Genotypic and phenotypic diversity of Bacillus spp. isolated from steel plant waste

Background: Molecular studies of Bacillus diversity in various environments have been reported. However, there have been few investigations concerning Bacillus in steel plant environments. In this study, genotypic and phenotypic diversity and phylogenetic relationships among 40 bacterial isolates recovered from steel plant waste were investigated using classical and molecular methods. Results: 16S rDNA partial sequencing assigned all the isolates to the Bacillus genus, with close genetic relatedness to the Bacillus subtilis and Bacillus cereus groups, and to the species Bacillus sphaericus. tDNA-intergenic spacer length polymorphisms and the 16S–23S intergenic transcribed spacer region failed to identify the isolates at the species level. Genomic diversity was investigatedby molecular typing with rep (repetitive sequence) based PCR using the primer sets ERIC2 (enterobacterial repetitive intergenic consensus), (GTG)5, and BOXAIR. Genotypic fingerprinting of the isolates reflected high intraspecies and interspecies diversity. Clustering of the isolates using ERIC-PCR fingerprinting was similar to that obtained from the 16S rRNA gene phylogenetic tree, indicating the potential of the former technique as a simple and useful tool for examining relationships among unknown Bacillus spp. Physiological, biochemical and heavy metal susceptibility profiles also indicated considerable phenotypic diversity. Among the heavy metal compounds tested Zn, Pb and Cu were least toxic to the bacterial isolates, whereas Ag inhibited all isolates at 0.001 mM. Conclusion: Isolates with identical 16S rRNA gene sequences had different genomic fingerprints and differed considerably in their physiological capabilities, so the high levels of phenotypic diversity found in this study are likely to have ecological relevance

Medida de função motora, corticoterapia e pacientes com distrofia muscular de Duchenne

Objective: To assess the evolution of motor function in patients with Duchenne muscular dystrophy (DMD) treated with steroids (prednisolone or deflazacort) through the Motor Function Measure (MFM), which evaluates three dimensions of motor performance (D1, D2, D3). Methods: Thirty-three patients with DMD (22 ambulant, 6 non-ambulant and 5 who lost the capacity to walk during the period of the study) were assessed using the MFM scale six times over a period of 18 months. Results: All the motor functions remained stable for 14 months in all patients, except D1 for those who lost their walking ability. In ambulant patients, D2 (axial and proximal motor capacities) motor functions improved during six months; an improvement in D3 (distal motor capacity) was noted during the total follow-up. D1 (standing posture and transfers) and total score were useful to predict the loss of the ability to walk. Conclusions: The use of the MFM in DMD patients confirms the benefits of the steroid treatment for slowing the progression of the disease.OBJETIVO: Avaliar a evolução da função motora de pacientes com distrofia muscular de Duchenne (DMD) em corticoterapia (predinisolona e deflazacort), por meio da escala Medida da Função Motora (MFM), que avalia três dimensões de funções motoras (D1, D2, D3). \ud MÉTODOS: Trinta e três pacientes com DMD (22 deambulantes, seis cadeirantes e cinco que perderam a capacidade de andar ao longo do estudo) foram avaliados pela escala MFM em seis momentos durante 18 meses. \ud RESULTADOS: Todas as funções motoras mantiveram-se estáveis durante 14 meses, exceto D1 para os pacientes que perderam a marcha. Nos pacientes deambulantes, a D2 (função motora axial e proximal) apresentou melhora durante seis meses. Melhora em D3 (função motora distal) também foi observada durante o seguimento. A D1 (postura em pé e transferências) e o escore total foram importantes para predizer a perda de marcha. \ud CONCLUSÕES: O uso da MFM nos pacientes com DMD confirma os benefícios do tratamento com corticoides na diminuição da velocidade de progressão da doença

DETERMINAÇÃO SIMULTÂNEA DE METAIS EM GASOLINA, ATRAVÉS DE MÉTODO ELETROANALÍTICO ALTERNATIVO, COM O USO DE CALIBRAÇÃO MULTIVARIADA / REDES NEURAIS

