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We conducted a multicentre test-negative case-control study covering the period from October 2023 to January 2024 among adult patients aged ≥ 18 years hospitalised with severe acute respiratory infection in Europe. We provide early estimates of the effectiveness of the newly adapted XBB.1.5 COVID-19 vaccines against PCR-confirmed SARS-CoV-2 hospitalisation. Vaccine effectiveness was 49% overall, ranging between 69% at 14-29 days and 40% at 60-105 days post vaccination. The adapted XBB.1.5 COVID-19 vaccines conferred protection against COVID-19 hospitalisation in the first 3.5 months post vaccination, with VE > 70% in older adults (≥ 65 years) up to 1 month post vaccination., (© 2024 The Author(s). Influenza and Other Respiratory Viruses published by John Wiley & Sons Ltd.)

We conducted a multicentre hospital-based test-negative case-control study to measure the effectiveness of adapted bivalent COVID-19 mRNA vaccines against PCR-confirmed SARS-CoV-2 infection during the Omicron XBB lineage-predominant period in patients aged ≥ 60 years with severe acute respiratory infection from five countries in Europe. Bivalent vaccines provided short-term additional protection compared with those vaccinated > 6 months before the campaign: from 80% (95% CI: 50 to 94) for 14-89 days post-vaccination, 15% (95% CI: -12 to 35) at 90-179 days, and lower to no effect thereafter.

Background: Participation rates in health surveys, recognized as an important quality dimension, have been declining over the years, which may affect representativeness and confidence in results. The Portuguese national health examination survey INSEF (2015) achieved a participation rate of 43.9%, which is in line with participation rates from other similar health examination surveys. The objective of this article is to describe how local teams of survey personnel conducted the survey, describing strategies used to solve practical survey problems and to try to increase the participation rate., Methods: After a literature search, informal interviews were conducted with 14 public health officials from local health examination teams, regional and central authorities. Forty-one of the local staff members (survey personnel) also filled in a short questionnaire anonymously. The interviews and self-administered questionnaires were analysed using mixed methods, informed by thematic analysis., Results: The local teams believed that the detailed manual, described as a "cookbook for making a health examination survey", made it possible to maintain high scientific standards while allowing for improvising solutions to problems in the local context. The quality of the manual, supported by a series of training workshops with the central research and support team, gave the teams the confidence and knowledge to implement local solutions. Motivation and cohesion within the local teams were among the goals of the training process. Local teams felt empowered by being given large responsibilities and worked hard to incite people to attend the examination through a close and persuasive approach. Local teams praised their INSA contacts for being available for assistance throughout the survey, and said they were inspired to try harder to reach participants to please their contacts for interpersonal reasons., Conclusions: The theory of organizational improvisation or bricolage, which means using limited resources to solve problems, was useful to discuss and understand what took place during INSEF. A detailed manual covering standard procedures, continuous monitoring of the data collection and face-to-face workshops, including role-play, were vital to assure high scientific standards and high participation rates in this health examination survey. Close contacts between the central team and local focal points in all regions and all survey sites were key to accommodating unexpected challenges and innovative solutions., (© 2022. The Author(s).)

BackgroundNon-pharmaceutical interventions (NPIs) were implemented worldwide to control the spread of SARS-CoV-2.AimTo evaluate the impact of tiered NPIs and a nationwide lockdown on reduction of COVID-19 incidence during the second and third epidemic waves in Portugal.MethodsSurveillance data on laboratory-confirmed COVID-19 cases were used to conduct an interrupted time series analysis to estimate changes in daily incidence during a second wave tiered NPI period (9 November-18 December 2020), and a third wave lockdown period without (15-21 January 2021) and with school closure (22 January-10 February 2021).ResultsSignificant changes in trends were observed for the overall incidence rate; declining trends were observed for tiered NPIs (-1.9% per day; incidence rate ratio (IRR): 0.981; 95% confidence interval (CI): 0.973-0.989) and a lockdown period without (-3.4% per day; IRR: 0.966; 95% CI: 0.935-0.998) and with school closure (-10.3% per day, IRR: 0.897; 95% CI: 0.846-0.951). Absolute effects associated with tiered NPIs and a lockdown on a subsequent 14-day period yielded 137 cases and 437 cases per 100,000 population potentially averted, respectively.ConclusionOur results indicate that tiered NPIs implemented during the second wave caused a decline in COVID-19 incidence, although modest. Moreover, a third wave lockdown without school closure was effective in reducing COVID-19 incidence, but the addition of school closure provided the strongest effect. These findings emphasise the importance of early and assertive decision-making to control the pandemic.

