Avaliacao de quatro metodos diferentes de extracao de DNA em isolados clinicos de estafilococos coagulase negativos (SCoN)

Currently there are several methods to extract bacterial DNA based on different principles. However, the amount and the quality of the DNA obtained by each one of those methods is highly variable and microorganism dependent, as illustrated by coagulase-negative staphylococci (CoNS) which have a thick cell wall that is difficult to lyse. This study was designed to compare the quality and the amount of CoNS DNA, extracted by four different techniques: two in-house protocols and two commercial kits. DNA amount and quality determination was performed through spectrophotometry. The extracted DNA was also analyzed using agarose gel electrophoresis and by PCR. 267 isolates of CoNS were used in this study. The column method and thermal lyses showed better results with regard to DNA quality (mean ratio of A260/280 = 1.95) and average concentration of DNA (), respectively. All four methods tested provided appropriate DNA for PCR amplification, but with different yields. DNA quality is important since it allows the application of a large number of molecular biology techniques, and also it's storage for a longer period of time. In this sense the extraction method based on an extraction column presented the best results for CoNS.Atualmente, para extrair o DNA bacteriano, existem diversos métodos baseados em diferentes princípios. Entretanto, a quantidade e qualidade do DNA obtido por cada um destes métodos é variável e depende do tipo de micro-organismo em questão; os estafilococos coagulase-negativos (CoNS), por exemplo, possuem parede celular espessa difícil de lisar. O objetivo deste estudo foi comparar a quantidade e a qualidade do DNA extraído de isolados clínicos de CoNS utilizando quatro metodologias diferentes: dois protocolos caseiros e dois kits comerciais. A determinação da quantidade e da qualidade do DNA foi realizada por espectrofotometria. O DNA extraído também foi analisado em eletroforese em gel de agarose e por PCR. A concentração média de DNA foi mais alta no método de lise térmica (). Entretanto, com relação à qualidade do DNA, o kit comercial que utiliza um método de extração baseado em uma coluna de separação apresentou melhor resultado (média da relação A260/280 = 1,95). As quatro técnicas testadas forneceram DNA passível de amplificação por PCR, porém com diferentes rendimentos. A qualidade do DNA extraído de bactérias é importante, pois possibilita a realização de maior número de técnicas de biologia molecular e também armazenamento do material por maior período de tempo. Neste sentido, a técnica de extração por coluna de separação apresentou melhor desempenho frente aos CoNS

Avaliacao de quatro metodos diferentes de extracao de DNA em isolados clinicos de estafilococos coagulase negativos (SCoN)

Currently there are several methods to extract bacterial DNA based on different principles. However, the amount and the quality of the DNA obtained by each one of those methods is highly variable and microorganism dependent, as illustrated by coagulase-negative staphylococci (CoNS) which have a thick cell wall that is difficult to lyse. This study was designed to compare the quality and the amount of CoNS DNA, extracted by four different techniques: two in-house protocols and two commercial kits. DNA amount and quality determination was performed through spectrophotometry. The extracted DNA was also analyzed using agarose gel electrophoresis and by PCR. 267 isolates of CoNS were used in this study. The column method and thermal lyses showed better results with regard to DNA quality (mean ratio of A260/280 = 1.95) and average concentration of DNA (), respectively. All four methods tested provided appropriate DNA for PCR amplification, but with different yields. DNA quality is important since it allows the application of a large number of molecular biology techniques, and also it's storage for a longer period of time. In this sense the extraction method based on an extraction column presented the best results for CoNS.Atualmente, para extrair o DNA bacteriano, existem diversos métodos baseados em diferentes princípios. Entretanto, a quantidade e qualidade do DNA obtido por cada um destes métodos é variável e depende do tipo de micro-organismo em questão; os estafilococos coagulase-negativos (CoNS), por exemplo, possuem parede celular espessa difícil de lisar. O objetivo deste estudo foi comparar a quantidade e a qualidade do DNA extraído de isolados clínicos de CoNS utilizando quatro metodologias diferentes: dois protocolos caseiros e dois kits comerciais. A determinação da quantidade e da qualidade do DNA foi realizada por espectrofotometria. O DNA extraído também foi analisado em eletroforese em gel de agarose e por PCR. A concentração média de DNA foi mais alta no método de lise térmica (). Entretanto, com relação à qualidade do DNA, o kit comercial que utiliza um método de extração baseado em uma coluna de separação apresentou melhor resultado (média da relação A260/280 = 1,95). As quatro técnicas testadas forneceram DNA passível de amplificação por PCR, porém com diferentes rendimentos. A qualidade do DNA extraído de bactérias é importante, pois possibilita a realização de maior número de técnicas de biologia molecular e também armazenamento do material por maior período de tempo. Neste sentido, a técnica de extração por coluna de separação apresentou melhor desempenho frente aos CoNS

Análise retrospectiva comparativa do tratamento cirúrgico da fratura de clavícula com haste versus placa

Objective: Studies confirm the benefit of surgical treatment for fixation of displaced midshaft clavicle fractures. Plate fixation and intramedullary nail are the two most used techniques. The purpose of this study is to compare these two surgical techniques. Method: Retrospective study, performed through the evaluation of patients treated for displaced midshaft clavicle fracture with intramedullary nail and plate and screws. Socioeconomic variables were collected, a visual pain scale questionnaire was applied, the shoulder function was measured using CONSTANT and UCLA scores, and radiography was performed to verify the consolidation and evaluation of the final clavicle shortening. Results: Sixty-five patients were evaluated, 36 (55.4%) of which were submitted to clavicle fixation with plate and screws and 29 (44.6%) with intramedullary nail. The median shortening was 0.1mm for plate and 5.8mm for nail (p = 0.001). The UCLA score shows an average of 35 in the plate group and 35 in the intramedullary group. The median CONSTANT scores were 96.5 for plate and 95 for nail, without significance. In all, 13 (20%) complications were registered, 9 fixed with plate and 4  fixed with intramedullary nail. The most common complication was skin erosion with exposure of the synthetic material. Conclusion: The two techniques present satisfactory results for the treatment of displaced midshaft clavicle fractures.Objetivo: Trabalhos confirmam o benefício do tratamento cirúrgico para fixação de fraturas desviadas do corpo da clavícula. A fixação com placa e haste intramedular são as duas técnicas mais utilizadas. O objetivo deste estudo é comparar as duas técnicas cirúrgicas. Método: Estudo retrospectivo realizado por meio da avaliação de pacientes submetidos à fixação com haste intramedular e com placa e parafusos para o tratamento de fraturas desviadas do corpo da clavícula. Neste caso, foram avaliadas variáveis socioeconômicas, aplicado questionário da escala visual da dor, mensurado a função do ombro pelo escore de CONSTANT e UCLA, e realizada radiografia para verificação da consolidação e avaliação do encurtamento final da clavícula. Resultados: foram avaliados 65 pacientes, destes 36 (55,4%) foram submetidos à fixação da clavícula com placa e parafusos e 29 (44,6%) com haste intramedular. A mediana do encurtamento foi 0,1mm para placa e 5,8mm para haste (p=0,001). O escore de UCLA apresentou mediana de 35 no grupo placa e 35 no grupo haste. O escore de Constant mostrou mediana de 96,5 no grupo placa e 95 no grupo haste, sem diferença significativa entre os grupos. Ao todo, foram registradas 13 (20%) complicações, 9 do grupo fixado com placa e 4 do grupo fixado com haste.  A complicação mais comum foi a erosão da pele com exposição do material de síntese. Conclusão: as duas técnicas apresentam resultados satisfatórios para o tratamento das fraturas do corpo da clavícula. Nível III: estudo terapêutico retrospectiv

The atypical chemokine receptor ACKR2 is protective against sepsis

Sepsis is a systemic inflammatory response as a result of uncontrolled infections. Neutrophils are the first cells to reach the primary sites of infection and chemokines play a key role in recruiting neutrophils. However, in sepsis chemokines could also contribute to neutrophil infiltration to vital organs leading to multiple organ failure. ACKR2 is an atypical chemokine receptor, which can remove and degrade inflammatory CC chemokines. The role of ACK2 in sepsis is unknown. Using a model of cecal ligation and puncture (CLP), we demonstrate here that ACKR2 deficient (−/−) mice exhibited a significant reduction in the survival rate compared to similarly treated wild type (WT) mice. However, neutrophil migration to the peritoneal cavity and bacterial load were similar between WT and ACKR2−/− mice during CLP. In contrast, ACKR2−/− mice showed increased neutrophil infiltration and elevated CC chemokine levels in the lung, kidney and heart compared to the WT mice. In addition, ACKR2−/− mice also showed more severe lesions in the lung and kidney than those in the WT mice. Consistent with these results, WT mice under non-severe sepsis (90% survival) had higher expression of ACKR2 in these organs than mice under severe sepsis (no survival). Finally, the lungs from septic patients showed increased number of ACKR2+ cells compared to those of non-septic patients. Our data indicate that ACKR2 may have a protective role during sepsis, and the absence of ACKR2 leads to exacerbated chemokine accumulation, neutrophil infiltration and damage to vital organs

Enhanced Optical Properties of Germanate and Tellurite Glasses Containing Metal or Semiconductor Nanoparticles

Germanium- and tellurium-based glasses have been largely studied due to their recognized potential for photonics. In this paper, we review our recent studies that include the investigation of the Stokes and anti-Stokes photoluminescence (PL) in different glass systems containing metallic and semiconductor nanoparticles (NPs). In the case of the samples with metallic NPs, the enhanced PL was attributed to the increased local field on the rare-earth ions located in the proximity of the NPs and/or the energy transfer from the metallic NPs to the rare-earth ions. For the glasses containing silicon NPs, the PL enhancement was mainly due to the energy transfer from the NPs to the Er3+ ions. The nonlinear (NL) optical properties of PbO-GeO2 films containing gold NPs were also investigated. The experiments in the pico- and subpicosecond regimes revealed enhanced values of the NL refractive indices and large NL absorption coefficients in comparison with the films without gold NPs. The reported experiments demonstrate that germanate and tellurite glasses, having appropriate rare-earth ions doping and NPs concentration, are strong candidates for PL-based devices, all-optical switches, and optical limiting

FRACTURE-DISLOCATIONS OF THE ELBOW: CAN THEY INFLUENCE THE PATTERN FRACTURE OF RADIAL HEAD?

