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BACKGROUND: The quality of the learning environment significantly impacts student engagement and professional identity formation in health professions education. Despite global recognition of its importance, research on student perceptions of learning environments across different health education programs is scarce. This study aimed to explore how health professional students perceive their learning environment and its influence on their professional identity development. METHODS: An explanatory mixed-methods approach was employed. In the quantitative phase, the Dundee Ready Education Environment Measure [Minimum-Maximum possible scores = 0-200] and Macleod Clark Professional Identity Scale [Minimum-Maximum possible scores = 1-45] were administered to Qatar University-Health students (N = 908), with a minimum required sample size of 271 students. Data were analyzed using SPSS, including descriptive statistics and inferential analysis. In the qualitative phase, seven focus groups (FGs) were conducted online via Microsoft Teams. FGs were guided by a topic guide developed from the quantitative results and the framework proposed by Gruppen et al. (Acad Med 94:969-74, 2019), transcribed verbatim, and thematically analyzed using NVIVO®. RESULTS: The questionnaire response rate was 57.8% (525 responses out of 908), with a usability rate of 74.3% (390 responses out of 525) after excluding students who only completed the demographic section. The study indicated a "more positive than negative" perception of the learning environment (Median [IQR] = 132 [116-174], Minimum-Maximum obtained scores = 43-185), and a "good" perception of their professional identity (Median [IQR] = 24 [22-27], Minimum-Maximum obtained scores = 3-36). Qualitative data confirmed that the learning environment was supportive in developing competence, interpersonal skills, and professional identity, though opinions on emotional support adequacy were mixed. Key attributes of an ideal learning environment included mentorship programs, a reward system, and measures to address fatigue and boredom. CONCLUSIONS: The learning environment at QU-Health was effective in developing competence and interpersonal skills. Students' perceptions of their learning environment positively correlated with their professional identity. Ideal environments should include mentorship programs, a reward system, and strategies to address fatigue and boredom, emphasizing the need for ongoing improvements in learning environments to enhance student satisfaction, professional identity development, and high-quality patient care.
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Identificação Social , Humanos , Masculino , Feminino , Catar , Estudantes de Ciências da Saúde/psicologia , Adulto Jovem , Grupos Focais , Aprendizagem , Inquéritos e Questionários , Adulto , Atitude do Pessoal de SaúdeRESUMO
In some countries, pharmacists have obtained prescribing rights to improve quality and accessibility of care and reduce physician workload. This case study explored pharmacists' current roles in and potential for prescribing in primary care in the Netherlands, where prescribing rights for pharmacists do not exist. Participatory observations of pharmacists working in either general practice or community pharmacy were conducted, as were semi-structured interviews about current and potential practice. The latter were extended to patients and other healthcare professionals, mainly general practitioners, resulting in 34 interviews in total. Thematic analyses revealed that pharmacists, in all cases, wrote prescriptions that were then authorized by a physician before dispensing. General practice-based pharmacists often prescribed medications during patient consultations. Community pharmacists mainly influenced prescribing through (a) medication reviews where the physician and/or practice nurse often were consulted to make treatment decisions, and (b) collaborative agreements with physicians to start or substitute medications in specific situations. These findings imply that the pharmacists' current roles in prescribing in the Netherlands resemble collaborative prescribing practices in other countries. We also identified several issues that should be addressed before formally introducing pharmacist prescribing, such as definitions of tasks and responsibilities and prescribing-specific training for pharmacists.
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OBJECTIVES: This study aims to assess Qatar community pharmacists' practices in advocating and promoting adult vaccination and to characterize and quantify potential determinants of participation in adult vaccination as vaccine administrators, based on the theoretical domains framework (TDF). METHODS: A cross-sectional survey of a randomly selected sample of community pharmacists in Qatar was conducted using a self-administered validated questionnaire. Items in the questionnaire on potential determinants of participation in adult vaccination were based on TDF. TDF items were subjected to principal components analysis. RESULTS: In total, 271 respondents completed the questionnaire (67.7%). Most respondents (83.5%) did not have any previous training in vaccination administration and were not involved in any vaccine-related advocacy activities (78.9%). Principal components analysis of TDF items gave eight components: pharmacists' perceived knowledge and skills (median score of 22, interquartile range [IQR] 17-26, possible range: 7-35); perceived confidence (16; IQR 12-20, possible range: 5-25); perceived external support (9; IQR 7-11, possible range: 3-15); professional role identity (38; IQR 33-42, possible range: 11-55); emotions (10; IQR 9-12, possible range: 3-15); perceived consequences (22; IQR 18-24, possible range: 6-30); perceived usefulness (16; IQR 14-18, possible range: 4-20); and behaviour control (6; IQR 4-8, possible range: 2-10). CONCLUSION: Pharmacists' perceived knowledge, skills, confidence and behavioural control are potentially important factors to address to facilitate participation in vaccination administration in Qatar. Along with providing vaccination training to community pharmacists, there is a need to change the current pharmacy practice structure to improve managerial and government support for and to equip pharmacies with resources for this role.
