Purpose: To investigate the presence of the variants of lysyl oxygenase (LOX) and superoxide dismutase 1 (SOD1) genes in Brazilian patients with advanced keratoconus. Methods: Donor genomic DNA extracted from blood samples was screened for 5’UTR, exonic LOX, and SOD1 variants in a subset of 26 patients presenting with advanced keratoconus (KISA > 1000% and I–S > 2.0) by Sanger sequencing. The impact of non-synonymous amino acid changes was evaluated by SIFT, PMUT, and PolyPhen algorithms. The Mutation Taster tool was used to evaluate the potential impact of formation of new donor and acceptor splice sites in the promoter region of affected volunteers carrying sequence variants. A 7-base SOD1 deletion (IVS2 + 50del7bp) previously associated with keratoconus was screened in 140 patients presenting classical keratoconus by gel fragment analysis, and positive samples were sequenced for confirmation. Results: We found an unreported missense variant in LOX exon 6 in one heterozygous patient, leading to substitution of proline with threonine at residue 392 (p. Thr392Pro) of LOX protein sequence. This mutation was predicted to be potentially damaging to LOX protein. Another LOX variant, Arg158Gln, was also detected in another patient but predicted to be non-pathogenic. Two additional new polymorphisms in LOX 5’UTR region (–116C > T and –58C > T) were found in two patients presenting with advanced keratoconus and were predicted to modulate or create donor/acceptor splice sites in LOX transcripts. Additionally, SOD1 deletion was detected in one patient presenting with severe keratoconus, not in control samples. Conclusion: We described three novel LOX polymorphisms identified for the first time in Brazilian patients with advanced keratoconus, as well as a previously described SOD1 deletion strongly associated with keratoconus. A possible role of these variants in modulating transcript levels in the cornea of affected individual requires further investigation.

Purpose:To screen visual system homeobox 1 (VSX1) gene in Brazilian subjects affected with keratoconus (KCN).Methods:Seventy-three patients with KCN and 106 healthy controls were enrolled in this study. Patients were diagnosed with KCN based on eye examination and corneal topographic features according to Rabinowitz's criteria (K > 47.2, I-S > 1.4 and KISA > 100%). DNA from blood samples was extracted from donors, and the exons and exon-intron boundaries of VSX1 were sequenced. The potential impact of the identified amino acid changes was assessed with Poly-Phen2, SIFT, and PMUT analysis tools. Genotyping was confirmed by RLFP technique, which was also applied to genotype non-affected individuals.Results:We found three non-synonymous substitutions (L68H, R131S, and D105E) in VSX1 exon 1, with L68H mutation as a novel variation in this gene. In silico analysis indicated that all variations found were predicted to be probably damaging to VSX1 structure and function. Examination of R131S and L68H variations segregating in one family suggested a strong effect of these variations in increasing disease severity in the proband, which presented bilateral KCN leading to corneal grafting before the age of sixteen. We found a novel synonymous substitution (P79P) and two previously described exonic polymorphisms, with unknown roles in VSX1 pathogenesis.Conclusion:VSX1 polymorphisms found in the Brazilian population support a genetic component in KCN pathogenesis. L68H is a novel mutation, and the phenotypic data suggest that this mutation might enhance disease severity when combined with other polymorphisms. However, further investigations are needed.

O trabalho teve por objetivo constatar, mediante o reagente produzido, a eficácia deste em curva de concentração de nitrogênio amoniacal pré-estabelecida e em amostras de água bruta, a fim de avaliar a qualidade de água dos reservatórios quanto à eutrofização e, portanto, o risco oferecido pelo consumo dessas águas. Para isso, foi realizado um ensaio de caráter semiquantitativo, direcionado à avaliação da qualidade de água, através da utilização de um padrão de concentrações por escala colorimétrica de nitrogênio amoniacal como indicador trófico. Com isso obtivemos o perfil de dos mananciais dos açudes de Bodocongó, Manancial Epitácio Pessoa e do centro universitário Unifacisa, referentes às classes de água doce propostas pelo CONAMA. Dessa forma, os resultados obtidos através da curva colorimétrica demonstraram a eficiência do reagente, havendo variação da intensidade da cor gerada a partir das variações de concentração da substância utilizada para construção da curva. Por conseguinte, verificou-se a inadequação das águas dos reservatórios para ingestão humana direta, ou seja, antes da realização de tratamentos adequados. O presente estudo pôde sugerir a situação na qual cada ambiente lêntico em estudo se apresenta na realidade, com exceção do açude do centro universitário, que necessita da realização de mais testes para se traçar o perfil do mesmo.

Resumo -O trabalho teve por objetivo constatar, mediante o reagente produzido, a eficácia deste em curva de concentração de nitrogênio amoniacal pré-estabelecida e em amostras de água bruta, a fim de avaliar a qualidade de água dos reservatórios quanto à eutrofização e, portanto, o risco oferecido pelo consumo dessas águas. Para isso, foi realizado um ensaio de caráter semiquantitativo, direcionado à avaliação da qualidade de água, através da utilização de um padrão de concentrações por escala colorimétrica de nitrogênio amoniacal como indicador trófico. Com isso obtivemos o perfil de dos mananciais dos açudes de Bodocongó, Manancial Epitácio Pessoa e do centro universitário Unifacisa, referentes às classes de água doce propostas pelo CONAMA. Dessa forma, os resultados obtidos através da curva colorimétrica demonstraram a eficiência do reagente, havendo variação da intensidade da cor gerada a partir das variações de concentração da substância utilizada para construção da curva. Por conseguinte, verificou-se a inadequação das águas dos reservatórios para ingestão humana direta, ou seja, antes da realização de tratamentos adequados. O presente estudo pôde sugerir a situação na qual cada ambiente lêntico em estudo se apresenta na realidade, com exceção do açude do centro universitário, que necessita da realização de mais testes para se traçar o perfil do mesmo. Palavras-chave: Nitrogênio amoniacal; Colorimetria; Eutrofização; Manancial Epitácio Pessoa.Abstract -The objective of this study was to verify, through the reagent produced, the efficacy of this in the concentration curve of ammoniacal nitrogen pre-established and in raw water samples, in order to evaluate of the reservoirs for eutrophication and, so, the risk offered by the consumption of these waters. For this, was performed an essay semiquantitative, directing to evaluation of water quality by using a concentration standard of ammoniacal nitrogen's colorimetric scale as trophic indicator. Through this we obtained the source profile of Bodocongó, Epitácio Pessoa and university center Unifacisa, about classes of freshwater proposed by the CONAMA. In this way, the obtained results through the colorimetric curve demonstrated reagent efficiency, there being color intensity varying generated from the concentra-Revista Ambientale Revista da Universidade Estadual de Alagoas/UNEAL