Differences in firing efficiency, chromatin, and transcription underlie the developmental plasticity of the Arabidopsis DNA replication origins.

Eukaryotic genome replication depends on thousands of DNA replication origins (ORIs). A major challenge is to learn ORI biology in multicellular organisms in the context of growing organs to understand their developmental plasticity. We have identified a set of ORIs of Arabidopsis thaliana and their chromatin landscape at two stages of post-embryonic development. ORIs associate with multiple chromatin signatures including transcription start sites (TSS) but also proximal and distal regulatory regions and heterochromatin, where ORIs colocalize with retrotransposons. In addition, quantitative analysis of ORI activity led us to conclude that strong ORIs have high GC content and clusters of GGN trinucleotides. Development primarily influences ORI firing strength rather than ORI location. ORIs that preferentially fire at early developmental stages colocalize with GC-rich heterochromatin, but at later stages with transcribed genes, perhaps as a consequence of changes in chromatin features associated with developmental processes. Our study provides the set of ORIs active in an organism at the post-embryo stage that should allow us to study ORI biology in response to development, environment, and mutations with a quantitative approach. In a wider scope, the computational strategies developed here can be transferred to other eukaryotic systems.

Contraceptive counselling experiences in Spain in the process of creating a web-based contraceptive decision support tool: a qualitative study.

BACKGROUND: The choice of contraceptive method is a complex decision, and professionals should offer counselling based on the preferences, values and personal situation of the user(s). Some users are unsatisfied with the counselling received, which may, among other consequences, adversely affect method use adherence. In view of this situation, we propose exploring the experiences and needs of users and professionals for contraceptive counselling, in the context of creating a web-based contraceptive decision support tool. METHODS/DESIGN: Qualitative research was conducted through focus group discussions (64 users split into eight groups, and 19 professionals in two groups, in Tarragona, Spain) to explore the subjects' experiences and needs. The data were categorized and the categories were defined and classified based on the three-step protocol or framework for Quality on Contraceptive Counseling (QCC), created by experts, which reviews the quality of interactions between user and professional during the counselling process. RESULTS: In counselling, users demand more information about the different methods, in an environment of erroneous knowledge and misinformation, which lead to false beliefs and myths in the population that are not contrasted by the professional in counselling. They complain that the method is imposed on them and that their views regarding the decision are not considered. Professionals are concerned that their lack of training leads to counselling directed towards the methods they know best. They acknowledge that a paternalistic paradigm persists in the healthcare they provide, and decision support tools may help to improve the situation. CONCLUSIONS: Users feel unsatisfied and/or demand more information and a warmer, more caring approach. Professionals are reluctant to assume a process of shared decision-making. The use of a contraception DST website may solve some shortcomings in counselling detected in our environment.

Histone H3 Dynamics Reveal Domains with Distinct Proliferation Potential in the Arabidopsis Root.

A coordinated transition from cell proliferation to differentiation is crucial for organogenesis. We found that extensive chromatin reorganization, shown here for histone H3 proteins, characterizes cell population dynamics in the root developmental compartments. The canonical H3.1 protein, incorporated during S-phase, is maintained at high levels in cells dividing at a high rate but is massively evicted in cells undergoing their last cell cycle before exit to differentiation. A similar pattern was observed in the quadruple mutant for the H3.1-encoding genes HTR1, HTR2, HTR3, and HTR9 (htr1,2,3,9), in which H3.1 is expressed only from the HTR13 gene. H3 eviction is a fast process occurring within the G2 phase of the last cell cycle, which is longer than G2 in earlier cell cycles. This longer G2 likely contributes to lower the H3.1/H3.3 ratio in cells leaving the root meristem. The high H3.1/H3.3 ratio and H3.1 eviction process also occurs in endocycling cells before differentiation, revealing a common principle of H3 eviction in the proliferating and endocycling domains of the root apex. Mutants in the H3.1 chaperone CAF-1 (fas1-4) maintain a pattern similar to that of wild-type roots. Our studies reveal that H3 incorporation and eviction dynamics identify cells with different cell division potential during organ patterning.

Retrotransposons are specified as DNA replication origins in the gene-poor regions of Arabidopsis heterochromatin.

