Analysis of Polymorphisms in Genes Differentially Expressed in the Enamel of Mice with Different Genetic Susceptibilities to Dental Fluorosis

2018 ◽  
Vol 53 (2) ◽  
pp. 228-233 ◽  
Author(s):  
Senda Charone ◽  
Erika Calvano Küchler ◽  
Aline de Lima Leite ◽  
Mileni Silva Fernandes ◽  
Vinicius Taioqui Pelá ◽  
...  

Genes expressed during amelogenesis are candidates to increase the risk of dental fluorosis (DF). Thus, this study aimed to evaluate the association between polymorphisms in enamel development genes and susceptibility to DF in mice. Mice of both sexes, representing strains 129P3/J (n = 20; resistant to DF) and A/J (n = 20; susceptible to DF), were divided into 2 groups. Each strain received a diet with a low concentration of fluoride (F) and drinking water containing 0 or 50 mg/L of F for 6 weeks. Clinical evaluation and analysis of Vickers enamel microhardness of the incisors were performed. Livers were collected for genomic DNA extraction. Seventeen genetic polymorphisms in Amelx, Ambn, Ambn, Col14a1, Col1a1, Col5a2, Enam, Fam20a, Fam83h, Foxo1, Klk4, Mmp20, Serpinf1, Serpinh1, Smad3, Tuft1, and Wdr72 were genotyped by real-time PCR using Taqman chemistry. Overrepresentation of alleles and genotypes in DF was evaluated using the χ2 test with an alpha of 5%. The clinical aspects of the enamel and the surface enamel microhardness confirmed the DF condition. In the polymorphisms rs29569969, rs13482592, and rs13480057 in Ambn, Col14a1, and Mmp20, respectively, genotype and allele distributions were statistically significantly different between A/J and 129P3/J strains (p < 0.05). In conclusion, polymorphisms in Ambn, Col14a1, and Mmp20 are associated with the susceptibility to DF.

2017 ◽  
Vol 52 (1-2) ◽  
pp. 1-6 ◽  
Author(s):  
Erika Calvano Küchler ◽  
Carolina Dea Bruzamolin ◽  
Marjorie Ayumi Omori ◽  
Marcelo C. Costa ◽  
Leonardo Santos Antunes ◽  
...  

The aim of this study was to evaluate whether genetic polymorphisms in AMELX, AMBN, ENAM, TFIP11, and TUFT1 genes are associated with dental fluorosis (DF). A total of 1,017 children from 2 Brazilian cohorts were evaluated. These populations lived in cities with fluoridation of public water supplies. DF was assessed in erupted permanent teeth using the modified Dean index. The polymorphisms rs946252, rs12640848, rs4694075, rs5997096, and rs4970957 were analyzed by real-time PCR from genomic DNA. Associations between DF, genotype, and allele distribution were evaluated using the χ2 test, with an alpha of 5%. The polymorphisms rs4694075, rs5997096, and rs4970957 in AMBN, TFIP11, and TUFT1 were associated with DF (p < 0.05). In conclusion, enamel matrix genes are associated with DF.


2016 ◽  
Vol 44 (1) ◽  
pp. 97-108 ◽  
Author(s):  
Linda Koshy ◽  
A. L. Anju ◽  
S. Harikrishnan ◽  
V. R. Kutty ◽  
V. T. Jissa ◽  
...  

2015 ◽  
Vol 15 (6) ◽  
pp. 1295-1303
Author(s):  
Gina H. Kimble ◽  
Vincent R. Hill ◽  
James E. Amburgey

USEPA Method 1623 is the standard method in the United States for the detection of Cryptosporidium in water samples, but quantitative real-time polymerase chain reaction (qPCR) is an alternative technique that has been successfully used to detect Cryptosporidium in aqueous matrices. This study examined various modifications to a commercial nucleic acid extraction procedure in order to enhance PCR detection sensitivity for Cryptosporidium. An alternative DNA extraction buffer allowed for qPCR detection at lower seed levels than a commercial extraction kit buffer. In addition, the use of a second spin column cycle produced significantly better detection (P = 0.031), and the volume of Tris–EDTA buffer significantly affected crossing threshold values (P = 0.001). The improved extraction procedure was evaluated using 10 L of tap water samples processed by ultrafiltration, centrifugation and immunomagnetic separation. Mean recovery for the sample processing method was determined to be 41% using microscopy and 49% by real-time PCR (P = 0.013). The results of this study demonstrate that real-time PCR can be an effective alternative for detecting and quantifying Cryptosporidium parvum in drinking water samples.


2012 ◽  
Vol 20 (4) ◽  
pp. 467-471 ◽  
Author(s):  
Erika Calvano Küchler ◽  
Patricia Nivoloni Tannure ◽  
Priscila Falagan-Lotsch ◽  
Taliria Silva Lopes ◽  
Jose Mauro Granjeiro ◽  
...  

2008 ◽  
Vol 57 (3) ◽  
pp. 296-303 ◽  
Author(s):  
L. Metwally ◽  
D. J. Fairley ◽  
P. V. Coyle ◽  
R. J. Hay ◽  
S. Hedderwick ◽  
...  

The limitations of classical diagnostic methods for invasive Candida infections have led to the development of molecular techniques such as real-time PCR to improve diagnosis. However, the detection of low titres of Candida DNA in blood from patients with candidaemia requires the use of extraction methods that efficiently lyse yeast cells and recover small amounts of DNA suitable for amplification. In this study, a Candida-specific real-time PCR assay was used to detect Candida albicans DNA in inoculated whole blood specimens extracted using seven different extraction protocols. The yield and quality of total nucleic acids were estimated using UV absorbance, and specific recovery of C. albicans genomic DNA was estimated quantitatively in comparison with a reference (Qiagen kit/lyticase) method currently in use in our laboratory. The extraction protocols were also compared with respect to sensitivity, cost and time required for completion. The TaqMan PCR assay used to amplify the DNA extracts achieved high levels of specificity, sensitivity and reproducibility. Of the seven extraction protocols evaluated, only the MasterPure yeast DNA extraction reagent kit gave significantly higher total nucleic acid yields than the reference method, although nucleic acid purity was highest using either the reference or YeaStar genomic DNA kit methods. More importantly, the YeaStar method enabled C. albicans DNA to be detected with highest sensitivity over the entire range of copy numbers evaluated, and appears to be an optimal method for extracting Candida DNA from whole blood.


2013 ◽  
Vol 19 (6) ◽  
pp. 1068-1073
Author(s):  
Xiaolan KONG ◽  
Zuozhi CHEN ◽  
Lin LIN ◽  
Chunhou LI ◽  
Peiwen LIANG

2002 ◽  
Vol 301 (1) ◽  
pp. 151-153 ◽  
Author(s):  
Adrián Vilalta ◽  
Vanessa Whitlow ◽  
Terrie Martin

2012 ◽  
Vol 29 ◽  
pp. S220
Author(s):  
Karlygash Aubakirova ◽  
Madina Omasheva ◽  
Natalya Ryabushkina ◽  
Laura Yerbolova ◽  
Tolepbergen Tazhibaev ◽  
...  

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