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Serviços Comunitários de Farmácia , Assistência Farmacêutica , Humanos , Adulto , Farmacêuticos/psicologia , Estudos Transversais , Inquéritos e Questionários , Papel Profissional , Atitude do Pessoal de SaúdeRESUMO
Objective Structured Clinical Examinations (OSCEs) and Work Based Assessments (WBAs) are the mainstays of assessing clinical competency in health professions' education. Underpinned by the extrapolation inference in Kane's Validity Framework, the purpose of this study is to determine whether OSCEs translate to real life performance by comparing students' OSCE performance to their performance in real-life (as a WBA) using the same clinical scenario, and to understand factors that affect students' performance. A sequential explanatory mixed methods approach where a grade comparison between students' performance in their OSCE and WBA was performed. Students were third year pharmacy undergraduates on placement at a community pharmacy in 2022. The WBA was conducted by a simulated patient, unbeknownst to students and indistinguishable from a genuine patient, visiting the pharmacy asking for health advice. The simulated patient was referred to as a 'mystery shopper' and the process to 'mystery shopping' in this manuscript. Community pharmacy is an ideal setting for real-time observation and mystery shopping as staff can be accessed without appointment. The students' provision of care and clinical knowledge was assessed by the mystery shopper using the same clinical checklist the student was assessed from in the OSCE. Students who had the WBA conducted were then invited to participate in semi-structured interviews to discuss their experiences in both settings. Overall, 92 mystery shopper (WBA) visits with students were conducted and 36 follow-up interviews were completed. The median WBA score was 41.7% [IQR 28.3] and significantly lower compared to the OSCE score 80.9% [IQR 19.0] in all participants (p < 0.001). Interviews revealed students knew they did not perform as well in the WBA compared to their OSCE, but reflected that they still need OSCEs to prepare them to manage real-life patients. Many students related their performance to how they perceived their role in OSCEs versus WBAs, and that OSCEs allowed them more autonomy to manage the patient as opposed to an unfamiliar workplace. As suggested by the activity theory, the performance of the student can be driven by their motivation which differed in the two contexts.
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BACKGROUND: A rigorous learning needs assessment (LNA) is a crucial initial step in the Continuing Professional Development (CPD) process. This scoping review aimed to collate, summarize, and categorize the reported LNA approaches adopted to inform healthcare professional CPD and highlight the gaps for further research. METHOD: In August 2020, nine bibliographic databases were searched for studies conducted with any health professional grouping, reporting the utilized LNA to inform CPD activities. Two reviewers independently screened the articles for eligibility and charted the data. A descriptive analytical approach was employed to collate, summarize, and categorize the literature. RESULTS: 151 studies were included in the review; the majority adopted quantitative methods in the form of self-assessment surveys. Mixed-methods approaches were reported in only 35 studies. Descriptions of LNA development lacked detail of measures taken to enhance their rigor or robustness. DISCUSSION: These findings do not reflect recommendations offered by the CPD literature. Further investigations are required to evaluate more recently advocated LNA approaches and add to their limited evidence-base. Similarly, the existing support afforded to CPD developers warrants further study in order to identify the necessary resource, infrastructure and expertise essential to design and deliver effective CPD programs.
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Pessoal de Saúde , Aprendizagem , Humanos , Avaliação das Necessidades , Pessoal de Saúde/educação , Desenvolvimento de Pessoal/métodosRESUMO
INTRODUCTION: Objective structured clinical examinations (OSCEs) are an essential examination tool within undergraduate dental education. Fear of spread of the COVID-19 virus led to dental institutions exploring alternative means of conducting OSCEs. The aim of this scoping review was to investigate what structures, processes and outcomes of dental OSCEs were reported during the COVID-19 pandemic. MATERIALS AND METHODS: This scoping review was conducted and reported adhering to the Preferred Reporting Items for Systematic Reviews and Meta-analyses extension for scoping review guidelines (PRISMA-ScR). Published literature was identified through a systematic search of PubMed, Embase, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Education Resources Information Center (Eric), ProQuest and Google Scholar. Identified articles were independently reviewed by two authors (KS, AD), followed by synthesis in terms of the reported structures, processes and outcomes. Articles reporting cancellation or rescheduling were also included, extracting data on reasons and any suggestions/recommendations. RESULTS: The search yielded a total of 290 studies of which 239 sources were excluded after removal of duplicates, leaving 51 studies for title and abstract evaluation. Thirty-four articles were excluded as they did not report on the topic of interest, leaving 17 for full-text evaluation, of which nine were analysed according to the pre-set themes. All dental OSCEs taking place (n = 6) were conducted online whilst the remaining (n = 3) were either cancelled or rescheduled. Data on structures reported specific online videoconferencing software used and provision of staff and student training. Processes on the execution of online OSCEs varied significantly from one study to the other, providing rich data on how dental institutions may carry out such assessments tailored to their need. Information regarding outcomes was sparse, as little attention was paid to the results of the students compared to pre-pandemic, lacking investigation into reliability and validity of online dental OSCEs. CONCLUSION: Dental OSCEs could be conducted online implementing well-planned structures and processes; however, further evidence is needed to prove its reliability and validity based on outcomes. Dental institutions may need to consider alternative methods to assess practical competencies if online OSCEs are to take place.
