Cloning, expression, molecular modelling and docking analysis of glutathione transferase from Saccharum officinarum

Sugarcane yield and quality are affected by a number of biotic and abiotic stresses. In response to such stresses, plants may increase the activities of some enzymes such as glutathione transferase (GST), which are involved in the detoxification of xenobiotics. Thus, a sugarcane GST was modelled and molecular docked using the program LIGIN to investigate the contributions of the active site residues towards the binding of reduced glutathione (GSH) and 1-chloro-2,4-dinitrobenzene (CDNB). As a result, W13 and I119 were identified as key residues for the specificity of sugarcane GSTF1 (SoGSTF1) towards CDNB. To obtain a better understanding of the catalytic specificity of sugarcane GST (SoGSTF1), two mutants were designed, W13L and I119F. Tertiary structure models and the same docking procedure were performed to explain the interactions between sugarcane GSTs with GSH and CDNB. An electron-sharing network for GSH interaction was also proposed. The SoGSTF1 and the mutated gene constructions were cloned and expressed in Escherichia coli and the expressed protein purified. Kinetic analyses revealed different Km values not only for CDNB, but also for GSH. The Km values were 0.2, 1.3 and 0.3 mM for GSH, and 0.9, 1.2 and 0.5 mM for CDNB, for the wild type, W13L mutant and I119F mutant, respectively. The V(max) values were 297.6, 224.5 and 171.8 mu mol min(-1) mg(-1) protein for GSH, and 372.3, 170.6 and 160.4 mu mol min(-1) mg(-1) protein for CDNB.

Large-scale ecological restoration of high-diversity tropical forests in SE Brazil

The complex interactions among endangered ecosystems, landowners` interests, and different models of land tenure and use, constitute an important series of challenges for those seeking to maintain and restore biodiversity and augment the flow of ecosystem services. Over the past 10 years, we have developed a data-based approach to address these challenges and to achieve medium and large-scale ecological restoration of riparian areas on private lands in the state of Sao Paulo, southeastern Brazil. Given varying motivations for ecological restoration, the location of riparian areas within landholdings, environmental zoning of different riparian areas, and best-practice restoration methods were developed for each situation. A total of 32 ongoing projects, covering 527,982 ha, were evaluated in large sugarcane farms and small mixed farms, and six different restoration techniques have been developed to help upscale the effort. Small mixed farms had higher portions of land requiring protection as riparian areas (13.3%), and lower forest cover of riparian areas (18.3%), than large sugarcane farms (10.0% and 36.9%, respectively for riparian areas and forest cover values). In both types of farms, forest fragments required some degree of restoration. Historical anthropogenic degradation has compromised forest ecosystem structure and functioning, despite their high-diversity of native tree and shrub species. Notably, land use patterns in riparian areas differed markedly. Large sugarcane farms had higher portions of riparian areas occupied by highly mechanized agriculture, abandoned fields, and anthropogenic wet fields created by siltation in water courses. In contrast, in small mixed crop farms, low or non-mechanized agriculture and pasturelands were predominant. Despite these differences, plantations of native tree species covering the entire area was by far the main restoration method needed both by large sugarcane farms (76.0%) and small mixed farms (92.4%), in view of the low resilience of target sites, reduced forest cover, and high fragmentation, all of which limit the potential for autogenic restoration. We propose that plantations should be carried out with a high-diversity of native species in order to create biologically viable restored forests, and to assist long-term biodiversity persistence at the landscape scale. Finally, we propose strategies to integrate the political, socio-economic and methodological aspects needed to upscale restoration efforts in tropical forest regions throughout Latin America and elsewhere. (C) 2010 Elsevier BA/. All rights reserved.

Ultrasound Irradiation Promoted Large-Scale Preparation in Aqueous Media and Antioxidant Activity of Azoles

The scaled-up preparation of 1H-pyrazole, 1-phenylpyrazole and isoxazole via sonocatalysis is reported. The products were isolated in good yields in short time reaction. These compounds had been assayed for antioxidant activity by ORAC and DPPH methodologies. The results showed that only 1-phenylpyrazole presented good antioxidant activity compared with Trolox(R).

