Development and evaluation of a recombinant DNA vaccine candidate expressing porcine circovirus 2 structural protein

Porcine circovirus 2 (PCV2) is generally associated with the porcine circovirosis syndrome, which is considered an important disease of swine and has potentially serious economic impact on the swine industry worldwide. This article describes the construction of a recombinant plasmid expressing the PCV2 structural protein and the evaluation of cellular and humoral immune responses produced by this recombinant vaccine in BALB/c mice. The vaccine candidate was obtained and analyzed in vivo, in an effort to determine the ability to induce a specific immune response in mice. DNA was extracted from a Brazilian PCV2 isolate and the gene coding for Cap protein was amplified by PCR and inserted into an expression plasmid. Groups of BALB/c mice were inoculated intra-muscularly and intradermally in a 15-day interval, with 100 µg and 50 µg of the vaccine construct, respectively. Another group was inoculated intramuscularly with 100 µg of empty plasmid, corresponding to the control group. Seroconversion and cellular response in BALB/c mice were compared and used for vaccine evaluation. Seroconversion was analyzed by ELISA. After a series of 3 immunizations the spleen cells of the immunized animals were used to perform lymphocyte proliferation assays. Seroconversion to PCV2 was detected by ELISA in the animals inoculated with the vaccine construct when compared with control groups. Lymphocyte proliferation assays showed a stronger cell proliferation in the inoculated animals compared with the control group. Thus, the vaccine candidate construct demonstrated to be able to induce both humoral and cellular responses in inoculated mice.

Object-oriented Programming Applied to the Development of Structural Analysis and Optimization Software

This paper presents the development of a two-dimensional interactive software environment for structural analysis and optimization based on object-oriented programming using the C++ language. The main feature of the software is the effective integration of several computational tools into graphical user interfaces implemented in the Windows-98 and Windows-NT operating systems. The interfaces simplify data specification in the simulation and optimization of two-dimensional linear elastic problems. NURBS have been used in the software modules to represent geometric and graphical data. Extensions to the analysis of three-dimensional problems have been implemented and are also discussed in this paper.

Reshaping European Union development policy: collective choices and the new global order

Abstract The European Union (EU) is one of the world´s leading donors in official development assistance (ODA) to give a strong weight in the relationship with recipient partner countries, in particular with those that are more dependent on it. Besides the material weight of its funding, the EU has retained historical ties and influence in diplomatic, political and economic terms in many of its ODA recipient partner countries (particular in Sub-Saharan Africa). Since the 2000s, the EU development policy has not only undergone major structural changes in its institutional framework but also has started to face a new international aid scenario. This paper explores why a normative-based EU development policy is being challenged by reformed EU institutions and a new global order, and how the EU is attempting to respond to this context in face of the deepest recession since the end of the Second World War.

Explicit discrimination and health: development and psychometric properties of an assessment instrument

OBJECTIVE: To develop an instrument to assess discrimination effects on health outcomes and behaviors, capable of distinguishing harmful differential treatment effects from their interpretation as discriminatory events. METHODS: Successive versions of an instrument were developed based on a systematic review of instruments assessing racial discrimination, focus groups and review by a panel comprising seven experts. The instrument was refined using cognitive interviews and pilot-testing. The final version of the instrument was administered to 424 undergraduate college students in the city of Rio de Janeiro, Southeastern Brazil, in 2010. Structural dimensionality, two types of reliability and construct validity were analyzed. RESULTS: Exploratory factor analysis corroborated the hypothesis of the instrument's unidimensionality, and seven experts verified its face and content validity. The internal consistency was 0.8, and test-retest reliability was higher than 0.5 for 14 out of 18 items. The overall score was higher among socially disadvantaged individuals and correlated with adverse health behaviors/conditions, particularly when differential treatments were attributed to discrimination. CONCLUSIONS: These findings indicate the validity and reliability of the instrument developed. The proposed instrument enables the investigation of novel aspects of the relationship between discrimination and health.