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COVID-19 , Humanos , Pandemias , Reprodutibilidade dos Testes , Educação em Odontologia , EstudantesRESUMO
AIMS: This study aimed to review systematically all available prediction tools identifying adult hospitalized patients at risk of drug-related problems, and to synthesize the evidence on performance and clinical usefulness. METHODS: PubMed, Scopus, Web of Science, Embase, and CINAHL databases were searched for relevant studies. Titles, abstracts and full-text studies were sequentially screened for inclusion by two independent reviewers. The Prediction Model Risk of Bias Assessment Tool (PROBAST) and the Revised Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2) checklists were used to assess risk of bias and applicability of prediction tools. A narrative synthesis was performed. RESULTS: A total of 21 studies were included, 14 of which described the development of new prediction tools (four risk assessment tools and ten clinical prediction models) and six studies were validation based and one an impact study. There were variations in tool development processes, outcome measures and included predictors. Overall, tool performance had limitations in reporting and consistency, with the discriminatory ability based on area under the curve receiver operating characteristics (AUROC) ranging from poor to good (0.62-0.81), sensitivity and specificity ranging from 57.0% to 89.9% and 30.2% to 88.0%, respectively. The Medicines Optimisation Assessment tool and Assessment of Risk tool were prediction tools with the lowest risk of bias and low concern for applicability. Studies reporting external validation and impact on patient outcomes were scarce. CONCLUSION: Most prediction tools have limitations in development and validation processes, as well as scarce evidence of clinical usefulness. Future studies should attempt to either refine currently available tools or apply a rigorous process capturing evidence of acceptance, usefulness, performance and outcomes.
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Modelos Estatísticos , Adulto , Viés , Humanos , Prognóstico , Medição de Risco , Sensibilidade e EspecificidadeRESUMO
OBJECTIVES: This systematic review (SR) reviews the evidence on use of theory in developing and evaluating behaviour change interventions (BCIs) to improve clinicians' antimicrobial prescribing (AP). METHODS: The SR protocol was registered with PROSPERO. Eleven databases were searched from inception to October 2018 for peer-reviewed, English-language, primary literature in any healthcare setting and for any medical condition. This included research on changing behavioural intentions (e.g. in simulated scenarios) and research measuring actual AP. All study designs/methodologies were included. Excluded were: grey literature and/or those which did not state a theory. Two reviewers independently extracted and quality assessed the data. The Theory Coding Scheme (TCS) evaluated the extent of the use of theory. RESULTS: Searches found 4227 potentially relevant papers after removal of duplicates. Screening of titles/abstracts led to dual assessment of 38 full-text papers. Ten (five quantitative, three qualitative and two mixed-methods) met the inclusion criteria. Studies were conducted in the UK (n = 8), Canada (n = 1) and Sweden (n = 1), most in primary care settings (n = 9), targeting respiratory tract infections (n = 8), and medical doctors (n = 10). The most common theories used were Theory of Planned Behaviour (n = 7), Social Cognitive Theory (n = 5) and Operant Learning Theory (n = 5). The use of theory to inform the design and choice of intervention varied, with no optimal use as recommended in the TCS. CONCLUSIONS: This SR is the first to investigate theoretically based BCIs around AP. Few studies were identified; most were suboptimal in theory use. There is a need to consider how theory is used and reported and the systematic use of the TCS could help.
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Anti-Infecciosos , Atenção à Saúde , Canadá , SuéciaRESUMO
Edible berries are considered to be among nature's treasure chests as they contain a large number of (poly)phenols with potentially health-promoting properties. However, as berries contain complex (poly)phenol mixtures, it is challenging to associate any interesting pharmacological activity with a single compound. Thus, identification of pharmacologically interesting phenols requires systematic analyses of berry extracts. Here, raspberry (Rubus idaeus, var Prestige) extracts were systematically analyzed to identify bioactive compounds against pathological processes of neurodegenerative diseases. Berry extracts were tested on different Saccharomyces cerevisiae strains expressing disease proteins associated with Alzheimer's, Parkinson's, or Huntington's disease, or amyotrophic lateral sclerosis. After identifying bioactivity against Huntington's disease, the extract was fractionated and the obtained fractions were tested in the yeast model, which revealed that salidroside, a glycosylated phenol, displayed significant bioactivity. Subsequently, a metabolic route to salidroside was reconstructed in S cerevisiae and Corynebacterium glutamicum The best-performing S cerevisiae strain was capable of producing 2.1 mm (640 mg L-1) salidroside from Glc in shake flasks, whereas an engineered C glutamicum strain could efficiently convert the precursor tyrosol to salidroside, accumulating up to 32 mm (9,700 mg L-1) salidroside in bioreactor cultivations (yield: 0.81 mol mol-1). Targeted yeast assays verified that salidroside produced by both organisms has the same positive effects as salidroside of natural origin.
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Glucosídeos/biossíntese , Proteína Huntingtina/química , Doença de Huntington/metabolismo , Extratos Vegetais/química , Rubus/química , Vias Biossintéticas , Fracionamento Químico , Glucosídeos/química , Glucosídeos/metabolismo , Modelos Biológicos , Fenóis/química , Fenóis/metabolismo , Extratos Vegetais/isolamento & purificação , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismoRESUMO
INTRODUCTION: Commercially, blackcurrants (Ribes nigrum L.) are grown mainly for processing, especially for juice production. They are valued for their high levels of polyphenols, especially anthocyanins, which contribute to their characteristic deep colour, but also as a good source of vitamin C. Recently, evidence has accrued that polyphenols, such as anthocyanins, may have specific human health benefits. OBJECTIVE: The aims of this study were to investigate the genetic control of polyphenols and other key juice processing traits in blackcurrants. METHODS: The levels, over 2 years, of vitamin C, citrate, malate, succinate, total organic acids, total anthocyanins and total phenolics together with 46 mainly polyphenol metabolites were measured in a blackcurrant biparental mapping population. Quantitative trait loci (QTLs) for these traits were mapped onto a high-density SNP linkage map. RESULTS: At least one QTL was detected for each trait, with good consistency between the 2 years. Clusters of QTLs were found on each of the eight linkage groups (LG). For example, QTLs for the major anthocyanidin glucosides, delphinidin-3-O-glucoside and cyanidin-3-O-glucoside, co-localised with a QTL for total anthocyanin content on LG3 whereas the major anthocyanidin rutinosides, delphinidin-3-O-rutinoside and cyanidin-3-O-rutinoside, had QTLs on LG1 and LG2. Many of the QTLs explained a high proportion of the trait variation, with the most significant region, on LG3 at ~ 35 cM, explaining more than 60% of the variation in the coumaroylated metabolites, Cyanidin-coumaroyl-glucose, Delphinidin-coumaroyl-glucose, Kaempferol-coumaroyl-glucose and Myricetin-coumaroyl-glucose. CONCLUSION: The identification of robust QTLs for key polyphenol classes and individual polyphenols in blackcurrant provides great potential for marker-assisted breeding for improved levels of key components.
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Polifenóis/genética , Polifenóis/metabolismo , Locos de Características Quantitativas/genética , Ribes/genética , Ribes/metabolismo , Frutas/química , Frutas/genética , Frutas/metabolismo , Extratos Vegetais/química , Extratos Vegetais/metabolismoRESUMO
PURPOSE: There is a lack of knowledge about factors that influence the performance of comprehensive medication reviews (CMRs) by multiprofessional teams in hospital practice. This study aimed to explore the facilitators and barriers for performing CMRs and post-discharge follow-up in older hospitalised patients from the healthcare professional perspective. METHODS: Physicians and ward-based pharmacists were recruited from an ongoing trial at four hospitals in Sweden. Semi-structured interviews were conducted with 16 physicians and 7 pharmacists. Interview topics were working processes, resources, competences, medication-related problems, intervention effects and collaboration. The interviews were audio-recorded, transcribed verbatim and thematically analysed using the Consolidated Framework for Implementation Research (CFIR). Identified subthemes were categorised as facilitators or barriers and grouped into overarching main themes. RESULTS: In total, 21 facilitators and 25 barriers were identified across all CFIR domains and grouped in 6 main themes: (a) CMRs and follow-up are needed, but not in all patients; (b) there is a general belief in positive effects; (c) lack of resources is an issue, although the performance of CMRs may save time; (d) pharmacists' knowledge and skills are valuable, but they need more clinical competence; (e) compatibility with hospital practice is challenging, and roles and responsibilities are unclear and (f) personal contact at the ward is essential for physician-pharmacist collaboration. CONCLUSION: Multiple facilitators and barriers for performing CMRs and post-discharge follow-up in older hospitalised patients exist. These factors should be addressed in future initiatives with similar interventions by multiprofessional teams to ensure successful implementation and performance in hospital practice.
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Assistência ao Convalescente , Assistência Integral à Saúde/organização & administração , Conduta do Tratamento Medicamentoso/organização & administração , Idoso , Seguimentos , Humanos , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Although there is evidence of suboptimal outcomes in older people with chronic pain, little emphasis has been placed on those in remote and rural settings. OBJECTIVE: To describe the perspectives of older people in the Scottish Highlands on their chronic pain management. DESIGN: Cross-sectional survey. SETTING: NHS Highland, the most remote and rural geographical health board in Scotland. SUBJECTS: Home-dwelling members of the public aged ≥70 years. METHODS: Anonymised questionnaires were mailed to a random sample of 1800 older people. Questionnaire items were demographics, nature of any chronic pain, management regimens and perceived effectiveness. Validated scales were the Pain Disability Questionnaire and the Tampa Scale for Kinesiophobia. RESULTS: Adjusted response rate was 39.3% (709/1755). One-quarter (25.0%, n = 177) were experiencing chronic pain, being more likely to live in deprived areas (P < 0.05). Median pain intensity was 6 (IQR 4-7, 10 high), causing distress (median 5, IQR 3-7). Respondents largely consulted GPs (66.1%, n = 117) with a minority (16.4%, n = 29) referred to a specialist pain clinic and few consulting other health professionals. Over three quarters (78.0%, n = 138) were receiving prescribed medicines, most commonly paracetamol, alone (35.6%, n = 63) or in combination with opioids (16.4%, n = 29). One-third (31.6%, n = 56) expressed a desire for more effective medicines; few reported using any non-pharmacological therapies. The median scores for the Pain Disability Questionnaire and Tampa Scale for Kinesiophobia were 74 (IQR 34-104.5, 150 high) and 40 (IQR 35-45, 68 high). CONCLUSIONS: Evidence of provision of appropriate integrated and person-centred chronic pain care is lacking.
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Dor Crônica , Idoso , Dor Crônica/diagnóstico , Dor Crônica/tratamento farmacológico , Dor Crônica/epidemiologia , Estudos Transversais , Humanos , População Rural , Escócia/epidemiologia , Inquéritos e QuestionáriosRESUMO
BACKGROUND: In Scotland, there has been significant investment in pharmacy teams in general medical practices over recent years, aligned to current government policy. OBJECTIVES: To characterize the national pharmacy workforce including activities undertaken, perceived competence and confidence, as well as perception of integration of the intervention. METHODS: A cross-sectional survey of all pharmacists and pharmacy technicians in general practices. Survey items were demographics, activities undertaken and experiences. The NoMAD tool (Improving the Normalization of Complex Interventions) was included as a measure of perspectives of implementation. Post-piloting, a questionnaire link was sent to all pharmacists (n = 471) and pharmacy technicians (n = 112). A total NoMAD score was obtained by assigning 1 (strongly disagree) to 5 (strongly agree) to each item. RESULTS: Responses were received from 393 (83.4%) pharmacists and 101 (91.8%) pharmacy technicians. Three quarters of pharmacists (74.6%) and pharmacy technicians (73.3%) had been qualified for over 10 years. Two-thirds of pharmacists (68.4%) were independent prescribers, with three quarters (72.3%) currently prescribing. Respondents worked in a median of two practices and were providing a range of activities including medication/polypharmacy reviews, medicines reconciliation, prescribing efficiencies and training. Respondents reported high levels of competence and confidence (median 8, scale 0-10 highest). Median NoMAD total score (scale 20-100 highest, Cronbach's alpha 0.89) was 80 for pharmacists and 75 for pharmacy technicians, P ≤ 0.001. CONCLUSIONS: The general practice pharmacy workforce in Scotland is experienced, well-qualified and integrated within general practices, delivering a range of activities. These findings have implications for workforce planning and future education and training.
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Medicina Geral/estatística & dados numéricos , Recursos Humanos/estatística & dados numéricos , Adulto , Estudos Transversais , Feminino , Medicina Geral/organização & administração , Humanos , Masculino , Pessoa de Meia-Idade , Farmacêuticos/estatística & dados numéricos , Técnicos em Farmácia/estatística & dados numéricos , Escócia , Inquéritos e QuestionáriosRESUMO
BACKGROUND: In 2017, the World Health Organization published "Medication Without Harm, WHO Global Patient Safety Challenge," to reduce patient harm caused by unsafe medication use practices. While the five objectives emphasise the need to create a framework for action, engaging key stakeholders and others, most published research has focused on the perspectives of health professionals. The aim was to explore the views and experiences of decision-makers in Qatar on organisational safety culture, medication errors and error reporting. METHOD: Qualitative, semi-structured interviews were conducted with healthcare decision-makers (policy-makers, professional leaders and managers, lead educators and trainers) in Qatar. Participants were recruited via purposive and snowball sampling, continued to the point of data saturation. The interview schedule focused on: error causation and error prevention; engendering a safety culture; and initiatives to encourage error reporting. Interviews were digitally recorded, transcribed and independently analysed by two researchers using the Framework Approach. RESULTS: From the 21 interviews conducted, key themes were the need to: promote trust within the organisation through articulating a fair blame culture; eliminate management, professional and cultural hierarchies; focus on team building, open communication and feedback; promote professional development; and scale-up successful initiatives. There was recognition that the current medication error reporting processes and systems were suboptimal, with suggested enhancements in themes of promoting a fair blame culture and open communication. CONCLUSION: These positive and negative aspects of organisational culture can inform the development of theory-based interventions to promote patient safety. Central to these will be the further development and sustainment of a "fair" blame culture in Qatar and beyond.
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Erros Médicos/prevenção & controle , Erros de Medicação/prevenção & controle , Segurança do Paciente/normas , Gestão da Segurança/normas , Pessoal de Saúde/normas , Humanos , Relações Interprofissionais , Cultura Organizacional , Catar , Qualidade da Assistência à Saúde/normasRESUMO
BACKGROUND: Flavonoids are produced in all flowering plants in a wide range of tissues including in berry fruits. These compounds are of considerable interest for their biological activities, health benefits and potential pharmacological applications. However, transcriptomic and genomic resources for wild and cultivated berry fruit species are often limited, despite their value in underpinning the in-depth study of metabolic pathways, fruit ripening as well as in the identification of genotypes rich in bioactive compounds. RESULTS: To access the genetic diversity of wild and cultivated berry fruit species that accumulate high levels of phenolic compounds in their fleshy berry(-like) fruits, we selected 13 species from Europe, South America and Asia representing eight genera, seven families and seven orders within three clades of the kingdom Plantae. RNA from either ripe fruits (ten species) or three ripening stages (two species) as well as leaf RNA (one species) were used to construct, assemble and analyse de novo transcriptomes. The transcriptome sequences are deposited in the BacHBerryGEN database (http://jicbio.nbi.ac.uk/berries) and were used, as a proof of concept, via its BLAST portal (http://jicbio.nbi.ac.uk/berries/blast.html) to identify candidate genes involved in the biosynthesis of phenylpropanoid compounds. Genes encoding regulatory proteins of the anthocyanin biosynthetic pathway (MYB and basic helix-loop-helix (bHLH) transcription factors and WD40 repeat proteins) were isolated using the transcriptomic resources of wild blackberry (Rubus genevieri) and cultivated red raspberry (Rubus idaeus cv. Prestige) and were shown to activate anthocyanin synthesis in Nicotiana benthamiana. Expression patterns of candidate flavonoid gene transcripts were also studied across three fruit developmental stages via the BacHBerryEXP gene expression browser (http://www.bachberryexp.com) in R. genevieri and R. idaeus cv. Prestige. CONCLUSIONS: We report a transcriptome resource that includes data for a wide range of berry(-like) fruit species that has been developed for gene identification and functional analysis to assist in berry fruit improvement. These resources will enable investigations of metabolic processes in berries beyond the phenylpropanoid biosynthetic pathway analysed in this study. The RNA-seq data will be useful for studies of berry fruit development and to select wild plant species useful for plant breeding purposes.
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Flavonoides/biossíntese , Frutas/genética , Genes de Plantas , Rubus/genética , Transcriptoma , Antocianinas/biossíntese , Vias Biossintéticas/genética , Frutas/crescimento & desenvolvimento , Frutas/metabolismo , Magnoliopsida/classificação , Magnoliopsida/genética , Fenóis/análise , Filogenia , Proteínas de Plantas/genética , RNA-Seq , Rubus/química , Rubus/crescimento & desenvolvimento , Rubus/metabolismoRESUMO
Plant material rich in anthocyanins has been historically used in traditional medicines, but only recently have the specific pharmacological properties of these compounds been the target of extensive studies. In addition to their potential to modulate the development of various diseases, coloured anthocyanins are valuable natural alternatives commonly used to replace synthetic colourants in food industry. Exploitation of microbial hosts as cell factories is an attractive alternative to extraction of anthocyanins and other flavonoids from plant sources or chemical synthesis. In this study, we present the lactic acid bacterium Lactococcus lactis as an ideal host for the production of high-value plant-derived bioactive anthocyanins using green tea as substrate. Besides the anticipated red-purple compounds cyanidin and delphinidin, orange and yellow pyranoanthocyanidins with unexpected methylation patterns were produced from green tea by engineered L. lactis strains. The pyranoanthocyanins are currently attracting significant interest as one of the most important classes of anthocyanin derivatives and are mainly formed during the aging of wine, contributing to both colour and sensory experience.
