Molecular analysis of PKU-associated PAH mutations: a fast and simple genotyping test

Abstract: Neonatal screening for phenylketonuria (PKU, OMIM: 261600) was introduced at the end of the 1960s. We developed a rapid and simple molecular test for the most frequent phenylalanine hydroxylase (PAH, Gene ID: 5053) mutations. Using this method to detect the 18 most frequent mutations, it is possible to achieve a 75% detection rate in Italian population. The variants selected also reach a high detection rate in other populations, for example, 70% in southern Germany, 68% in western Germany, 76% in Denmark, 68% in Sweden, 63% in Poland, and 60% in Bulgaria. We successfully applied this confirmation test in neonatal screening for hyperphenylalaninemias using dried blood spots and obtained the genotype in approximately 48 h. The method was found to be suitable as second tier test in neonatal screening for hyperphenylalaninemias in neonates with a positive screening test. This test can also be useful for carrier screening because it can bypass the entire coding sequence and intron–exon boundaries sequencing, thereby overcoming the questions that this approach implies, such as new variant interpretations

Editorial: Bioaerosol emission characteristics and the epidemiological, occupational, and public health risk assessment of waste and wastewater management

Waste management industries are vital to achieving the Sustainable Development Goals suggested by World Health Organization. Besides SDGs achievement, also the circular economy is dependent on the waste sector. The circular economy intends to reuse existing resources instead of disposing of materials that are no longer useful. Thus, the circular economy is very dependent on an increased workforce dedicated to waste management. Nonetheless, while better waste management is being predicted as a critical contributor to reducing health outcomes and environmental negative impacts, the bioaerosols´ occupational exposure in the waste sector is being neglected leading to negative outcomes on workers’ health. The scientific community and stakeholders should be also aware of the foreseen increased exposure to microbes and antimicrobial resistance in different environments that will lead to a boost of exposure to bioaerosols in the waste sector, mainly due to climate change.info:eu-repo/semantics/publishedVersio

Biochemical data from the characterization of a new pathogenic mutation of human pyridoxine-5'-phosphate oxidase (PNPO)

PNPO deficiency is responsible of severe neonatal encephalopathy, responsive to pyridoxal-5’-phosphate (PLP) or pyridoxine. Recent studies widened the phenotype of this condition and detected new genetic variants on PNPO gene, whose pathogenetic role and clinical expression remain to be established. One of these mutations, Arg116Gln, is of particular interest because of its later onset of symptoms (beyond the first months of life) and its peculiar epileptic manifestations in patients. This protein variant was expressed as recombinant protein in E coli, purified to homogeneity, and characterized with respect to structural and kinetic properties, stability, binding constants of cofactor flavin mononucleotide (FMN) and product (PLP) in order to define the molecular and structural bases of its pathogenicity. For interpretation and discussion of reported data, together with the description of clinical studies, refer to the article [7][1] (doi: 10.1016/j.ymgme.2017.08.003)

In vitro study of uptake and synthesis of creatine and its precursors by cerebellar granule cells and astrocytes suggests some hypotheses on the physiopathology of the inherited disorders of creatine metabolism

