Regulation of antibiotic production in root-colonizing Peudomonas spp. and relevance for biological control of plant disease.

Certain strains of fluorescent pseudomonads are important biological components of agricultural soils that are suppressive to diseases caused by pathogenic fungi on crop plants. The biocontrol abilities of such strains depend essentially on aggressive root colonization, induction of systemic resistance in the plant, and the production of diffusible or volatile antifungal antibiotics. Evidence that these compounds are produced in situ is based on their chemical extraction from the rhizosphere and on the expression of antibiotic biosynthetic genes in the producer strains colonizing plant roots. Well-characterized antibiotics with biocontrol properties include phenazines, 2,4-diacetylphloroglucinol, pyoluteorin, pyrrolnitrin, lipopeptides, and hydrogen cyanide. In vitro, optimal production of these compounds occurs at high cell densities and during conditions of restricted growth, involving (i) a number of transcriptional regulators, which are mostly pathway-specific, and (ii) the GacS/GacA two-component system, which globally exerts a positive effect on the production of extracellular metabolites at a posttranscriptional level. Small untranslated RNAs have important roles in the GacS/GacA signal transduction pathway. One challenge in future biocontrol research involves development of new strategies to overcome the broad toxicity and lack of antifungal specificity displayed by most biocontrol antibiotics studied so far.

Performance of Poly-Carb, Inc. Flexogrid Bridge Overlay System, MLR-86-04, 2001

Intrusion of deicing materials and surface water into concrete bridge decks is a main contributor in deck reinforcing steel corrosion and concrete delamination. Salt, spread on bridge decks to melt ice, dissolves in water and permeates voids in the concrete deck. When the chloride content of the concrete in contact with reinforcing steel reaches a high enough concentration, the steel oxidizes. In Iowa, the method used to reduce bridge deck chloride penetration is the application of a low slump dense concrete overlay after the completion of all Class A and Class B floor repairs. A possible alternative to the use of dense concrete overlays, developed by Poly-Carb, Inc., is the MARK-163 FLEXOGRID Overlay System. FLEXOGRID is a two component system of epoxy and urethane which is applied on a bridge deck to a minimum thickness of ¼ inch. An aggregate mixture of silica quartz and aluminum oxide is broadcast onto the epoxy at a prescribed rate to provide deck protection and superior friction properties. The material is mixed on site and applied to the deck in a series of lifts (usually two) until the desired overlay thickness has been attained.

Mechanism of hcnA mRNA recognition in the Gac/Rsm signal transduction pathway of Pseudomonas fluorescens.

In the plant-beneficial bacterium Pseudomonas fluorescens CHA0, the expression of antifungal exoproducts is controlled by the GacS/GacA two-component system. Two RNA binding proteins (RsmA, RsmE) ensure effective translational repression of exoproduct mRNAs. At high cell population densities, GacA induces three small RNAs (RsmX, RsmY, RsmZ) which sequester both RsmA and RsmE, thereby relieving translational repression. Here we systematically analyse the features that allow the RNA binding proteins to interact strongly with the 5' untranslated leader mRNA of the P. fluorescens hcnA gene (encoding hydrogen cyanide synthase subunit A). We obtained evidence for three major RsmA/RsmE recognition elements in the hcnA leader, based on directed mutagenesis, RsmE footprints and toeprints, and in vivo expression data. Two recognition elements were found in two stem-loop structures whose existence in the 5' leader region was confirmed by lead(II) cleavage analysis. The third recognition element, which overlapped the hcnA Shine-Dalgarno sequence, was postulated to adopt either an open conformation, which would favour ribosome binding, or a stem-loop structure, which may form upon interaction with RsmA/RsmE and would inhibit access of ribosomes. Effective control of hcnA expression by the Gac/Rsm system appears to result from the combination of the three appropriately spaced recognition elements.

Brittle to ductile transition in a fiber bundle with strong heterogeneity

We analyze the failure process of a two-component system with widely different fracture strength in the framework of a fiber bundle model with localized load sharing. A fraction 0≤α≤1 of the bundle is strong and it is represented by unbreakable fibers, while fibers of the weak component have randomly distributed failure strength. Computer simulations revealed that there exists a critical composition αc which separates two qualitatively different behaviors: Below the critical point, the failure of the bundle is brittle, characterized by an abrupt damage growth within the breakable part of the system. Above αc, however, the macroscopic response becomes ductile, providing stability during the entire breaking process. The transition occurs at an astonishingly low fraction of strong fibers which can have importance for applications. We show that in the ductile phase, the size distribution of breaking bursts has a power law functional form with an exponent μ=2 followed by an exponential cutoff. In the brittle phase, the power law also prevails but with a higher exponent μ=92. The transition between the two phases shows analogies to continuous phase transitions. Analyzing the microstructure of the damage, it was found that at the beginning of the fracture process cracks nucleate randomly, while later on growth and coalescence of cracks dominate, which give rise to power law distributed crack sizes.

Monooxygenase Diversity and Oxygen Activation within Polyketide Antibiotic Biosynthesis

Molecular oxygen (O2) is a key component in cellular respiration and aerobic life. Through the redox potential of O2, the amount of free energy available to organisms that utilize it is greatly increased. Yet, due to the nature of the O2 electron configuration, it is non-reactive to most organic molecules in the ground state. For O2 to react with most organic compounds it must be activated. By activating O2, oxygenases can catalyze reactions involving oxygen incorporation into organic compounds. The oxygen activation mechanisms employed by many oxygenases to have been studied, and they often include transition metals and selected organic compounds. Despite the diversity of mechanisms for O2 activation explored in this thesis, all of the monooxygenases studied in the experimental part activate O2 through a transient carbanion intermediate. One of these enzymes is the small cofactorless monooxygenase SnoaB. Cofactorless monooxygenases are unusual oxygenases that require neither transition metals nor cofactors to activate oxygen. Based on our biochemical characterization and the crystal structure of this enzyme, the mechanism most likely employed by SnoaB relies on a carbanion intermediate to activate oxygen, which is consistent with the proposed substrate-assisted mechanism for this family of enzymes. From the studies conducted on the two-component system AlnT and AlnH, both the functions of the NADH-dependent flavin reductase, AlnH, and the reduced flavin dependent monooxygenase, AlnT, were confirmed. The unusual regiochemistry proposed for AlnT was also confirmed on the basis of the structure of a reaction product. The mechanism of AlnT, as with other flavin-dependent monooxygenases, is likely to involve a caged radical pair consisting of a superoxide anion and a neutral flavin radical formed from an initial carbanion intermediate. In the studies concerning the engineering of the S-adenosyl-L-methionine (SAM) dependent 4-O-methylase DnrK and the homologous atypical 10-hydroxylase RdmB, our data suggest that an initial decarboxylation of the substrate is catalyzed by both of these enzymes, which results in the generation of a carbanion intermediate. This intermediate is not essential for the 4-O-methylation reaction, but it is important for the 10-hydroxylation reaction, since it enables substrate-assisted activation of molecular oxygen involving a single electron transfer to O2 from a carbanion intermediate. The only role for SAM in the hydroxylation reaction is likely to be stabilization of the carbanion through the positive charge of the cofactor. Based on the DnrK variant crystal structure and the characterizations of several DnrK variants, the insertion of a single amino acid in DnrK (S297) is sufficient for gaining a hydroxylation function, which is likely caused by carbanion stabilization through active site solvent restriction. Despite large differences in the three-dimensional structures of the oxygenases and the potential for multiple oxygen activation mechanisms, all the enzymes in my studies rely on carbanion intermediates to activate oxygen from either flavins or their substrates. This thesis provides interesting examples of divergent evolution and the prevalence of carbanion intermediates within polyketide biosynthesis. This mechanism appears to be recurrent in aromatic polyketide biosynthesis and may reflect the acidic nature of these compounds, propensity towards hydrogen bonding and their ability to delocalize π-electrons.

Efectos de la simpatectomía torácica sobre la pletismografía de los miembros superiores en sujetos con hiperhidrosis palmar

Introducción: La simpatectomía es el tratamiento de la hiperhidrosis palmar y consiste en denervación simpática de miembros superiores que produce un efecto en el flujo sanguíneo al impactar la respuesta vasoconstrictora. El cambio en el flujo sanguíneo se puede evaluar a través de la onda fotopletismográfica. Metodología: Se realizaron 2 sesiones (presimpatectomía y postsimpatectomía) de 10 minutos en cada miembro superior en 28 pacientes obteniendo 79 señales fotopletismográficas distribuidas así: 37 presimpatectomía y 42 señales postsimpatectomía. De cada señal se analizó 1.5 minutos donde se tienen 80 ondas de fotopletismografía y se miden 6 variables: 1. Componente AC (componente pulsátil), 2. Componente DC (componente no pulsátil), 3. Relación entre AC/DC (índice de perfusión), 4. Area bajo la curva (AUC), 5. Tiempo entre el inicio de la onda y pico sistólico (T_DA) y 6. Tiempo entre cada onda de pulso (T_DD). Resultados: Aumentó 120% el componente AC y disminuyó 78% en DC del miembro superior derecho (MSD) con 99% de confiabilidad (p<0.001) entre presimpatectomía (n=18) y postsimpatectomía (n=21). AC/DC aumentó 55% con 95% de confiabilidad entre presimpatectomía (n=19) y postsimpatectomía (n=21) en el miembro superior izquierdo (p<0.05). No se encontró diferencia para T_DA, T_DD ni AUC. Discusión y Conclusión: La simpatectomía en pacientes con hiperhidrosis palmar produce un cambio en el flujo sanguíneo de los miembros superiores evidenciado por los cambios en el componente no pulsátil (DC) y pulsátil (AC) que es secundario a la vasodilatación consecuencia del bloqueo simpático por la denervación quirúrgica de los ganglios simpáticos torácicos.

A direct comparison of one- and two-component dendritic self-assembled materials: elucidating molecular recognition pathways

This paper compares and contrasts, for the first time, one- and two-component gelation systems that are direct structural analogues and draws conclusions about the molecular recognition pathways that underpin fibrillar self-assembly. The new one-component systems comprise L-lysine-based dendritic headgroups covalently connected to an aliphatic diamine spacer chain via an amide bond, One-component gelators with different generations of headgroup (from first to third generation) and different length spacer chains are reported. The self-assembly of these dendrimers in toluene was elucidated using thermal measurements, circular dichroism (CD) and NMR spectroscopies, scanning electron microscopy (SEM), and small-angle X-ray scattering (SAXS). The observations are compared with previous results for the analogous two-component gelation system in which the dendritic headgroups are bound to the aliphatic spacer chain noncovalently via acid-amine interactions. The one-component system is inherently a more effective gelator, partly as a consequence of the additional covalent amide groups that provide a new hydrogen bonding molecular recognition pathway, whereas the two-component analogue relies solely on intermolecular hydrogen bond interactions between the chiral dendritic headgroups. Furthermore, because these amide groups are important in the assembly process for the one-component system, the chiral information preset in the dendritic headgroups is not always transcribed into the nanoscale assembly, whereas for the two-component system, fiber formation is always accompanied by chiral ordering because the molecular recognition pathway is completely dependent on hydrogen bond interactions between well-organized chiral dendritic headgroups.

A study of the relaxation dynamics in a quadrupolar NMR system using Quantum State Tomography

This article reports a relaxation study in an oriented system containing spin 3/2 nuclei using quantum state tomography (QST). The use of QST allowed evaluating the time evolution of all density matrix elements starting from several initial states. Using an appropriated treatment based on the Redfield theory, the relaxation rate of each density matrix element was measured and the reduced spectral densities that describe the system relaxation were determined. All the experimental data could be well described assuming pure quadrupolar relaxation and reduced spectral densities corresponding to a superposition of slow and fast motions. The data were also analyzed in the context of Quantum Information Processing, where the coherence loss of each qubit of the system was determined using the partial trace operation. (C) 2008 Elsevier Inc. All rights reserved.

Human pulp response after an adhesive system application in deep cavities

Objectives: To evaluate the pulpo-dentin complex response to a dentin adhesive application in deep cavities performed in human teeth.Methods: Deep class V cavities were prepared on the buccal surface of 46 premolars. The remaining dentin of the axial wall received 10% phosphoric acid and dentin adhesive (group DA), or was protected before the acid and dentin adhesive application with calcium hydroxide cement (group CH). Half of the teeth, which received the acid application directly over the axial wall, were contaminated prior to the procedures with dental plaque collected from the patient's own teeth (group DAC). The plaque was placed on the dentin for 5 min and then the cavity was washed. All teeth were restored with a light-cured composite resin. The teeth were extracted after 7, 30 or 60 days and prepared according to normal histologic techniques. Serial sections were stained with WE, Masson's trichrome and Brown & Brenn technique for demonstration of bacteria.Results: the histopathologic evaluation showed that in groups DA and DAC, the inflammatory response was more evident than in group CH. Also, the intensity of the pulp reaction increased as the remaining dentin thickness decreased. There was no statistical difference in the inflammatory response between the groups DA and DAC.Conclusion: Based on the experimental conditions, we concluded that the All Bond 2 adhesive system, when applied on dentin in deep cavities, showed an acceptable biocompatibility. However, the intensity of the pulpo-dentin complex response depends on the remaining dentin thickness. (C) 1999 Elsevier B.V. Ltd. All rights reserved.

On the N=2 superstring BRST operator

We show that the BRST charge for the N = 2 superstring system can be written as Q = e(-R)(phi dz/2 pi ib gamma(+)gamma(-))e(R), when b and gamma(+/-) are super-reparametrizations ghosts. This provides a trivial proof of the nilpotence of this operator. (C) 2000 Published by Elsevier B.V. B.V. All rights reserved.

