Immunohistochemical and mRNA expression of RANK, RANKL, OPG, TLR2 and MyD88 during apical periodontitis progression in mice

Objective: To evaluate and correlate, in the same research, the mRNA expression and the staining of RANK, RANKL, OPG, TLR2 and MyD88 by immunohistochemistry in the apical periodontitis (AP) progression in mice. Material and Methods: AP was induced in the lower first molars of thirty-five C57BL/6 mice. They were assigned to four groups according to their euthanasia periods (G0, G7, G21 and G42). The jaws were removed and subjected to histotechnical processing, immunohistochemistry and real-time reverse transcription-PCR (qRT-PCR). Data were analyzed with parametric and nonparametric tests (α=0.05). Results: An increase of positive immunoreactivity for RANK, RANKL, OPG, TLR2 and MyD88 was observed over time (p<0.05). The RANKL expression was different between the groups G0 and G42, G21 and G42 (p=0.006), with G42 presenting the higher expression in both comparations. The OPG expression was statistically different between the groups G0 and G7, G7 and G21 and G7 and G42 (p<0.001), with G7 presenting higher expression in all the time points. The TLR2 expression was different between the groups G0 and G42 (p=0.03), with G42 showing the higher expression. The MyD88 expression presented a statistical significant difference between groups G7, G21 and G42 compared with G0 (p=0.01), with G0 presenting the smallest expression in all the comparisons. The Tnfrsf11/Tnfrsf11b (RANKL/OPG) ratio increased with the AP progression (p=0.002). A moderate positive correlation between MyD88 and RANKL (r=0.42; p=0.03) and between MyD88 and TLR2 (r=0.48; p<0.0001) was observed. Conclusion:The expression of the RANK, RANKL, OPG, MyD88 and TLR2 proteins as well as the ratio Tnfrsf11/Tnfrsf11b (RANKL/OPG) increased with AP progression. There was also a moderate positive correlation between the expression Myd88-Tnfrsf11 and Tlr2-Myd88, suggesting the relevance of Tlr2-Myd88 in bone loss due to bacterial infection

Assessing the association between hypoxia during craniofacial development and oral clefts

Objectives: To evaluate the association between hypoxia during embryo development and oral clefts in an animal model, and to evaluate the association between polymorphisms in the HIF-1A gene with oral clefts in human families. Material and Methods: The study with the animal model used zebrafish embryos at 8 hours post-fertilization submitted to 30% and 50% hypoxia for 24 hours. At 5 days post-fertilization, the larvae were fixed. The cartilage structures were stained to evaluate craniofacial phenotypes. The family-based association study included 148 Brazilian nuclear families with oral clefts. The association between the genetic polymorphisms rs2301113 and rs2057482 in HIF-1A with oral clefts was tested. We used real time PCR genotyping approach. ANOVA with Tukey's post-test was used to compare means. The transmission/disequilibrium test was used to analyze the distortion of the inheritance of alleles from parents to their affected offspring. Results: For the hypoxic animal model, the anterior portion of the ethmoid plate presented a gap in the anterior edge, forming a cleft. The hypoxia level was associated with the severity of the phenotype (p<0.0001). For the families, there was no under-transmitted allele among the affected progeny (p>0.05). Conclusion: Hypoxia is involved in the oral cleft etiology, however, polymorphisms in HIF-1A are not associated with oral clefts in humans

Desafios para a integralidade da assistência à pessoa idosa nos serviços da atenção primária à saúde