Pedochronology and development of peat bog in the environmental protection area pau-de-fruta - Diamantina, Brazil

In the region of the Serra do Espinhaço Meridional, peat bog is formed in hydromorphic environments developed in sunken areas on the plain surfaces with vegetation adapted to hydromorphic conditions, favoring the accumulation and preservation of organic matter. This pedoenvironment is developed on the regionally predominant quartzite rocks. Peat bog in the Environmental Protection Area - APA Pau-de-Fruta, located in the watershed of Córrego das Pedras, Diamantina,Brazil, was mapped and three representative profiles were morphologically characterized and sampled for physical, chemical and microbiological analyses. The organic matter was fractionated into fulvic acid (FA), humic acids (HA) and humin (H). Two profiles were sampled to determine the radiocarbon age and δ13C. The structural organization of the three profiles is homogeneous. The first two layers consist of fibric, the two subsequent of hemic and the four deepest of sapric peat, showing that organic matter decomposition advances with depth and that the influence of mineral materials in deeper layers is greater. Physical properties were homogeneous in the profiles, but varied in the sampled layers. Chemical properties were similar in the layers, but the Ca content, sum of bases and base saturation differed between profiles. Contents of H predominated in the more soluble organic matter fractions and were accumulated at a higher rate in the surface and deeper layers, while HA levels were higher in the intermediate and FA in the deeper layers. Microbial activity did not vary among profiles and was highest in the surface layers, decreasing with depth. From the results of radiocarbon dating and isotope analysis, it was inferred that bog formation began about 20 thousand years ago and that the vegetation of the area had not changed significantly since then.

Structural sustainability of cambisol under different land use system

Incongruous management techniques have been associated with some significant loss of agricultural land to degradation in many parts of the world. Land degradation results in the alteration of physical, chemical and biological properties of the soil, thereby posing a serious threat to sustainable agricultural development. In this study, our objective is to evaluate the changes in a Cambisol structure under six land use systems using the load bearing capacity model. Sampling was conducted in Amazonas Region, Brazil, in the following land use: a) young secondary forest; b) old secondary forest; c) forest; d) pasture; e) cropping, and f) agroforestry. To obtain the load bearing capacity models the undisturbed soil samples were collected in those land use systems and subjected to the uniaxial compression test. These models were used to evaluate which land use system preserved or degraded the Cambisol structure. The results of the bulk density and total porosity of the soil samples were not adequate to quantify structural degradation in Cambisol. Using the forest topsoil level (0-0.03 m) as a reference, it was observed that pasture land use system was most severe in the degradation of the soil structure while the structure were most preserved under old secondary forest, cropping system and forest. At the subsoil level (0.10-0.13 m depth), the soil structure was most degraded in the cropping land use system while it was most preserved in young secondary forest and pasture. At the 0.20-0.23 m depth, soil structure degradation was most severe in the old secondary forest system and well preserved in young secondary forest, cropping and agroforestry.

Ovule ontogeny of Tabebuia pulcherrima Sandwith (Bignoniaceae): megasporogenesis and integument development

The ovule of Tabebuia pulcherrima is anatropous, unitegmic and tenuinucellate. The nucellus shows a trizonate structural organization. The integument is initiated by periclinal divisions in the dermal layer (zone I), around the base of the archesporium. Subsequently, cells derived from the subdermal layer (zone II) start to push the dermal cells, so that they shift toward the micropylar region. The archesporial cell differentiates directly into the megaspore mother cell, undergoes meiosis, and originates a linear tetrad of megaspores. The mature embryo sac mother cell is elongated, possess a conspicuous central nucleus, and a characteristic bipolar vacuome with fibrous-granulated content. The inner cell layers of the integument differentiate into an amyloplast-rich endothelium. Patterns of callose deposition in the tetrad and selection of the functional megaspore, as well as the taxonomic value of some characters are discussed.

Structural, functional and immunological studies on a polymeric bacterial protein

The characterization of proteins from Brucella spp, the causative agent of brucellosis, has been the subject of intensive research. We have described an 18-kDa cytoplasmic protein of Brucella abortus and shown the potential usefulness of this protein as an antigen for the serologic diagnosis of brucellosis. The amino acid sequence of the protein showed a low but significant homology with that of lumazine synthases. Lumazine is an intermediate product in bacterial riboflavin biosynthesis. The recombinant form of the 18-kDa protein (expressed in E. coli) folds like the native Brucella protein and has lumazine-synthase enzymatic activity. Three-dimensional analysis by X-ray crystallography of the homolog Bacillus subtilis lumazine synthase has revealed that the enzyme forms an icosahedral capsid. Recombinant lumazine synthase from B. abortus was crystallized, diffracted X rays to 2.7-Å resolution at room temperature, and the structure successfully solved by molecular replacement procedures. The macromolecular assembly of the enzyme differs from that of the enzyme from B. subtilis. The Brucella enzyme remains pentameric (90 kDa) in its crystallographic form. Nonetheless, the active sites of the two enzymes are virtually identical at the structural level, indicating that inhibitors of these enzymes could be viable pharmaceuticals across a broad species range. We describe the structural reasons for the differences in their quaternary arrangement and also discuss the potential use of this protein as a target for the development of acellular vaccines.

