On the origin of the selectivity of plasmidic H-NS towards horizontally acquired DNA: Linking H-NS oligomerization and cooperative DNA binding

The nucleoid-associated protein H-NS is a global modulator of the expression of genes associated with adaptation to environmental changes. A variant of H-NS expressed in the R27 plasmid was previously shown to selectively modulate the expression of horizontally acquired genes, with minimal effects on core genes that are repressed by the chromosomal form of H-NS. Both H-NS proteins are formed by an oligomerization domain and a DNA-binding domain, which are connected by a linker that is highly flexible in the absence of DNA. We studied DNA binding by means of oligomer-forming chimeric proteins in which domains of the chromosomal and plasmidic variants are exchanged, as well as in monomeric truncated forms containing the DNA-binding domain and variable portions of the linker. Point mutations in the linker were also examined in full-length and truncated H-NS constructs. These experiments show that the linker region contributes to DNA binding affinity and that it is a main component of the distinct DNA binding properties of chromosomal and plasmidic H-NS. We propose that interactions between the linker and DNA limit the flexibility of the connection between H- NS oligomerization and DNA binding and provide an allosteric indirect readout mechanism to detect long- range distortions of DNA, thus enabling discrimination between core and horizontally acquired DNA.

Congenital Nystagmus Gene FRMD7 Is Necessary for Establishing a Neuronal Circuit Asymmetry for Direction Selectivity.

Neuronal circuit asymmetries are important components of brain circuits, but the molecular pathways leading to their establishment remain unknown. Here we found that the mutation of FRMD7, a gene that is defective in human congenital nystagmus, leads to the selective loss of the horizontal optokinetic reflex in mice, as it does in humans. This is accompanied by the selective loss of horizontal direction selectivity in retinal ganglion cells and the transition from asymmetric to symmetric inhibitory input to horizontal direction-selective ganglion cells. In wild-type retinas, we found FRMD7 specifically expressed in starburst amacrine cells, the interneuron type that provides asymmetric inhibition to direction-selective retinal ganglion cells. This work identifies FRMD7 as a key regulator in establishing a neuronal circuit asymmetry, and it suggests the involvement of a specific inhibitory neuron type in the pathophysiology of a neurological disease. VIDEO ABSTRACT.

Combining T2-preparation with spectral and spatial selectivity for improved coronary MRA

Cardiovascular disease is the leading cause of death worldwide. Within this subset, coronary artery disease (CAD) is the most prevalent. Magnetic resonance angiography (MRA) is an emerging technique that provides a safe, non-invasive way of assessing CAD progression. To generate contrast between tissues, MR images are weighted according to the magnetic properties of those tissues. In cardiac MRI, T2 contrast, which is governed by the rate of transverse signal loss, is often created through the use of a T2-Preparation module. T2-Preparation, or T2-Prep, is a magnetization preparation scheme used to improve blood/myocardium contrast in cardiac MRI. T2-Prep methods generally use a non-selective +90°, 180°, 180°, -90° train of radiofrequency (RF) pulses (or variant thereof), to tip magnetization into the transverse plane, allow it to evolve, and then to restore it to the longitudinal plane. A key feature in this process is the combination of a +90° and -90° RF pulse. By changing either one of these, a mismatch occurs between signal excitation and restoration. This feature can be exploited to provide additional spectral or spatial selectivity. In this work, both of these possibilities are explored. The first - spectral selectivity - has been examined as a method of improving fat saturation in coronary MRA. The second - spatial selectivity - has been examined as a means of reducing imaging time by decreasing the field of view, and as a method of reducing artefacts originating from the tissues surrounding the heart. Two additional applications, parallel imaging and self-navigation, are also presented. This thesis is thus composed of four sections. The first, "A Fat Signal Suppression for Coronary MRA at 3T using a Water-Selective Adiabatic T2-Preparation Technique", was originally published in the journal Magnetic Resonance in Medicine (MRM) with co-authors Ruud B. van Heeswijk and Matthias Stuber. The second, "Combined T2-Preparation and 2D Pencil Beam Inner Volume Selection", again with co-authors Ruud van Heeswijk and Matthias Stuber, was also published in the journal MRM. The third, "A cylindrical, inner volume selecting 2D-T2-Prep improves GRAPPA-accelerated image quality in MRA of the right coronary artery", written with co-authors Jerome Yerly and Matthias Stuber, has been submitted to the "Journal of Cardiovascular Magnetic Resonance", and the fourth, "Combined respiratory self-navigation and 'pencil-beam' 2D-T2 -Prep for free-breathing, whole-heart coronary MRA", with co¬authors Jerome Chaptinel, Giulia Ginami, Gabriele Bonanno, Simone Coppo, Ruud van Heeswijk, Davide Piccini, and Matthias Stuber, is undergoing internal review prior to submission to the journal MRM. -- Les maladies cardiovasculaires sont la cause principale de décès dans le monde : parmi celles-ci, les maladies coronariennes sont les plus répandues. L'angiographie par résonance magnétique (ARM) est une technique émergente qui fournit une manière sûre, non invasive d'évaluer la progression de la coronaropathie. Pour obtenir un contraste entre les tissus, les images d'IRM sont pondérées en fonction des propriétés magnétiques de ces tissus. En IRM cardiaque, le contraste en T2, qui est lié à la décroissance du signal transversal, est souvent créé grâce à l'utilisàtion d'un module de préparation T2. La préparation T2, ou T2-Prep, est un système de préparation de l'aimantation qui est utilisé pour améliorer le contraste entre le sang et le myocarde lors d'une IRM cardiaque. Les méthodes de T2-Prep utilisent généralement une série non-sélective d'impulsions de radiofréquence (RF), typiquement [+ 90°, 180°, 180°, -90°] ou une variante, qui bascule l'aimantation dans le plan transversal, lui permet d'évoluer, puis la restaure dans le plan longitudinal. Un élément clé de ce processus est la combinaison des impulsions RF de +90° et -90°. En changeant l'une ou l'autre des impulsions, un décalage se produit entre l'excitation du signal et de la restauration. Cette fonction peut être exploitée pour fournir une sélectivité spectrale ou spatiale. Dans cette thèse, les deux possibilités sont explorées. La première - la sélectivité spectrale - a été examinée comme une méthode d'améliorer la saturation de la graisse dans l'IRM coronarienne. La deuxième - la sélectivité spatiale - a été étudiée comme un moyen de réduire le temps d'imagerie en diminuant le champ de vue, et comme une méthode de réduction des artefacts provenant des tissus entourant le coeur. Deux applications supplémentaires, l'imagerie parallèle et la self-navigation, sont également présentées. Cette thèse est ainsi composée de quatre sections. La première, "A Fat Signal Suppression for Coronary MRA at 3T using a Water-Selective Adiabatic T2-Preparation Technique", a été publiée dans la revue médicale Magnetic Resonance .in Medicine (MRM) avec les co-auteurs Ruud B. van Heeswijk et Matthias Stuber. La deuxième, Combined T2-Preparation and 2D Pencil Beam Inner Volume Selection", encore une fois avec les co-auteurs Ruud van Heeswijk et Matthias Stuber, a également été publiée dans le journal MRM. La troisième, "A cylindrical, inner volume selecting 2D-T2-Prep improves GRAPPA- accelerated image quality in MRA of the right coronary artery", écrite avec les co-auteurs Jérôme Yerly et Matthias Stuber, a été présentée au "Journal of Cardiovascular Magnetic Resonance", et la quatrième, "Combined respiratory self-navigation and 'pencil-beam' 2D-T2 -Prep for free-breathing, whole-heart coronary MRA", avec les co-auteurs Jérôme Chaptinel, Giulia Ginami, Gabriele Bonanno , Simone Coppo, Ruud van Heeswijk, Davide Piccini, et Matthias Stuber, subit un examen interne avant la soumission à la revue MRM.

Ion-selective electrodes: historical, mechanism of response, selectivity and concept review

This paper presents a review of the concepts involved in the working mechanism of the ion-selective electrodes, searching a historical overview, moreover to describe the new advances in the area.

Chemical study of leaves of Chrysophyllum marginatum (Hook. & Arn.) Radlk (Sapotaceae)

The fractionation of the antioxidant ethyl acetate extract obtained from the dried leaves of Chrysophyllum marginatum afforded six substances identified as: alpha-amirin, gallic acid, myricitrin, quercitrin, (-)-epigallocatechin and (-)-epigallocatechin-3-O-gallate. This study contributes to the knowledge of the secondary metabolites produced by one more species of the Brazilian Flora, until now not investigated. Moreover, this study allowed the identification of three substances with antioxidant activity previously detected in this species.

Specificity and selectivity improvement in doping analysis using comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry

Comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry was used for the identification of forty doping agents. The improvement in the specificity was remarkable, allowing the resolution of analytes that could not be done by one-dimensional chromatographic systems. The sensitivity observed for different classes of prohibited substances was clearly below the value required by the World Anti-Doping Agency. In addition time-of-flight mass spectrometry gives full spectrum for all analytes without any interference from the matrix, resulting in selectivity improvements. These results could support the implementation of an exhaustive monitoring approach for hundreds of doping agents in a single injection.

Resistência natural da madeira de Corymbia maculata (Hook.) K.D.Hill & L.A.S. Johnson a fungos e cupins xilófagos, em condições de laboratório

A pesquisa teve o objetivo de avaliar a resistência natural da madeira de Corymbia maculata a fungos e a cupins xilófagos, em condições de laboratório. De peças radiais (tábuas) que continham o cerne e o alburno intactos foram retirados corpos-de-prova de 2,00 x 2,00 x 1,00 cm, com a menor dimensão na direção tangencial (ensaio com fungos), e de 2,54 x 2,00 x 0,64 cm, com a maior dimensão na direção das fibras (ensaio com cupins), em quatro posições na direção medula-casca. As amostras foram submetidas à ação dos fungos Postia placenta, Neolentinus lepideus e Polyporus fumosus por 12 semanas, ou à ação de cupins do gênero Nasutitermes por 30 dias. Constatou-se que a resistência da madeira ao apodrecimento foi dependente da posição na direção medula-casca e dos fungos utilizados. As amostras retiradas nas posições mais externas do tronco foram mais deterioradas que as internas. Dentro de cada posição, os fungos causaram deterioração semelhante à madeira, exceto para a posição mais externa (alburno), em que o fungo P. fumosus causou menos deterioração que os demais. De modo geral, a madeira de C. maculata foi altamente resistente (posições internas) ou resistente (posições externas) aos fungos ensaiados. Somente para o fungo N. lepideus a posição mais externa foi moderadamente resistente. Quanto aos cupins, a resistência da madeira não foi afetada pela posição na direção medula-casca e apresentou uma baixa perda de massa para as posições analisadas. Além disto, os cupins causaram somente desgaste superficial à madeira, e morreram durante o ensaio, o que permitiu classificar a madeira de C.maculata como resistente aos cupins ensaiados.

Caracterização morfológica, viabilidade e vigor de sementes de Tabebuia aurea (Silva Manso) Benth. & Hook. f. ex. S. Moore

A espécie Tabebuia aurea é encontrada em áreas de Cerrado, Caatinga, Floresta Amazônica e Pantanal, sendo utilizada para fins ornamentais, medicinais, construção civil, carpintaria e produção de carvão, entre outros. Por apresentar ampla utilização e distribuição, os objetivos deste estudo foram caracterizar morfologicamente e analisar a viabilidade e o vigor das sementes dessa espécie quando colhidas e armazenadas por diferentes períodos. As sementes foram germinadas recém-colhidas, 30, 60 e 90 dias após o armazenamento em laboratório e em campo, sendo também submetidas ao teste de tetrazólio. As sementes mediram, em média, 57,8 x 20,6 x 3,1 mm (com alas) e 17,3 x 13,3 x 1,7 mm (sem alas), sendo assimétricas. Seu peso médio foi de 13,18 g (com alas) e 11,55 g (sem alas). As sementes apresentaram maior viabilidade quando recém-colhidas, maior porcentagem de germinação em laboratório após 30 dias de armazenamento e maior porcentagem de germinação acumulada a campo quando semeadas na superfície. A viabilidade das sementes, medida através do teste de tetrazólio, apresentou resultados similares aos obtidos por meio do teste de germinação em laboratório, indicando ser esse teste adequado para medir a viabilidade desta espécie.

Superação da dormência das sementes de Delonix regia (Bojer ex Hook.) Raf.

Delonix regia (flamboyant) é uma espécie arbórea amplamente utilizada no Brasil, de alto valor ornamental. Sua propagação ocorre por meio de sementes, que apresentam dormência pela impermeabilidade do tegumento à água. Com o objetivo de determinar a melhor metodologia para a superação da dormência, as sementes da espécie foram submetidas a diferentes tratamentos: 1) imersão em água fervente por 1 min; 2) imersão em água a 80 °C e a 90 °C por 1 e 3 min; 3) escarificação manual com lixa; e 4) imersão em ácido sulfúrico concentrado por 15, 30, 45 e 60 min, além da testemunha. Após cada tratamento, as sementes foram colocadas para germinar em rolo de papel, na temperatura de 25 ºC. Em uma segunda etapa, os tratamentos mais efetivos na quebra de dormência das sementes foram aplicados e a germinação, testada à temperatura de 30 °C. Em ambas as temperaturas, as contagens de germinação (plântulas normais) foram realizadas diariamente. O tratamento das sementes de flamboyant em água quente a 90 ºC por 1 min foi o mais eficiente na promoção da germinação, sendo prático e dispensando o uso de tratamentos químicos. O teste de germinação realizado à temperatura de 30 °C não forneceu resultados satisfatórios, com menores porcentagens de plântulas normais e IVG que os testes a 25 ºC. Foram descritas como plântulas normais as que possuíam visíveis dois cotilédones semiabertos, hipocótilo alongado, raiz primária bem desenvolvida e raízes adventícias curtas. Conclui-se que sementes de Delonix regia germinam melhor a 25 ºC após a imersão por 1 min em água a 90 ºC, sendo este indicado como tratamento para superação da dormência da espécie.

VALORES LIMÍTROFES PARA TRANSPIRAÇÃO, DESENVOLVIMENTO E CRESCIMENTO DE Corymbia citriodora (Hook.) K.D. Hill & L.A.S. Johnson EM RESPOSTA À DEFICIÊNCIA HÍDRICA NO SOLO1

RESUMO Quando a disponibilidade de água no solo é reduzida, as plantas respondem diminuindo a taxa de transpiração, o crescimento e o desenvolvimento, na tentativa de aclimatação à deficiência hídrica. Com o objetivo de quantificar essas respostas em mudas de Corymbia citriodora (Hook.) K.D. Hill & L.A.S. Johnson, foi utilizada a metodologia da fração de água transpirável no solo (FATS), em um experimento conduzido em casa de vegetação, sob o delineamento inteiramente casualizado, sendo dois níveis de suplementação hídrica, duas épocas de aplicação da deficiência hídrica e nove repetições. A FATS e os parâmetros de transpiração, crescimento e desenvolvimento foram medidos diariamente durante a aplicação da deficiência hídrica. A FATS crítica, em que a transpiração, o crescimento e desenvolvimento começam a ser reduzidos, foi distinta nas épocas de aplicação da deficiência hídrica devido à diferença das condições meteorológicas. Os valores elevados de FATS indicaram que a espécie possuiu boa aclimatação à deficiência hídrica no solo, quando comparada com as espécies anuais e outras perenes.

Linfadenectomia no câncer gástrico com hook laparotômico

O câncer gástrico é uma neoplasia frequente e de tratamento cirúrgico. Gastrectomia com linfadenectomia D2 é o procedimento padrão com intuito curativo. Os autores relatam o uso do hook laparotômico como alternativa técnica para linfadenectomia.

Selectivity of glyphosate in tank mixtures for RR soybean in sequential applications with mixtures only in the first or second application

Tank mixtures among herbicides of different action mechanisms might increase weed control spectrum and may be an important strategy for preventing the development of resistance in RR soybean. However, little is known about the effects of these herbicide combinations on soybean plants. Hence, two experiments were carried out aiming at evaluating the selectivity of glyphosate mixtures with other active ingredients applied in postemergence to RR soybean. The first application was carried out at V1 to V2 soybean stage and the second at V3 to V4 (15 days after the first one). For experiment I, treatments (rates in g ha-1) evaluated were composed by two sequential applications: the first one with glyphosate (720) in tank mixtures with cloransulam (30.24), fomesafen (125), lactofen (72), chlorimuron (12.5), flumiclorac (30), bentazon (480) and imazethapyr (80); the second application consisted of isolated glyphosate (480). In experiment II, treatments also consisted of two sequential applications, but tank mixtures as described above were applied as the second application. The first one in this experiment consisted of isolated glyphosate (720). For both experiments, sequential applications of glyphosate alone at 720/480, 960/480, 1200/480 and 960/720 (Expt. I) or 720/480, 720/720, 720/960 and 720/1200 (Expt. II) were used as control treatments. Applications of glyphosate tank mixtures with other herbicides are more selective to RR soybean when applied at younger stages whereas applications at later stages might cause yield losses, especially when glyphosate is mixed with lactofen and bentazon.

Selectivity of glyphosate tank mixtures for RR soybean

An active ingredients mixture of different action mechanisms is an essential tool to prevent or manage areas with resistant weeds. However, it is important that such a mixture provides adequate selectivity to the crop. The aim of this work was to evaluate glyphosate selectivity to glyphosate-resistant (RR) soybean, and also verify if there is selectivity in mixtures with other active ingredients applied postemergence aimed at new control strategies, which might be used in RR soybean cultivation. The herbicides and respective rates (g ha-1) evaluated were: glyphosate (720, 960, 1,200, and 1,440), and the mixtures of glyphosate (960) with cloransulam-methyl (30.24), fomesafen (125), lactofen (72), chlorimuron-ethyl (12.5), flumiclorac-pentyl (30), bentazon (480), or imazethapyr (80). All treatments were applied in postemergence when the soybean crop was at V2 to V3 stage. Treatments with glyphosate or in mixtures with postemergent herbicides showed visual effects of phytotoxicity when applied to the glyphosate-resistant soybean. Effects such as reduction in plant height, crop closure, number of pods per plant, and hundred grain weight could be observed. However, the effects related to plant development were mostly transient and did not persist during the crop cycle. Among the studied treatments, only the mixture of glyphosate and lactofen was not selective to the crop, promoting negative effects on most characteristics analyzed and consequently reducing grain yield.

Herbicide selectivity in alfalfa crops

This study aimed to determine the selectivity of herbicides applied in pre- and post-emergence for alfalfa crops. Three separate experiments were carried out under greenhouse conditions. The first experiment was arranged in a completely randomized design with three replications in a 4 x 11 + 1 factorial scheme , with eleven herbicides (bentazon, chlorimuron-ethyl, fomesafen, fluazifop-p-butyl, saflufenacil, imazethapyr, clethodim, nicosulfuron, imazaquin, haloxyfop-methyl and MSMA), four doses of each herbicide (0.5 D, 0.75 D, 1.0 D and 1.25 D, where D = recommended dose), plus an untreated control. The products were applied to alfalfa plants at the stage of 4 to 5 leaf pairs. In the second experiment, the effect of pre-emergent herbicides on early alfalfa development was observed through a completely randomized design with five replications in a 3 x 4 x 2 factorial scheme, with three herbicides (hexazinone, atrazine + simazine, S-metolachlor), four doses (0.5 D, 0.75 D, 1.0 D and 1.25 D), and two types of soil texture (loamy and clay soil), plus an untreated control. The third experiment evaluated the action of atrazine, 2,550 g ha-1; clomazone - 600 g ha-1; diclosulam - 25 g ha-1; diuron+hexazinone - 936 + 264 g ha-1 and diuron+hexazinone +sulfometuron - 1,386 + 391 + 33.35 g ha-1 on alfalfa sown at different times after herbicide application. The effects of the treatments on alfalfa were evaluated according to visual phytotoxicity symptoms, plant height, and biomass of roots and shoots. Among the herbicides applied at post-emergence, imazethapyr, clethodim, haloxyfop-p-methyl and MSMA were selective for alfalfa, while among those applied at pre-emergence, none were selective, regardless of soil texture. The results of the third experiment showed that the herbicides diclosulam, hexazinone + diuron and atrazine caused less toxicity in alfalfa plants.