Devido a complexidade de amostras como a gasolina, a utilização de microemulsões, bem como de estratégias de calibração multivariada tem sido cada vez maior. Este trabalho apresenta o uso do programa STATISTICA 10.0 na aplicação de redes neurais artificiais (RNA) para prever a concentração dos íons metálicos Pb2+ e Cd2+ em amostras microemulsionadas de gasolina. Foi necessário descartar alguns dados experimentais de Cd2+, devido a baixa reprodutibilidade na determinação simultânea dos dois ions. Os dados foram complementados por interpolação a partir de medidas individuais de Cd2+, nas mesmas condições e no intervalo de 2x10-8 mol L-1 a 6x10-8mol L-1. Através da otimização multivariada, os seguintes parâmetros ideais permitiram a obtenção de resposta simultânea dos metais pela técnica de voltametria de redissolução anódica em DP modo (DPASV): Edep= -1200 mV; tdep = 240s; Amplitude = 50 mV. Para o estudo de predição no presente trabalho, o tipo de RNA usado foi o MLP (Multi Layer Perceptron). Os valores das concentrações (10-8 mol L-1 a 10-7 mol L-1) e as respectivas correntes, determinados, experimentalmente, em condições otimizadas, e obtidos a partir de padrões dos dois ions, suplementaram, por interpolação dos dados, e foram usados como dados de entrada da rede, para ambos os analitos. As saídas dos modelos foram as concentrações dos analitos em amostras reais de gasolina. Estas amostras reais apresentaram arranjos do tipo MLP 1-4-1 para Pb2+, e MLP 1-2-1 para o Cd2+, sendo estes capazes de estimar a concentração na determinação simultânea dos dois metais na amostra de gasolina. Os valores simulados obtidos com a rede neural corresponderam estreitamente aos resultados experimentais esperados. Os valores médios de correlações foram considerados satisfatórios na previsão das concentrações de Cd2+ (R2 = 0,998) e Pb2+ (R2 = 0,995). O eletrodo de filme de bismuto foi usado devido a sua baixa toxicidade e boa sensibilidade. Do ponto de vista analítico, o procedimento foi aplicado com sucesso em amostras reais de gasolina, no nível de 10-9 e 10-8 mol L-1 para os ions Pb2+ e Cd2+, respectivamente.Palavras-chave: Redes neurais artificiais. Planejamento experimental. Voltametria de redissolução anódica. Cobre. Chumbo. Gasolina.SIMULTANEOUS DETERMINATION OF METALS IN GASOLINE THROUGH ALTERNATIVE ELECTROANALYTICAL METHOD, WITH THE USE OF MULTIVARIATE CALIBRATION / NEURAL NETWORKSAbstract: Due to the complexity of samples such as gasoline, the use of microemulsions, as well as the multivariate calibration strategies have been used increasingly. This work presents the use of STATISTICASoftware 10.0 in the application of artificial neural networks (ANN) to predict the concentration of metal ions Pb2+ and Cd2+ in microemulsified samples of gasoline. It was necessary to dismiss some experimental data of Cd2+, due to low reproducibility in the simultaneous determination. The data were supplemented by interpolation from individual measures of Cd2+ in the same conditions and in the range of 2x10-8 mol L-1 to 6x10-8mol L-1. Through multivariate optimization, the following optimal parameters allowed to obtain simultaneous response of metals by anodic stripping voltammetry technique in DP mode: Edep = -1200 mV; tdep = 240s; Amp = 50 mV. For the prediction study in the present work, the type of ANN used was the MLP (Multi Layer Perceptron). Values of concentrations (10-8 mol L-1 to 10-7 mol L-1) and respective currents,determined experimentally in optimized conditions, and obtained from standard of the two metals, supplemented the data by interpolation, and were used as input data in the network, for both