Emulsified systems based on glyceryl monostearate and potassium cetyl phosphate: Scale-up and characterization of physical properties

introducing a pharmaceutical product on the market involves several stages of research. The scale-up stage comprises the integration of previous phases of development and their integration. This phase is extremely important since many process limitations which do not appear on the small scale become significant on the transposition to a large one. Since scientific literature presents only a few reports about the characterization of emulsified systems involving their scaling-up, this research work aimed at evaluating physical properties of non-ionic and anionic emulsions during their manufacturing phases: laboratory stage and scale-up. Prototype non-ionic (glyceryl monostearate) and anionic (potassium cetyl phosphate) emulsified systems had the physical properties by the determination of the droplet size (D[4,3 1, mu m) and rheology profile. Transposition occurred from a batch of 500-50,000 g. Semi-industrial manufacturing involved distinct conditions: intensity of agitation and homogenization. Comparing the non-ionic and anionic systems, it was observed that anionic emulsifiers generated systems with smaller droplet size and higher viscosity in laboratory scale. Besides that, for the concentrations tested, augmentation of the glyceryl monostearate emulsifier content provided formulations with better physical characteristics. For systems with potassium cetyl phosphate, droplet size increased with the elevation of the emulsifier concentration, suggesting inadequate stability. The scale-up provoked more significant alterations on the rheological profile and droplet size on the anionic systems than the non-ionic. (C) 2008 Elsevier B.V. All rights reserved.

Influence of Plasma Protein Binding on Pharmacodynamics: Estimation of In Vivo Receptor Affinities of beta Blockers Using a New Mechanism-Based PK-PD Modelling Approach

The objective of this investigation was to examine in a systematic manner the influence of plasma protein binding on in vivo pharmacodynamics. Comparative pharmacokinetic-pharmacodynamic studies with four beta blockers were performed in conscious rats, using heart rate under isoprenaline-induced tachycardia as a pharmacodynamic endpoint. A recently proposed mechanism-based agonist-antagonist interaction model was used to obtain in vivo estimates of receptor affinities (K(B),(vivo)). These values were compared with in vitro affinities (K(B),(vitro)) on the basis of both total and free drug concentrations. For the total drug concentrations, the K(B),(vivo) estimates were 26, 13, 6.5 and 0.89 nM for S(-)-atenolol, S(-)-propranolol, S(-)-metoprolol and timolol. The K(B),(vivo) estimates on the basis of the free concentrations were 25, 2.0, 5.2 and 0.56 nM, respectively. The K(B),(vivo)-K(B),(vitro) correlation for total drug concentrations clearly deviated from the line of identity, especially for the most highly bound drug S(-)-propranolol (ratio K(B),(vivo)/K(B),(vitro) similar to 6.8). For the free drug, the correlation approximated the line of identity. Using this model, for beta-blockers the free plasma concentration appears to be the best predictor of in vivo pharmacodynamics. (C) 2008 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 98:3816-3828, 2009

Spatial aspects of trade liberalization in Colombia: A general equilibrium approach

This paper offers some preliminary steps in the marriage of some of the theoretical foundations of new economic geography with spatial computable general equilibrium models. Modelling the spatial economy of Colombia using the traditional assumptions of computable general equilibrium (CGE) models makes little sense when one territorial unit, Bogota, accounts for over one quarter of GDP and where transportation costs are high and accessibility low compared to European or North American standards. Hence, handling market imperfections becomes imperative as does the need to address internal spatial issues from the perspective of Colombia`s increasing involvement with external markets. The paper builds on the Centro de Estudios de Economia Regional (CEER) model, a spatial CGE model of the Colombian economy; non-constant returns and non-iceberg transportation costs are introduced and some simulation exercises carried out. The results confirm the asymmetric impacts that trade liberalization has on a spatial economy in which one region, Bogota, is able to more fully exploit scale economies vis--vis the rest of Colombia. The analysis also reveals the importance of different hypotheses on factor mobility and the role of price effects to better understand the consequences of trade opening in a developing economy.

Assessing the ex ante economic impacts of transportation infrastructure policies in Brazil

This paper uses a fully operational inter-regional computable general equilibrium (CGE) model implemented for the Brazilian economy, based on previous work by Haddad and Hewings, in order to assess the likely economic effects of road transportation policy changes in Brazil. Among the features embedded in this framework, modelling of external scale economies and transportation costs provides an innovative way of dealing explicitly with theoretical issues related to integrated regional systems. The model is calibrated for 109 regions. The explicit modelling of transportation costs built into the inter-regional CGE model, based on origin-destination flows, which takes into account the spatial structure of the Brazilian economy, creates the capability of integrating the inter-regional CGE model with a geo-coded transportation network model enhancing the potential of the framework in understanding the role of infrastructure on regional development. The transportation model used is the so-called Highway Development and Management, developed by the World Bank, implemented using the software TransCAD. Further extensions of the current model specification for integrating other features of transport planning in a continental industrialising country like Brazil are discussed, with the goal of building a bridge between conventional transport planning practices and the innovative use of CGE models. In order to illustrate the analytical power of the integrated system, the authors present a set of simulations, which evaluate the ex ante economic impacts of physical/qualitative changes in the Brazilian road network (for example, a highway improvement), in accordance with recent policy developments in Brazil. Rather than providing a critical evaluation of this debate, they intend to emphasise the likely structural impacts of such policies. They expect that the results will reinforce the need to better specifying spatial interactions in inter-regional CGE models.

Hierarchical risk scale of agribusiness activities

This study aims to elaborate a hierarchical risk scale (HRS) of agricultural and cattle breeding activities and to classify the main agricultural crops and cattle breeding activities according to their risk levels. The research is characterized as exploratory and quantitative and was based on previous risk assessment (MARKOWITZ, 1952) and capital cost calculation (SHARPE, 1964) work for other business segments. The calculations on agricultural and cattle breeding data were processed for the period from 2000 to 2006. The used methods considers simplifications and adaptations needed to achieve the proposed objective. The final result, pioneering and embryonic, provides support to improve the management of these activities that are so essential to produce food for society.

Portuguese version of the Delirium Rating Scale-Revised-98: reliability and validity

Objective To assess the validity and the reliability of the Portuguese version of the Delirium Rating Scale-Revised-98 (DRS-R-98). Methods The scale was translated into Portuguese and back-translated into English. After assessing its face validity, five diagnostic groups (n = 64; delirium, depression, dementia, schizophrenia and others) were evaluated by two independent researchers blinded to the diagnosis. Diagnosis and severity of delirium as measured by the DRS-R-98 were compared to clinical diagnosis, Mini-Mental State Exam, Confusion Assessment Method, and Clinical Global Impressions scale (CGI). Results Mean and rnedian DRS-R-98 total scores significantly distinguished delirium from the other groups (p < 0.001). Inter-rater reliability (ICC between 0.9 and 1) and internal consistency (alpha = 0.91) were very high. DRS-R-98 severity scores correlated highly with the CGI. Mean DRS-R-98 severity scores during delirium differed significantly (p < 0.01) from the post-treatment values. The area under the curve established by ROC analysis was 0.99 and using the cut-off Value of 20 the scale showed sensitivity and specificity of 92.6% and 94.6%, respectively. Conclusion The Portuguese version of the DRS-R-98 is a valid and reliable measure of delirium that distinguishes delirium from other disorders and is sensitive to change in delirium severity, which may be of great value for longitudinal studies. Copyright (c) 2007 John Wiley & Sons, Ltd.

VALIDATION OF THE LONDON CHEST ACTIVITY OF DAILY LIVING SCALE IN PATIENTS WITH HEART FAILURE

Background: The Minnesota Living with Heart Failure Questionnaire (MLHFQ) is a well-validated, commonly-used tool to assess quality of life in patients with heart failure. However, it lacks specific information concerning breathlessness during daily activities. Objective: To determine the validity of the London Chest Activity of Daily Living (LCADL) scale for use in patients with heart failure. Methods: Forty-seven patients with heart failure (57% males, mean age 50 years (standard deviation 9), mean left ventricle ejection fraction 29% (SD 6), New York Heart Association (NYHA) functional class I-III) were included. All subjects first performed a cardiopulmonary exercise test and then responded to the LCADL and the MLHFQ, with guidance from the same investigator. The re-test for the LCADL was applied one week later. Results: LCADL was correlated with MLHFQ (r=0.88; p < 0.0001). LCADL and MLHFQ were also correlated with exercise capacity (r=-0.75 and r=-0.73, respectively; both p < 0.0001). The LCADL was shown to be reproducible (r(i)=0.98). There was a significant difference (p < 0.05) in the LCADL scores between NYHA functional classes I and II, as well as classes I and III, hut not between classes II and III. Conclusion: The LCADL was shown to be a valid measurement of dyspnoea during daily activities in patients with heart failure. This scale could be an additional useful tool for the assessment of patients` dyspnoea during activities of daily living.

Wavelet correlation between subjects: A time-scale data driven analysis for brain mapping using fMRI

Functional magnetic resonance imaging (fMRI) based on BOLD signal has been used to indirectly measure the local neural activity induced by cognitive tasks or stimulation. Most fMRI data analysis is carried out using the general linear model (GLM), a statistical approach which predicts the changes in the observed BOLD response based on an expected hemodynamic response function (HRF). In cases when the task is cognitively complex or in cases of diseases, variations in shape and/or delay may reduce the reliability of results. A novel exploratory method using fMRI data, which attempts to discriminate between neurophysiological signals induced by the stimulation protocol from artifacts or other confounding factors, is introduced in this paper. This new method is based on the fusion between correlation analysis and the discrete wavelet transform, to identify similarities in the time course of the BOLD signal in a group of volunteers. We illustrate the usefulness of this approach by analyzing fMRI data from normal subjects presented with standardized human face pictures expressing different degrees of sadness. The results show that the proposed wavelet correlation analysis has greater statistical power than conventional GLM or time domain intersubject correlation analysis. (C) 2010 Elsevier B.V. All rights reserved.

