Genome-Scale Characterization of Mycobacterium abscessus Complex Isolates from Portugal.

The Mycobacterium abscessus complex (MABC) is an emerging, difficult to treat, multidrug-resistant nontuberculous mycobacteria responsible for a wide spectrum of infections and associated with an increasing number of cases worldwide. Dominant circulating clones (DCCs) of MABC have been genetically identified as groups of strains associated with higher prevalence, higher levels of antimicrobial resistance, and worse clinical outcomes. To date, little is known about the genomic characteristics of MABC species circulating in Portugal. Here, we examined the genetic diversity and antimicrobial resistance profiles of 30 MABC strains isolated between 2014 and 2022 in Portugal. The genetic diversity of circulating MABC strains was assessed through a gene-by-gene approach (wgMLST), allowing their subspecies differentiation and the classification of isolates into DCCs. Antimicrobial resistance profiles were defined using phenotypic, molecular, and genomic approaches. The majority of isolates were resistant to at least two antimicrobials, although a poor correlation between phenotype and genotype data was observed. Portuguese genomes were highly diverse, and data suggest the existence of MABC lineages with potential international circulation or cross-border transmission. This study highlights the genetic diversity and antimicrobial resistance profile of circulating MABC isolates in Portugal while representing the first step towards the implementation of a genomic-based surveillance system for MABC at the Portuguese NIH.

Microevolution of a Mycobacteroides abscessus subsp. bolletii strain in a clinical persistent infection.

Mycobacteroides abscessus complex (MAB), a fast-growing nontuberculous mycobacterium, is emerging as a significant infectious disease threat, due to both intrinsic and acquired resistance mechanisms to antibiotics and disinfectants and the need for extensive and multidrug regimens for treatment. Despite the prolonged regimens, outcomes are poor and persistence cases have been reported. Here, we describe clinical, microbiologic and genomic features of a M. abscessus subsp. bolletii (M. bolletii) strain consecutively isolated from a patient within an eight-year infection period. From April 2014 to September 2021, the National Reference Laboratory for Mycobacteria received eight strains isolated from a male patient. Species identification, molecular resistance profile and phenotypic drug susceptibility were determined. Five of these isolates were recovered for further in-depth genomic analysis. Genomic analysis confirmed the multidrug resistant pattern of the strain and also other genetic changes associated with adaptation to environment and defence mechanisms. We highlight the identification of new mutations in locus MAB_1881c and in locus MAB_4099c (mps1 gene), already described as associated with macrolides resistance and morphotype switching, respectively. Additionally, we also observed the emergence and fixation of a mutation in locus MAB_0364c that appeared at a frequency of 36% for the 2014 isolate, 57% for the 2015 isolate and 100% for the 2017 and 2021 isolates, clearly illustrating a fixation process underlying a microevolution of the MAB strain within the patient. Altogether these results suggest that the observed genetic alterations are a reflection of the bacterial population's continuous adaptation and survival to the host environment during infection, contributing to persistence and treatment failure.

Bioinspired Silk Fibroin-Based Composite Grafts as Bone Tunnel Fillers for Anterior Cruciate Ligament Reconstruction.

Anterior cruciate ligament (ACL) replacement is still a big challenge in orthopedics due to the need to develop bioinspired implants that can mimic the complexity of bone-ligament interface. In this study, we propose biomimetic composite tubular grafts (CTGs) made of horseradish peroxidase (HRP)-cross-linked silk fibroin (SF) hydrogels containing ZnSr-doped ß-tricalcium phosphate (ZnSr-ß-TCP) particles, as promising bone tunnel fillers to be used in ACL grafts (ACLGs) implantation. For comparative purposes, plain HRP-cross-linked SF hydrogels (PTGs) were fabricated. Sonication and freeze-drying methodologies capable of inducing crystalline ß-sheet conformation were carried out to produce both the CTGs and PTGs. A homogeneous microstructure was achieved from microporous to nanoporous scales. The mechanical properties were dependent on the inorganic powder's incorporation, with a superior tensile modulus observed on the CTGs (12.05 ± 1.03 MPa) as compared to the PTGs (5.30 ± 0.93 MPa). The CTGs presented adequate swelling properties to fill the space in the bone structure after bone tunnel enlargement and provide a stable degradation profile under low concentration of protease XIV. The in vitro studies revealed that SaOs-2 cells adhered, proliferated and remained viable when cultured into the CTGs. In addition, the bioactive CTGs supported the osteogenic activity of cells in terms of alkaline phosphatase (ALP) production, activity, and relative gene expression of osteogenic-related markers. Therefore, this study is the first evidence that the developed CTGs hold adequate structural, chemical, and biological properties to be used as bone tunnel fillers capable of connecting to the ACL tissue while stimulating bone tissue regeneration for a faster osteointegration.