In this work we use simple mathematical models to study the impact of vaccination against COVID-19 in Portugal. First, we fit a SEIR type model without vaccination to the Portuguese data on confirmed cases of COVID-19 by the date of symptom onset, from the beginning of the epidemic until the 23rd January of 2021, to estimate changes in the transmission intensity. Then, by including vaccination in the model we develop different scenarios for the fade-out of the non pharmacological intervention (NPIs) as vaccine coverage increases in the population according to Portuguese vaccination goals. We include a feedback function to mimic the implementation and relaxation of NPIs, according to some disease incidence thresholds defined by the Portuguese health authorities.

As SARS-CoV-2 and its related clinical syndrome (COVID-19) became a pandemic worldwide, questions regarding its clinical presentation, infectivity, and immune response have been the subject of investigation. We present a case of a patient previously considered recovered from nosocomially transmitted asymptomatic COVID-19 illness, who presented with new respiratory, radiological, and RT-PCR findings consistent with COVID-19, while on high-dose prednisolone due to a suspected secondary demyelinating disease. Importantly, it led to three subsequent cases within patient's household after discharge from the hospital. After reviewing this case in light of current evidence and debates surrounding SARS-CoV-2 RT-PCR results, we hypothesize that patients on corticosteroids may have particular viral shedding dynamics and should prompt a more conservative approach in regard to isolation discontinuation and monitoring., Competing Interests: Conflict of InterestThe authors declare that they have no conflict of interest., (© Springer Nature Switzerland AG 2020.)

Background: In Health Examination Surveys interview information is complemented with objective information, providing more accurate indicators than self-reported data. We report the study design, planning and implementation of the first Portuguese Health Examination Survey (INSEF)., Methods: INSEF was a cross-sectional population-based study representative at regional and national level. Individuals aged between 25 and 74 years old, residing in Portugal were selected from the national health users' registry through multi-stage stratified probabilistic sampling. Sample size was set at 4200 individuals. Data was collected in primary care units and included blood pressure, height, weight, hip and waist measurements, blood collection for lipid profile, HbA1c and blood count and a general health questionnaire. European HES procedures were followed., Results: A total of 4911 individuals agreed to participate (43.9% participation rate). Participation rate varied by region, sex and age group, being lower in Lisbon and Tagus Valley (32.8%), for men (41.8%) and for those aged 25-34 years old (36%)., Conclusions: INSEF has set up an experienced national and regional structure for HES implementation. Nationally representative quality epidemiological data is now available for public health monitoring, planning and research., (© The Author(s) 2018. Published by Oxford University Press on behalf of Faculty of Public Health. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Introduction: Cardiovascular disease is an important cause of death and disability worldwide, and hypertension is responsible for at least 45% of all deaths due to heart disease and 51% of deaths due to stroke. This study aimed to estimate and describe the distribution of prevalence, awareness, treatment and control of hypertension in the Portuguese population in 2015., Methods: A national survey using a representative sample of 4911 individuals residing in Portugal and aged between 25 and 74 years was implemented. Trained nurses performed a health interview and a physical examination, including blood pressure measurement (right arm, three measurements at 1-min intervals). The prevalence of hypertension was stratified by gender, age group, marital status, education, occupation and type of residential area. Associations between hypertension prevalence and sociodemographic factors were assessed using bivariate and multivariate Poisson regression., Results: The overall hypertension prevalence was 36.0%. The highest rates were observed in males (39.6%), in individuals aged between 65 and 74 years (71.3%), and in those with low levels of education (62.6%) and with no formal occupation (64.5%). Among hypertensive individuals, 69.8% were aware of their condition and 69.4% were under treatment, of whom 71.3% were controlled. Rates of awareness and medical treatment were significantly higher among women and older individuals., Conclusions: A large majority of the adult Portuguese population are likely to reach blood pressure levels defined as hypertension in adulthood. Significant differences in hypertension prevalence were found according to gender, age and socioeconomic status, which highlights the importance of population strategies in public health policies., (Copyright © 2019 Sociedade Portuguesa de Cardiologia. Publicado por Elsevier España, S.L.U. All rights reserved.)

