Subsite substrate specificity of midgut insect chymotrypsins

Insect chymotrypsins are distinctively sensitive to plant protein inhibitors, suggesting that they differ in subsite architecture and hence in substrate specificities. Purified digestive chymotrypsins from insects of three different orders were assayed with internally quenched fluorescent oligopeptides with three different amino acids at P1 (Tyr, Phe, and Leu) and 13 amino acid replacements in positions P1`, P2, and P3. The binding energy (Delta G(s), calculated from Km values) and the activation energy (Delta G(T)(double dagger), determined from k(cat)/K-m values) were calculated. The hydrophobicities of each subsite were calculated from the efficiency of hydrolysis of the different amino acid replacements at that subsite. The results showed that except for S1, the other subsites (S2, S3, and S1`) vary among chymotrypsins. This result contrasts with insect trypsin data that revealed a trend along evolution, putatively associated with resistance to plant inhibitors. In spite of those differences, the data suggested that in lepidopteran chymotrypsins S2 and S1` bind the substrate ground state, whereas only S1` binds the transition state, supporting aspects of the present accepted mechanism of catalysis. 2008 Elsevier Ltd. All rights reserved.

The role in the substrate specificity and catalysis of residues forming the substrate aglycone-binding site of a beta-glycosidase

The relative contributions to the specificity and catalysis of aglycone, of residues E190, E194, K201 and M453 that form the aglycone-binding site of a beta-glycosidase from Spodoptera frugiperda (EC 3.2.1.21), were investigated through site-directed mutagenesis and enzyme kinetic experiments. The results showed that E190 favors the binding of the initial portion of alkyl-type aglycones (up to the sixth methylene group) and also the first glucose unit of oligosaccharidic aglycones, whereas a balance between interactions with E194 and K201 determines the preference for glucose units versus alkyl moieties. E194 favors the binding of alkyl moieties, whereas K201 is more relevant for the binding of glucose units, in spite of its favorable interaction with alkyl moieties. The three residues E190, E194 and K201 reduce the affinity for phenyl moieties. In addition, M453 favors the binding of the second glucose unit of oligosaccharidic aglycones and also of the initial portion of alkyl-type aglycones. None of the residues investigated interacted with the terminal portion of alkyl-type aglycones. It was also demonstrated that E190, E194, K201 and M453 similarly contribute to stabilize ES double dagger. Their interactions with aglycone are individually weaker than those formed by residues interacting with glycone, but their joint catalytic effects are similar. Finally, these interactions with aglycone do not influence glycone binding.

Flexibility and inhibitor binding in Cdc25 phosphatases

Cdc25 phosphatases involved in cell cycle checkpoints are now active targets for the development of anti-cancer therapies. Rational drug design would certainly benefit from detailed structural information for Cdc25s. However, only apo- or sulfate-bound crystal structures of the Cdc25 catalytic domain have been described so far. Together with previously available crystalographic data, results from molecular dynamics simulations, bioinformatic analysis, and computer-generated conformational ensembles shown here indicate that the last 30-40 residues in the C-terminus of Cdc25B are partially unfolded or disordered in solution. The effect of C-terminal flexibility upon binding of two potent small molecule inhibitors to Cdc25B is then analyzed by using three structural models with variable levels of flexibility, including an equilibrium distributed ensemble of Cdc25B backbone conformations. The three Cdc25B structural models are used in combination with flexible docking, clustering, and calculation of binding free energies by the linear interaction energy approximation to construct and validate Cdc25B-inhibitor complexes. Two binding sites are identified on top and beside the Cdc25B active site. The diversity of interaction modes found increases with receptor flexibility. Backbone flexibility allows the formation of transient cavities or compact hydrophobic units on the surface of the stable, folded protein core that are unexposed or unavailable for ligand binding in rigid and densely packed crystal structures. The present results may help to speculate on the mechanisms of small molecule complexation to partially unfolded or locally disordered proteins.

Managing Dual Enrollment Programs: Review of Program Models, Operations, and Outcomes

Universities that offer dual enrollment programs for high school students must manage increasing student demand in the face of changing state legislation. This brief examines how institutions finance their dual enrollment programs, and how they ensure academic rigor as their programs continue to grow. The report also considers how dual enrollment programs ensure that students can transfer credits, and what strategies they use to encourage DE students to matriculate as undergraduates.