<p>Abstract</p> <p>Background</p> <p>The discovery of the inherited disorders of creatine (Cr) synthesis and transport in the last few years disclosed the importance of blood Cr supply for the normal functioning of the brain. These putatively rare diseases share a common pathogenetic mechanism (the depletion of brain Cr) and similar phenotypes characterized by mental retardation, language disturbances, seizures and movement disorders. In the effort to improve our knowledge on the mechanisms regulating Cr pool inside the nervous tissue, Cr transport and synthesis and related gene transcripts were explored in primary cultures of rat cerebellar granule cells and astrocytes.</p> <p>Methods</p> <p>Cr uptake and synthesis were explored in vitro by incubating monotypic primary cultures of rat type I astrocytes and cerebellar granule cells with: a) D₃-Creatine (D₃Cr) and D3Cr plus β-guanidinopropionate (GPA, an inhibitor of Cr transporter), and b) labelled precursors of Guanidinoacetate (GAA) and Cr (Arginine, Arg; Glycine, Gly). Intracellular D3Cr and labelled GAA and Cr were assessed by ESI-MS/MS. Creatine transporter (<it>CT1</it>), L-arginine:glycine amidinotransferase (<it>AGAT</it>), and S-adenosylmethionine:guanidinoacetate N-methyltransferase (<it>GAMT</it>) gene expression was assessed in the same cells by real time PCR.</p> <p>Results</p> <p>D3Cr signal was extremely high in cells incubated with this isotope (labelled/unlabelled Cr ratio reached about 10 and 122, respectively in cerebellar granule cells and astrocytes) and was reduced by GPA. Labelled Arg and Gly were taken up by the cells and incorporated in GAA, whose concentration paralleled that of these precursors both in the extracellular medium and inside the cells (astrocytes). In contrast, the increase of labelled Cr was relatively much more limited since labelled Cr after precursors' supplementation did not exceed 2,7% (cerebellar granule cells) and 21% (astrocytes) of unlabelled Cr. Finally, <it>AGAT, GAMT </it>and <it>SLC6A8 </it>were expressed in both kind of cells.</p> <p>Conclusions</p> <p>Our results confirm that both neurons and astrocytes have the capability to synthesize and uptake Cr, and suggest that at least in vitro intracellular Cr can increase to a much greater extent through uptake than through <it>de novo </it>synthesis. Our results are compatible with the clinical observations that when the Cr transporter is defective, intracellular Cr is absent despite the brain should be able to synthesize it. Further research is needed to fully understand to what extent our results reflect the in vivo situation.</p

Case report: Childhood epilepsy and borderline intellectual functioning hiding an AADC deficiency disorder associated with compound heterozygous DDC gene pathogenic variants

Aromatic L-amino acid decarboxylase (AADC) deficiency is a rare autosomal recessive neurometabolic disorder leading to severe combined serotonin, dopamine, norepinephrine, and epinephrine deficiency. We report on a female patient with borderline functioning and sporadic clear-cut focal to bilateral seizures from age 10 years. A neuropsychological assessment highlighted a mild impairment in executive functions, affecting attention span and visual-spatial abilities. Following the diagnosis of epilepsy with a presumed genetic etiology, we applied a diagnostic approach inclusive of a next-generation sequencing (NGS) gene panel, which uncovered two variants in trans in the DOPA decarboxylase (DDC) gene underlying an AADC deficiency. This compound heterozygous genotype was associated with a mild reduction of homovanillic acid, a low level of the norepinephrine catabolite, and a significant reduction of 5-hydroxyindoleacetic acid in cerebrospinal fluid. Remarkably, 3-O-methyldopa (3-OMD) and 5-hydroxytryptophan were instead increased. During the genetically guided re-evaluation process, some mild signs of dysautonomic dysfunction (nasal congestion, abnormal sweating, hypotension and fainting, excessive sleepiness, small hands and feet, and increased levels of prolactin, tiredness, and fatigue), more typical of AADC deficiency, were evaluated with new insight. Of the two AADC variants, the R347Q has already been characterized as a loss-of-function with severe catalytic impairments, while the novel L391P variant has been predicted to have a less severe impact. Bioinformatic analyses suggest that the amino acid substitution may affect affinity for the PLP coenzyme. Thus, the genotype corresponds to a phenotype with mild and late-onset symptoms, of which seizures were the clinical sign, leading to medical attention. This case report expands the spectrum of AADC deficiency phenotypes to encompass a less-disabling clinical condition including borderline cognitive functioning, drug-responsive epilepsy, and mild autonomic dysfunction