Oral health is a determinant for quality of life and preventive behaviours such as regular tooth brushing can reduce the risk of a wide spectrum of oral diseases. Adopting preventive behaviours increases the likelihood of being healthy and can be conditioned by demographic and socio-economic factors., Objective: This study aims to describe preventive oral hygiene behaviours in the Portuguese population and assess their association with sociodemographic and socioeconomic factors., Material and Methods: A cross-sectional epidemiologic study was developed using data from the first Portuguese National Health Examination Survey. The target population comprised Portuguese community-dwelling residents aged between 25 and 74 years old. The percentage of individuals who brushed their teeth at least twice a day, provided that once was before sleeping, was considered the indicator showing a preventive behaviour, as this is recommended by the General Directorate of Health in Portugal. Poisson regression was used to identify factors independently associated with this behaviour., Results: Sixty-five per cent of the participants reported tooth brushing as recommended. The prevalence of adoption of this preventive behaviour was higher among those living in urban areas and those who have higher educational level. Results show an association between being male and having low educational level with lesser adoption of preventive oral health behaviours., Conclusion: These findings suggest a need for integrated approaches, from measures tackling social inequalities to actions focused on improving health literacy. It is also important to expand dental healthcare services and improve effective coverage to increase access for rural population.

We recently derived mouse expanded potential stem cells (EPSCs) from individual blastomeres by inhibiting the critical molecular pathways that predispose their differentiation. EPSCs had enriched molecular signatures of blastomeres and possessed developmental potency for all embryonic and extra-embryonic cell lineages. Here, we report the derivation of porcine EPSCs, which express key pluripotency genes, are genetically stable, permit genome editing, differentiate to derivatives of the three germ layers in chimeras and produce primordial germ cell-like cells in vitro. Under similar conditions, human embryonic stem cells and induced pluripotent stem cells can be converted, or somatic cells directly reprogrammed, to EPSCs that display the molecular and functional attributes reminiscent of porcine EPSCs. Importantly, trophoblast stem-cell-like cells can be generated from both human and porcine EPSCs. Our pathway-inhibition paradigm thus opens an avenue for generating mammalian pluripotent stem cells, and EPSCs present a unique cellular platform for translational research in biotechnology and regenerative medicine.