Genomic stability depends on faithful genome replication. This is achieved by the concerted activity of thousands of DNA replication origins (ORIs) scattered throughout the genome. The DNA and chromatin features determining ORI specification are not presently known. We have generated a high-resolution genome-wide map of 3230 ORIs in cultured Arabidopsis thaliana cells. Here, we focused on defining the features associated with ORIs in heterochromatin. In pericentromeric gene-poor domains ORIs associate almost exclusively with the retrotransposon class of transposable elements (TEs), in particular of the Gypsy family. ORI activity in retrotransposons occurs independently of TE expression and while maintaining high levels of H3K9me2 and H3K27me1, typical marks of repressed heterochromatin. ORI-TEs largely colocalize with chromatin signatures defining GC-rich heterochromatin. Importantly, TEs with active ORIs contain a local GC content higher than the TEs lacking them. Our results lead us to conclude that ORI colocalization with retrotransposons is determined by their transposition mechanism based on transcription, and a specific chromatin landscape. Our detailed analysis of ORIs responsible for heterochromatin replication has implications on the mechanisms of ORI specification in other multicellular organisms in which retrotransposons are major components of heterochromatin and of the entire genome.

The effectiveness of a digital shared decision-making tool in hormonal contraception during clinical assessment: study protocol of a randomized controlled trial in Spain.

BACKGROUND: Decision-making tools represent a paradigm shift in the relationship between the clinician and the user/patient. Some of their advantages include patient commitment, the promotion of preferences and values, and increased treatment adherence. This study protocol aims to assess the effectiveness of a decision-making tool in contraception (SHARECONTRACEPT) concerning: a) Improvement in counselling on hormonal contraception at the medical consultation, measured in terms of decreasing decisional conflict and improving knowledge of available contraceptive options; b) Improvement in adherence to treatment measured in terms of: persistence in the chosen treatment, compliance with dose or procedure of use, and ability to deal with incidents related to the use of the contraceptive method; and decreasing unwanted pregnancies and voluntary interruption of pregnancy. The SHARECONTRACEPT tool, developed by previous phases of this project, is available at: http://decisionscompartides.gencat.cat/en/decidir-sobre/anticoncepcio_hormonal/ METHODS/DESIGN: A longitudinal, prospective-type, randomized, controlled community clinical trial, carried out in the clinical contraceptive counselling units of 6 autonomous regions in Spain, with an experimental group and a control group. Description of the intervention: The health professionals participating will be randomly assigned to one of the two groups. Clinicians assigned to the experimental group will perform contraceptive counselling assisted by SHARECONTRACEPT, and those of the control group will follow the conventional contraceptive counselling provided in their clinical unit. It is planned to study 1708 users (control group n = 854 and intervention group n = 854), recruited from women who attend the consultations of the health professionals. The selected users will be followed up for one year. The data will be collected through ad-hoc questionnaires, and validated instruments for measuring decisional conflict and adherence to treatment. DISCUSSION: The results of this study protocol will offer evidence of the effectiveness of a shared decision-making tool, SHARECONTRACEPT, which may prove a useful tool for users and professionals to promote adherence to contraceptive methods. TRIAL REGISTRATION: Clinical Register number ISRCTN5827994 . Date: 15/04/2019 (Retrospectively registered).

Basolateral sorting and transcytosis define the Cu+-regulated translocation of ATP7B to the bile canaliculus.

The Cu(+) pump ATP7B plays an irreplaceable role in the elimination of excess Cu(+) by the hepatocyte into the bile. The trafficking and site of action of ATP7B are subjects of controversy. One current proposal is that an increase in intracellular Cu(+) results in the translocation of ATP7B to the lysosomes and excretion of excess Cu(+) through lysosomal-mediated exocytosis at the bile canaliculus. Here, we show that ATP7B is transported from the trans-Golgi network (TGN) to the bile canaliculus by basolateral sorting and endocytosis, and microtubule-mediated transcytosis through the subapical compartment. Trafficking ATP7B is not incorporated into lysosomes, and addition of Cu(+) does not cause relocalization of lysosomes and the appearance of lysosome markers in the bile canaliculus. Our data reveal the pathway of the Cu(+)-mediated transport of ATP7B from the TGN to the bile canaliculus and indicates that the bile canaliculus is the primary site of ATP7B action in the elimination of excess Cu(.)