Price discovery in dual-class shares across multiple markets

We extend the standard price discovery analysis to estimate the information share of dual-class shares across domestic and foreign markets. By examining both common and preferred shares, we aim to extract information not only about the fundamental value of the rm, but also about the dual-class premium. In particular, our interest lies on the price discovery mechanism regulating the prices of common and preferred shares in the BM&FBovespa as well as the prices of their ADR counterparts in the NYSE and in the Arca platform. However, in the presence of contemporaneous correlation between the innovations, the standard information share measure depends heavily on the ordering we attribute to prices in the system. To remain agnostic about which are the leading share class and market, one could for instance compute some weighted average information share across all possible orderings. This is extremely inconvenient given that we are dealing with 2 share prices in Brazil, 4 share prices in the US, plus the exchange rate (and hence over 5,000 permutations!). We thus develop a novel methodology to carry out price discovery analyses that does not impose any ex-ante assumption about which share class or trading platform conveys more information about shocks in the fundamental price. As such, our procedure yields a single measure of information share, which is invariant to the ordering of the variables in the system. Simulations of a simple market microstructure model show that our information share estimator works pretty well in practice. We then employ transactions data to study price discovery in two dual-class Brazilian stocks and their ADRs. We uncover two interesting ndings. First, the foreign market is at least as informative as the home market. Second, shocks in the dual-class premium entail a permanent e ect in normal times, but transitory in periods of nancial distress. We argue that the latter is consistent with the expropriation of preferred shareholders as a class.

Impacto da emissão dual-class na estrutura de capital

A tese propõe que empresas que emitem apenas ações com direitos de voto utilizam mais capitais de terceiros do que empresas que emitem tanto ações votantes quanto não votantes. No desenvolvimento do trabalho, foi demonstrada a relevância de relacionar endividamento ao fato de uma empresa emitir ou não ações sem direito a voto, considerando as principais teorias de estrutura de capital e a realidade brasileira. Como os modelos teóricos que explicam o nível de endividamento das empresas ainda carecem de capacidade explicativa, a busca por novos determinantes está presente na literatura de estrutura de capital. A ocorrência da emissão de ações em classes diferenciadas (dual-class) como fator impactante no nível de endividamento foi analisada sob três prismas: de mercado, dos setores e das empresas que unificaram suas ações. Pelas três investigações ficou evidenciada a perspectiva de que o endividamento seja menor nos casos de emissão de ações preferenciais, considerando o ambiente de negociação e regulamentação do Brasil. A aceitação da tese tem reflexos teóricos na identificação de um fator que deve ser levado em consideração nos modelos de estrutura de capital, bem como suscita a importância de gestores, investidores e credores reconhecerem que o fato de uma empresa ser dual-class impacta não apenas na sua estrutura de controle, mas principalmente em sua estrutura de capital. Entre as considerações da aceitação da tese, estaria o reconhecimento de que empresas que ingressam no Novo Mercado na prática estão, no longo prazo, trocando o uso de ações preferenciais como forma de financiamento pela emissão de dívida.

Dual resource transfers and interruption in external debt service

Externai debt service requires a dual resource transfer. Trade surpluses have to be generated in order to make foreign exchange revenues available for debt repayment. In addition, with developing countries' externai debt being largely a public liability, debt service requires that resources can be effectively transferred from the private to the public sector. This paper derives a statistical model for dealing with dual constraints in the presence of binary dependent variables and applies it to the dual resource transfer problem. The results from the estimation of the model for a sample of 31 middle-income developing countries in the period of 1980 to 1990, strongly support the hypothesis that both externai and fiscal constraints are important in explaining externai debt service disruptions.

Insights into the Specificity of Thioredoxin Reductase-Thioredoxin Interactions. A Structural and Functional Investigation of the Yeast Thioredoxin System

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

Host specificity of a Brazilian isolate of Alternaria cassiae (Cenargen CG593) under greenhouse conditions

Several Alternaria cassiae isolates were recovered from diseased sicklepod plants (Senna obtusifolia) in the southern regions of Brazil. A representative isolate (Cenargen CG593) was tested for its host range under greenhouse conditions. The fungus promoted symptoms in sicklepod, cassava (Manihot dulce), tomato (Lycopersicon esculentum) and eggplant (Solanum melongena) when tested at a spore concentration of 10(6) spores ml(-1). When the plants were inoculated with a suspension of 10(5) spores ml(-1) and held at a dew period of 12 h (cassava) or 18 h (tomato and eggplant), the plants showed symptoms of the disease, but they recovered and continued their normal vegetative growth. These results show that the fungus A. cassiae is safe to use for the control of S. obtusifolia under Brazilian conditions, because it did not cause excessive damage in the three plants tested.