O acelerado crescimento demográfico resulta em impacto financeiro nas políticas sociais e de saúde, emergindo preocupações acerca dos cuidados dispensados às pessoas idosas. O aumento da longevidade resulta na elevação da ocorrência de doenças e agravos não transmissíveis (DANT) e figuram como principais causas de adoecimento e morte. Com isso, o cuidado voltado às pessoas idosas se configura como um desafio no campo das políticas de saúde vigente, entre elas, a Atenção Primária à Saúde (APS). Objetivo: discorrer sobre desafios enfrentados pelas equipes de Saúde da Família (eSF) para a integralidade da assistência à pessoa idosa no âmbito da APS. Método: trata-se de uma revisão sistemática de literatura, para a qual foram estabelecidos os seguintes métodos: delimitação do objetivo e questão da pesquisa, seleção das bases de consulta, estratégias para a busca do conteúdo, elaboração de estratégias para busca avançada, seleção de estudos e sistematização das informações encontradas. As bases selecionadas para a busca dos estudos foram LILACS e MEDLINE, a partir dos seguintes critérios: artigos, publicados entre 2017 a 2022, nos idiomas inglês e português, com textos disponíveis. Foram identificados 59 estudos que após a leitura dos seus títulos e objetivos excluiu-se 40 deles, restando 19, para compor a amostra dessa revisão. Resultados: falta de qualificação, rotatividade, falta de humanização, falta de priorização do idoso, inobservância, incipiência e fragmentação na oferta dos serviços, baixo desempenho, ausência de ações abrangentes, barreiras na criação de vínculo, falta de autonomia, dificuldades no atendimento integral e longitudinal, insuficiência no quantitativo de ACS, dificuldade de acesso, insatisfação do idoso, busca por outros níveis de atenção, baixa adesão ao tratamento, sexo, idade, renda, distância residência-unidade, baixa escolaridade, memória fraca, desemprego, baixa compreensão do controle medicamentoso, dependência em atividades instrumentais de vida diária, dependência de cuidados de terceiros, baixa qualidade de vida, comorbidades, polifarmácia, queixas cognitivas, sinais de alterações de humor e dificuldades de compreensão foram identificados como desafios enfrentados pelas eSF para a integralidade da assistência ao idoso no âmbito da APS. Conclusão: o investimento em ações educativas e intervenções de apoio que visem estreitar o vínculo e promover o desenvolvimento de competências e habilidades para a promoção do cuidado integral ao idoso são imprescindíveis para diminuir as dificuldades frente à assistência e prática profissional. É imperativo que os profissionais que constituem as eSF da APS repensem criticamente a sua prática em relação à assistência ao idoso, conheçam a realidade a que eles estão inseridos e possuam entendimento da significância do envelhecimento saudável e seu impacto na qualidade de vida dessa população. Para isso, é preciso fortalecer a prática profissional no campo da atenção à saúde do idoso, formulando estratégias que visem à (re)organização do processo de trabalho das eSF para o alcance de ações e metas que promovam uma abordagem sistematizada para o atendimento integral ao idoso na APS

Lifetime prevalence of dental pain and associated factors in two live birth cohorts in Brazil: Ribeirão Preto - SP and São Luís - MA