The Functional Topography of the Arabidopsis Genome Is Organized in a Reduced Number of Linear Motifs of Chromatin States.

Chromatin is of major relevance for gene expression, cell division, and differentiation. Here, we determined the landscape of Arabidopsis thaliana chromatin states using 16 features, including DNA sequence, CG methylation, histone variants, and modifications. The combinatorial complexity of chromatin can be reduced to nine states that describe chromatin with high resolution and robustness. Each chromatin state has a strong propensity to associate with a subset of other states defining a discrete number of chromatin motifs. These topographical relationships revealed that an intergenic state, characterized by H3K27me3 and slightly enriched in activation marks, physically separates the canonical Polycomb chromatin and two heterochromatin states from the rest of the euchromatin domains. Genomic elements are distinguished by specific chromatin states: four states span genes from transcriptional start sites (TSS) to termination sites and two contain regulatory regions upstream of TSS. Polycomb regions and the rest of the euchromatin can be connected by two major chromatin paths. Sequential chromatin immunoprecipitation experiments demonstrated the occurrence of H3K27me3 and H3K4me3 in the same chromatin fiber, within a two to three nucleosome size range. Our data provide insight into the Arabidopsis genome topography and the establishment of gene expression patterns, specification of DNA replication origins, and definition of chromatin domains.

Variation of 45S rDNA intergenic spacers in Arabidopsis thaliana.

Approximately seven hundred 45S rRNA genes (rDNA) in the Arabidopsis thaliana genome are organised in two 4 Mbp-long arrays of tandem repeats arranged in head-to-tail fashion separated by an intergenic spacer (IGS). These arrays make up 5 % of the A. thaliana genome. IGS are rapidly evolving sequences and frequent rearrangements inside the rDNA loci have generated considerable interspecific and even intra-individual variability which allows to distinguish among otherwise highly conserved rRNA genes. The IGS has not been comprehensively described despite its potential importance in regulation of rDNA transcription and replication. Here we describe the detailed sequence variation in the complete IGS of A. thaliana WT plants and provide the reference/consensus IGS sequence, as well as genomic DNA analysis. We further investigate mutants dysfunctional in chromatin assembly factor-1 (CAF-1) (fas1 and fas2 mutants), which are known to have a reduced number of rDNA copies, and plant lines with restored CAF-1 function (segregated from a fas1xfas2 genetic background) showing major rDNA rearrangements. The systematic rDNA loss in CAF-1 mutants leads to the decreased variability of the IGS and to the occurrence of distinct IGS variants. We present for the first time a comprehensive and representative set of complete IGS sequences, obtained by conventional cloning and by Pacific Biosciences sequencing. Our data expands the knowledge of the A. thaliana IGS sequence arrangement and variability, which has not been available in full and in detail until now. This is also the first study combining IGS sequencing data with RFLP analysis of genomic DNA.

Impact of maternity care policy in Catalonia: a retrospective cross-sectional study of service delivery in public and private hospitals.

BACKGROUND: As a result of the growing number of interventions that are now performed in the context of maternity care, health authorities have begun to examine the possible repercussions for service provision and for maternal and neonatal health. In Spain the Strategy Paper on Normal Childbirth was published in 2008, and since then the authorities in Catalonia have sought to implement its recommendations. This paper reviews the current provision of maternity care in Catalonia. METHODS: This was a descriptive study. Hospitals were grouped according to their source of funding (public or private) and were stratified (across four strata) on the basis of the annual number of births recorded within their respective maternity service. Data regarding the distribution of obstetric professionals were taken from an official government survey of hospitals published in 2010. The data on obstetric interventions (caesarean, use of forceps, vacuum or non-specified instruments) performed in 2007, 2010 and 2012 were obtained by consulting discharge records of 44 public and 20 private hospitals, which together provide care in 98% of all births in Catalonia. Proportions and confidence intervals were calculated for each intervention performed in all full-term (37-42 weeks) singleton births. RESULTS: Analysis of staff profiles according to the stratification of hospitals showed that almost all the hospitals had more obstetricians than midwives among their maternity care staff. Public hospitals performed fewer caesareans [range between 19.20% (CI 18.84-19.55) and 28.14% (CI 27.73-28.54)] than did private hospitals [range between 32.21% (CI 31.78-32.63) and 39.43% (CI 38.98-39.87)]. The use of forceps has decreased in public hospitals. The use of a vacuum extractor has increased and is more common in private hospitals. CONCLUSIONS: Caesarean section is the most common obstetric intervention performed during full-term singleton births in Catalonia. The observed trend is stable in the group of public hospitals, but shows signs of a rise among private institutions. The number of caesareans performed in accredited public hospitals covers a limited range with a stable trend. Among public hospitals the highest rate of caesareans is found in non-accredited hospitals with a lower annual number of births.