Annual influenza vaccination is recommended to people with chronic conditions. This study aimed to estimate the proportion of chronically ill adults vaccinated against influenza in consecutive seasons and to identify associated factors. We used data from the first National Health Examination Survey (INSEF), a cross-sectional study conducted in 2015 on a probabilistic sample of individuals aged 25-74 years. The population was restricted to individuals who self-reported diabetes, a respiratory, cardiovascular, liver or kidney disease. Self-reported vaccination in 4 consecutive seasons was categorized in 3 levels: unvaccinated, occasionally (vaccinated 1-3 seasons) and repeatedly vaccinated (in all 4 seasons). A multinomial logistic regression was applied to estimate odds-ratio (OR) of influenza vaccination according to sociodemographic factors, chronic condition, health care use and status. In the target population, the 2014/15 influenza vaccine coverage was 33.8% (95% CI: 29.8-38.1). The higher coverage was found in individuals reporting renal disease (66.7%) and diabetes (43.8%). The coverage decreased to 32.6%, 26.0% and 20.8% for individuals with respiratory, cardiovascular and liver diseases, respectively. The probability of being repeatedly vaccinated, compared to unvaccinated, was higher in males (OR = 2.14: 95% CI: 1.31-3.52); aged 65 and 74 (OR = 4.39; 95% CI: 1.99-9.69); whom had an appointment with a general practitioner (OR = 2.77; 95% CI: 1.00-7.66) or other physician (OR = 3.95: 95% CI: 2.53-6.16); with no smoking habits (OR = 1.58; 95% I: 1.02-2.46) and reporting diabetes (OR = 2.13; 95% CI: 1.02-4.45). Finally, having a self-reported cardiovascular condition decreased the likelihood of being occasionally (OR = 0.38; 95% CI = 0.22-0.65) vaccinated against influenza. Younger individuals, females and the ones with a self-reported cardiovascular condition were identified as more likely of non-compliance to the vaccine uptake recommendation. Future vaccination strategies should focus on the previous identified population subgroups. Also, the medical recommendation of the influenza vaccine uptake should continue and be reinforced particularly in individuals with a cardiovascular condition., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Aims: Diabetes Mellitus is a major public health threat worldwide and continues to increase in numbers and significance. Estimates of diabetes prevalence, awareness, treatment and control are essential to effectively monitor its trends, plan and evaluate interventions., Methods: We conducted a nationwide health examination survey in the population residing in Portugal aged between 25 and 74 years old in 2015. It consisted in a cross sectional prevalence study which included the measurement of HbA1c, a physical examination and a general health interview of a probabilistic sample of 4911 individuals (Authorization n°9348/2010 of the National Committee for Data Protection)., Results: The overall prevalence of diabetes was 9.9% (95%CI: 8.4; 11.5). It was higher in males than in females (12.1% vs 7.8%). Diabetes was more prevalent among individuals of lower education and without any professional activity. The majority of persons with diabetes was aware of their condition (87.1%) and was taking antidiabetic medication (79.7%). Of these, 63.2% had glycated hemoglobin levels lower than 7.0% (53 mmol/mol), but the majority failed to comply with the LDL and blood pressure recommended clinical targets (71.9% and 59.0%). Similarly, the prevalence of prediabetes was 16%, higher among women than men (17.5% vs 14.4%)., Conclusion: The prevalence of diabetes and prediabetes remains higher than the global and European estimates, although there is increasing awareness of this disorder., (Copyright © 2018. Published by Elsevier B.V.)

Influenza epidemics are a serious global public health and economic problem. The IFITM3 allele (rs12252-C) was suggested as a population-based genetic risk factor for severe influenza virus infection by A(H1N1)pdm09. We analyzed the population genetics of IFITM3 variants in the Portuguese general population (n = 200) and Central Africans (largely Angolan) (n = 148) as well as its association to influenza severity in Portuguese patients (n = 41). Seven SNPs, within the 352 bp IFITM3 amplicon around rs12252, were identified. SNP distributions in the Portuguese appeared at an intermediate level between the Africans and other Europeans. According to HapMap, rs34481144 belongs to the same linkage disequilibrium (LD) block as rs12252 and is in strong LD with rs6421983. A negative association with severe relative to mild disease was observed for allele rs34481144-A, indicating a protective effect under the dominant model. Moreover, haplotype Hap4 with rs34481144-A, not including rs12252-C, was significantly associated to mild influenza. Conversely, although with borderline significance, haplotype Hap1 with rs34481144-G, not including rs12252-C, was associated to severe disease. Moreover, in comparison to the general Portuguese population, statistical significant differences in the frequencies of the protective allele rs34481144-A in the severe disease group, the deleterious Hap1 in the mild disease group, and the protective Hap4 in the severe disease group were observed. The population attributable risk (PAR) for the targeted rs34481144 allele or genotype was of 55.91 and 64.44% in the general population and the mildly infected individuals, respectively. Implication of these variants in disease phenotype needs further validation, namely through functional analysis as is discussed.

Obesity is recognised as a serious public health issue, due to its associated morbidity and mortality. This study aimed at estimating the prevalence of overweight and obesity in the Portuguese population through direct measurements obtained by the 1st National Health Examination Survey (INSEF 2015) and to identify its associated sociodemographic factors. INSEF was a nationally representative cross-sectional prevalence study conducted on 4911 Portuguese adults aged 25-74 years, in 2015. Height and weight were measured according to the European Health Examination Survey procedures. Poisson regression was used to estimate the adjusted prevalence ratios of overweight and obesity according to age, marital status, occupational activity, education, urbanization of living area and smoking status. Overall prevalences of overweight and obesity were 39.1% and 28.6%, respectively. The prevalence of overweight was higher among men (45.5% vs 33.2%) while the prevalence of obesity was higher among women (32% vs 25%). The sociodemographic factors associated with overweight and obesity were age, marital status and education. Smoking status was associated with overweight and obesity but only in women. INSEF suggests that a high prevalences of overweight and obesity are found in older individuals, married, with lower education levels and non-smoking women. Public health interventions are urgently required for obesity prevention, namely throughout health literacy strategies., (Copyright © 2017 Asia Oceania Association for the Study of Obesity. Published by Elsevier Ltd. All rights reserved.)

