Resumo A homeostasia entre cálcio e fósforo salivares é importante para a manutenção da saúde bucal. O objetivo deste estudo foi avaliar se polimorfismos genéticos no receptor de estrógeno alfa (ESR1), receptor de estrógeno beta (ESR2) e no microRNA17 (microRNA17) estão associados com os níveis salivares de cálcio e fósforo. Saliva de 276 crianças com 12 anos de idade foi coletada com estímulo mastigatório e os níveis de cálcio e fósforo foram determinados por Espectrofotometria de Massa. O DNA genômico foi extraído da saliva remanescente e os polimorfismos genéticos em ESR1 (rs12154178, rs1884051, rs9340799 e rs2234693), em ESR2 (rs4986938 e rs1256049) e no miRNA17 (rs4284505) foram genotipados pela reação em cadeia da polimerase em tempo real utilizando sondas TaqMan. As diferenças estatísticas nas distribuições alélicas e genotípicas entre “baixo” e “alto” níveis de cálcio e fósforo foram determinadas usando os testes qui-quadrado e teste exato de Fisher. As análises foram ajustadas por sexo (alfa de 5%). O polimorfismo rs9340799 em ESR1 foi o genótipo menos comum associado com altos níveis de cálcio (p=0,03). O alelo menos comum em ESR1 rs1884051 foi associado com baixos níveis de fósforo (p=0,005) e houve um excesso de heterozigotos para miRNA17 rs4284505 entre os indivíduos com baixos níveis de cálcio salivar (p=0,002), ambos ajustados pelo sexo. Este estudo fornece evidências de que polimorfismos genéticos em ESR1 e miRNA17 estão envolvidos na determinação dos níveis de cálcio e fósforo salivares.

Abstract Homeostasis between salivary calcium and phosphorus is important for maintaining oral health. The aim of this study was to evaluate if polymorphisms in ESR1 (Estrogen Receptor Alpha), ESR2 (Estrogen Receptor Beta) and miRNA17 (microRNA17) are associated with calcium and phosphorus levels in saliva. Saliva from 276 12-year-old children were collected by masticatory stimulation and calcium and phosphorus levels were determined by Mass Spectrometry. Genomic DNA was extracted from remaining saliva and genetic polymorphisms in ESR1 (rs12154178, rs1884051, rs9340799 and rs2234693), in ESR2 (rs4986938 and rs1256049) and in miRNA17 (rs4284505) were genotyped using TaqMan chemistry and a real-time PCR equipment. Statistical differences in genotype and allele distributions between ‘low’ and ‘high’ calcium and phosphorus levels were determined using chi-square or Fisher´s exact tests. The analysis was also adjusted by sex (alpha of 5%). ESR1 rs9340799 had the less common genotype associated with higher calcium levels (p=0.03). The less common allele of ESR1 rs1884051 was associated with lower phosphorus levels (p=0.005) and there was an excess of heterozygotes for miRNA17 rs4284505 among individuals with lower calcium levels (p=0.002), both adjusted by sex. This study provides evidence that genetic polymorphisms in ESR1 and miRNA17 are involved in determining salivary calcium and phosphorus levels.

RESUMEN Objetivo. Identificar los principales puntos donde mueren animales para proponer planes de mitigación. Materiales y métodos. Se recorrieron las vías alrededor de una área protegida de Cerrado (São Paulo, Brasil) entre Mayo de 2012 y Agosto de 2013. Se registró el lugar del atropellamiento, datos biométricos y morfológicos de los animales y se colectaron muestras de tejido para la confirmación molecular de la especie. Resultados. Se registraron 31 animales atropellados (muertos), incluyendo especies amenazadas: Leopardus pardalis; Cuniculus paca y Chrysocyon brachyurus. La mayoría de los atropellamientos fueron representados por mamíferos (54.8%) y reptiles (38.7%) y la tasa de mortalidad fue de 1.46 animales/km/año. Fueron detectados tres hotspots de atropellamiento, sugiriendo que son puntos importantes en la probabilidad de cruce de animales, debido a un remanente de vegetación natural y la intercepción de la carretera con bosques de galería. Conclusiones. Este trabajo proporciona fuerte evidencia de los puntos más críticos donde las estrategias de mitigación deben ser implementadas inmediatamente y resalta la importancia de detectar hotspots de atropellamiento, las especies y los grupos taxonómicos más afectados ayudando a elaborar acciones efectivas que pueden mejorar la conservación de la fauna.

ABSTRACT Objective. Here we aimed to identify the main points of animal death by roadkill in the view of helping mitigation plans and reducing the impact over the local fauna of a protected area. Materials and methods. We surveyed the roads around a protected area of Cerrado (São Paulo, Brazil) from May 2012 to August 2013. We recorded the local of roadkills, biometric and morphologic data of the animals, and collected samples of tissue for molecular species confirmation. Results. Thirty-one roadkilled animals were registered, including threatened species: Leopardus pardalis; Cuniculus paca and Chrysocyon brachyurus. Most roadkills were represented by mammals (54.8%) and reptiles (38.7%), and the mortality rate was 1.46 animals/km/year. Three roadkill hotspots were detected, suggesting that they were important points of animal crossing, probably because of the existence of natural remnant vegetation and intersection of roads by riparian vegetation. Conclusions. This work provided strong evidence of the most critical points where mitigation strategies should be immediately implemented and highlighted the importance of detecting roadkill hotspots and the species or taxonomic groups more affected, helping to elaborate effective actions that can improve fauna conservation.