after electrical or chemical (glutamate) dlPAG stimulations. Both electrical and chemical stmulations evoked a significant decrease of laryngeal resistance (subglottal pressure) (p<0.001) accompanied with an increase in respiratory rate together with a pressor and tachycardic response. The results of our study contribute with new data on the role of the mesencephalic neuronal circuits in the control mechanisms of subglottic pressure and laryngeal activity.Funding for open access charge: Universidad de Málaga / CBU

Service-learning educational approach for undergraduate students: development of an outreach workshop for high school students

In the last decade, research institutes and universities have strengthened the development of outreach activities in the biomedical field, involving researchers and professors as well as graduate students, but with little or no implication of undergraduate students. However, the development of this type of activities, using the Service-Learning educational approach, could be a valuable tool that would manage the acquisition of learning competencies by undergraduate students of Health Science Degrees and would put science at the service of society. In this project, we present the development of the workshop entitled “Exploring the human body”, in which 205 students in their first and second year of a Degree in Nursing or Medicine (University of Málaga, Spain) acted as mentors of 753 high school students (15 to 16 years old) in several school years (since 2016-2017, excluding 2019-2020 and 2020-2021 due to the COVID-19 pandemic). The workshop consisted of five work stations. Each station featured a set of different experiments and activities that were designed to teach the multiple levels by which the human body, and particularly the nervous system, can be studied: biomolecules, cells, tissues, organs and systems. Both high school and undergraduate students gave an evaluation of the workshop via questionnaires (Likert scale-based and short-answer questions) and a debriefing with the university professors. Data showed an overall score of 4.6 out of 5 points for the workshop by both high school and undergraduate students. In addition, undergraduate students pointed out that their participation had a positive impact on their academic background (4.8 out of 5 points), mainly due to the improvement of their oral communication skills (78 students) and self-confidence (58 students).Universidad de Málaga. Servicio de Publicaciones y Divulgación Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Laryngeal effects of stimulation of the dorsolateral Periaqueductal Grey Matter in spontaneously breathing anaesthetized rats

The stimulation of the Periaqueductal Gray matter (PAG) and nucleus retroambiguus (nRA) produces vocalization. A high expression of FOXP2 protein at mesencephalic (PAG) and pontine regions involved in cardiorespiratory control has been described. The aim of this study was to characterize the possible role of the dlPAG in modulating laryngeal activity and their effects on vocalization. Experimental studies were carried out with non-inbred male rats (n=27), Sprague-Dawley (250-300 g). Animals were anesthetized with sodium pentobarbitone (60 mg/kg i.p., initial dose, supplemented 2mg/ kg, i.v., as necessary). Neuromorphological study (n=6) The pattern of staining for c-Fos and FOXP2 protein immunoreactivity (c-Fos-ir) were examinated throughout the rostrocaudal extent of the nRa/nA region during electrical stimulation of the dlPAG. Neuropharmacological study (n=21) A double tracheal cannulation was used to obtain an “isolated glottis in situ” and to record respiratory airflow. Bilateral parietostomy allowed access to the dlPAG. Electrical stimulations (n=7) of this region using concentric bipolar electrodes (1ms pulses, 20-40µA, 100Hz for 5s) were performed. Microinjections of PBS-Evans Blue (250nl, pH 7.4±0.1, 5-s duration) (n=7) or glutamate (0,25M, 250nl) (n=7) were performed. Respiratory flow, pleural pressure, blood pressure and heart rate were also recorded. Activation of the dlPAG elicited a selective increase in c-Fos-ir with an ipsilateral predominance in nRA/nA somatas (p<0.01) and confirm the expression of FOXP2 bilaterally in both nuclei. dlPAG PBS-Evans Blue microinjections did not produce any significant changes in any of the cardiorespiratory variables recorded. dlPAG stimulations evoked a decrease of laryngeal resistance (subglottal pressure) (p<0,001) accompanied with an inspiratory facilitatory response consisted of an increase in respiratory rate (p<0,001), together with a pressor (p<0,001) and tachycardic response (p<0,001).Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Exploring laryngeal effects of dorsolateral periaqueductal grey stimulation in anesthetized rats: implications for c-Fos and FOXP2 expression in the nucleus ambiguus subdivisions.