analytes. The outputs of the models were the concentrations of the analytes in real samples of gasoline. This real sample, presented a MLP arrangement of the type 1-4-1 for Pb2+, and 1-2-1 for Cd2+, being them able to estimate the concentrations of simultaneous determination of the two metals in gasoline sample. The simulated values obtained with the neural network, corresponded closely to the experimental results expected. Mean values of correlations, considered very satisfactory, were obtained in the sample gasoline, in the prediction of the concentrations of Cd2+ (R2 = 0.998) and Pb2+ (R2 = 0.995). The bismuth film electrode has been useddue to its low toxicity and good sensitivity. From the analytical point of view, the procedure was applied successfully to the real samples of gasoline at the level of 10-9 and 10-8 mol L-1 for the ions Pb2+ and Cd2+,respectively.Keywords: Artificial neural networks. Experimental design. Anodic stripping voltammetry. Copper. Lead. Gasoline.DETERMINACIÓN SIMULTÁNEA DE METALES EN GASOLINA, ATRAVES DE UN MÉTODO ELECTROANALÍTICO ALTERNATIVO, USANDO CALIBRACIÓN MULTIVARIADA / REDES NEURALESResumen: Debido a la complexidad de muestras, tales como la gasolina, la utilización de microemulsiones, asi como también de estrategias de calibración multivariada, están siendo cada vez mas comunes. Este trabajo presenta el empleo del programa STATISTICA 10.0 en la aplicación de redes neurales artificiales (RNA), para visualizar anticipadamente la concentración de los iones metálicos Pb2+ y Cd2+ en muestras microemulsionadas de gasolina. Fue necesario descartar algunos datos experimentales de Cd2+ debido a la baja reproductibilidad en la determinación simultánes de los dos iones. Los datos fueron complementados por interpolación a partir de mediciones individuales de Cd2+ , en las mismas condiciones, y en el intervalo de 2x10-8 mol L-1 a 6x10-8mol L-1. Atraves de la optimización multivariada, los siguientes parámetros ideales permitieron obtener la respuesta simultánea de los metales por la técnica de voltametría de redisolución anódica en DP modo (DPASV): Edep= -1200 mV; tdep = 240s; Amplitude = 50 mV. Para el estudio de visualización anticipada en este trabjo, el tipo de RNA usado fue el MLP (Multi Layer Perceptron). Los valores de concentraciones (10-8 mol L-1 a 10-7 mol L-1), y las respectivas corrientes, determinados experimentalmente en condiciones optimizadas, y obtenidos a partir de muestras standard de los dos iones, suplementaron por interpolación de los datos, y fueron usados como informaciones de entrada de la red, para ambos analitos. Las salidas de los modelos fueron las concentraciones de los analitos en muestras reales de gasolina. Estas muestras reales presentaron organizaciones del tipo MLP 1-4-1 para Pb2+, e MLP 1-2-1 para o Cd2+, lo que permite estimar la concentración en la determinación simultánea de los dos metales en la muestra de gasolina. Los valores simulados obtenidos con la red neural corriespondieron muy proximamente a los resultados experimentales esperados. Los valores medios de las correlaciones, considerados muy satisfactorios, fueron obtenidos en el gas de la muestra, la previsión de las concentraciones de Cd2+ (R2 = 0,998) e Pb2+ (R2 = 0,995). El electrodo de película de bismuto fue usado debido a sua baja toxicidad y buena sensibilidad. Del punto de vista analítico, el procedimiento fue aplicado con éxito en muestras reales de gasolina, en el nivel de 10-9 y 10-8 mol L-1 para los iones Pb2+ e Cd2+, respectivamente.Palabras clave: Redes neurales artificiales. Planeamento experimental. Voltametria de redisolução anódica.Cobre. Plomo. Gasolina

Human Skeletal myopathy myosin mutations disrupt myosin head sequestration

Myosin heavy chains encoded by MYH7 and MYH2 are abundant in human skeletal muscle, and important for muscle contraction. However, it is unclear how mutations in these genes disrupt myosin structure and function leading to skeletal muscle myopathies termed myosinopathies. Here, we used multiple approaches to analyse the effects of common MYH7 and MYH2 mutations in the light meromyosin region of myosin (LMM). Analyses of expressed and purified MYH7 and MYH2 LMM mutant proteins combined with in-silico modelling showed that myosin coiled-coil structure and packing of filaments in vitro are commonly disrupted. Using muscle biopsies from patients, and Mant-ATP chase protocols to estimate the proportion of myosin heads that were super-relaxed, together with X-ray diffraction measurements to estimate myosin head order we found that basal myosin ATP consumption was increased and the myosin super-relaxed state was decreased in vivo. In addition, myofibre mechanics experiments to investigate contractile function showed myofibre contractility was not affected. These findings indicate that the structural remodelling associated with LMM mutations induces a pathogenic state in which formation of shutdown heads is impaired, thus increasing myosin head ATP demand in the filaments, rather than affecting contractility. These key findings will help design future therapies for myosinopathies

Nemaline Myopathy in Brazilian Patients: Molecular and Clinical Characterization

Nemaline myopathy (NM), a structural congenital myopathy, presents a significant clinical and genetic heterogeneity. Here, we compiled molecular and clinical data of 30 Brazilian patients from 25 unrelated families. Next-generation sequencing was able to genetically classify all patients: sixteen families (64%) with mutation in NEB, five (20%) in ACTA1, two (8%) in KLHL40, and one in TPM2 (4%) and TPM3 (4%). In the NEB-related families, 25 different variants, 11 of them novel, were identified; splice site (10/25) and frame shift (9/25) mutations were the most common. Mutation c.24579 G>C was recurrent in three unrelated patients from the same region, suggesting a common ancestor. Clinically, the “typical” form was the more frequent and caused by mutations in the different NM genes. Phenotypic heterogeneity was observed among patients with mutations in the same gene. Respiratory involvement was very common and often out of proportion with limb weakness. Muscle MRI patterns showed variability within the forms and genes, which was related to the severity of the weakness. Considering the high frequency of NEB mutations and the complexity of this gene, NGS tools should be combined with CNV identification, especially in patients with a likely non-identified second mutation

Nemaline Myopathy in Brazilian Patients: Molecular and Clinical Characterization

Nemaline myopathy (NM), a structural congenital myopathy, presents a significant clinical and genetic heterogeneity. Here, we compiled molecular and clinical data of 30 Brazilian patients from 25 unrelated families. Next-generation sequencing was able to genetically classify all patients: sixteen families (64%) with mutation in NEB, five (20%) in ACTA1, two (8%) in KLHL40, and one in TPM2 (4%) and TPM3 (4%). In the NEB-related families, 25 different variants, 11 of them novel, were identified; splice site (10/25) and frame shift (9/25) mutations were the most common. Mutation c.24579 G>C was recurrent in three unrelated patients from the same region, suggesting a common ancestor. Clinically, the “typical” form was the more frequent and caused by mutations in the different NM genes. Phenotypic heterogeneity was observed among patients with mutations in the same gene. Respiratory involvement was very common and often out of proportion with limb weakness. Muscle MRI patterns showed variability within the forms and genes, which was related to the severity of the weakness. Considering the high frequency of NEB mutations and the complexity of this gene, NGS tools should be combined with CNV identification, especially in patients with a likely non-identified second mutation

Evaluation of surgical treatment of congenital heart disease in patients aged above 16 years

