Análise da saliva total de portadores de fibrose cística : composição inorgânica e a abordagem proteômica

Tese (doutorado)—Universidade de Brasília, Faculdade de Ciências da Saúde, 2008.A Fibrose Cística (FC) é uma doença hereditária autossômica, recessiva e letal que apresenta certas dificuldades na realização do diagnóstico precoce e preciso. Devido ao comprometimento patológico essa doença assume grande importância e o diagnóstico precoce é decisivo para um melhor prognóstico. Este estudo teve por objetivo determinar a composição inorgânica e análise proteômica da saliva total de indivíduos com FC e identificar biomarcadores para uso no diagnóstico da doença. Foram coletadas amostras de saliva de habitantes da região Centro Oeste do Brasil: 12 indivíduos homozigotos para a mutação ?•F 508 (4 mulheres, 8 homens), 12 heterozigoto para ?ýF 508 (5 mulheres, 7 homens), 13 indivíduos com FC com mutações não identificadas (6 mulheres, 7 homens), 8 indivíduos com diagnóstico duvidosos para FC ( 5 mulheres, 3 homens) e 43 indivíduos saudáveis (23 mulheres e 20 homens). Indivíduos com dúvida no diagnóstico para FC ou que não cumpriram os requisitos para compor o grupo controle foram excluídos das análises. Os teores dos elementos traço Al, As, B, Ba, Ca, Cd, Co, Cr, Cu, Fe, Hg, K, Mg, Mn, Mo, Na, Ni, Pb, Se, Si, Sr, Ti, V e Zn na saliva foram avaliados por espectrometria de massa com plasma de argônio acoplado indutivamente (ICP-MS), espectrometria de emissão óptica com plasma de argônio acoplado indutivamente (ICP-OES) e espectrometria de absorção atômica (AAS). O perfil protéico foi estudado por eletroforese em gel de poliacrilamida na presença de Sodium Dodecyl Sulfate (SDS-PAGE), por eletroforese bidimensional (2-DE) e análise de bases peptídicas por espectrometria de massa (MS/MS). Os elementos Na e K apresentaram, em média, teores mais elevados, enquanto, V, Cr e Se menos elevado quando comparados a indivíduos saudáveis, o elemento As foi detectado apenas no grupo de indivíduos saudáveis (p<0,05). Nas análises por eletroforese, o grupo com FC apresentou uma banda nítida de massa molecular de 38 kDa, não visualizada nos indivíduos sem a doença. A espectrometria de massa identificou a uma imunoglobulina (IgA), a qual esta visivelmente expressa nos indivíduos com FC. Existe diferença significante entre a saliva de indivíduos com FC e às de indivíduos saudáveis. Foi também identificado diferenças no perfil protéico. Com os resultados deste trabalho podemos inferir que a saliva total estimulada é uma fonte potencial para estudos que possam corroborar no desenvolvimento de um método de diagnóstico precoce e preciso da FC. ______________________________________________________________________________________ ABSTRACTCystic Fibrosis (CF) is an autossomic recessive lethal inherited disease that presents certain difficulties for an early and accurate diagnosis. Due to its pathological character this disease assumes great importance and early diagnosis is of paramount importance for a good prognostic. The objective of this study was to determine the inorganic composition and the proteomic analysis of stimulated whole saliva obtained from CF and identify biomarkers for CF diagnosis. Samples were collected from individuals living in the Brazilian West Central Region: 12 homozygous for mutation ?•F 508 (4 females; 8 males); 12 heterogeneous for mutation ?•F 508 (4 females; 8 males); 13 with unidentified CF mutations (6 females; 7 males); 8 with doubtful CF diagnosis (5 females; 3 males); and 43 healthy individuals (23 females; 20 males) for the control group. Samples from individuals with doubtful diagnosis as well as from those that did not fulfill the basic requirements to compose the control group were discarded. Concentration of trace elements Al, As, B, Ba, Ca, Cd, Co, Cr, Cu, Fe, Hg, K, Mg, Mn, Mo, Na, Ni, Pb, Se, Si, Sr, Ti, V e Zn were evaluated using mass spectrometer with inductively coupled argonium plasma (ICP-MS), optical emission spectrometer with inductively coupled argonium plasma (ICP-OES), and atomic absorption spectrometer (AAS). Protein profile was accessed by polyacrylamide gel electrophoresis 15% in presence of Sodium Dodecyl Sulfate (SDS-PAGE), by bidimensional electrophoresis (2D), and analysis of the peptide bases by mass spectrometry (MS) In every analyses, results obtained for CF individuals were compared to those for healthy subjects. Average concentration values for Na and K were higher in CF patients as compared to healthy individuals. In contrast, V, Cr, and Se, were lower. Element As was detected only in the healthy group (p<0,05). In the SDS-PAGE analysis CF patients presented a distinct band with molecular mass between 30 and 45 kDa not visualized in the healthy subjects. By 2D electrophoresis associated with mass spectrometry it was possible to identify a heavy chain immunoglobulin (C region) of Iga¥(IgA) clearly visible in CF patients. There were significant differences between saliva from healthy and CF individuals as well as in their protein profile. Results obtained in the present study indicate that stimulated whole saliva is a potential source for studies that may corroborate the development of an early and accurate screening method