Level of Job Burnout among Midwives Working in Labour Rooms in Barcelona Region: A Cross-Sectional Study.

BACKGROUND: Nowadays, burnout syndrome (BS) symptoms appear to have increased in healthcare workers, specifically midwives, but there are no studies on burnout among midwives in Catalonia. The present study aimed to assess and describe the prevalence of BS in midwives working in labour rooms. METHODS: A cross-sectional descriptive study was conducted on 122 midwives working in 24 maternity hospitals in the Barcelona (region) which were selected using purposive sampling from January to March 2017. Data were collected using two questionnaires (demographic information, job burnout using Spanish Burnout Inventory with 20 items and four subscales). Data analysis was performed using SPSS software version 21 and Chi-Square, U Mann-Whitney, and Kruskall-Wallis. P<0.05 was considered statistically significant. RESULTS: None of the participants obtained a critical level of BS. 37 (30.33%) participants scored medium burnout and 47 (38.52%) recorded low burnout. Statistically, work stress (P=0.01), marital status (P=0.006), attendance of more than three women per shift (P=0.001), the number of children (P=0.01), parity (P=0.005), health status (P=0.04), and being on sick leave over last year (P=0.04) were significantly correlated with medium-high levels of burnout. Burnout scores were higher in midwives having a life partner and those without children. CONCLUSION: Following the results, no participant obtained a critical level of BS; about one-third of them scored medium-high burnout. However, specific interventions are suggested to be conducted to maintain the midwives' motivation and prevent burnout development.

Validation and psychometric properties of the Spanish version of the questionnaire for assessing the childbirth experience (QACE).

OBJECTIVE: When evaluating childbirth experience, some of the factors considered by women include their previous births experience, pain management during birth, and companion and healthcare professional support received. The objective of this paper is to validate the Questionnaire for Assessing the Childbirth Experience (QACE) into the Spanish population by assessing its psychometric properties. METHODS: Due to the differences between the Spanish and English languages, a careful translation process was the first step to making the QACE useable to Spanish speaking cohorts, once thoroughly translated their conceptual equivalence was evaluated by a group of experts and tested later via interviews with postpartum women for comprehensibility evaluation. Secondly, the validation process was obtained throughout the factorial analysis, internal consistency, test-retest evaluation and convergent and discriminant validity. RESULTS: A total of 268 postpartum women participated in the validity study. The KMO (0.84) and Bartlett test (p < 0.001) confirmed the adequacy of factor analysis and the Screen plot showed four factors with the predictive power of 52.63%, which supported total variance. Confirmatory factor analysis indicated an adequate/good fitness for the new model (χ2/df = 1.47, GFI = 0.979, RMSEA = 0.052, CFI = 0.889, NFI = 0.727, NNFI = 0.873, and SRMR = 0.155). Internal consistency was confirmed with McDonal's Omega level of 0.818. Test-retest evaluation supported test stability (r = 0.79, p < 0.01). Convergent and discriminant validity were obtained with 0.803 and 0.475 Pearson coefficients respectively. CONCLUSIONS: The Spanish version of QACE is a relevant tool for measuring childbirth experience into the Spanish context with acceptable validity and stability.

Into the Thermus Mobilome: Presence, Diversity and Recent Activities of Insertion Sequences Across Thermus spp.

