Active-site properties of Phrixotrix railroad worm green and red bioluminescence-eliciting luciferases

The luciferases of the railroad worm Phrixotrix (Coleoptera: Phengodidae) are the only beetle luciferases that naturally produce true red bioluminescence. Previously, we cloned the green- (PxGR) and red-emitting (PxRE) luciferases of railroad worms Phrixotrix viviani and P. hirtus[OLE1]. These luciferases were expressed and purified, and their active-site properties were determined. The red-emitting PxRE luciferase displays flash-like kinetics, whereas PxGR luciferase displays slow-type kinetics. The substrate affinities and catalytic efficiency of PxRE luciferase are also higher than those of PxGR luciferase. Fluorescence studies with 8-anilino-1-naphthalene sulfonic acid and 6-p-toluidino-2-naphthalene sulfonic acid showed that the PxRE luciferase luciferin-binding site is more polar than that of PxGR luciferase, and it is sensitive to guanidine. Alutagenesis and modelling studies suggest that several invariant residues in the putative luciferin-binding site of PxRE luciferase cannot interact with excited oxyluciferin. These results suggest that one portion of the luciferin-binding site of the red-emitting luciferase is tighter than that of PxGR luciferase, whereas the other portion could be more open and polar.

The influence of the region between residues 220 and 344 and beyond in Phrixotrix railroad worm luciferases green and red bioluminescence

To find the regions having a major influence on the bioluminescence spectra of railroad worm luciferases, we constructed new chimeric luciferases switching the fragments from residues 1-219 and from 220-545 between Phrixotrix viviani (PxvGR; λmax = 548 nm) green light-emitting luciferase and Phrixothrix hirtus (PxhRE; λmax = 623 nm) red light-emitting luciferases. The emission spectrum (λmax = 571 nm) and KM for luciferin in the chimera PxRE220GR (1-219, PxhRE; 220-545, PxvGR) suggested that the region above residue 220 of PxvGR had a major effect on the active site. However, switching the sequence between the residues 226-344 from PxvGR luciferase into PxhRE (PxREGRRE) luciferase resulted in red light emission (λmax = 603 nm), indicating that the region 220-344 by itself does not determine the emission spectrum. Furthermore, the sequence before residue 220 of the green-emitting luciferase is incompatible for light emission with the sequence above residue 220 of PxhRE. These results suggest that the fragments before and after residue 220, which correspond to distinct subdomains, may fold differently in the green- and red-emitting luciferases, affecting the active site conformation.

Method for estimating railroad track settlements due to dynamic traffic loads

This paper is the result of real-scale physical modeling study designed to simulate the load-deformation characteristics of railroad foundation systems that include the railroad ties, the ballast, and the sub-base layers of a railroad embankment. The study presents comparisons of the application of dynamic loads of 100kN on the rails, and the resulting deformations during a 500,000 cycle testing period for three rail support systems; wood, concrete and steel. The results show that the deformation curve has an exponential shape, with the larger portion of the deformation occurring during the first 50,000 load cycles followed by a tendency to stabilize between 100,000 to 500,000 cycles. These results indicate that the critical phase of deformations of a new railroad is within the first 50,000 cycles of loading, and after that, it slowly attenuates as it approaches a stable value. The paper also presents empirically derived formulations for the estimation of the deformations of the rail supports as a result of rail traffic.

Benchmarking nos processos de gestão de qualidade entre dois serviços de hemmoterapia

Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB

Avaliação não-destrutiva de madeiras para dormentes ferroviários

Pós-graduação em Agronomia (Energia na Agricultura) - FCA

Tecnologias de Informação e Comunicação no cotidiano dos adolescentes: enfoque no comportamento e na competências informacionais da geração Google

Introduction: Individuals born from the 1990’s to accompany technological change course and learn how to use the technological resources that have access before they even learn to read or write. In this sense, the literature considers the digital natives or even 'Google generation'. However, it has been questioned their skills to identify information needs and seeking and use of information in the most varied environments digital information available on the Web. Objectives: To characterize the behavior of information seeking in everyday life of a group of teenage students from a private school in Marilia, Sao Paulo. Methodology: A qualitative study with 30 adolescent students of a private school in Marilia, Sao Paulo. Data collection was accomplished at first by means of a questionnaire, based on the research of Hughes-Hussell and August (2007), containing questions about age, gender, hobbies and everyday tasks, and issues the use of computers and the internet, which allowed determination of the social, economic and cultural rights of respondents. Results: have focused on the use of Information and Communication Technologies (ICT) to search access and use information. It was suggested that the "Brotherhood of Informatics', an incentive for students to (re) learn the tools of digital communication and make appropriate use of the resources, products and services on the web to search and effective use of information compatible with their needs and that is tied to the ethical and responsible use of technology environments. Conclusions: The results arising from this study will continue through the continuation of the Confraternity of Computing, which became a "thermometer" on the use of digital information environments and the attitude of the students in front of Information and Communication Technologies in the College “Cristo Rei”.

Carimbó do Santo Preto: a presença negra na performance musical da festividade do Glorioso São Benedito em Santarém Novo (PA)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Molecular insights on the evolution of the lateral and head lantern luciferases and bioluminescence colors in Mastinocerini railroad-worms (Coleoptera: Phengodidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Geotechnical and structural behavior of the railroad track in large-scale apparatus

This paper analyses the static and dynamic behavior of the railroad track model in laboratory. Measurements of stresses and strains on a large-scale railroad track apparatus were studied. The model includes: compacted soil, representing the final layers of platform, ballast layer, and ties (steel, wooden, and pre-stressed concrete). The soil and soil ballast interface were instrumented with pneumatic stress gauge. Settlement measurement device were positioned at the same levels as the load cells. Loads were applied by hydraulic actuators, statically and dynamically. After the prescribed number of load cycles, in pre-determined intervals, stresses and strains were measured. Observations indicate that stress and strain distributions, transmitted by wooden or steel ties, behave similarly. A more favorable behavior was observed with pre-stressed concrete mono block ties. Non-linear response was observed after a threshold numbers of cycles were surpassed, showing that the strain modulus increases with the numbers of cycles. © 2009 IOS Press.

Few substitutions affect the bioluminescence spectra of Phrixotrix (Coleoptera: Phengodidae) luciferases: a site-directed mutagenesis survey

Phrixotrix (railroad worm) luciferases produce bioluminescence in the green and red regions of the spectrum, depending on the location of the lanterns, and are the only luciferases naturally producing red bioluminescence. Comparison of the luciferase sequences showed a set of substitutions that could be involved in bioluminescence colour determination: (a) unique substitutions in the red luciferase replacing otherwise invariant residues; (b) conserved basic residues in the green-yellow emitting luciferases; and (c) an additional R353 residue in red-emitting luciferase (Viviani et al., 1999). To investigate whether these sites have a functional role in bioluminescence colour determination, we performed a site-directed mutagenesis. Natural substitutions in the region 220-344 and residues in the putative luciferin-binding site were also investigated. With the exception of the previously identified substitution of R215 and T226 (Viviani et al., 2002), which display dramatic red-shift effects on the spectrum of green-yellow-emitting luciferases, only a few substitutions had a moderate effect on the spectrum of the green-emitting luciferase. In contrast, no single substitution affected the spectrum of the red-emitting luciferase. The results suggest that the identity of the active site residues is not so critical for determining red bioluminescence in PxRE luciferase. Rather, the conformation assumed during the emitting step could be critical to set up proper interactions with excited oxyluciferin. Copyright ©2007 John Wiley & Sons, Ltd.

Estimating hardness of eucalyptus wood with a portable hardness tester

More than 80% of the 29600 km of the Brazilian railroad mesh employs wooden sleepers. The problem of hard availability of native wood for this purpose leads to the alternative use of reforestation species to produce sleepers. Considering the great difficulty to, in field condition, evaluate characteristics that are of major importance to define its suitability to sleeper production the Research Group on Forest Products from FCA/UNESP - Brazil had developed equipment for field evaluation of hardness in wood - Portable Hardness Tester. This paper reports the functional validation tests, performed with different species of Eucalyptus. Results revealed the equipment great functionality, easy-to-use characteristics and applicability to Eucalyptus wood. Moderate to strong relationships between laboratory and validated values of hardness were found. The best validation model was obtained using the data provided by the experimental dispositive 3 (R2=0.74 and SSE= 7.71 kJ/m2) while the experimental dispositive 1 gave the worse validation (R2=0.55 and SSE= 13.46 kJ/m2).