Physical education teachers' knowledge of physical activity recommendations for health promotion in children and adolescents.

This study aimed to analyse the knowledge of Portuguese physical education (PE) teachers according to the health recommendations for physical activity (PA) for children and adolescents. A total of 764 teachers participated (55.2% men) with a mean age of 48.2 years. Data was collected using answering to an online survey. Teachers were asked about PA's frequency, duration and intensity to achieve the recommended level PA. Chi-square was applied to analyse the associations. The main finding is that PE teachers have a lack of knowledge of the health recommendations of PA. Specifically, only 7.5% of the PE teachers in our study could identify the PA recommendations correctly. The intensity component in the health recommendation is the one in which most PE teachers demonstrate correct knowledge of (60.5%), with significant differences in gender and teaching level. However, significantly fewer PE teachers knew the frequency component in the health recommendations for PA (25%), with significant differences in education level. For the duration component, only 37.6% of teachers knew the recommendations. In light of our findings, it is a concern that PE teachers generally lack knowledge according to children's fulfilment of health recommendations for PA.

Legionella maioricensis sp. nov., a new species isolated from the hot water distribution systems of a hospital and a shopping center during routine sampling.

Two Legionella-like strains isolated from hot water distribution systems in 2012 have been characterized phenotypically, biochemically and genomically in terms of DNA relatedness. Both strains, HCPI-6T and EUR-108, exhibited biochemical phenotypic profiles typical of Legionella species. Cells were Gram-negative motile rods which grew on BCYEα agar but not on blood agar and displayed phenotypic characteristics typical of the family Legionellaceae, including a requirement for l-cysteine and testing catalase positive. Both strains were negative for oxidase, urease, nitrate reduction and hippurate negative, and non-fermentative. The major ubiquinone was Q12 (59.4 % HCPI-6T) and the dominant fatty acids were C16 : 1 ω7c (28.4 % HCPI-6T, ≈16 % EUR-108), C16 : 0 iso (≈22.5 % and ≈13 %) and C15 : 0 anteiso (19.5 % and ≈23.5 %, respectively). The percent G+C content of genomic DNA was determined to be 39.3 mol %. The 16S rRNA gene, mip sequence and comparative genome sequence-based analyses (average nucleotide identity, ANI; digital DNA-DNA hybridization, dDDH; and phylogenomic treeing) demonstrated that the strains represent a new species of the genus Legionella. The analysis based on the 16S rRNA gene sequences showed that the sequence similarities for both strains ranged from 98.8-90.1 % to other members of the genus. The core genome-based phylogenomic tree (protein-concatemer tree based on concatenation of 418 proteins present in single copy) revealed that these two strains clearly form a separate cluster within the genus Legionella. ANI and dDDH values confirmed the distinctiveness of the strains. Based on the genomic, genotypic and phenotypic findings from a polyphasic study, the isolates are considered to represent a single novel species, for which the name Legionella maioricensis sp. nov. is proposed. The type strain is HCPI-6T (=CCUG 75071T=CECT 30569T).

Knockout of Targeted Plasmid-Borne ß-Lactamase Genes in an Extended-Spectrum-ß-Lactamase-Producing Escherichia coli Strain: Impact on Resistance and Proteomic Profile.

Resistance to ß-lactams is known to be multifactorial, although the underlying mechanisms are not well established. The aim of our study was to develop a system for assessing the phenotypic and proteomic responses of bacteria to antibiotic stress as a result of the loss of selected antimicrobial resistance genes. We applied homologous recombination to knock out plasmid-borne ß-lactamase genes (blaOXA-1, blaTEM-1, and blaCTX-M15) in Escherichia coli CCUG 73778, generating knockout clone variants lacking the respective deleted ß-lactamases. Quantitative proteomic analyses were performed on the knockout variants and the wild-type strain, using bottom-up liquid chromatography tandem mass spectrometry (LC-MS/MS), after exposure to different concentrations of cefadroxil. Loss of the blaCTX-M-15 gene had the greatest impact on the resulting protein expression dynamics, while losses of blaOXA-1 and blaTEM-1 affected fewer proteins' expression levels. Proteins involved in antibiotic resistance, cell membrane integrity, stress, and gene expression and unknown function proteins exhibited differential expression. The present study provides a framework for studying protein expression in response to antibiotic exposure and identifying the genomic, proteomic, and phenotypic impacts of resistance gene loss. IMPORTANCE The critical situation regarding antibiotic resistance requires a more in-depth effort for understanding underlying mechanisms involved in antibiotic resistance, beyond just detecting resistance genes. The methodology presented in this work provides a framework for knocking out selected resistance factors, to study the adjustments of the bacterium in response to a particular antibiotic stress, elucidating the genetic response and proteins that are mobilized. The protocol uses MS-based determination of the proteins that are expressed in response to an antibiotic, enabling the selection of strong candidates representing putative resistance factors or mechanisms and providing a basis for future studies to understand their implications in antibiotic resistance. This allows us to better understand how the cell responds to the presence of the antibiotic when a specific gene is lost and, consequently, identify alternative targets for possible future treatment development.