Sickle cell anemia (SCA) is characterized by chronic hemolysis, severe vasoocclusive crises (VOCs), and recurrent often severe infections. A cohort of 95 SCA pediatric patients was the background for genotype-to-phenotype association of the patient's infectious disease phenotype and three non-coding polymorphic regions of the TLR2 gene, the -196 to -174 indel, SNP rs4696480, and a (GT)n short tandem repeat. The infectious subphenotypes included (A) recurrent respiratory infections and (B) severe bacterial infection at least once during the patient's follow-up. The absence of the haplotype [Del]-T-[n ≥ 17] (Hap7) in homozygocity protected against subphenotype (B), in a statistically significant association, resisting correction for multiple testing. For the individual loci, the same association tendencies were observed as in the haplotype, including a deleterious association between the SNP rs4696480 T allele and subphenotype (A), whereas the A/A genotype was protective, and a deleterious effect of the A/T genotype with subphenotype (B), as well as including the protective effect of -196 to -174 insert (Ins) and deleterious effect of the deletion (Del) in homozygocity, against subphenotype (B). Moreover, a reduction in the incidence rate of severe bacterial infection was associated to a rise in the hemolytic score, fetal hemoglobin levels (prior to hydroxyurea treatment), and 3.7-kb alpha-thalassemia. Interestingly, differences between the effects of the two latter covariables favoring a reduction in the incidence rate of subphenotype (B) contrast with a resulting increase in relation to subphenotype (A). These results could have practical implications in health care strategies to lower the morbidity and mortality of SCA patients.

Mouse embryonic stem cells derived from the epiblast contribute to the somatic lineages and the germline but are excluded from the extra-embryonic tissues that are derived from the trophectoderm and the primitive endoderm upon reintroduction to the blastocyst. Here we report that cultures of expanded potential stem cells can be established from individual eight-cell blastomeres, and by direct conversion of mouse embryonic stem cells and induced pluripotent stem cells. Remarkably, a single expanded potential stem cell can contribute both to the embryo proper and to the trophectoderm lineages in a chimaera assay. Bona fide trophoblast stem cell lines and extra-embryonic endoderm stem cells can be directly derived from expanded potential stem cells in vitro. Molecular analyses of the epigenome and single-cell transcriptome reveal enrichment for blastomere-specific signature and a dynamic DNA methylome in expanded potential stem cells. The generation of mouse expanded potential stem cells highlights the feasibility of establishing expanded potential stem cells for other mammalian species.

The potential of using a synthetic cardosin-based rennet in cheese manufacturing was recently demonstrated with the development and optimization of production of a recombinant form of cardosin B in Kluyveromyces lactis. With the goal of providing a more detailed characterization of this rennet, we herein evaluate the impact of the plant-specific insert (PSI) on cardosin B secretion in this yeast, and provide a thorough analysis of the specificity requirements as well as the biochemical and structural properties of the isolated recombinant protease. We demonstrate that the PSI domain can be substituted by different linker sequences without substantially affecting protein secretion and milk clotting activity. However, the presence of small portions of the PSI results in dramatic reductions of secretion yields in this heterologous system. Kinetic characterization and specificity profiling results clearly suggest that synthetic cardosin B displays lower catalytic efficiency and is more sequence selective than native cardosin B. Elucidation of the structure of synthetic cardosin B confirms the canonical fold of an aspartic protease with the presence of two high mannose-type, N-linked glycan structures; however, there are some differences in the conformation of the flap region when compared to cardosin A. These subtle variations in catalytic properties and the more stringent substrate specificity of synthetic cardosin B help to explain the observed suitability of this rennet for cheese production.