O objetivo do estudo foi estimar a prevalência de dor de dente devido à cárie dentária e fatores associados em duas coortes de nascidos vivos de diferentes níveis socioeconômicos. Os dados foram coletados em 2004/6, durante o acompanhamento das coortes de Ribeirão Preto - SP (1994) e São Luís - MA (1997). O estudo incluiu todas as crianças de baixo peso e prematuros a partir das coortes originais e 1/3 dos nascidos a termo e com peso normal. As informações foram obtidas por meio de entrevistas com as mães de 673 crianças (7 a 9 anos) em São Luís - MA e 790 (11 a 12 anos) em Ribeirão Preto - SP. A variável desfecho foi considerada o histórico de dor de dente devido a cárie dentária relatado pelas mães. As covariáveis incluíram informações sobre as mães no início do estudo (1994/97) - fumar durante a gravidez, estado civil e escolaridade - e nos seguimentos da coorte (2004/5) poder de compra (ABEP), número de irmãos e hábitos de fumar dos pais. Em relação às crianças as informações incluídas foram: cor da pele, sexo, se frequentou a creche, idade da primeira visita ao dentista, posse de plano de saúde e odontológico e percepção de saúde bucal. Toda a análise estatística levou em consideração o desenho amostral do seguimento. A análise bivariada foi seguida por análise estratificada e multivariável usando modelo de equações generalizadas com distribuição de Poisson e estimadores robustos. Toda a análise foi realizada utilizando os pacotes estatísticos SAS e SUDAAN. Embora as crianças de Ribeirão Preto apresentassem idade superior (11-12 anos) às crianças de São Luís (7-9 anos), a prevalência de dor de dente em São Luís foi maior (56,4%; Intervalo de Confiança - IC95% 52,6 - 60,3%) do que em Ribeirão Preto (32,3%; IC95% 28,8 - 35,9). O modelo final revelou que para Ribeirão Preto os fatores associados à experiência de dor de dente foram as características do nível socioeconômico e das famílias. Para São Luís, os melhores preditores foram as variáveis que caracterizavam o poder de compra, a posse de planos de saúde além da idade da primeira visita ao dentista.The objective of the study was to estimate the lifetime prevalence of dental pain due to dental caries and associated factors in two live births cohorts of different socioeconomic status. Data was collected in 2004/5 during the follow up of the cohorts of Ribeirão Preto - SP (1994) and São Luís - MA (1997). The follow up study included all low birthweight and preterm children from original cohorts and a sample of 1/3 those who had born at term and with normal weight. At total information was collected through interview with children´s mothers from 666 children in São Luís - MA, and 808 in Ribeirão Preto - SP. Outcome variable was defined as a lifetime experience of dental pain due to caries reported by the mothers and covariates included information from mothers during the baseline (smoking during pregnancy, marital status and education) and from the follow-up study (socioeconomic position, number of siblings parents smoking habits). Also information about children included skin color, sex, daycare utilization, age at the first visit to a dentist, general and oral health insurance and oral health perception of mothers). All statistical analysis considered the sampling design of the follow-up. Bivariate analysis was followed by stratified and multivariable analysis using generalized estimation equation model with Poisson distribution and robust estimators. Models were built using stepway like analysis including known covariates. All analysis was performed using SAS and SUDAAN statistical packages. Although children from Ribeirão Preto were older (11/12 yrs old) than children from São Luís (7- 9 years old), the prevalence of dental pain in São Luís was higher 56,4% (Confidence Interval -IC 95% 52,6 - 60,3%) than from Ribeirão Preto 32.3% (IC 95% 28,8 - 35,9). The final model revealed that for Ribeirão Preto factors associated with the experience of toothache were the characteristics of the socioeconomic and families. For St. Louis, the best predictors were the variables that characterized the purchasing power, possession of health insurance beyond the age of first visit to the dentist

Audiovisual distraction methods for anxiety in children during dental treatment: A systematic review and meta-analysis

Background: Dental treatment can cause symptoms such as fear and anxiety. Audiovisual distraction (AD) is a simple and low-cost technique that does not interfere with the dental treatment. Aim: The aim of this study was to perform a systematic review and meta-analysis to evaluate the effectiveness of AD methods for children who experience anxiety during dental treatment. Materials and Methods: Two reviewers performed a database search of the studies published between January 1950 and November 2015. The inclusion criteria were papers published in the English language, child samples aged 4–10 years, and use of AD. All potentially relevant studies were identified by the title and the abstract. After the full-text analysis of the potentially relevant studies, the selected studies were included in the systematic review. A meta-analysis calculation was performed for the overall data and the subgroup data. Results: Thirty-seven nonduplicated studies were found. However, after reviewing the articles, only five were included. A high variability was observed among the papers. Tools and questionnaires used to measure the anxiety during dental treatment presented the most common variability. Meta-analysis demonstrated a lower anxiety level in AD method groups when Modified Child Dental Anxiety Scale was used (P = 0.02) with a mean difference (confidence interval) of −8.72 (−16.7, −1.38). Conclusion: The AD method is effective for controlling dental anxiety in children

Immunohistochemical and mRNA expression of RANK, RANKL, OPG, TLR2 and MyD88 during apical periodontitis progression in mice