Corynebacterium genitalium sp. nov., nom. rev. and Corynebacterium pseudogenitalium sp. nov., nom. rev., two old species of the genus Corynebacterium described from clinical and environmental samples.

Two Corynebacterium species were proposed decades ago, isolated from clinical samples and divided into biovars: "Corynebacterium genitalium" biovars I-V and "Corynebacterium pseudogenitalium" biovars C1-C6. Several biovars have been re-classified as new species. Nevertheless, biovar I and C5, together with their respective specific epithets "Corynebacterium genitalium" and "Corynebacterium pseudogenitalium", remained not validly published after more than 40 years. Several more strains, temptatively classified as "C. genitalium" biovar I and "Corynebacterium pseudogenitalium" C5, have been isolated from clinical and environmental samples. Both species presented Gram-positive, non-spore forming rod-shaped cells, able to grow aerobically with CO2. Core-genome analysis identified "C. genitalium" to be most closely related to Corynebacterium tuscaniense, Corynebacterium urinipleomorphum, Corynebacterium aquatimens and C appendicis, and Corynebacterium gottingense as the most closely related species to "C. pseudogenitalium". Comprehensive genomic, genotypic, phenotypic analyses, as well as chemotaxonomic, support the proposal for "C. genitalium" and "C. pseudogenitalium" as distinct species within the genus Corynebacterium. The designated type strains of the two species are Furness 392-1T = ATCC 33030T = CCUG 38989T = CCM 9178T = DSM 113155T for C. genitalium sp. nov., nom. rev., and Furness 162-C2T = ATCC 33039T = CCUG 27540T = CCM 9177T = DSM 113154T for C. pseudogenitalium sp. nov., nom. rev.

The Effect of a Sleep Intervention on Sleep Quality in Nursing Students: Study Protocol for a Randomized Controlled Trial.

We develop a protocol for assessing the impact of an intervention aimed at improving sleep quality among university nursing students. The study is designed as a pilot randomized controlled trial to be applied during the 2022-23 academic year and is registered at Clinical Trials Gov website (NCT05273086). A total of 60 nursing students will be recruited from a Spanish university. They will be divided into two groups: (30) intervention group and (30) control group. The intervention group will attend two cognitive-behavioural therapy sleep programme sessions focused on knowledge of anatomical structures involved in sleep, chronotype, synchronization, and good sleeping habits. Subjective and objective sleep quality will be assessed before and after the intervention for both groups. In addition to sleep quality, socio-demographic parameters, physical activity, lifestyle habits, and anthropometric measures will be considered prior to intervention. Finally, a satisfaction questionnaire will be applied for posterior analysis. This study is an innovative, relevant intervention that aims to improve sleep quality among university nursing students. Both the approach and the use of objective and subjective validated outcome measurements are key features of this study.

Sports Injuries of a Portuguese Professional Football Team during Three Consecutive Seasons.

Professional football players are exposed to high injury risk due to the physical demands of this sport. The purpose of this study was to characterize the injuries of a professional football team in the First Portuguese League over three consecutive sports seasons. Seventy-one male professional football players in the First Portuguese Football League were followed throughout the sports seasons of 2019/2020, 2020/2021, and 2021/2022. In total, 84 injuries were recorded. Each player missed an average of 16.6 days per injury. Lower limbs were massively affected by injuries across all three seasons, mainly with muscular injuries in the quadriceps and hamstrings and sprains in the tibiotarsal structure. Overall, the injury incidence was considerably higher in matches than in training. The two times of the season that proved most conducive to injuries were the months of July and January. Our results emphasize the importance of monitoring sports performance, including injury occurrence, and assisting in identifying risk factors in professional football. Designing individualized training programs and optimizing prevention and recovery protocols are crucial for maximizing this global process.

Perceived Neighborhood Safety and Active Transportation in Adults from Eight Latin American Countries.

Neighborhood built environment is associated with domain-specific physical activity. However, few studies with representative samples have examined the association between perceived neighborhood safety indicators and domain-specific active transportation in Latin America. This study aimed to examine the associations of perceived neighborhood safety with domain-specific active transportation in adults from eight Latin American countries. Data were obtained from the Latin American Study of Nutrition and Health (n = 8547, aged 18-65). Active transportation (walking and cycling) was assessed using the long form of the International Physical Activity Questionnaire. Specifically, traffic density and speed as well as street lightening, visibility of residents regarding pedestrians and bicyclists, traffic lights and crosswalks, safety of public spaces during the day and at night, crime rate during the day and at night were used to evaluate perceived neighborhood safety. Slow traffic speeds, unsafe public spaces during the day, and crime during the day were associated with ≥10 min/week vs. <10 min/week of walking. Furthermore, drivers exceeding the speed limit and crime rate during the day were associated with reporting ≥10 min/week vs. <10 min/week of cycling. These results indicate a stronger association of the perceived neighborhood safety with walking compared to cycling.

Identification of Antibiotic Resistance Proteins via MiCId's Augmented Workflow. A Mass Spectrometry-Based Proteomics Approach.