Introduction: Diabetes is a major public health problem and it is related to socioeconomic factors. The aim of this study is to describe socioeconomic inequalities in the distribution of diabetes in the population with 25 years or more, resident in Portugal in 2014., Material and Methods: Data from the Health National Survey 2014 was analysed, n = 16 786. We estimated the prevalence of diabetes in the population and stratified by socioeconomic variables namely educational level and income. The extent of socioeconomic inequalities was assessed using concentration index and the relative index of inequality., Results: Diabetes was found to be concentrated among the people with lower educational levels (concentration index = -0.26) and lower income quintiles (concentration index = -0.14). Relative index of inequality also showed a lower degree of inequality among the most educated (0,20; CI 95% = [0,12; 0,32]) and with higher income (0,59; CI 95% = [0,48; 0,74])., Discussion: Distribution of diabetes is associated with education and income. Previous studies have shown that although income might reflect lifestyle patterns, education reflects better social factors that are important for establishing healthier behaviours. Also, the National Health Service, of universal coverage and free of charge, might have contributed to reduce inequalities in the access to health by those with the lowest income., Conclusion: Supporting 'Health in All Policies' might reduce inequalities, namely by improving population educational level and actions that promote health literacy.

We present the case of a 63-year-old man, admitted for hand cellulitis and acute kidney injury. A Streptococcus dysgalactiae subsp equisimilis (SDSE) was isolated in blood cultures and despite directed intravenous antibiotherapy, the patient evolved unfavourably, with dorsolumbar spondylodiscitis, multifocal septic arthritis and abscesses. CT also showed densification of the gluteal muscles, multiple air bubbles in the psoas, paraspinal muscles and spinal canal that were associated with an intramuscular injection administered 1 week earlier for a backache. After escalation of the antibiotherapy and intensive supportive measures, the patient showed improvement and was discharged after 8 weeks of antibiotherapy.The incidence of invasive SDSE infections has been growing, especially in immunosupressed patients. In this case, despite no predisposing factor identified, it evolved to severe sepsis. The intramuscular injection, a trivialised but not harmless procedure, was the assumed port of entry, as previously described in another case report., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Ltd (unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.)

CRISPR-Cas9 technology has accelerated biological research becoming routine for many laboratories. It is rapidly replacing conventional gene editing techniques and has high utility for both genome-wide and gene-focussed applications. Here we present the first individually cloned CRISPR-Cas9 genome wide arrayed sgRNA libraries covering 17,166 human and 20,430 mouse genes at a complexity of 34,332 sgRNAs for human and 40,860 sgRNAs for the mouse genome. For flexibility in generating stable cell lines the sgRNAs have been cloned in a lentivirus backbone containing PiggyBac transposase recognition elements together with fluorescent and drug selection markers. Over 95% of tested sgRNA induced specific DNA cleavage as measured by CEL-1 assays. Furthermore, sgRNA targeting GPI anchor protein pathway genes induced loss of function mutations in human and mouse cell lines measured by FLAER labelling. These arrayed libraries offer the prospect for performing screens on individual genes, combinations as well as larger gene sets. They also facilitate rapid deconvolution of signals from genome-wide screens. This set of vectors provide an organized comprehensive gene editing toolbox of considerable scientific value.

It is well known that meteorological conditions influence the comfort and human health. Southern European countries, including Portugal, show the highest mortality rates during winter, but the effects of extreme cold temperatures in Portugal have never been estimated. The objective of this study was the estimation of the effect of extreme cold temperatures on the risk of death in Lisbon and Oporto, aiming the production of scientific evidence for the development of a real-time health warning system. Poisson regression models combined with distributed lag non-linear models were applied to assess the exposure-response relation and lag patterns of the association between minimum temperature and all-causes mortality and between minimum temperature and circulatory and respiratory system diseases mortality from 1992 to 2012, stratified by age, for the period from November to March. The analysis was adjusted for over dispersion and population size, for the confounding effect of influenza epidemics and controlled for long-term trend, seasonality and day of the week. Results showed that the effect of cold temperatures in mortality was not immediate, presenting a 1-2-day delay, reaching maximum increased risk of death after 6-7 days and lasting up to 20-28 days. The overall effect was generally higher and more persistent in Lisbon than in Oporto, particularly for circulatory and respiratory mortality and for the elderly. Exposure to cold temperatures is an important public health problem for a relevant part of the Portuguese population, in particular in Lisbon.

Organisms use circadian clocks to generate 24-h rhythms in gene expression. However, the clock can interact with other pathways to generate shorter period oscillations. It remains unclear how these different frequencies are generated. Here, we examine this problem by studying the coupling of the clock to the alternative sigma factor sigC in the cyanobacterium Synechococcus elongatus Using single-cell microscopy, we find that psbAI, a key photosynthesis gene regulated by both sigC and the clock, is activated with two peaks of gene expression every circadian cycle under constant low light. This two-peak oscillation is dependent on sigC, without which psbAI rhythms revert to one oscillatory peak per day. We also observe two circadian peaks of elongation rate, which are dependent on sigC, suggesting a role for the frequency doubling in modulating growth. We propose that the two-peak rhythm in psbAI expression is generated by an incoherent feedforward loop between the clock, sigC and psbAI Modelling and experiments suggest that this could be a general network motif to allow frequency doubling of outputs., (© 2016 The Authors. Published under the terms of the CC BY 4.0 license.)

Background: Portuguese community pharmacies provide pharmaceutical services, such as therapeutic outcomes follow-up, supplemented by relevant point-of-care testing that require continuity of provision to be effective., Objectives: To identify factors of technical and communication nature that during a patient interview contribute to patients' loyalty., Methods: A cross-sectional descriptive study, with a purposive sample of community pharmacies providing pharmaceutical care, was conducted. Patient interviews were taped and transcribed verbatim. Duration, segments and utterances were identified and time stamped, using a previously validated coding scheme. To identify predictors of loyalty, logistic regression analyses were performed., Results: From 59 interviews, participants' average age was 65.7 years and 42 (71.2%) were female; 45 (76.3%) interviews were classified as outcomes measurements and 14 (23.7%) as pharmaceutical consultations, with 33.2% of the patients booking a following appointment. The significant items to explain loyalty were associated with lifestyle and psychosocial exchange, age of the patient, and the presence of all interview segments (i.e. a complete consultation)., Conclusion: Contrary to common professional beliefs and practice orientation it would appear that pharmacists' technical skills are not the essential factors that promote patients' loyalty needed for continuity of care, at least in the same extent as the social and lifestyle-related content of the exchange. Pharmaceutical care education should focus on relational skills as much as on medication-related competencies., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Background: Nontuberculous mycobacteria (NTM) are a heterogeneous group of microorganisms with distinct clinical relevance. The treatment of NTM infections depends significantly upon the crucial identification of species at this level. The steady increase of mycobacteria species, the use of time-consuming techniques and the lack of standardized identification methods makes the achievement of this goal a demanding challenge. Additionally, inaccurate diagnosis can lead to therapeutic approaches consistent with Mycobacterium tuberculosis infection that are useless. In the present study, the performance of public databases in the accurate identification of NTM by sequence analysis of 16S rRNA and hsp65 genes were evaluated and compared. An algorithm is proposed to achieve an accurate classification of NTM in the geographic region of Portugal (Western Europe)., Methods: Partial sequencing of 16S rRNA and hsp65 genes of 22 reference strains and 54 clinical isolates was performed. The resulting sequences were analysed by public web databases since their performance is evaluated statistically. The phenotypic characteristics of the isolates were also evaluated., Results: The use of commercial kits allowed the accurate identification of 57.4% of the clinical isolates. This result was improved either by the use of 16S rRNA (75.9%) and hsp65 (88.9%) genes analysis alone or combined (96.3%)., Conclusions: Analysis of 16S rRNA gene alone is insufficient for the accurate identification of NTM. A stepwise algorithm combining 16S rRNA and hsp65 gene analysis by multiple public databases is proposed to identify NTM at the species' level., (Copyright © 2014 Asian-African Society for Mycobacteriology. Published by Elsevier Ltd. All rights reserved.)