X chromosome inactivation does not necessarily determine the severity of the phenotype in Rett syndrome patients

Rett syndrome (RTT) is a severe neurological disorder usually caused by mutations in the MECP2 gene. Since the MECP2 gene is located on the X chromosome, X chromosome inactivation (XCI) could play a role in the wide range of phenotypic variation of RTT patients; however, classical methylation-based protocols to evaluate XCI could not determine whether the preferentially inactivated X chromosome carried the mutant or the wild-type allele. Therefore, we developed an allele-specific methylation-based assay to evaluate methylation at the loci of several recurrent MECP2 mutations. We analyzed the XCI patterns in the blood of 174 RTT patients, but we did not find a clear correlation between XCI and the clinical presentation. We also compared XCI in blood and brain cortex samples of two patients and found differences between XCI patterns in these tissues. However, RTT mainly being a neurological disease complicates the establishment of a correlation between the XCI in blood and the clinical presentation of the patients. Furthermore, we analyzed MECP2 transcript levels and found differences from the expected levels according to XCI. Many factors other than XCI could affect the RTT phenotype, which in combination could influence the clinical presentation of RTT patients to a greater extent than slight variations in the XCI pattern

Calibração de extratores providos de cápsula porosa para monitoramento da salinidade e da concentração de íons Calibration of porous ceramic cup extractors in monitoring soil salinity and ion concentration

Os resultados satisfatórios obtidos no monitoramento da salinidade e concentração de íons na solução do solo, usando extratores de cápsulas porosas, motivaram a realização deste trabalho, o qual teve como objetivo a obtenção de curvas de calibração para estes extratores monitorarem a condutividade elétrica e a concentração de nitrato e de potássio na solução de dois tipos de solo (Latossolo Vermelho-Amarelo Argissólico franco arenoso, e o outro como Cambissolo Eutrófico), em Mossoró - RN. As concentrações de nitrato utilizadas foram as mesmas nos dois solos (0; 84; 168; 252; 336; 420 e 504 mg L-1); quanto ao potássio, foram utilizadas concentrações diferentes no solo arenoso (0; 117; 234; 351; 468; 585 e 702 mg L-1) e no argiloso (0; 117; 234; 351; 468; 585; 702; 1.170; 1.755; 2.340; 2.925 e 3.510 mg L-1). A condutividade elétrica da solução do solo e a concentração de nitrato e de potássio foram estimadas com precisão satisfatória a partir da solução coletada com extratores providos de cápsulas porosas, sendo necessária uma calibração prévia para cada tipo de solo.<br>The satisfactory results obtained in monitoring soil salinity and ion concentration, by means of porous ceramic cup extractors, motivated the realization of this study, which objective was obtaining calibration curves for these extractors to monitor soil electrical conductivity and concentration of nitrate and potassium in two different soils. The nitrate concentrations tested were the same for the two soils (0; 84; 168; 252; 336; 420 and 504 mg L-1) but, for potassium, different concentrations were used for sandy soil (0; 117; 234; 351; 468; 585 and 702 mg L-1) and clay soil (0; 117; 234; 351; 468; 585; 702; 1,170; 1,755; 2,340; 2,925 and 3,510 mg L-1). Electrical conductivity of soil solution and concentrations of nitrate and potassium were estimated with satisfactory precision from soil solution collected with porous ceramic cup extractors, but this task must be preceded by a calibration for each soil type

The utility of Next Generation Sequencing for molecular diagnostics in Rett syndrome

Rett syndrome (RTT) is an early-onset neurodevelopmental disorder that almost exclusively affects girls and is totally disabling. Three genes have been identified that cause RTT: MECP2, CDKL5 and FOXG1. However, the etiology of some of RTT patients still remains unknown. Recently, next generation sequencing (NGS) has promoted genetic diagnoses because of the quickness and affordability of the method. To evaluate the usefulness of NGS in genetic diagnosis, we present the genetic study of RTT-like patients using different techniques based on this technology. We studied 1577 patients with RTT-like clinical diagnoses and reviewed patients who were previously studied and thought to have RTT genes by Sanger sequencing. Genetically, 477 of 1577 patients with a RTT-like suspicion have been diagnosed. Positive results were found in 30% by Sanger sequencing, 23% with a custom panel, 24% with a commercial panel and 32% with whole exome sequencing. A genetic study using NGS allows the study of a larger number of genes associated with RTT-like symptoms simultaneously, providing genetic study of a wider group of patients as well as significantly reducing the response time and cost of the study