SEQUESTRO DE CARBONO DE UM LATOSSOLO CAULINÍTICO SOB CULTIVO DE LINHAÇA EM MANEJO CONSERVACIONISTA

Agriculture that employs soil conservation practices promotes ecosystem services such as the mitigation of greenhouse gases. The objective of this work was to evaluate the carbon sequestration from a Latossolo Vermelho Distroférrico (Haplustox) under a conservationist management system for flaxseed production over three years yield. The experiment was carried out for three years in a soil conservationist management system (no-tillage) for the production of three brown flaxseed varieties: Caburé, Aguará and UFSC cultivated in Latossolo Vermelho. Undesturbed and deformed soil samples were collected in two contrasting layers: 0-0.15 and 0.15-0.30 m depth. Soil bulk density, total organic carbon, stocks and CO2 sequestration -IPCC were determined. Data were submitted to variance analysis and Tukey's test (p&lt;0.05). The soil management used contributed to the soil carbon stock, regardless of the variety used, on average 27 Mg C ha was stored in the 0-0.15m layer and 29.8 Mg C ha-1 at 0.15- 0.30 m in depth, in addition to mitigating an average of 10.48 Mg CO2eq ha-1 in just three years of cultivation, which makes this management system environmentally sustainable with yields equivalent to those obtained in South region of Brazil (0,71 Mg ha-1).La agricultura que emplea prácticas de conservación del suelo promueve los servicios ecosistémicos, como la mitigación de gases de efecto invernadero.  El objetivo de este trabajo fue evaluar el secuestro de carbono de un Latosol Rojo Distrófico bajo sistema de manejo de conservación para la producción de cerda cerda durante tres años de cultivo. El experimento se llevó a cabo durante tres años en un sistema de conservación de manejo para la producción de tres variedades de linaza parda: Caburé, Aguará y UFSC cultivadas en latosol rojo distrófico caolinítico. Las muestras preservadas y deformadas se recogieron en dos capas constratantes: 0-0,15 y 0,15-0,30 m. Se determinó la densidad global del suelo, el carbono orgánico total, las reservas y el secuestro de CO2 -IPCC. Los datos fueron sometidos a análisis de varianza y prueba de Tukey (p&lt;0,05). El manejo utilizado contribuyó al stock de carbono en el suelo, independientemente de la variedad utilizada, en promedio, se sembraron 27 Mg C ha-1 en la capa de 0-0.15m y 29.8 Mg C ha-1 a 0.15-0.30 m de profundidad, además de mitigar en promedio 10.48 Mg CO2 eq ha-1 en solo tres años de cultivo, lo que hace que este sistema de manejo sea ambientalmente sostenible con producciones equivalentes a las obtenidas en el sur de Brasil (0.71 Mg ha-1).A agricultura que emprega práticas de conservação do solo promove serviços ecossistêmicos como a mitigação dos gases de efeito estufa.&nbsp; O objetivo desse trabalho foi avaliar o sequestro de carbono de um Latossolo Vermelho Distroférrico sob sistema de manejo conservacionista para a produção de linhaça ao longo de três anos de cultivo. O experimento foi desenvolvido durante três anos em sistema conservacionista de manejo para produção de três variedades de linhaça marrom: Caburé, Aguará e UFSC cultivados em Latossolo Vermelho Distroférrico caulinítico. Foram coletadas amostras preservadas e deformadas em duas camadas constratantes: 0-0,15 e 0,15-0,30 m. Determinou-se a densidade global do solo, o carbono orgânico total, os estoques e o sequestro de CO2 -IPCC. Os dados foram submetidos a análise de variância e ao teste de Tukey (p&lt;0,05). O manejo utilizado contribuiu com o estoque de carbono no solo, independente da variedade utilizada, em média, estocou-se 27 Mg C ha-1 na camada de 0-0,15m e 29,8 Mg C ha-1 a 0,15-0,30 m de profundidade, além de mitigar em média 10,48 Mg CO2 eq ha-1 em apenas três anos de cultivo, o que torna esse sistema de manejo ambientalmente sustentável com produções equivalente as obtidas no Sul do Brasil (0,71 Mg ha-1)