ABSTRACT Introduction: Radial head fractures are consistently part of a terrible triad of the elbow and can occur in association with Monteggia fracture-dislocations, transolecranon fractures, and their variations. Understanding the degree of comminution of the radial head fracture and the location of fragments determines the course of action to be taken. Objectives: To correlate fracture-dislocations with the pattern of radial head fracture (number of fragments) and involvement in the proximal radioulnar region. Materials and Methods: A retrospective study (level II) of patients undergoing surgery for radial head fractures associated with fracture-dislocations. Patients had radiographs in anteroposterior and lateral views, as well as tomography. The number of radial head fracture fragments and the presence of fractures in the proximal radioulnar region were correlated with the type of fracture-dislocation and demographic variables. Conclusion: Elbow fracture-dislocation types could not predict the number of fragments and the location of radial head fractures. However, most injuries presented three or more fragments in the radial head, and many had involvement of the proximal radioulnar region, suggesting high-energy trauma. Level of Evidence II; Retrospective Study

Artesunate ameliorates experimental autoimmune encephalomyelitis by inhibiting leukocyte migration to the central nervous system

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOExperimental autoimmune encephalomyelitis (EAE) is T-celldependent disease of the central nervous system (CNS) of mice. This model resembles multiple sclerosis (MS) in many aspects. Therapies that focus in the modulation of the immune response and cellular infiltration in the CNS present best effects in the clinics. Artesunate (Art) is a semi-synthetic sesquiterpene derivative from artemisinin and has been shown to reduce the clinical signs of autoimmune disease models through mechanisms not yet understood. In this study, we aimed to evaluate whether administration of Art would ameliorate EAE. Methods and Results: C57BL6 mice were immunized with MOG(35-55) peptide to induce EAE. At the same time, Art treatment started (3 mg/kg/day via i.p.) for five consecutive days. We found that Art treatment reduced the clinical signs of EAE and that correlated with a reduced infiltration of cells in the CNS. Disease amelioration did not correlate with immunomodulation as recall responses, leukocyte subpopulations, and gene expression analysis were similar among treated and untreated mice. Ultimately, further analysis provided data indicating that a possible mechanism of action for Art is dependent on the cellular migration to the CNS. Conclusions: Artesunate reduces the severity of EAE by inhibiting migration of pathogenic T cells to the CNS.Experimental autoimmune encephalomyelitis (EAE) is T-celldependent disease of the central nervous system (CNS) of mice. This model resembles multiple sclerosis (MS) in many aspects. Therapies that focus in the modulation of the immune response and cellula228707714FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO2014/19492-3; 2014/02631-0; 2014/11588-

An overview of the ant fauna (Hymenoptera: Formicidae) of the state of Maranhão, Brazil

The state of Maranhão, located in northeastern Brazil, comprises three biomes: Amazonian, Caatinga, and the Cerrado. To date, 99 ant species have been recorded in the literature from the state. In the present work, we provide for the first time a profile of the ant fauna in the state based on data from the historical literature and Brazilian institutional collections. The updated records on ant diversity for the state of Maranhão revealed a total of 279 species, belonging to 71 genera and 10 subfamilies. In total, 180 species are recorded for the first time in the state, of which four species recorded for the first time in Brazil. In summary, apart from documenting the ant fauna of the region, these results provide a basis for further studies and may contribute to future conservation efforts for the biomes present in this complex landscape

Construindo o conceito de derivada por meio do software Geogebra como estratégia de ensino e aprendizagem na educação superior

Este trabalho apresenta uma sequência de atividades para o ensino do conceito de derivada por meio do software Geogebra. A metodologia utilizada foram os princípios da engenharia didática. A sequencia de atividade foi desenvolvida a partir de uma proposta construtivista, diferente do ensino tradicional. A proposta foi aplicada a 11 alunos dos cursos de graduação da área de exatas. Os resultados apontaram que o desempenho dos alunos não foi o esperado. Apesar dos mesmos terem conseguido desenvolver as atividades, os alunos não conseguiram perceber o conceito de derivada, e acreditamos que isso ocorreu porque os participantes não estavam acostumados a este tipo de proposta de ensino, pois é muito diferente do ensino tradicional

Morphological evaluation of macrophage infected with Toxoplasma Gondii / Avaliação morfológica de macrófagos infectados com Toxoplasma Gondii

O Toxoplasma gondii é o parasita causador da toxoplasmose, doença negligenciada que ainda carece de estudos que visem elucidar a relação entre a parasitemia e o sistema imunológico. Uma das principais células do sistema imunológico são os macrófagos e estes possuem linhagens celulares imortalizadas que fornecem um modelo de estudo acessível para experimentos in vitro . Com isso, este trabalho pretende enfatizar a importância da linhagem de macrófagos J774G8 em estudos com protozoários, destacando a viabilidade e alterações morfológicas da cultura de células infectadas com Toxoplasma gondii