Development of an Evaluation Scale for Sitting and Standing From the Ground for Children With Duchenne Muscular Dystrophy

The authors developed an evaluation scale for sit-stand from the ground for children with Duchenne muscular dystrophy (DMD) and tested its reliability. The construction occurred in stages: (a) the characterization of the movement in healthy children, (b) the characterization of the movement in children with DMD, (c) the elaboration of the 1st version of the scale and the manual, (d) the evaluation by experts and readjustments, and (e) the analysis of inter- and intraexaminer reliability and correlation with the Vignos Scale, age, and time for the execution of the activity. The scale comprehended 3 phases for sitting and 5 for the standing. A very good repeatability of the measures of sitting and standing (ICC = 0.89 and 0.84, respectively) and excellent reproducibility (ICC = 0.93 and 0.92, respectively) was demonstrated. The Kappa coefficient for the 8 phases in the interexaminer analysis varied from 0.77 to 1.00 (excellent reliability), and in the intraexaminer analysis varied from 0.80 to 1.00 (excellent reliability). Good correlation was found between the variables on the Vignos Scale (age: r = 0.58; stand: r = 0.56). The scale is a reliability instrument that allows evaluation of the activity of sitting and standing in children with DMD.

Modelling congenital transmission of Chagas` disease

The successful elimination of vectorial and transfusional transmission of Chagas` disease from some countries is a result of the reduction of domestic density of the primary vector Triatoma infestans, of almost 100% of coverage in blood serological selection and to the fact that the basic reproductive number of Chagas` disease is very close to one (1.25). Therefore, congenital transmission is currently the only way of acquiring Chagas` Disease in such regions. In this paper we propose a model of congenital transmission of Chagas` disease. Its aim is to provide an estimation of the time period it will take to eliminate this form of transmission in regions where vetorial transmission was reduced to close to zero, like in Brazil. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

Scale-free network of a dengue epidemic

In this work we show that the dengue epidemic in the city of Singapore organized itself into a scale-free network of transmission as the 2000-2005 outbreaks progressed. This scale-free network of cluster comprised geographical breeding places for the aedes mosquitoes, acting as super-spreaders nodes in a network of transmission. The geographical organization of the network was analysed by the corresponding distribution of weekly number of new cases. Therefore, our hypothesis is that the distribution of dengue cases reflects the geographical organization of a transmission network, which evolved towards a power law as the epidemic intensity progressed until 2005. (c) 2007 Elsevier Inc. All rights reserved.

Validation of the University of Sao Paolo Sensory Phenomena Scale: Initial Psychometric Properties

Introduction: Although obsessions and compulsions comprise the main features of obsessive-compulsive disorder (OCD), many patients report that their compulsions are preceded by a sense of ""incompleteness"" or other unpleasant feelings such as premonitory urges or a need perform action`s until feeling ""just right."" These manifestations have been characterized as Sensory Phenomena (SP). The current study presents initial psychometric data for a new scale designed to measure SP. Methods: Seventy-six adult OCD subjects were probed twice. Patients were assessed with an open clinical interview (considered as the ""gold standard"") and with the following standardized instruments: Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition Axis I Disorders, Yale-Brown Obsessive-Compulsive Scale, Dimensional Yale-Brown Obsessive-Compulsive Scale, Yale Global Tic Severity Scale, Beck Anxiety Inventory, and Beck Depression Inventory. Results: SP were present in 51 OCD patients (67.1%). Tics were present in 16 (21.1%) of the overall sample. The presence of SP was significantly higher in early-onset OCD patients. There were no significant differences in the presence of SP according to comorbidity with tics or gender. The comparison between the results from the open clinical interviews and the University of Sao Paulo Sensory Phenomena Scale (USP-SPS) showed an excellent concordance between them, with no significant differences between interviewers. The inter-rater reliability between the expert raters for the USP-SPS was high, with K=.92. The Pearson correlation coefficient between the SP severity scores given by the two raters was .89. Conclusion: Preliminary results suggest that the USP-SPS is a valid and reliable instrument for assessing the presence and severity of SP in OCD subjects. CNS Spectr. 2009;14(6):315-323