Controlador em modo dual adaptativo robusto - DMARC

The so-called Dual Mode Adaptive Robust Control (DMARC) is proposed. The DMARC is a control strategy which interpolates the Model Reference Adaptive Control (MRAC) and the Variable Structure Model Reference Adaptive Control (VS-MRAC). The main idea is to incorporate the transient performance advantages of the VS-MRAC controller with the smoothness control signal in steady-state of the MRAC controller. Two basic algorithms are developed for the DMARC controller. In the first algorithm the controller's adjustment is made, in real time, through the variation of a parameter in the adaptation law. In the second algorithm the control law is generated, using fuzzy logic with Takagi-Sugeno s model, to obtain a combination of the MRAC and VS-MRAC control laws. In both cases, the combined control structure is shown to be robust to the parametric uncertainties and external disturbances, with a fast transient performance, practically without oscillations, and a smoothness steady-state control signal

Uma abordagem indireta para o Controlador em Modo Dual Adaptativo Robusto

In this work is proposed an indirect approach to the DualMode Adaptive Robust Controller (DMARC), combining the typicals transient and robustness properties of Variable Structure Systems, more specifically of Variable Structure Model Reference Adaptive Controller (VS-MRAC), with a smooth control signal in steady-state, typical of conventional Adaptive Controllers, as Model Reference Adaptive Controller (MRAC). The goal is to provide a more intuitive controller design, based on physical plant parameters, as resistances, inertia moments, capacitances, etc. Furthermore, with the objective to follow the evolutionary line of direct controllers, it will be proposed an indirect version for the Binary Model Reference Adaptive Controller (B-MRAC), that was the first controller attemptting to act as MRAC as well as VS-MRAC, depending on a pre-defined fixed parameter

A influência das injunções de orientação relativa na calibração de um sistema dual de câmeras digitais

Image acquisition systems based on multi-head arrangement of digital frame cameras, such as the commercial systems DMC, UltraCam, besides others, are attractive alternatives enabling larger imaging area when compared to a single frame camera. Considering that in these systems, cameras are tightly attached to an external mount, it is assumed that relative position and orientation between cameras are stable during image acquisition and, consequently, these constraint can be included in the calibration step. This constraint is acceptable because estimates of the relative orientation (RO) parameters between cameras, from previously estimated exterior orientation parameters, present higher and significant deviations than the expected physical variations, due to error propagation. In order to solve this problem, this work presents an approach based on simultaneous calibration of two or more cameras using constraints that state that the relative rotation matrix and the distance between the cameras head are stable. Experiments with images acquired by an arrangement of two Hasselblad H2D cameras were accomplished, without and with the mentioned constraints. The experiments showed that the calibration process with RO constraints allows better results than the approach based on single camera calibration, provided that the estimation has included only images with good target distribution.

Molecular Modeling Suggests Cruzain Specificity for Peptide Primaquine Prodrugs

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

Efeitos do treinamento de dupla tarefa no controle do equilíbrio dinâmico e estático de idosas pré-frágeis: um estudo piloto

The increasing world population of older individuals has become a subject of growing research for prevention and reversibility of the frailty because it is a major risk factor for the occurrence of falls, especially when it involves everyday situations of dual task. Some rehabilitation programs have already used the methods of dual-task with general exercises for improving gait and postural control, but has been reported that these interventions have little specificity with limited success to improve certain aspects of static and dynamic position during the performance of functional tasks. This study aimed to verify the measures of postural control in a group of elderly women with fragility phenotype after physical therapy intervention program based on dual-task treadmill training. We selected six pre-frail elderly subjects, with a minimum age of 65, female, living in the community and randomly assigned to two groups. The survey was conducted twice a week for 45 minutes, for four weeks. The simple task intervention consisted only in training on a treadmill and the dual task consisted of in treadmill training associated with visual stimuli. The assessments were made with the use of the Berg Balance Scale (BBS) and the Balance Master® computerized posturography, static and dynamically. The effects of retention were observed after one month, using the same instruments earlier used. The results showed a tendency toward improvement or maintenance of the balance after training on a treadmill, especially with respect to static equilibrium. Both groups showed the most notable changes in the variables related to gait, as the length and speed. The BBS scores and the baropodometric variables showed that the experimental group could keep all values similar or better even one month after completion of training unlike the control group. We concluded that the dual-task performance had no additional value in relation to the improvement of balance in general, but we observed that the effectiveness of visual stimulation occurred in the maintenance of short term balancevariables

Um método primal-dual aplicado na resolução do problema de fluxo de potência ótimo

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