Fast and accurate identifications of pathogenic bacteria along with their associated antibiotic resistance proteins are of paramount importance for patient treatments and public health. To meet this goal from the mass spectrometry aspect, we have augmented the previously published Microorganism Classification and Identification (MiCId) workflow for this capability. To evaluate the performance of this augmented workflow, we have used MS/MS datafiles from samples of 10 antibiotic resistance bacterial strains belonging to three different species: Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. The evaluation shows that MiCId's workflow has a sensitivity value around 85% (with a lower bound at about 72%) and a precision greater than 95% in identifying antibiotic resistance proteins. In addition to having high sensitivity and precision, MiCId's workflow is fast and portable, making it a valuable tool for rapid identifications of bacteria as well as detection of their antibiotic resistance proteins. It performs microorganismal identifications, protein identifications, sample biomass estimates, and antibiotic resistance protein identifications in 6-17 min per MS/MS sample using computing resources that are available in most desktop and laptop computers. We have also demonstrated other use of MiCId's workflow. Using MS/MS data sets from samples of two bacterial clonal isolates, one being antibiotic-sensitive while the other being multidrug-resistant, we applied MiCId's workflow to investigate possible mechanisms of antibiotic resistance in these pathogenic bacteria; the results showed that MiCId's conclusions agree with the published study. The new version of MiCId (v.07.01.2021) is freely available for download at https://www.ncbi.nlm.nih.gov/CBBresearch/Yu/downloads.html.

Mass Spectrometry Proteotyping-Based Detection and Identification of Staphylococcus aureus, Escherichia coli, and Candida albicans in Blood.

Bloodstream infections (BSIs), the presence of microorganisms in blood, are potentially serious conditions that can quickly develop into sepsis and life-threatening situations. When assessing proper treatment, rapid diagnosis is the key; besides clinical judgement performed by attending physicians, supporting microbiological tests typically are performed, often requiring microbial isolation and culturing steps, which increases the time required for confirming positive cases of BSI. The additional waiting time forces physicians to prescribe broad-spectrum antibiotics and empirically based treatments, before determining the precise cause of the disease. Thus, alternative and more rapid cultivation-independent methods are needed to improve clinical diagnostics, supporting prompt and accurate treatment and reducing the development of antibiotic resistance. In this study, a culture-independent workflow for pathogen detection and identification in blood samples was developed, using peptide biomarkers and applying bottom-up proteomics analyses, i.e., so-called "proteotyping". To demonstrate the feasibility of detection of blood infectious pathogens, using proteotyping, Escherichia coli and Staphylococcus aureus were included in the study, as the most prominent bacterial causes of bacteremia and sepsis, as well as Candida albicans, one of the most prominent causes of fungemia. Model systems including spiked negative blood samples, as well as positive blood cultures, without further culturing steps, were investigated. Furthermore, an experiment designed to determine the incubation time needed for correct identification of the infectious pathogens in blood cultures was performed. The results for the spiked negative blood samples showed that proteotyping was 100- to 1,000-fold more sensitive, in comparison with the MALDI-TOF MS-based approach. Furthermore, in the analyses of ten positive blood cultures each of E. coli and S. aureus, both the MALDI-TOF MS-based and proteotyping approaches were successful in the identification of E. coli, although only proteotyping could identify S. aureus correctly in all samples. Compared with the MALDI-TOF MS-based approaches, shotgun proteotyping demonstrated higher sensitivity and accuracy, and required significantly shorter incubation time before detection and identification of the correct pathogen could be accomplished.

Complete Multipartite Genome Sequence of the Cupriavidus basilensis Type Strain, a 2,6-Dichlorophenol-Degrading Bacterium.

We report the complete 8.94-Mb genome sequence of the type strain of Cupriavidus basilensis (DSM 11853T = CCUG 49340T = RK1T), formed by two chromosomes and six putative plasmids, which offers insights into its chloroaromatic-biodegrading capabilities.

Staphylococcus borealis sp. nov., isolated from human skin and blood.

When analysing a large cohort of Staphylococcus haemolyticus, using whole-genome sequencing, five human isolates (four from the skin and one from a blood culture) with aberrant phenotypic and genotypic traits were identified. They were phenotypically similar with yellow colonies, nearly identical 16S rRNA gene sequences and initially speciated as S. haemolyticus based on 16S rRNA gene sequence and MALDI-TOF MS. However, compared to S. haemolyticus, these five strains demonstrate: (i) considerable phylogenetic distance with an average nucleotide identity <95 % and inferred DNA-DNA hybridization <70  %; (ii) a pigmented phenotype; (iii) urease production; and (iv) different fatty acid composition. Based on the phenotypic and genotypic results, we conclude that these strains represent a novel species, for which the name Staphylococcus borealis sp. nov. is proposed. The novel species belong to the genus Staphylococcus and is coagulase- and oxidase-negative and catalase-positive. The type strain, 51-48T, is deposited in the Culture Collection University of Gothenburg (CCUG 73747T) and in the Spanish Type Culture Collection (CECT 30011T).