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.

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)

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.

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.

O segredo da longevidade segundo as percepções dos próprios longevos

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

Ressignificação existencial do pretérito e longevidade humana

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

Analise espacial de populacao: o caso da depressao demografica na Regiao da Alta Paulistra (Marilia-SP).

The population redistribution processes considered at a micro level, such as that for the Upper-Paulista Railroad micro-region (Marilia) in Brazil, shows that for nearly 50 years there was accelerated demographic growth followed immediately afterwards by varied declines involving the rural and urban populations.-from English summary

Fadiga em dormentes de concreto reforçados com fibras de aço

A prestressed concrete monoblock railroad tie was designed to attend the characteristics of a Brazilian railroad track with 1,600 mm gauge and 320 kN axle load. Concrete ties specimens were manufactured without and with steel fibers in a volume fraction of 60 kg/m3 (0.76 % by volume), and three different initial prestress forces. Static and fatigue tests were carried out on the ties. Static tests showed that steel fibers increase the first crack and ultimate bending moments, increase significantly the slip force of the prestressing tendons, reduce crack width, add higher ductility to the ties and decrease the stress in the prestressing tendons. Under dynamic loading the steel fibers decreased the stress in the prestressing tendons by about 50%, which improved significantly the fatigue strength of the tie. A tie without fibers failed after only 150,000 cycles. However, a similar tie, with fibers, resisted 3,000,000 cycles without suffering fatigue failure.

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.

Para além do Rio Grande: Os impactos da economia paulista sobre O Triângulo Mineiro

In this work our intention was to investigate the impact of Sao Paulo's coffee economy on the Minas Gerais Triangle Region, during its penetration into the area because of the extension of the Mogiana Railroad Company in 1889. We attempt to understand the impact caused along this coffee railroad in a region where this product almost did not exist, and the changes that occurred to the economic panorama of the region. Furthermore we have taken as an example, the city of Uberaba, the most important, in this region at that time.

Análise geoambiental da região de marília, SP: Suscetibilidade a processos erosivos frente ao histórico de ocupação da área

The study area comprises the western portion of the Marília Plateau, State of São Paulo, southeastern Brazil. The geological substrate encompasses Cretaceous sedimentary rocks of the Bauru Group and local Neocenozoic units like colluvium aprons, fluvial terrace deposits, in situ regoliths, and modern alluvial deposits. In a geomorphologic sense, the study area might be characterized as showing two main pediplanes, viz. P1 Surface (upper) and P2 Surface (lower), both surfaces are presently under dissecation processes. After the 1920's the expansion of the railroad system fostered the removal of the natural vegetation, which in turn was followed by the introduction of coffee, peanut, corn and cotton crops. This intense exploitation was conducted without respect to the soil carrying capacity and its natural susceptibility to erosional processes, including an aggressive form known as voçoroca. As a result, the study shows that the most susceptible material includes colluvium aprons, in situ regoliths, and colluvium-alluvium or alluvium deposits. The most critical situation is in the P2 Surface context, near the bottom of the Marília Plateau scarpment, where surface runoff can be very strong. Another point of active erosion is represented by the exposed walls of gullies and voçorocas, mainly in fluvial reaches subjected to talweg lowering. In a general sense, this study shows current evidence of erosional stability due to the ìntroduction of pastures as a predominant type of land occupation and to a series of erosion control procedures. Among these actions are terrace implantation, construction of small pits for runoff control, natural or induced reforestation by land owners. Despite these efforts, some erosion points remain chiefly in steeper country roads and trail stretches, in areas of concentration of cattle tracks (e.g., near cattle ponds), gullies or voçorocas exposed walls, and badly planned urbanization. The permanence of these erosion points demonstrates the necessity of a continuous monitoring of surface dynamics as well as a rapid and effective intervining measures of erosion and/or silting control.

Avaliação preliminar do potencial turístico do município de rifaina (SP) e os impactos decorrentes do uso público de seus atrativos paisagísticos

This paper attempts to assess both the touristic potential of scenic attractions and the environmental impacts resulting from the public use of such attractions in the Rifaina Municipality, State of São Paulo, Southeastern Brazil. In the recent past, this municipality had its economy concentrated in rural activities (livestock, corn, rice and bean crops) and in the ceramic industry (production of bricks, tiles and shackles),which was largely developed in the floodplain of the Rio Grande before the construction of the Jaguara Power Plant Dam. The impacts resulting from touristic activities were evaluated by applying the VIM - Visitor Impact Management method, with emphasis on biophysical indicators. The main touristic attractions analyzed were: (1) Rifaina artificial beach; (2) Jaguara Reservoir; (3) Morro do Chapéu (scenic hill); (4) Touristic railroad; and (5) Cuestas cliffs. Attractions (1) and (2) are the most exploited ones, including, in the second case, nautical sports, sportive and craft fishing, and summer lodging. Although currently disabled, the touristic railroad presents potential conditions to be re-explored. Attractions (3) and (5), despite keeping high landscape and ecological ecotouristic potential, are the least visited places. In the present paper, a preliminary characterization of the major issues of tourism industry in the Rifaina municipality is made, including suggestions for the sustainable exploitation of tourism resources. The final results of the environmental impact evaluation of the tourism activities include small impacts in attractions (4) and (5) and moderate impacts to the three first attractions. In order to keep the sustainable touristic potential for that municipality this study proposes the improvement of the following measures: to foster research projects on technical or scientific aspects of the touristic attractions, improvement of tourism infrastructure and the creation of an Environmental Protection Area (APA) of the cuestas' scarps.

Uso do AHP na avaliação do desempenho do sistema de gestão da qualidade, meio ambiente e segurança e saúde ocupacional integrados

The paper analyses the application of the AHP method together with the attach A of ISO 9004:2000 in order to evaluate the performance of two industrial organizations, both possessing quality management systems with ISO 9001:2000 certifications, based on the specific requirements of quality, environment, occupational health and management systems. The evaluated organizations were a government-owned company from the science and technology sector that works in the nuclear area, and a private company that works in the automotive and railroad area. The open software Web-Hipre version 1.22 from Helsinki University of Technology, Finland, was used. The results demonstrated that the AHP method together with the attach A of ISO 9004:2000 is adequate to evaluate organizational performances. Both organizations presented similar performance levels.

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).