Seroprevalence of meningococcal serogroup C bactericidal antibodies in the Portuguese population, a decade after vaccine introduction in the National Immunisation Programme.

BACKGROUND: The incidence of invasive meningococcal disease due to serogroup C (MenC) decreased in Portugal since the introduction of the conjugate vaccine (MCC) in the free market in 2001 and in the National Immunisation Plan in 2006. Considering the potential waning of the antibody response reported in the literature, the different vaccination schemes that were used in our country over the past decade, and that Neisseria meningitidis serogroup C continues to circulate, the Portuguese population may currently be at increased risk of infection. In the absence of national data, we evaluated the seroprotection level of the Portuguese population against MenC, in order to identify the protected fraction of the population and ponder on the necessity of a booster dose of the MCC vaccine. METHODS: We measured serum bactericidal antibody levels against MenC in a representative sample of the population (n = 1500) aged 2-64 years who participated in the 2015/2016 National Serological Survey. RESULTS: A total of 31.1% (466/1500, 95%CI: 29-33%) of the individuals studied were protected against MenC. The geometric mean titre was 6.5. The proportion of seroprotected was particularly low in children aged 2-4 years (<16%) who received a single dose of the vaccine at 12 months of age (vaccination strategy since 2012). The proportion of seroprotected was higher (44.7% to 53.5%) in adolescent and young adults (15-24 years of age), resulting from vaccination during the catch-up campaign at 5-15 years of age. The highest protection rates were observed when the vaccine was administered during adolescence. CONCLUSION: The small fraction of population seroprotected, combined with the already known waning effect of the antibody response over time, may indicate that the Portuguese population will become progressively more exposed to the risk of infection. Taking in consideration our results, we recommend to change the current vaccination strategy and introduce a booster dose of the MCC vaccine during adolescence.

Dental trauma with intrusive luxation and six-year monitoring: clinical case report / Traumatismo dentário com luxação intrusiva com acompanhamento de seis anos: relato de caso clínico

Objective: this study is designed to report a clinical intrusion case, which exhibited enamel hypoplasia on the crown of tooth 11 and which underwent clinical and X-ray monitoring for six years. Case report: A threeyear- old female patient exhibited intrusion of deciduous teeth 51, 52, 61, and 62. After X-ray, it was detected that teeth 51 and 61 had perforated the nasal cavity floor, but all four teeth had re-erupted and began to be monitored. After six months, teeth 51 and 61 exhibited bone loss in the interproximal area, as well as mobility, which led to tooth extraction. Teeth 52 and 62 re-erupted in a satisfactory position, and exhibited no signs and symptoms of infection, periapical alteration, or mobility. These teeth were therefore maintained in the arch. After six years, it was detected that tooth 11 had erupted with enamel hypoplasia. Final considerations: Longterm clinical and X-ray control proved essential, as it allowed for the early diagnosis of potential alterations, and minimized potential sequelae to the permanent teeth.(AU)

Objetivo: relatar um caso clínico de intrusão com acompanhamento clínico e radiográfico de 6 anos, que apresentou hipoplasia de esmalte na coroa do dente 11. Relato de caso: paciente de 3 anos, sexo feminino, apresentou intrusão dos dentes decíduos 51, 52, 61 e 62. Após radiografia, foi detectado que os dentes 51 e 61 tinham perfurado o soalho da fossa nasal, porém, ocorreu re-erupção dos 4 dentes e foi realizado acompanhamento. Após 6 meses, nos dentes 51 e 61, foram observadas perda óssea na região interproximal e mobilidade, optando-se pelas exodontias. Os dentes 52 e 62 re-erupcionaram em posição satisfatória e com ausência de sinais e sintomas de infecções, alterações periapicais e mobilidade, sendo mantidos no arco. Após 6 anos, foi observado que o dente 11 erupcionou apresentando hipoplasia de esmalte. Considerações finais: mostrou-se essencial o controle, clínico e radiográfico, em longo prazo, diagnosticando precocemente possíveis alterações e minimizando sequelas que podem acometer os dentes permanentes.(AU)