Neighborhood contexts and oral health outcomes in a pediatric population: An exploratory study

Aims: This study aimed to explore the impacts of neighborhood-level socioeconomic contexts on the therapeutic and preventative dental quality outcome of children under 16 years. Materials and Methods: Anonymized billing data of 842 patients reporting to a university children’s dental over three years (March 2017–2020) met the inclusion criteria. Their access to care (OEVCH-A), topical fluoride application (TFL-CH-A) and dental treatment burden (TRT-CH-A) were determined by dental quality alliance (DQA) criteria. The three oral health variables were aggregated at the neighborhood level and analyzed with Canadian census data. Their partial postal code (FSA) was chosen as a neighborhood spatial unit and maps were created to visualize neighborhood-level differences. Results: The individual-level regression models showed significant negative associations between OEV-CH-A (p = 0.027) and TFL-CH-A (p = 0.001) and the cost of dental care. While there was no significant association between neighborhood-level sociodemographic variables and OEV-CH-A, TRT-CH-A showed a significant negative association at the neighborhood level with median household income and significant positive association with percentage of non-official first language (English or French) speakers. Conclusion: Initial analysis suggests differences exist in dental outcomes according to neighborhood-level sociodemographic variables, even when access to dental care is similar

Intranasal sedation using ketamine and midazolam for pediatric dental treatment (NASO): study protocol for a randomized controlled trial

Abstract Background Uncooperative children may need to receive dental treatment under sedation, which is indicated when nonpharmacological behavior guidance is unsuccessful. There are randomized controlled trials (RCTs) comparing different sedative protocols for dental procedures; however, the evidence for superiority of one form over another is weak. The primary aim of this study is to investigate the efficacy of intranasally administered ketamine plus midazolam for the dental treatment of children. Methods We have designed a three-armed, parallel RCT to assess intranasal sedation using ketamine/midazolam in terms of the following measures: efficacy, safety, and cost-effectiveness. Two- to 6-year-old healthy children, referred for dental treatment in a dental sedation center in Brazil due to uncooperative behavior and requiring restorative dental procedures, will be recruited. Each child will be randomly assigned to one of the three groups: A – Intranasal administration of ketamine (4.0 mg/kg, maximum 100 mg) and midazolam (0.2 mg/kg, maximum 5.0 mg); B – Oral administration of ketamine (4.0 mg/kg, maximum 100 mg) and midazolam (0.5 mg/kg, maximum 20 mg); and C – Oral administration of midazolam (1.0 mg/kg, maximum 20 mg). The primary outcome is the child’s behavior assessed through an observational scale using digital videos of the restorative dental treatment under sedation. The secondary outcomes are as follows: acceptance of sedative administration; memory of intraoperative events; the child’s stress; adverse events; the child’s pain during the procedure; the parent’s, dentists’, and child’s perceptions of sedation; and economic analysis. Measures will be taken at baseline and drug administration and during and after the dental procedure. The necessary sample size was estimated to be 84 children after a blinded interim analysis of the first 30 cases. Discussion This study will provide data that can substantially add to science and pediatric dentistry as it examines the effect of sedative regimes from different perspectives (outcomes). Trial registration ClinicalTrials.gov, identifier: NCT02447289 . Registered on 11 May 2015, named “Midazolam and Ketamine Effect Administered Through the Nose for Sedation of Children for Dental Treatment (NASO).