A high level of transposon-mediated genome rearrangement is a common trait among microorganisms isolated from thermal environments, probably contributing to the extraordinary genomic plasticity and horizontal gene transfer (HGT) observed in these habitats. In this work, active and inactive insertion sequences (ISs) spanning the sequenced members of the genus Thermus were characterized, with special emphasis on three T. thermophilus strains: HB27, HB8, and NAR1. A large number of full ISs and fragments derived from different IS families were found, concentrating within megaplasmids present in most isolates. Potentially active ISs were identified through analysis of transposase integrity, and domestication-related transposition events of ISTth7 were identified in laboratory-adapted HB27 derivatives. Many partial copies of ISs appeared throughout the genome, which may serve as specific targets for homologous recombination contributing to genome rearrangement. Moreover, recruitment of IS1000 32 bp segments as spacers for CRISPR sequence was identified, pointing to the adaptability of these elements in the biology of these thermophiles. Further knowledge about the activity and functional diversity of ISs in this genus may contribute to the generation of engineered transposons as new genetic tools, and enrich our understanding of the outstanding plasticity shown by these thermophiles.

Optimal settings for childbirth.

Many studies highlight how health is influenced by the settings in which people live, work, and receive health care. In particular, the setting in which childbirth takes place is highly influential. The physiological processes of women's labor and birth are enhanced in optimal ("salutogenic," or health promoting) environments. Settings can also make a difference in the way maternity staff practice. This paper focuses on how positive examples of Italian birth places incorporate principles of healthy settings. The "Margherita" Birth Center in Florence and the Maternity Home "Il Nido" in Bologna were purposively selected as cases where the physical-environmental setting seemed to reflect an embedded model of care that promotes health in the context of childbirth. Narrative accounts of the project design were collected from lead professional and direct inspections performed to elicit the key salutogenic components of the physical layout. Comparisons between cases with a standard hospital labor ward layout were performed. Cross-case similarities emerged. The physical characteristics mostly related to optimal settings were a result of collaborative design decisions with stakeholders and users, and the resulting local intention to maximize safe physiological birth, psychosocial wellbeing, facilitate movement and relaxation, prioritize space for privacy, intimacy, and favor human contact and relationships. The key elements identified in this paper have the potential to inform further investigations for the design or renovation of all birth places (including hospitals) in order to optimize the salutogenic component of any setting in any country.

Optimal outcomes and women's positive pregnancy experience: a comparison between the World Health Organization guideline and recommendations in European national antenatal care guidelines.

BACKGROUND: The publication of the World Health Organization (WHO) recommendations on antenatal care in 2016 introduced the perspective of women as a necessary component of clinical guidelines in maternity care. WHO highlights the crucial role played by evidence-based recommendations in promoting and supporting normal birth processes and a positive experience of pregnancy. This paper aims to explore and critically appraise recommendations of national antenatal care guidelines across European countries in comparison with the WHO guideline. METHODS: We collected guidelines from country partners of the EU COST Action IS1405. Components of the documents structure and main recommendations within and between them were compared and contrasted with the WHO guideline on antenatal care with a particular interest in exploring whether and how women's experience was included in the recommendations. RESULTS: Eight out of eleven countries had a single national guideline on antenatal care while three countries did not. National guidelines mostly focused on care of healthy women with a straightforward pregnancy. The level of concordance between the national and the WHO recommendations varied along a continuum from almost total concordance to almost total dissonance. Women's views and experiences were accounted for in some guidelines, but mostly not placed at the same level of importance as clinical items. CONCLUSIONS: Findings outline convergences and divergences with the WHO recommendations. They highlight the need for considering women's views more in the development of evidence-based recommendations and in practice for positive impacts on perinatal health at a global level, and on the experiences of each family.

Organización de la evidencia científica: sinopsis / Organisation of Scientific Evidence: Synopsis

Desde la recopilación de información hasta la creación de conocimientoexiste un proceso complejo. La organización de la evidenciacientífica nos ayuda a entenderlo y a identificar las fuentes que nosofrecen la información ya tratada. Mantener el conocimiento actualizadoes necesario pero también cada vez más complicado por la grancantidad de información y escasa disponibilidad de tiempo entre losprofesionales. El artículo presenta la sinopsis dentro de la organizaciónde la evidencia científica, explica su utilidad como instrumentopara la actualización y difusión del conocimiento, y expone los aspectosnecesarios para la elaboración de un resumen estructurado enforma de sinopsis. También se exponen los CAT (critical appraisal topics)como instrumento útil y rápido (AU)

A complex process takes place between the collection of informationand the creation of knowledge. The organisation of scientific evidencehelps us to understand that process and to identify the sources that providealready processed information. Keeping knowledge up-to-date is anecessary but increasingly complicated task because of the large quantitiesof information and the lack of time among professionals. The articlepresents the synopsis within the organisation of scientific evidence,explains its use as a tool for the updating and spreading of knowledgeand sets out the necessary aspects for the drafting of a structured summaryin the form of a synopsis. The CATs (Critical Appraisal Topics) arealso presented as a useful and quick tool (AU)

Acceso a la contracepción de emergencia. Experiencia en Mollet del Vallés / Access to emergency contraception: experience in Mollet del Vallés (Barcelona, Spain)

Objetivo: Identificar si existen diferencias en el perfil de las mujeres y la demanda de contracepción de emergencia antes y después de la implantación de la guía clínica del Departamento de Salud, en el Servicio de Salud Sexual y Reproductiva de Mollet del Vallés. Personas y método: Se planteó un estudio transversal en el que se revisaron todos los registros de demanda de contracepción de emergencia (CE) en dos periodos: antes del comienzo de la campaña (2000- 2004) y tras la implantación de la guía clínica y la campaña «Mejor sin Riesgo» (2005-2006). Se estudiaron los registros de todas las mujeres que acudieron al Servicio mencionado solicitando CE. Las variables se estratificaron y se realizó un análisis descriptivo. Resultados: Se recogieron un total de 1.259 registros válidos, de un total de 1.537 en ambos periodos de tiempo. En ambos periodos, más de la mitad de las solicitudes registradas correspondieron a jóvenes de hasta 21 años. La demanda de CE aumentó un 71% durante el segundo periodo del estudio La mayoría de solicitudes de CE se dieron los lunes, 43,6% en el primer periodo y 37,3% en el segundo. Aproximadamente la mitad de las mujeres que la solicitaron tenían menos de 21 años, 65% (1.º periodo) y 59,5% (2.º periodo). La causa más frecuente por la que se solicitó la CE en ambos periodos fue la rotura o mal uso del preservativo 81,3% (1.º periodo) y 76,8% (2.º periodo). Conclusiones: El aumento de la demanda en el segundo periodo evidencia una mejora de la accesibilidad a la CE que puede atribuirse a la campaña «Mejor sin riesgo». No se apreciaron diferencias importantes en las variables analizadas entre el primer y segundo periodo (AU)

Objective: To determine whether there were changes in the demand for emergency contraception (EC) in the Sexual and Reproductive Health service of Mollet del Vallés, and in the profiles of the women requesting it, after the introduction of the clinical guidelines issued by the Health Department. Subjects and methods: A cross-sectional study was designed to review all the records of the demand for EC during two periods: prior to the start of the campaign (2000-2004) and after the introduction of the clinical guidelines and the “Better without Risk” campaign (2005-2006). The Records of all the women who came to the aforementioned service requesting EC were studied. They were stratified according to the variables and a descriptive analysis was carried out. Results: In all, 1,259 records were collected, out of a total of 1,537 corresponding to the two time periods. In both periods, more than half of the requests registered had been made by girls aged 21 years or younger. The demand for EC increased 71% during the second phase of the study. Most of the requests for EC took place on Monday, 43.6% during the first period and 37.3% during the second. Approximately half of the women who requested it were under 21 years of age (65% in the First phase and 59.5% in the second). The most common reason for requesting EC during both periods was the breakage or incorrect use of the condom (81.3% in the first period and 76.8% in the second). Conclusion: The increase in demand in the second period is evidence of an improvement in the accessibility of EC, which can be attributed to the “Better without Risk” campaign. There were no important differences between the first and second periods in terms of the variables analyzed (AU)