Avaliação dos Efeitos da Educação em Saúde sobre o Conhecimento e Comportamento de Higiene Bucal de Escolares / Evaluation of the effects of health education on oral hygiene knowledge and behavior of schoolchildren

Objetivo: Avaliar a eficácia de uma atividade de educação em saúde bucal realizada na Escola de Ensino Fundamental Nemésio Bezerra no Município de Quixadá ­ Ceará sobre o conhecimento e o comportamento de higiene bucal de escolares. Material e Métodos: Foi realizado um estudo descritivo e prospectivo com abordagem quantitativa. A amostra na primeira etapa foi de 86 escolares de 12 a 16 anos e de 79 na segunda etapa do estudo. Na primeira etapa do estudo, foi aplicado um questionário estruturado antes da atividade educativa e na segunda etapa, o questionário foi aplicado um mês depois, com a finalidade de avaliar se ocorreu mudanças nos hábitos e conhecimentos sobre a saúde bucal dos alunos. Os dados coletados por meio dos questionários foram submetidos à análise estatística descritiva. Resultados: Antes da atividade educativa, somente 78% (n=67) dos alunos tinham ouvido falar em saúde bucal, enquanto um mês depois, 96% (n=76) dos estudantes responderam já terem ouvido falar. Antes da atividade educativa, somente 17% (n=15) dos estudantes responderam que a hora mais importante de escovar os dentes era antes de dormir. Depois de um mês, aumentou 43% (n=34). Conclusão: Após a atividade proposta, verificou-se um aumento expressivo na quantidade de alunos que melhoram seu conhecimento e seus hábitos de saúde bucal. O estudo mostra o quão importante se configuram as atividades de promoção de saúde bucal que podem ser capazes de proporcionar mudanças consideráveis no perfil de saúde e na qualidade de vida das populações.(AU)

Objective: To evaluate the efficiency of an oral health education activity carried out at Nemésio Bezerra Elementary School in the city of Quixadá - Ceará on oral hygiene knowledge and behavior of school children. Material and Methods: We carried out a descriptive and prospective study with a quantitative approach. The sample in the first stage was 86 students aged 12 to 16 years and 79 in the second stage of the study. In the first stage of the study, a structured questionnaire was applied before the educational activity and in the second stage the questionnaire was applied one month later with the purpose of evaluating if there were changes in the habits and knowledge about the oral health of the students. The data collected through the questionnaires were submitted to descriptive statistical analysis. Results: Before the educational activity, only 78% (n = 67) of the students had heard about oral health, while a month later, 96% (n = 76) of the students answered they had heard. Before the educational activity, only 17% (n = 15) of the students answered that the most important time to brush their teeth was before bed. After one month, it increased 43% (n = 34). Conclusion: After the proposed activity, there was an expressive increase in the number of students who improved their knowledge and their oral health habits. The study shows how important are the activities of oral health promotion that may be able to provide considerable changes in the health profile and quality of life of populations.(AU)

Assessment of Extruded Debris in Primary Molars Comparing Manual and Reciprocating Instrumentation

Objective: To compare the amount of extruded debris in primary molars after manual instrumentation or WaveOne system. Material and Methods: Twenty-five primary molar roots with mild and moderate root angulation (between 10 to 20º), having at least two thirds of root length, no pathological reabsorption (internal or external) or furcation perforation, were selected for this study. Roots were standardized at 6 mm in length, inserted and stabilized in individual acrylic resin blocks with random distribution into two groups: G1 (n=12): Instrumentation with crown-down manual technique; and G2 (n=13): instrumentation with WaveOne system. Blocks with roots were weighed before and after instrumentation, allowing the calculation of extruded debris using both mechanical preparations. Data analysis was performed by Bioestat 4.0 statistical software using Mann Whitney test. Results: The mean weight of extruded debris in manual instrumentation and WaveOne system have no significant difference (p=0.8704). Conclusion: Manual crown-down instrumentation or WaveOne system does not influence the amount of extruded debris after mechanical preparation, suggesting that both techniques were effective in canals clean causing similar extrused debris, however more studies with largest sample should be done to better validate this evidence.