FUNDAMENTO: O número crescente de crianças com cardiopatias congênitas em evolução demanda maior preparo dos profissionais e das instituições que as manuseiam. OBJETIVO: Descrever o perfil dos pacientes com idade superior a 16 anos com cardiopatia congênita operados e analisar os fatores de risco preditivos de mortalidade hospitalar. MÉTODOS: Mil, quinhentos e vinte pacientes (idade média 27 ± 13 anos) foram operados entre janeiro de 1986 e dezembro de 2010. Foram realizadas análise descritiva do perfil epidemiológico da população estudada e análise dos fatores de risco para mortalidade hospitalar, considerando escore de complexidade, ano em que a cirurgia foi realizada, procedimento realizado pelo cirurgião pediátrico ou não e presença de reoperação. RESULTADOS: Ocorreu um crescimento expressivo no número de casos a partir do ano 2000. A média do escore de complexidade foi 5,4 e os defeitos septais corresponderam a 45% dos casos. A mortalidade geral foi 7,7% e o maior número de procedimentos (973 ou 61,9%) com maior complexidade foi realizado por cirurgiões pediátricos. Complexidade (OR 1,5), reoperação (OR 2,17) e cirurgião pediátrico (OR 0,28) foram fatores de risco independentes que influenciaram a mortalidade. A análise multivariada mostrou que o ano em que a cirurgia foi realizada (OR 1,03), a complexidade (OR 1,44) e o cirurgião pediátrico (OR 0,28) influenciaram no resultado. CONCLUSÃO: Observa-se um número crescente de pacientes com idade superior a 16 anos e que, apesar do grande número de casos simples, os mais complexos foram encaminhados para os cirurgiões pediátricos, que apresentaram menor mortalidade, em especial nos anos mais recentes. (Arq Bras Cardiol. 2012; [online].ahead print, PP.0-0

Lung function monitoring in patients with duchenne muscular dystrophy on steroid therapy

Background\ud Duchenne muscular dystrophy (DMD) is a sex-linked inherited muscle disease characterized by a progressive loss in muscle strength and respiratory muscle involvement. After 12 years of age, lung function declines at a rate of 6 % to 10.7 % per year in patients with DMD. Steroid therapy has been proposed to delay the loss of motor function and also the respiratory involvement.\ud \ud Method\ud In 21 patients with DMD aged between seven and 16 years, the forced vital capacity (FVC) and the forced expiratory volume in one second (FEV1) were evaluated at three different times during a period of two years.\ud \ud Results\ud We observed in this period of evaluation the maintenance of the FVC and the FEV1 in this group of patients independently of chronological age, age at onset of steroid therapy, and walking capacity.\ud \ud Conclusion\ud The steroid therapy has the potential to stabilize or delay the loss of lung function in DMD patients even if they are non-ambulant or older than 10 years, and in those in whom the medication was started after 7 years of age.DLM was supported by CAPES (Coordenação de Aperfeiçoamento Pessoal de Ensino Superior)

The BINGO Project IX: Search for Fast Radio Bursts -- A Forecast for the BINGO Interferometry System

The Baryon Acoustic Oscillations (BAO) from Integrated Neutral Gas Observations (BINGO) radio telescope will use the neutral Hydrogen emission line to map the Universe in the redshift range $0.127 \le z \le 0.449$, with the main goal of probing BAO. In addition, the instrument optical design and hardware configuration support the search for Fast Radio Bursts (FRBs). In this work, we propose the use of a BINGO Interferometry System (BIS) including new auxiliary, smaller, radio telescopes (hereafter \emph{outriggers}). The interferometric approach makes it possible to pinpoint the FRB sources in the sky. We present here the results of several BIS configurations combining BINGO horns with and without mirrors ($4$ m, $5$ m, and $6$ m) and 5, 7, 9, or 10 for single horns. We developed a new {\tt Python} package, the {\tt FRBlip}, which generates synthetic FRB mock catalogs and computes, based on a telescope model, the observed signal-to-noise ratio (S/N) that we used to compute numerically the detection rates of the telescopes and how many interferometry pairs of telescopes (\emph{baselines}) can observe an FRB. FRBs observed by more than one baseline are the ones whose location can be determined. We thus evaluate the performance of BIS regarding FRB localization. We found that BIS will be able to localize 23 FRBs yearly with single horn outriggers in the best configuration (using 10 outriggers of 6 m mirrors), with redshift $z \leq 0.96$; the full localization capability depends on the number and the type of the outriggers. Wider beams are best to pinpoint FRB sources because potential candidates will be observed by more baselines, while narrow beams look deep in redshift. The BIS can be a powerful extension of the regular BINGO telescope, dedicated to observe hundreds of FRBs during Phase 1. Many of them will be well localized with a single horn + 6 m dish as outriggers.(Abridged)Comment: 12 pages, 9 figures, 5 tables, submitted to A&