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Antocianinas , Lactococcus lactis , Engenharia Metabólica , Chá/química , Antocianinas/biossíntese , Antocianinas/genética , Lactococcus lactis/genética , Lactococcus lactis/metabolismoRESUMO
INTRODUCTION: Oats (Avena sativa L.) are a whole grain cereal recognised for their health benefits and which are cultivated largely in temperate regions providing both a source of food for humans and animals, as well as being used in cosmetics and as a potential treatment for a number of diseases. Oats are known as being a cereal source high in dietary fibre (e.g. ß-glucans), as well as being high in antioxidants, minerals and vitamins. Recently, oats have been gaining increased global attention due to their large number of beneficial health effects. Consumption of oats has been proven to lower blood LDL cholesterol levels and blood pressure, thus reducing the risk of heart disease, as well as reducing blood-sugar and insulin levels. OBJECTIVES: Oats are seen as a low input cereal. Current agricultural guidelines on nitrogen application are believed to be suboptimal and only consider the effect of nitrogen on grain yield. It is important to understand the role of both variety and of crop management in determining nutritional quality of oats. In this study the response of yield, grain quality and grain metabolites to increasing nitrogen application to levels greater than current guidelines were investigated. METHODS: Four winter oat varieties (Mascani, Tardis, Balado and Gerald) were grown in a replicated nitrogen response trial consisting of a no added nitrogen control and four added nitrogen treatments between 50 and 200 kg N ha-1 in a randomised split-plot design. Grain yield, milling quality traits, ß-glucan, total protein and oil content were assessed. The de-hulled oats (groats) were also subjected to a rapid Ultra High Performance Liquid Chromatography-Mass Spectrometry (UHPLC-MS) metabolomic screening approach. RESULTS: Application of nitrogen had a significant effect on grain yield but there was no significant difference between the response of the four varieties. Grain quality traits however displayed significant differences both between varieties and nitrogen application level. ß-glucan content significantly increased with nitrogen application. The UHPLC-MS approach has provided a rapid, sub 15 min per sample, metabolite profiling method that is repeatable and appropriate for the screening of large numbers of cereal samples. The method captured a wide range of compounds, inclusive of primary metabolites such as the amino acids, organic acids, vitamins and lipids, as well as a number of key secondary metabolites, including the avenanthramides, caffeic acid, and sinapic acid and its derivatives and was able to identify distinct metabolic phenotypes for the varieties studied. Amino acid metabolism was massively upregulated by nitrogen supplementation as were total protein levels, whilst the levels of organic acids were decreased, likely due to them acting as a carbon skeleton source. Several TCA cycle intermediates were also impacted, potentially indicating increased TCA cycle turn over, thus providing the plant with a source of energy and reductant power to aid elevated nitrogen assimilation. Elevated nitrogen availability was also directed towards the increased production of nitrogen containing phospholipids. A number of both positive and negative impacts on the metabolism of phenolic compounds that have influence upon the health beneficial value of oats and their products were also observed. CONCLUSIONS: Although the developed method has broad applicability as a rapid screening method or a rapid metabolite profiling method and in this study has provided valuable metabolic insights, it still must be considered that much greater confidence in metabolite identification, as well as quantitative precision, will be gained by the application of higher resolution chromatography methods, although at a large expense to sample throughput. Follow up studies will apply higher resolution GC (gas chromatography) and LC (reversed phase and HILIC) approaches, oats will be also analysed from across multiple growth locations and growth seasons, effectively providing a cross validation for the results obtained within this preliminary study. It will also be fascinating to perform more controlled experiments with sampling of green tissues, as well as oat grains, throughout the plants and grains development, to reveal greater insight of carbon and nitrogen metabolism balance, as well as resource partitioning into lipid and secondary metabolism.
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Avena/metabolismo , Produção Agrícola/métodos , Nitrogênio/metabolismo , Agricultura/métodos , Animais , Antioxidantes , Ácidos Cafeicos , Cromatografia Líquida de Alta Pressão/métodos , Ácidos Cumáricos , Grão Comestível/metabolismo , Cromatografia Gasosa-Espectrometria de Massas/métodos , Humanos , Espectrometria de Massas/métodos , Metabolômica/métodos , Fenótipo , Solo/químicaRESUMO
INTRODUCTION: Blackcurrant (Ribes nigrum L.) is an excellent example of a "super fruit" with potential health benefits. Both genotype and cultivation environment are known to affect the chemical composition of blackcurrant, especially ascorbic acid and various phenolic compounds. Environmental conditions, like temperature, solar radiation and precipitation can also have significant impact on fruit chemical composition. The relevance of the study is further accentuated by the predicted and ongoing changes in global climate. OBJECTIVES: The aim of the present study was to provide new knowledge and a deeper understanding of the effects of post flowering environmental conditions, namely temperature and day length, on fruit quality and chemical composition of blackcurrant using an untargeted high performance liquid chromatography-photo diode array-mass spectrometry (HPLC-PDA-MS) metabolomics approach. METHODS: A phytotron experiment with cultivation of single-stemmed potted plants of blackcurrant cv. Narve Viking was conducted using constant temperatures of 12, 18 or 24 °C and three different photoperiods (short day, short day with night interruption, and natural summer daylight conditions). Plants were also grown under ambient outdoor conditions. Ripe berries were analysed using an untargeted HPLC-PDA-MS metabolomics approach to detect the presence and concentration of molecules as affected by controlled climatic factors. RESULTS: The untargeted metabolomics dataset contained a total of 7274 deconvolved retention time-m/z pairs across both electrospray ionisation (ESI) positive and negative polarities, from which 549 metabolites were identified or minimally annotated based upon accurate mass MS. Conventional principal component analysis (PCA) in combination with the Friedman significance test were applied to first identify which metabolites responded to temperature in a linear fashion. Multi-block hierarchical PCA in combination with the Friedman significance test was secondly applied to identify metabolites that were responsive to different day length conditions. Temperature had significant effect on a total of 365 metabolites representing a diverse range of chemical classes. It was observed that ripening of the blackcurrant berries under ambient conditions, compared to controlled conditions, resulted in an increased accumulation of 34 annotated metabolites, mainly anthocyanins and flavonoids. 18 metabolites were found to be regulated differentially under the different daylength conditions. Moreover, based upon the most abundant anthocyanins, a comparison between targeted and untargeted analyses, revealed a close convergence of the two analytical methods. Therefore, the study not just illustrates the value of non-targeted metabolomics approaches with respect to the huge diversity and numbers of significantly changed metabolites detected (and which would be missed by conventional targeted analyses), but also shows the validity of the non-targeted approach with respect to its precision compared to targeted analyses. CONCLUSIONS: Blackcurrant maturation under controlled ambient conditions revealed a number of insightful relationships between environment and chemical composition of the fruit. A prominent reduction of the most abundant anthocyanins under the highest temperature treatments indicated that blackcurrant berries in general may accumulate lower total anthocyanins in years with extreme hot summer conditions. HPLC-PDA-MS metabolomics is an excellent method for broad analysis of chemical composition of berries rich in phenolic compounds. Moreover, the experiment in controlled phytotron conditions provided additional knowledge concerning plant interactions with the environment.