Abstract Objective To evaluate and correlate, in the same research, the mRNA expression and the staining of RANK, RANKL, OPG, TLR2 and MyD88 by immunohistochemistry in the apical periodontitis (AP) progression in mice. Material and Methods AP was induced in the lower first molars of thirty-five C57BL/6 mice. They were assigned to four groups according to their euthanasia periods (G0, G7, G21 and G42). The jaws were removed and subjected to histotechnical processing, immunohistochemistry and real-time reverse transcription-PCR (qRT-PCR). Data were analyzed with parametric and nonparametric tests (α=0.05). Results An increase of positive immunoreactivity for RANK, RANKL, OPG, TLR2 and MyD88 was observed over time (p<0.05). The RANKL expression was different between the groups G0 and G42, G21 and G42 (p=0.006), with G42 presenting the higher expression in both comparations. The OPG expression was statistically different between the groups G0 and G7, G7 and G21 and G7 and G42 (p<0.001), with G7 presenting higher expression in all the time points. The TLR2 expression was different between the groups G0 and G42 (p=0.03), with G42 showing the higher expression. The MyD88 expression presented a statistical significant difference between groups G7, G21 and G42 compared with G0 (p=0.01), with G0 presenting the smallest expression in all the comparisons. The Tnfrsf11/Tnfrsf11b (RANKL/OPG) ratio increased with the AP progression (p=0.002). A moderate positive correlation between MyD88 and RANKL (r=0.42; p=0.03) and between MyD88 and TLR2 (r=0.48; p<0.0001) was observed. Conclusion The expression of the RANK, RANKL, OPG, MyD88 and TLR2 proteins as well as the ratio Tnfrsf11/Tnfrsf11b (RANKL/OPG) increased with AP progression. There was also a moderate positive correlation between the expression Myd88-Tnfrsf11 and Tlr2-Myd88, suggesting the relevance of Tlr2-Myd88 in bone loss due to bacterial infection

MMP2 and MMP9 are Associated with Apical Periodontitis Progression and Might be Modulated by TLR2 and MyD88

<div><p>Abstract The aim of this study was to evaluate the expression of MMP2 and MMP9 during apical periodontitis (AP) progression in TLR2 (TLR2 KO) and in MyD88 (MyD88 KO) knockout mice compared to wild type (WT) mice. AP was induced in mandibular first molars of TLR2 KO (n= 18), MyD88 KO (n= 18), and WT mice (n= 18). After 7, 21, and 42 days, the animals were euthanized and the jaws were dissected and subjected to histotechnical processing. Subsequent sections were stained by immunohistochemistry and evaluated for detection of MMP2 and MMP9. Statistical analysis of the semi-quantitative analysis of immunohistochemistry was performed using chi-square test (α = 0.05). In the initial periods of AP progression, an increased expression of MMP9 in the TLR2 KO and MyD88 KO mice was observed. In the final periods of AP progression, a reduction of MMP2 expression and an increase of MMP9 expression in the TLR2 KO mice were observed. MMP2 and MMP9 production was modulated for TLR2 and MyD88 during apical periodontitis progression.</p></div

Assessing the association between hypoxia during craniofacial development and oral clefts

<div><p>Abstract Objectives To evaluate the association between hypoxia during embryo development and oral clefts in an animal model, and to evaluate the association between polymorphisms in the HIF-1A gene with oral clefts in human families. Material and Methods The study with the animal model used zebrafish embryos at 8 hours post-fertilization submitted to 30% and 50% hypoxia for 24 hours. At 5 days post-fertilization, the larvae were fixed. The cartilage structures were stained to evaluate craniofacial phenotypes. The family-based association study included 148 Brazilian nuclear families with oral clefts. The association between the genetic polymorphisms rs2301113 and rs2057482 in HIF-1A with oral clefts was tested. We used real time PCR genotyping approach. ANOVA with Tukey's post-test was used to compare means. The transmission/disequilibrium test was used to analyze the distortion of the inheritance of alleles from parents to their affected offspring. Results For the hypoxic animal model, the anterior portion of the ethmoid plate presented a gap in the anterior edge, forming a cleft. The hypoxia level was associated with the severity of the phenotype (p<0.0001). For the families, there was no under-transmitted allele among the affected progeny (p>0.05). Conclusion Hypoxia is involved in the oral cleft etiology, however, polymorphisms in HIF-1A are not associated with oral clefts in humans.</p></div