SEQUESTRO DE CARBONO DE UM LATOSSOLO CAULINÍTICO SOB CULTIVO DE LINHAÇA EM MANEJO CONSERVACIONISTA

Agriculture that employs soil conservation practices promotes ecosystem services such as the mitigation of greenhouse gases. The objective of this work was to evaluate the carbon sequestration from a Latossolo Vermelho Distroférrico (Haplustox) under a conservationist management system for flaxseed production over three years yield. The experiment was carried out for three years in a soil conservationist management system (no-tillage) for the production of three brown flaxseed varieties: Caburé, Aguará and UFSC cultivated in Latossolo Vermelho. Undesturbed and deformed soil samples were collected in two contrasting layers: 0-0.15 and 0.15-0.30 m depth. Soil bulk density, total organic carbon, stocks and CO2 sequestration -IPCC were determined. Data were submitted to variance analysis and Tukey's test (p&lt;0.05). The soil management used contributed to the soil carbon stock, regardless of the variety used, on average 27 Mg C ha was stored in the 0-0.15m layer and 29.8 Mg C ha-1 at 0.15- 0.30 m in depth, in addition to mitigating an average of 10.48 Mg CO2eq ha-1 in just three years of cultivation, which makes this management system environmentally sustainable with yields equivalent to those obtained in South region of Brazil (0,71 Mg ha-1).La agricultura que emplea prácticas de conservación del suelo promueve los servicios ecosistémicos, como la mitigación de gases de efecto invernadero.  El objetivo de este trabajo fue evaluar el secuestro de carbono de un Latosol Rojo Distrófico bajo sistema de manejo de conservación para la producción de cerda cerda durante tres años de cultivo. El experimento se llevó a cabo durante tres años en un sistema de conservación de manejo para la producción de tres variedades de linaza parda: Caburé, Aguará y UFSC cultivadas en latosol rojo distrófico caolinítico. Las muestras preservadas y deformadas se recogieron en dos capas constratantes: 0-0,15 y 0,15-0,30 m. Se determinó la densidad global del suelo, el carbono orgánico total, las reservas y el secuestro de CO2 -IPCC. Los datos fueron sometidos a análisis de varianza y prueba de Tukey (p&lt;0,05). El manejo utilizado contribuyó al stock de carbono en el suelo, independientemente de la variedad utilizada, en promedio, se sembraron 27 Mg C ha-1 en la capa de 0-0.15m y 29.8 Mg C ha-1 a 0.15-0.30 m de profundidad, además de mitigar en promedio 10.48 Mg CO2 eq ha-1 en solo tres años de cultivo, lo que hace que este sistema de manejo sea ambientalmente sostenible con producciones equivalentes a las obtenidas en el sur de Brasil (0.71 Mg ha-1).A agricultura que emprega práticas de conservação do solo promove serviços ecossistêmicos como a mitigação dos gases de efeito estufa.&nbsp; O objetivo desse trabalho foi avaliar o sequestro de carbono de um Latossolo Vermelho Distroférrico sob sistema de manejo conservacionista para a produção de linhaça ao longo de três anos de cultivo. O experimento foi desenvolvido durante três anos em sistema conservacionista de manejo para produção de três variedades de linhaça marrom: Caburé, Aguará e UFSC cultivados em Latossolo Vermelho Distroférrico caulinítico. Foram coletadas amostras preservadas e deformadas em duas camadas constratantes: 0-0,15 e 0,15-0,30 m. Determinou-se a densidade global do solo, o carbono orgânico total, os estoques e o sequestro de CO2 -IPCC. Os dados foram submetidos a análise de variância e ao teste de Tukey (p&lt;0,05). O manejo utilizado contribuiu com o estoque de carbono no solo, independente da variedade utilizada, em média, estocou-se 27 Mg C ha-1 na camada de 0-0,15m e 29,8 Mg C ha-1 a 0,15-0,30 m de profundidade, além de mitigar em média 10,48 Mg CO2 eq ha-1 em apenas três anos de cultivo, o que torna esse sistema de manejo ambientalmente sustentável com produções equivalente as obtidas no Sul do Brasil (0,71 Mg ha-1)

A NOVEL MISSENSE MUTATION PATTERN OF THE GCH1 GENE IN DOPA-RESPONSIVE DYSTONIA

Dopa-responsive dystonia (DRD) is an inherited metabolic disorder now classified as DYT5 with two different biochemical defects: autosomal dominant GTP cyclohydrolase 1 (GCH1) deficiency or autosomal recessive tyrosine hydroxylase deficiency. We report the case of a 10-years-old girl with progressive generalized dystonia and gait disorder who presented dramatic response to levodopa. The phenylalanine to tyrosine ratio was significantly higher after phenylalanine loading test. This condition had two different heterozygous mutations in the GCH1 gene: the previously reported P23L mutation and a new Q182E mutation. The characteristics of the DRD and the molecular genetic findings are discussed