Assuntos
Ribes/crescimento & desenvolvimento , Ribes/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Ritmo Circadiano/fisiologia , Flavonoides/metabolismo , Frutas/química , Frutas/genética , Frutas/metabolismo , Interação Gene-Ambiente , Metabolômica/métodos , Fenóis/metabolismo , Ribes/genética , Espectrometria de Massas por Ionização por Electrospray/métodos , TemperaturaRESUMO
AIMS: A recent systematic review highlighted the lack of robust studies on prescribers' perspectives of direct-acting oral anticoagulants (DOACs) for nonvalvular atrial fibrillation. The aim was to determine prescribers' views and experiences of prescribing DOACs. METHODS: A cross-sectional survey of prescribers in a remote and rural area of Scotland. Survey items were: demographics; prescribing of DOACs; views of potential influences on DOAC prescribing; knowledge of prescribing guidelines; and experiences. Items on potential influences were based on the Theoretical Domains Framework. Data were analysed using descriptive and inferential statistics, and content analysis of responses to open questions. Principal component analysis was performed on the items of potential influences. RESULTS: In total, 154 responses were received, 120 (77.9%) from doctors, 18 (11.7%) from nurse prescribers and 10 (6.4%) from pharmacist prescribers (6 missing). Principal component analysis of the Theoretical Domains Framework items of potential influences gave 4 components. Component scores for (i) role of professionals, their knowledge and skills and (ii) influences on prescribing were positive. Those for (iii) consequences of prescribing and (iv) monitoring for safety and effectiveness were more neutral. There were low levels of agreement for statements relating to DOACs being more effective, safer and cost-effective than warfarin. There were similar responses around the complexity of bleeding management and detection of over and under-anticoagulation. CONCLUSION: This study has identified several key issues of DOAC prescribing (e.g. bleeding management) hence further emphasis is required in continuing professional development and during guideline implementation and evaluation.
Assuntos
Anticoagulantes/administração & dosagem , Fibrilação Atrial/tratamento farmacológico , Inibidores do Fator Xa/administração & dosagem , Padrões de Prática Médica/estatística & dados numéricos , Adulto , Anticoagulantes/efeitos adversos , Estudos Transversais , Inibidores do Fator Xa/efeitos adversos , Feminino , Hemorragia/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Enfermeiras e Enfermeiros/estatística & dados numéricos , Farmacêuticos/estatística & dados numéricos , Médicos/estatística & dados numéricos , Serviços de Saúde Rural , Escócia , Inquéritos e Questionários , Varfarina/administração & dosagemRESUMO
PURPOSE: Epidemiological and intervention studies have attempted to link the health effects of a diet rich in fruits and vegetables with the consumption of polyphenols and their impact in neurodegenerative diseases. Studies have shown that polyphenols can cross the intestinal barrier and reach concentrations in the bloodstream able to exert effects in vivo. However, the effective uptake of polyphenols into the brain is still regarded with some reservations. Here we describe a combination of approaches to examine the putative transport of blackberry-digested polyphenols (BDP) across the blood-brain barrier (BBB) and ultimate evaluation of their neuroprotective effects. METHODS: BDP was obtained by in vitro digestion of blackberry extract and BDP major aglycones (hBDP) were obtained by enzymatic hydrolysis. Chemical characterization and BBB transport of extracts were evaluated by LC-MSn. BBB transport and cytoprotection of both extracts was assessed in HBMEC monolayers. Neuroprotective potential of BDP was assessed in NT2-derived 3D co-cultures of neurons and astrocytes and in primary mouse cerebellar granule cells. BDP-modulated genes were evaluated by microarray analysis. RESULTS: Components from BDP and hBDP were shown to be transported across the BBB. Physiologically relevant concentrations of both extracts were cytoprotective at endothelial level and BDP was neuroprotective in primary neurons and in an advanced 3D cell model. The major canonical pathways involved in the neuroprotective effect of BDP were unveiled, including mTOR signaling and the unfolded protein response pathway. Genes such as ASNS and ATF5 emerged as novel BDP-modulated targets. CONCLUSIONS: BBB transport of BDP and hBDP components reinforces the health benefits of a diet rich in polyphenols in neurodegenerative disorders. Our results suggest some novel pathways and genes that may be involved in the neuroprotective mechanism of the BDP polyphenol components.