Abstract Objectives To analyze the efficiency of a multigenic targeted massively parallel sequencing panel related to endocrine disorders for molecular diagnosis of patients assisted in a tertiary hospital involved in the training of medical faculty. Material and methods Retrospective analysis of the clinical diagnosis and genotype obtained from 272 patients in the Endocrine unit of a tertiary hospital was performed using a custom panel designed with 653 genes, most of them already associated with the phenotype (OMIM) and some candidate genes that englobes developmental, metabolic and adrenal diseases. The enriched DNA libraries were sequenced in NextSeq 500. Variants found were then classified according to ACMG/AMP criteria, with Varsome and InterVar. Results Three runs were performed; the mean coverage depth of the targeted regions in panel sequencing data was 249×, with at least 96.3% of the sequenced bases being covered more than 20-fold. The authors identified 66 LP/P variants (24%) and 27 VUS (10%). Considering the solved cases, 49 have developmental diseases, 12 have metabolic and 5 have adrenal diseases. Conclusion The application of a multigenic panel aids the training of medical faculty in an academic hospital by showing the picture of the molecular pathways behind each disorder. This may be particularly helpful in developmental disease cases. A precise genetic etiology provides an improvement in understanding the disease, guides decisions about prevention or treatment, and allows genetic counseling.

Resumo Entre outros fatores, os tipos de bisfosfonatos bem como os regimes de tratamento parecem estar diretamente associados com o sucesso ou falhas na instalação de implantes osseointegrados. Este estudo avaliou a influência de dois bisfosfonatos, o alendronato de sódio (AS) e o ácido zoledrônico (AZ), no metabolismo de osteoblastos. Osteoblastos humanos (Saos-2) foram cultivados sobre discos de titânio polidos ou submetidos a tratamento ácido superficial, previamente alocados em placas de 24 compartimentos, utilizando meio de cultura completo. Após 24 horas, as células foram expostas aos bisfosfonatos, nas concentrações de 0,5, 1 ou 5 µM, por 24 h, 48 h, ou 7 dias. Os efeitos do AZ e AZ sobre os osteoblastos foram determinados considerando a adesão destas células às superfícies de titânio, por meio de fluorescência direta, a viabilidade celular, produção de proteína total e síntese de colágeno. A atividade de fosfatase alcalina e a deposição de nódulos mineralizados também foram avaliadas. Os dados foram analisados por meio do teste ANOVA complementado por Tukey (α = 0.05). Menor adesão dos osteoblastos foi observada quando estas células foram expostas a ambos os bisfosfonatos, porém, esta falha na adesão foi mais evidente para as células tratadas com AZ. Além disso, a exposição dos osteoblastos ao AZ também resultou em diminuição da viabilidade, atividade de ALP e deposição de nódulos mineralizados, o que pode estar relacionado a uma pobre osseointegração após a instalação do implante.

Abstract Among other factors, types of bisphosphonates and treatment regimens seem to be strongly associated with the success or failure of installation of osseointegrated implants. This study investigated the influence of two bisphosphonates, sodium alendronate (SA) and zoledronic acid (ZA), on the metabolism of osteoblasts. Human osteoblasts (Saos-2) were seeded onto machined or acid-treated titanium discs previously placed on 24-well plates in complete culture medium. After 24 h, cells were exposed to bisphosphonates at 0.5, 1 or 5 µM for 24 h, 48 h or 7 days. The effects of SA and ZA on osteoblasts were assessed based on the adhesion of these cells to the titanium surfaces by direct fluorescence, cell viability, total protein and collagen synthesis. Alkaline phosphatase activity and mineral nodule deposition by these cells were also evaluated. Data were evaluated by ANOVA and Tukey tests (α=0.05). Decreased adhesion of cells to the titanium discs was observed when exposed to both bisphosphonates; however, this lack of cell adhesion was more evident for ZA-treated cells. In addition, the exposure of osteoblasts to ZA decreased the viability, ALP activity and mineral nodule deposition, which may be related to poor osseointegration after implant installation.

O objetivo deste estudo foi avaliar os níveis de estresse transitório e o estado de ansiedade em crianças submetidas à anestesia dental convencional e computadorizada. Vinte crianças (7 a 12 anos de idade) foram randomicamente designadas para receber anestesia dental convencional e computadorizada. Para investigar a hipótese que o estresse transitório poderia ser menor após a anestesia computadorizada, comparada à anestesia convencional, os níveis de cortisol na saliva foram medidos antes e depois de cada técnica. A ansiedade também foi avaliada individualmente por meio do State-Trait Anxiety Inventory for Children (STAIC). Os dados numéricos foram analisados estatisticamente pelo teste não paramétrico de Mann-Whitney (nível de significância de 5%). Os níveis salivares de cortisol aumentaram em 8 (40%) pacientes após anestesia convencional e em 9 (45%) pacientes após anestesia computadorizada, sem diferença estatisticamente significante entre os dois tipos (p=0,34). Da mesma maneira, não foi encontrada diferença estatisticamente significante entre as técnicas (p=0,39) com relação à análise psicológica baseada nos escores STAIC. Anestesia local usando tanto a anestesia convencional quanto o sistema de aplicação computadorizado produziu nível similar de estresse/ansiedade em pacientes infantis, utilizando análises quantitativa e qualitativa.

The aim of this study was to evaluate the transitory stress levels and the anxiety state in children submitted to conventional and computerized dental anesthesia. Twenty children (7 to 12 years) were randomly assigned to receive conventional and computerized dental anesthesia. To investigate the hypothesis that transitory stress could be lower after using computerized anesthesia compared to conventional anesthesia, cortisol levels in saliva were measured before and after each technique. Anxiety was also evaluated individually by answering the State-Trait Anxiety Inventory for Children (STAIC). Numerical data were analyzed statistically by the Mann-Whitney non-parametric test (5% significance level). Salivary cortisol levels increased in 8 (40%) patients after conventional anesthesia and in 9 (45%) patients after computerized anesthesia, with no statistically significant difference between the two types (p=0.34). In the same way, no statistically significant difference was found between the techniques (p=0.39) related to the psychological analysis based on the STAIC scores. Local anesthesia using either conventional anesthesia or a computerized delivery system produced similar level of stress/anxiety in pediatric patients, using both quantitative and qualitative analyses.