VIABILIDADE ECONÔMICA PARA A PRODUÇÃO DE LINHAÇA MARROM

The aim of this study was to evaluate the economic viability of brown linseed production in three crop years for the insertion of this agricultural activity in Dourados-MS. For this, primary productivity data were obtained through the experiment conducted in the experimental area of UFGD with three brown flaxseed genotypes: Aguará and Caburé-INTA/Argentina and UFSC/Brazil, grown in three crop years: 2016/2017, 2017/2018 and 2018/2019. The production cost was based on the data obtained in the experiment according to the production costs of winter crops from CONAB. The minimum attractiveness rate (TMA), the cash flow that involves the production of flaxseed, as well as the economic indicators: internal rate of return - IRR, net present value - NPV, time of return and the profitability index - IL. It was found that the production of brown linseed is economically viable for the farmer and may be a new alternative to compose the agricultural systems of the region of Dourados / MS.El objetivo de este estudio fue evaluar la viabilidad económica de la producción de sosa parda en tres años de cultivo para su inserción en la actividad agrícola en Dourados-MS. Para ello se utilizaron datos primarios de productividad, obtenidos a través del experimento realizado en el área experimental de UFGD con tres genotipos de línea parda: Aguará y Caburé-INTA/argentina y UFSC/Brasil, cultivados en tres años de cultivo: 2016/2017, 2017/2018 y 2018/2019. El costo de producción se realizó con base en los datos obtenidos en el experimento de acuerdo con los costos de producción de los cultivos de invierno de la CONAB. Evaluar la viabilidad de la inversión, la tasa mínima de atractivo (TMA), el flujo de caja que implica la producción de linaza, así como los indicadores económicos: tasa interna de retorno - TIR, valor presente neto-LPV, tiempo de retorno e índice de rentabilidad - IL. Se verificó que la producción de linaza marrón es económicamente viable para el agricultor y puede ser una nueva alternativa para conformar los sistemas agrícolas de Dourados/MS.O objetivo deste estudo foi avaliar a viabilidade econômica da produção de linhaça marrom em três anos safra para a inserção desta na atividade agrícola em Dourados-MS. Para isto, foram utilizados dados primários de produtividade, obtidos através do experimento conduzido na área experimental da UFGD com três genótipos de linhaça marrom: Aguará e Caburé-INTA/argentina e UFSC/Brasil, cultivados em três anos safra: 2016/2017, 2017/2018 e 2018/2019. O custo de produção foi realizado a partir dos dados obtidos no experimento conforme os custos de produção de culturas de Inverno da CONAB. Para avaliação da viabilidade do investimento, foi mensurada a taxa mínima de atratividade (TMA), o fluxo de caixa que envolve a produção de linhaça, bem como os indicadores econômicos: taxa interna de retorno - TIR, valor presente líquido-VPL, tempo de retorno e o índice de lucratividade - IL. Verificou-se que a produção de linhaça marrom é economicamente viável ao agricultor e pode ser uma nova alternativa para compor os sistemas agrícolas de Dourados/MS.O objetivo deste estudo foi avaliar a viabilidade econômica da produção de linhaça marrom em três anos safra para a inserção desta na atividade agrícola em Dourados-MS. Para isto, foram utilizados dados primários de produtividade, obtidos através do experimento conduzido na área experimental da UFGD com três genótipos de linhaça marrom: Aguará e Caburé-INTA/argentina e UFSC/Brasil, cultivados em três anos safra: 2016/2017, 2017/2018 e 2018/2019. O custo de produção foi realizado a partir dos dados obtidos no experimento conforme os custos de produção de culturas de Inverno da CONAB. Para avaliação da viabilidade do investimento, foi mensurada a taxa mínima de atratividade (TMA), o fluxo de caixa que envolve a produção de linhaça, bem como os indicadores econômicos: taxa interna de retorno - TIR, valor presente líquido-VPL, tempo de retorno e o índice de lucratividade - IL. Verificou-se que a produção de linhaça marrom é economicamente viável ao agricultor e pode ser uma nova alternativa para compor os sistemas agrícolas de Dourados/MS