Stimulation of the Periaqueductal Gray matter (PAG) and nucleus retroambiguus (nRA) evokes vocalization (1). The nRA serves as a key target for converting passive into active expiration by modulating the activity of laryngeal motoneurons located in the nucleus ambiguus (nA) (2). Previously, we have demonstrated the involvement of rostral and ventral pontine structures in altering laryngeal caliber (3). Furthermore, a heightened expression of the FOXP2 protein (a transcription factor closely linked to vocalization) has been observed in both mesencephalic (PAG) and pontine regions implicated in cardiorespiratory control (4). To investigate the potential interactions between the dlPAG and laryngeal motor neurons of the nA assessing the double staining patterns of c-Fos/FoxP2 protein immunoreactivity (c-Fos-ir/Fox-P2-ir) and Tyrosine Hydroxylase (TH) across the rostro-caudal extent of the nA. Experimental studies were carried out with non-inbred male rats (n=10), SPF, Sprague-Dawley (250-300 g). Animals were anesthetized with sodium pentobarbitone (60 mg/kg i.p., initial dose, supplemented 2mg/ kg, i.v.). The pattern of immunohistochemical and immunofluorescence staining for c-Fos and FOXP2 protein immunoreactivity (c-Fos-ir/FOXP2-ir) were examinated throughout the rostrocaudal extent of the nA region during electrical stimulation of the dlPAG. Guanethidine was administered to suppress sympathetically mediated cardiovascular responses. Stimulation of the dlPAG induced a specific increase in c-Fos-ir, with ipsilateral predominance in the somatas of both the loose (p<0.05) and compact formation (p<0.01) within the nA. Furthermore, we could confirm the bilateral expression of FoxP2 across all domains within the nA. Our study contributes with new data on the role of the mesencephalic neuronal circuits in the control mechanisms of subglottic pressure and laryngeal activity.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Dorsolateral Periaqueductal Grey Matter in the control of laryngeal activity and subglottic pressure in spontaneously breathing anaesthetized rats.

The stimulation of the dlPAG produces vocalization. The nRA turn passive into active expiration modifying the activity of laryngeal motoneurons located in the nA. We have shown that cPB and A5 Region are involved in changes of laryngeal caliber. A high expression of FOXP2 protein at mesencephalic and pontine regions involved in cardiorespiratory control has been described. The aim is to characterize the role of the dlPAG in modulating laryngeal activity and their effects on vocalization. Experimental studies were carried out with male Sprague-Dawley rats (n=25) (250-300g). Animals were anesthetized with sodium pentobarbitone (60 mg/kg i.p., initial dose, supplemented 2 mg/kg, i.v.). The pattern of staining for c-Fos and FOXP2 protein immunoreactivity were examinated throughout the rostrocaudal extent of the nRa/nA region during electrical stimulation of the dlPAG. Electrical stimulations (n=7) (1ms pulses, 20-40μA, 100Hz for 5s), microinjections of PBS-Evans Blue (250nl, pH 7.4±0.1, 5-s duration) (n=7) or glutamate (0,25M, 250nl) (n=7) were performed. Respiratory flow, pleural pressure, subglottic pressure, blood pressure and heart rate were recorded. Activation of the dlPAG elicited a selective increase in c-Fos-ir with an ipsilateral predominance in nRA/nA somatas (p<0.01) and confirm the expression of FOXP2 bilaterally in both nuclei. dlPAG PBS-Evans Blue microinjections did not produce any significant changes in any of the cardiorespiratory variables recorded. dlPAG electrical and chemical (glutamate) stimulations evoked a decrease of laryngeal resistance (subglottal pressure) (p<0,001) accompanied with an inspiratory facilitatory response consisted of an increase in respiratory rate (p<0,001), together with a pressor (p<0,001) and tachycardic response (p<0,001). Our study contributes with new data on the role of the mesencephalic neuronal circuits in the control mechanisms of subglottic pressure and laryngeal activity.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech