Multiple independent L-gulonolactone oxidase (GULO) gene losses and vitamin C synthesis reacquisition events in non-Deuterostomian animal species.

BACKGROUND: L-ascorbate (Vitamin C) is an important antioxidant and co-factor in eukaryotic cells, and in mammals it is indispensable for brain development and cognitive function. Vertebrates usually become L-ascorbate auxothrophs when the last enzyme of the synthetic pathway, an L-gulonolactone oxidase (GULO), is lost. Since Protostomes were until recently thought not to have a GULO gene, they were considered to be auxothrophs for Vitamin C. RESULTS: By performing phylogenetic analyses with tens of non-Bilateria and Protostomian genomes, it is shown, that a GULO gene is present in the non-Bilateria Placozoa, Myxozoa (here reported for the first time) and Anthozoa groups, and in Protostomians, in the Araneae family, the Gastropoda class, the Acari subclass (here reported for the first time), and the Priapulida, Annelida (here reported for the first time) and Brachiopoda phyla lineages. GULO is an old gene that predates the separation of Animals and Fungi, although it could be much older. We also show that within Protostomes, GULO has been lost multiple times in large taxonomic groups, namely the Pancrustacea, Nematoda, Platyhelminthes and Bivalvia groups, a pattern similar to that reported for Vertebrate species. Nevertheless, we show that Drosophila melanogaster seems to be capable of synthesizing L-ascorbate, likely through an alternative pathway, as recently reported for Caenorhabditis elegans. CONCLUSIONS: Non-Bilaterian and Protostomians seem to be able to synthesize Vitamin C either through the conventional animal pathway or an alternative pathway, but in this animal group, not being able to synthesize L-ascorbate seems to be the exception rather than the rule.

Improvement of yeast tolerance to acetic acid through Haa1 transcription factor engineering: towards the underlying mechanisms.

BACKGROUND: Besides being a major regulator of the response to acetic acid in Saccharomyces cerevisiae, the transcription factor Haa1 is an important determinant of the tolerance to this acid. The engineering of Haa1 either by overexpression or mutagenesis has therefore been considered to be a promising avenue towards the construction of more robust strains with improved acetic acid tolerance. RESULTS: By applying the concept of global transcription machinery engineering to the regulon-specific transcription factor Haa1, a mutant allele containing two point mutations could be selected that resulted in a significantly higher acetic acid tolerance as compared to the wild-type allele. The level of improvement obtained was comparable to the level obtained by overexpression of HAA1, which was achieved by introduction of a second copy of the native HAA1 gene. Dissection of the contribution of the two point mutations to the phenotype showed that the major improvement was caused by an amino acid exchange at position 135 (serine to phenylalanine). In order to further study the mechanisms underlying the tolerance phenotype, Haa1 translocation and transcriptional activation of Haa1 target genes was compared between Haa1 mutant, overproduction and wild-type strains. While the rapid Haa1 translocation from the cytosol to the nucleus in response to acetic acid was not affected in the Haa1S135F mutant strain, the levels of transcriptional activation of four selected Haa1-target genes by acetic acid were significantly higher in cells of the mutant strain as compared to cells of the wild-type strain. Interestingly, the time-course of transcriptional activation in response to acetic acid was comparable for the mutant and wild-type strain whereas the maximum mRNA levels obtained correlate with each strain's tolerance level. CONCLUSION: Our data confirms that engineering of the regulon-specific transcription factor Haa1 allows the improvement of acetic acid tolerance in S. cerevisiae. It was also shown that the beneficial S135F mutation identified in the current work did not lead to an increase of HAA1 transcript level, suggesting that an altered protein structure of the Haa1S135F mutant protein led to an increased recruitment of the transcription machinery to Haa1 target genes.

Conformational and mechanical changes of DNA upon transcription factor binding detected by a QCM and transmission line model.

A novel quartz crystal microbalance (QCM) analytical method is developed based on the transmission line model (TLM) algorithm to analyze the binding of transcription factors (TFs) to immobilized DNA oligoduplexes. The method is used to characterize the mechanical properties of biological films through the estimation of the film dynamic shear moduli, G and G, and the film thickness. Using the Saccharomyces cerevisiae transcription factor Haa1 (Haa1DBD) as a biological model two sensors were prepared by immobilizing DNA oligoduplexes, one containing the Haa1 recognition element (HRE(wt)) and another with a random sequence (HRE(neg)) used as a negative control. The immobilization of DNA oligoduplexes was followed in real time and we show that DNA strands initially adsorb with low or non-tilting, laying flat close to the surface, which then lift-off the surface leading to final film tilting angles of 62.9° and 46.7° for HRE(wt) and HRE(neg), respectively. Furthermore we show that the binding of Haa1DBD to HRE(wt) leads to a more ordered and compact film, and forces a 31.7° bending of the immobilized HRE(wt) oligoduplex. This work demonstrates the suitability of the QCM to monitor the specific binding of TFs to immobilized DNA sequences and provides an analytical methodology to study protein-DNA biophysics and kinetics.

Transmission line model analysis of transcription factors binding to oligoduplexes - differentiation of the effect of single nucleotide modifications.

Advanced impedance spectroscopy analysis based on the transmission line model (TLM) is explored as a novel QCM acoustic biosensing platform for the detection of the single point mutation effect on the binding of the transcription factors (TFs) to immobilized DNA oligoduplexes and the characterization of the protein-DNA mechanical properties.

Identification of a DNA-binding site for the transcription factor Haa1, required for Saccharomyces cerevisiae response to acetic acid stress.

The transcription factor Haa1 is the main player in reprogramming yeast genomic expression in response to acetic acid stress. Mapping of the promoter region of one of the Haa1-activated genes, TPO3, allowed the identification of an acetic acid responsive element (ACRE) to which Haa1 binds in vivo. The in silico analysis of the promoter regions of the genes of the Haa1-regulon led to the identification of an Haa1-responsive element (HRE) 5'-GNN(G/C)(A/C)(A/G)G(A/G/C)G-3'. Using surface plasmon resonance experiments and electrophoretic mobility shift assays it is demonstrated that Haa1 interacts with high affinity (K(D) of 2 nM) with the HRE motif present in the ACRE region of TPO3 promoter. No significant interaction was found between Haa1 and HRE motifs having adenine nucleotides at positions 6 and 8 (K(D) of 396 and 6780 nM, respectively) suggesting that Haa1p does not recognize these motifs in vivo. A lower affinity of Haa1 toward HRE motifs having mutations in the guanine nucleotides at position 7 and 9 (K(D) of 21 and 119 nM, respectively) was also observed. Altogether, the results obtained indicate that the minimal functional binding site of Haa1 is 5'-(G/C)(A/C)GG(G/C)G-3'. The Haa1-dependent transcriptional regulatory network active in yeast response to acetic acid stress is proposed.

Managerial Decision-Making of Nurses in Hospitals: creation and validation of a simulation scenario.

OBJECTIVE: to build and validate a clinical simulation scenario on hospital nurse managerial decision-making competence for undergraduate nursing students. METHOD: a descriptive and methodological study was carried out in a higher education institution, with the participation of 10 judges and five players. To do so, the conceptual simulation model proposed by Jeffries and standards of the International Nursing Association for Clinical Simulation and Learning were used to prepare the scenario and the checklist. RESULTS: the scenario was called "Managerial decision-making of nurses in the face of adverse events in a hospital". The scenario script and checklist were built for validation. The checklist was face- and content-validated. Afterward, judges used the checklist to validate the scenario, which, in its final version, was composed of Prebriefing (seven items), Scenario in Action (18 items) and Debriefing (seven items). CONCLUSION: the scenario proved to be a teaching strategy that anticipates the reality of future nurses, bringing them the self-confidence to perform their activities and helping them to act critically and reflectively during decision-making processes.

Surveying people with spinal cord injuries in Brazil to ascertain research priorities.

Scientists are concerned that the research they conduct accurately portrays the needs of people living with spinal cord injuries (SCI). As such, this study set out to investigate the main problems faced by people with SCI and their expectations for research. This quantitative, exploratory, analytical, and cross-sectional study was carried out online, with a non-probabilistic sample of 618 Brazilian adults with SCI who had registered voluntarily to participate in the research of the Neurorehab group. The virtual questionnaire consisted of 22 questions based on ISCOS Data Sets. The majority (68.9%) of participants were male, with higher education or a post-graduate qualification (49.5%). Most injuries had experienced traumatic injuries (78.5%) and 58.7% were paraplegic. The mean age was 38.04 years (SD = 9.85). The main difficulties faced after SCI were locomotion/accessibility (70.9%), neurogenic bladder (68.8%), neurogenic bowel (48.2%), and sexuality (36.1%). The highest demand was for experimental studies on stem cells (22.5%), rehabilitation (14.2%), and cures (13.9%). Most (84.3%) of those who reported sexuality problems after SCI were men (p = 0.013). The findings obtained empower people with SCI by enabling them to influence the agenda of scientific research based on their expectations and difficulties. This survey will also aid organizations to engage stakeholders to implement a comprehensive SCI management program.

Educational strategy to develop nursing students' management competencies in hospital practice.

OBJECTIVES: to apply and analyze an educational strategy to develop management skills in nursing students to work in hospital practice. METHODS: exploratory, intervention, qualitative study conducted from February 2020 to 2021. Fifty-four nursing students from a public higher education institution participated in this study, in which thirteen workshops were held to discuss management cases. Subsequently, semi-structured interviews were conducted using inductive thematic analysis. RESULTS: the case studies addressed the management competencies of communication, decision making, leadership, and interpersonal relationships. After the intervention, the strategy was evaluated through interviews, identifying positive aspects regarding knowledge acquisition; and other limiting aspects, such as limited time to discuss the cases. FINAL CONSIDERATIONS: the workshops proved to be effective as teaching strategies for students, adding new management knowledge that should help their performance as future nurses, capable of reflection, and the subjects of knowledge construction for the professional practice of nursing.

The diet-microbiome tango: how nutrients lead the gut brain axis.

Nutrients and the microbiome have a profound impact on the brain by influencing its development and function in health and disease. The mechanisms by which they shape brain function have only started to be uncovered. Here we propose that the interaction of diet with the microbiome is at the core of most mechanisms by which gut microbes affect host brain function. The microbiome acts on the host by altering the nutrients in the diet and by using them as precursors for synthetizing psychoactive metabolites. Diet is also a major modulator of gut microbiome composition making this another key mechanism by which they affect the host brain. Nutrient-microbiome-host interactions therefore provide an overarching framework to understand the function of the gut-brain axis.

Work-related psychosocial risks of the Family Health nurse and management strategies. / Riscos psicossociais relacionados ao trabalho do enfermeiro da Saúde da Família e estratégias de gerenciamento.

OBJECTIVE: To analyze the work-related psychosocial risks of Family Health nurses and the management strategies to minimize them. METHOD: Exploratory, qualitative study with nurses from teams of the Family Health Strategy (FHS). Data collection took place in July 2018 through semi-structured interviews and subsequently, an inductive thematic analysis was carried out. RESULTS: Participation of 18 nurses. Psychosocial risks related to the work context were identified, namely: insufficient professional training, compromised interpersonal relationships, work-family interface and psychological violence; as well as those related to work content: insufficient work equipment, lack of human resources and heavy workload. In addition, strategies for managing these risks were identified, such as using the family, spirituality, music and reading. CONCLUSION: The study should provoke the reflection of nurses about the eventual presence of risks in their work and contribute to fill knowledge production gaps in the area. It also provides subsidies for health managers' knowledge of the psychosocial risks to which these professionals are exposed, with a view to adopting measures.

Construction of the matrix of individual nursing competences in surgical units.

OBJECTIVE: to build and to describe an Individual Skills Matrix for nurses working in surgical units and their associated behaviors / attitudes. METHOD: Exploratory, qualitative study. 43 nurses from five surgical units participated, and data collection was carried out between April and September 2017. The focus group technique was used and for data interpretation an inductive thematic analysis was performed. The competence matrix was built from the testimonies of the participants plus a search in the literature for concepts directed to each competence and description of the expected behaviors and / or attitudes. RESULTS: For the Matrix, the following individual skills were identified: Planning; Communication, Relational Competence, Leadership, Decision Making and Ethics. FINAL CONSIDERATIONS: The construction of a Matrix should assist managers in recognizing the professional profile and assessing their performance, strengthening the achievement of professional and organizational objectives, as well as contributing to the quality and effectiveness of the care provided by nurses in these places.

Metabolic cross-feeding in imbalanced diets allows gut microbes to improve reproduction and alter host behaviour.

The impact of commensal bacteria on the host arises from complex microbial-diet-host interactions. Mapping metabolic interactions in gut microbial communities is therefore key to understand how the microbiome influences the host. Here we use an interdisciplinary approach including isotope-resolved metabolomics to show that in Drosophila melanogaster, Acetobacter pomorum (Ap) and Lactobacillus plantarum (Lp) a syntrophic relationship is established to overcome detrimental host diets and identify Ap as the bacterium altering the host's feeding decisions. Specifically, we show that Ap uses the lactate produced by Lp to supply amino acids that are essential to Lp, allowing it to grow in imbalanced diets. Lactate is also necessary and sufficient for Ap to alter the fly's protein appetite. Our data show that gut bacterial communities use metabolic interactions to become resilient to detrimental host diets. These interactions also ensure the constant flow of metabolites used by the microbiome to alter reproduction and host behaviour.

Burnout syndrome in university professors and academic staff members: psychometric properties of the Copenhagen Burnout Inventory-Brazilian version.

The aims of this study were to evaluate the psychometric properties of the Copenhagen Burnout Inventory-Brazilian version (CBI-Br) in a sample of university professors and academic staff members of Brazilian public universities, to estimate the level of burnout syndrome (BS) among these workers, and to assess the associations of BS with demographic and occupational determinants of the syndrome. A total of 676 workers participated in the study. Confirmatory factor analysis results supported a three-factor model with 18 items and an acceptable overall fit. Adequate convergent and discriminant validity of the CBI-Br's factors were observed, as well as adequate reliability of the instrument for the sample. In conclusion, the results of this study provide evidence of the validity and reliability of the CBI-Br for the measurement of BS in Brazilian university professors and academic staff members. In addition, the CBI-Br may be an important tool for the diagnosis of psychosocial risks related to BS in the academic environment.

Brazilian questionnaire of competencies of oncology nurses: Construction and test of content validation.

OBJECTIVE: This study aimed to describe construction and content validation of the Brazilian Questionnaire of Competencies of Oncology Nurses. METHODS: The methodological research was constructed based on the literature and observation in Brazilian hospitals searching to identify local evidence in the nursing practice. After, the construction of the 30 items distributed in eight sub-dimensions of competencies, the instrument was tested the content validation by 7 experts and 61 oncology nurses. Item responses were analyzed through content validity index and Cronbach's α were used in this test pilot. RESULTS: The content validity index of the scale was 0.90, and the content validity index of each item was 0.80-1.00. The mean of items varied between 4.98 (standard deviation = 0.13) and 4.70 (standard deviation = 0.53). The Cronbach's α of the instrument was 0.77. The Cronbach's α if each item was excluded ranged from 0.78 to 0.74, indicating consistency between items. CONCLUSION: The instrument is concise and clear, resulting in a valid content on test pilot in conducting a self-assessment the oncology nurses. It can be used to facilitate decision-making by identifying sub-dimensions that require attention in nursing education to improve patient care.

Psychosocial risks related to the nurse in the psychiatric hospital and management strategies.

OBJECTIVE: To characterize the presence of psychosocial risks related to the work of the nurse in a psychiatric hospital and the strategies for managing these risks. METHODS: Qualitative, in which 25 nurses from a psychiatric hospital participated using semi-structured interviews from November 2014 to January 2015. Data analysis was performed using the thematic method. RESULTS: The results showed psychosocial risks related to the work of psychiatric nurses, such as: insufficient academic training; lack of preparation and maintenance of equipment; poor relationship with colleagues; shortage of human resources and lack of capacity building; conflict between the demands of the home and work, as well as strategies for managing psychosocial risks such as family, cinema, music, reading, among others. FINAL CONSIDERATIONS: This study should provoke the reflection of managers and future nurses regarding the working conditions in a psychiatric hospital and possible psychosocial risks to which they are exposed.

Eliminação das hepatites virais: o que a enfermagem brasileira pode contribuir? / Elimination of viral hepatitis: what can brazilian nursing contribute? / Eliminación de las hepatitis virales: ¿qué puede aportar la enfermería brasileña?

Objetivo: Refletir acerca das contribuições da enfermagem para a eliminação das hepatites virais no Sistema Único de Saúde. Métodos: Estudo reflexivo de abordagem qualitativa sustentado na prática do enfermeiro em sistemas de saúde organizado em três categorias. Resultados: Na categoria Ampliação do escopo de prática: percurso para fortalecer a atuação do enfermeiro na Rede de Atenção à Saúde, aborda-se atuação na agenda estratégica do Ministério da Saúde e do Conselho Federal de Enfermagem em prover condições para ampliação do acesso por meio da gestão, assistência, ensino e pesquisa. Na categoria Subsídios da gestão e coordenação do cuidado para guiar a prática do enfermeiro aborda-se a dimensão individual e familiar, profissional, organizacional, sistêmica e societária com reconhecimento da necessidade de ampliar a reflexão. Na categoria O enfermeiro na gestão de programas de enfrentamento às hepatites virais pontua-se aspectos intrínsecos construídos desde a formação com ênfase para a gestão e gerência, em uma dinâmica real de responsabilidade técnica em programas, serviços e equipes. Conclusão: A reflexão da contribuição do enfermeiro possui relação com a gestão e coordenação do cuidado, prática já estabelecida ao profissional, mas que requer diretrizes e investimento para às hepatites por meio da educação permanente. (AU)

Objective: To reflect on the contributions of nursing to the elimination of viral hepatitis in the Unified Health System. Methods: This was a reflexive study with a qualitative approach based on the practice of nursing in health systems, organized into three categories. Results: In the category Expansion of the scope of practice: path to strengthen the role of nurses in the Health Care Network, the strategic agenda of the Ministry of Health and the Federal Council of Nursing is addressed in providing conditions for expanding access through management, care, teaching and research. In the category subsidies of care management and coordination to guide nursing practice, the individual and family, professional, organizational, systemic and societal dimensions are addressed, with recognition of the need to broaden reflection. In the category Nurses in the management of programs to combat viral hepatitis, intrinsic aspects built since training are highlighted, with emphasis on management and management, in a real dynamic of technical responsibility in programs, services and teams. Conclusion: The reflection on the contribution of nurses is related to the management and coordination of care, a practice already established for professionals, but which requires guidelines and investment for hepatitis through continuing education. (AU)

Objetivo: Reflexionar sobre las contribuciones de la enfermería a la eliminación de las hepatitis virales en el Sistema Único de Salud. Métodos: Se trata de un estudio reflexivo con abordaje cualitativo basado en la práctica de enfermería en los sistemas de salud, organizado en tres categorías. Resultados: En la categoría Ampliación del ámbito de la práctica: camino para fortalecer el papel de los enfermeros en la Red de Atención a la Salud, se aborda la agenda estratégica del Ministerio de Salud y del Consejo Federal de Enfermería en la provisión de condiciones para ampliar el acceso a través de la gestión, el cuidado, la enseñanza y la investigación. En la categoría Subsidios de gestión y coordinación del cuidado para orientar la práctica de enfermería, se abordan las dimensiones individual y familiar, profesional, organizacional, sistémica y social, reconociendo la necesidad de ampliar la reflexión. En la categoría Enfermeras en la gestión de programas de combate a las hepatitis virales, se destacan aspectos intrínsecos construidos desde la formación, con énfasis en la gestión y gestión, en una dinámica real de responsabilidad técnica en programas, servicios y equipos. Conclusión: La reflexión sobre la contribución de los enfermeros se relaciona con la gestión y coordinación de los cuidados, una práctica ya establecida para los profesionales, pero que requiere directrices e inversión para la hepatitis a través de la educación continua. (AU)

Bioinformatics Protocols for Quickly Obtaining Large-Scale Data Sets for Phylogenetic Inferences.

Useful insight into the evolution of genes and gene families can be provided by the analysis of all available genome datasets rather than just a few, which are usually those of model species. Handling and transforming such datasets into the desired format for downstream analyses is, however, often a difficult and time-consuming task for researchers without a background in informatics. Therefore, we present two simple and fast protocols for data preparation, using an easy-to-install, open-source, cross-platform software application with user-friendly, rich graphical user interface (SEDA; http://www.sing-group.org/seda/index.html ). The first protocol is a substantial improvement over one recently published (López-Fernández et al. Practical applications of computational biology and bioinformatics, 12th International conference. Springer, Cham, pp 88-96 (2019)[1]), which was used to study the evolution of GULO, a gene that encodes the enzyme responsible for the last step of vitamin C synthesis. In this paper, we show how the sequence data file used for the phylogenetic analyses can now be obtained much faster by changing the way coding sequence isoforms are removed, using the newly implemented SEDA operation "Remove isoforms". This protocol can be used to easily show that putative functional GULO genes are present in several Prostotomian groups such as Molluscs, Priapulida and Arachnida. Such findings could have been easily missed if only a few Protostomian model species had been used. The second protocol allowed us to identify positively selected amino acid sites in a set of 19 primate HLA immunity genes. Interestingly, the proteins encoded by MHC class II genes can show just as many positively selected amino acid sites as those encoded by classical MHC class I genes. Although a significant percentage of codons, which can be as high as 14.8%, are evolving under positive selection, the main mode of evolution of HLA immunity genes is purifying selection. Using a large number of primate species, the probability of missing the identification of positively selected amino acid sites is lower. Both projects were performed in less than one week, and most of the time was spent running the analyses rather than preparing the files. Such protocols can be easily adapted to answer many other questions using a phylogenetic approach.

Transcultural adaptation of the Competency Evaluation Questionnaire - Brazilian version. / Adaptação transcultural do Competency Evaluation Questionnaire - versão brasileira.

OBJECTIVE: Describe the process of translation and adaptation to Brazilian culture of the Competency Evaluation Questionnaire for hospital nurses. METHOD: Methodological study of translation and cross-cultural adaptation, including the steps translation of the original instrument into Portuguese, consensual evaluation of translated versions, back - translation, validation by judges and pilot with 29 hospital nurses. RESULTS: They presented the stages of process of cross-cultural adaptation of the instrument, with adjustments proposed by a Judges Committee, composed of 27 items distributed in five domains of competencies for hospital nurses. The instrument adapted to Portuguese presented semantic, idiomatic, experimental and conceptual equivalence with the original, and the total value of the Cronbach Alpha 0.932, in addition to the relative validation. CONCLUSIONS: The Competency Evaluation Questionnaire has been shown to be a reliable instrument that can be used but does not exclude the need for future statistical tests aiming at psychometric validity.

Competence-based performance evaluation in hospital nurses. / Avaliação de desempenho por competências em enfermeiros hospitalares.

OBJECTIVE: to evaluate the frequencies attributed to the professional competences of hospital nurses, discussing the ones that obtained higher and lower frequencies. METHOD: descriptive, cross-sectional, quantitative study with 45 nurses of a hospital of high complexity in the interior of São Paulo state. The study used the Competences Evaluation Questionnaire, translated and validated in Brazil, composed of 27 items in five domains: professionalism, communication, management, nursing process and problem solving. Data were analyzed using descriptive statistics. RESULTS: it was found that 80% of the participants were female and 20% were male. The age ranged between 25 and 63 years, with an average of 40.02 years, and the average length of professional experience was 13.39 years. The competences "Communication with the hospital's administrative staff" and "Participation in scientific research and / or application of results" had the lowest frequencies, while "Commitment to punctuality and workload" and "Commitment to the ethical principles of the profession" were evaluated with the highest frequencies. CONCLUSION: evaluating the performance by competences becomes essential for managers and training centers, since it contributes to the identification of gaps in knowledge, skills and attitudes of professionals, by promoting the elaboration and implementation of strategies for their development.

Access to viral hepatitis care: distribution of health services in the Northern region of Brazil.

OBJECTIVE: To analyze the distribution of health care services for viral hepatitis and reported cases of viral hepatitis according to the health regions of Northern Brazil. METHOD: It is an evaluative, descriptive and quantitative research considering viral hepatitis care services and reported cases in the Northern region of Brazil, using data collected from the National Registry of Health Establishments and the Notifiable Diseases Information System. Descriptive statistics and georeferencing, through software, were used to demonstrate the spatial distribution of services and reported cases. RESULTS: Viral hepatitis health services are distributed in a differentiated way; rapid tests are capillaries in the states; confirmatory tests and treatment are performed in some health regions, with a greater grouping of services in the capitals and their surroundings. Cases were reported across all regions, with areas of higher concentration near services. CONCLUSION: The availability of services can favor access to prevention, diagnosis and monitoring of cases. However, organizational peculiarities of the health system and services highlight fragilities that have repercussions on the access and entirety of viral hepatitis care.