Low-voltage varistor based on (Sn,Ti)O-2 ceramics

A description is given of the nonohmic behavior obtained in (SnxTi1-x)O-2-based systems. A matrix founded on (SnxTi1-x)O-2-based systems doped with Nb2O5 leads to a low-voltage varistor system with nonlinear coefficient values of similar to9. The presence of the back-to-back Schottky-type barrier is observed based on the voltage dependence of the capacitance. When doped with CoO, the (SnxTi1-x)O(2)(.)based system presents higher nonlinear coefficient values (>30) than does the SnO2-based varistor system.

Bond strength of adhesive systems to human tooth enamel

The purpose of this study was to evaluate in vitro three adhesive systems: a total etching single-component system (G1 Prime & Bond 2.1), a self-etching primer (G2 Clearfil SE Bond), and a self-etching adhesive (G3 One Up Bond F), through shear bond strength to enamel of human teeth, evaluating the type of fracture through stereomicroscopy, following the ISO guidance on adhesive testing. Thirty sound premolars were bisected mesiodistally and the buccal and lingual surfaces were embedded in acrylic resin, polished up to 600-grit sandpapers, and randomly assigned to three experimental groups (n = 20). Composite resin cylinders were added to the tested surfaces. The specimens were kept in distilled water (37°C/24 h), thermocycled for 500 cycles (5°C-55°C) and submitted to shear testing at a crosshead speed of 0.5 mm/min. The type of fracture was analyzed under stereomicroscopy and the data were submitted to Anova, Tukey and Chi-squared (5%) statistical analyses. The mean adhesive strengths were G1: 18.13 ± 6.49 MPa, (55% of resin cohesive fractures); G2: 17.12 ± 5.80 MPa (90% of adhesive fractures); and G3: 10.47 ± 3.14 MPa (85% of adhesive fractures). In terms of bond strength, there were no significant differences between G1 and G2, and G3 was significantly different from the other groups. G1 presented a different type of fracture from that of G2 and G3. In conclusion, although the total etching and self-etching systems presented similar shear bond strength values, the types of fracture presented by them were different, which can have clinical implications.

Influência in vitro da clorexidina e do chá verde na longevidade de união adesiva resina composta – dentina

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estratigrafia de seqüências do Grupo Itararé (Neocarbonifero-Eopermiano) na região de Rio Negro (PR) - Mafra (SC)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Metodologia baseada em sistema fuzzy intervalar do tipo-2 para detecção e identificação de faltas de incipientes em motores de indução

Desde a incorporação da automação no processo produtivo, a busca por sistemas mais eficientes, objetivando o aumento da produtividade e da qualidade dos produtos e serviços, direcionou os estudos para o planejamento de estratégias que permitissem o monitoramento de sistemas com o intuito principal de torna-los mais autônomos e robustos. Por esse motivo, as pesquisas envolvendo o diagnóstico de faltas em sistemas industriais tornaram-se mais intensivas, visto a necessidade da incorporação de técnicas para monitoramente detalhado de sistemas. Tais técnicas permitem a verificação de perturbações, falta ou mesmo falhas. Em vista disso, essa trabalho investiga técnicas de detecção e diagnostico de faltas e sua aplicação em motores de indução trifásicos, delimitando o seu estudo em duas situações: sistemas livre de faltas, e sobre atuação da falta incipiente do tipo curto-circuitoparcial nas espiras do enrolamento do estator. Para a detecção de faltas, utilizou-se analise paramétrica dos parâmetros de um modelo de tempo discreto, de primeira ordem, na estrutura autoregressivo com entradas exógenas (ARX). Os parâmetros do modelo ARX, que trazem informação sobre a dinâmica dominante do sistema, são obtidos recursivamente pela técnica dos mínimos quadrados recursivos (MQR). Para avaliação da falta, foi desenvolvido um sistema de inferência fuzzy (SIF) intervala do tipo-2, cuja mancha de incerteza ou footprint of uncertainty (FOU), características de sistema fuzzy tipo-2, é ideal como forma de representar ruídos inerentes a sistemas reais e erros numéricos provenientes do processo de estimação paramétrica. Os parâmetros do modelo ARX são entradas para o SIF. Algoritmos genéricos (AG’s) foram utilizados para otimização dos SIF intervalares tipo-2, objetivando reduzir o erro de diagnóstico da falta identificada na saída desses sistemas. Os resultados obtidos em teste de simulação computacional demonstram a efetividade da metodologia proposta.