Extracellular calcium-sensing receptor: structural and functional features and association with diseases

The recently cloned extracellular calcium-sensing receptor (CaR) is a G protein-coupled receptor that plays an essential role in the regulation of extracellular calcium homeostasis. This receptor is expressed in all tissues related to this control (parathyroid glands, thyroid C-cells, kidneys, intestine and bones) and also in tissues with apparently no role in the maintenance of extracellular calcium levels, such as brain, skin and pancreas. The CaR amino acid sequence is compatible with three major domains: a long and hydrophilic aminoterminal extracellular domain, where most of the activating and inactivating mutations described to date are located and where the dimerization process occurs, and the agonist-binding site is located, a hydrophobic transmembrane domain involved in the signal transduction mechanism from the extracellular domain to its respective G protein, and a carboxyterminal intracellular tail, with a well-established role for cell surface CaR expression and for signal transduction. CaR cloning was immediately followed by the association of genetic human diseases with inactivating and activating CaR mutations: familial hypocalciuric hypercalcemia and neonatal severe hyperparathyroidism are caused by CaR-inactivating mutations, whereas autosomal dominant hypoparathyroidism is secondary to CaR-activating mutations. Finally, we will comment on the development of drugs that modulate CaR function by either activating (calcimimetic drugs) or antagonizing it (calcilytic drugs), and on their potential therapeutic implications, such as medical control of specific cases of primary and uremic hyperparathyroidism with calcimimetic drugs and a potential treatment for osteoporosis with a calcilytic drug.

Specific structural features of syndecans and heparan sulfate chains are needed for cell signaling

The syndecans, heparan sulfate proteoglycans, are abundant molecules associated with the cell surface and extracellular matrix and consist of a protein core to which heparan sulfate chains are covalently attached. Each of the syndecan core proteins has a short cytoplasmic domain that binds cytosolic regulatory factors. The syndecans also contain highly conserved transmembrane domains and extracellular domains for which important activities are becoming known. These protein domains locate the syndecan on cell surface sites during development and tumor formation where they interact with other receptors to regulate signaling and cytoskeletal organization. The functions of cell surface heparan sulfate proteoglycan have been centered on the role of heparan sulfate chains, located on the outer side of the cell surface, in the binding of a wide array of ligands, including extracellular matrix proteins and soluble growth factors. More recently, the core proteins of the syndecan family transmembrane proteoglycans have also been shown to be involved in cell signaling through interaction with integrins and tyrosine kinase receptors.

Heparan sulphate, its derivatives and analogues share structural characteristics that can be exploited, particularly in inhibiting microbial attachment

Heparan sulphate (HS) and the related polysaccharide, heparin, exhibit conformational and charge arrangement properties, which provide a degree of redundancy allowing several seemingly distinct sequences to exhibit the same activity. This can also be mimicked by other sulphated polysaccharides, both in overall effect and in the details of interactions and structural consequences of interactions with proteins. Together, these provide a source of active compounds suitable for further development as potential drugs. These polysaccharides also possess considerable size, which bestows upon them an additional useful property: the capability of disrupting processes comprising many individual interactions, such as those characterising the attachment of microbial pathogens to host cells. The range of involvement of HS in microbial attachment is reviewed and examples, which include viral, bacterial and parasitic infections and which, in many cases, are now being investigated as potential targets for intervention, are identified.

States and Economic Development

Today the Washington Consensus on development lies in tatters. The recent history of the developing world has been unkind to the core claim that a nation that opens its economy and keeps government's role to a minimum invariably experiences rapid economic growth. The evidence against this claim is strong: the developing world as a whole grew faster during the era of state intervention and import substitution (1950-1980) than in the more recent era of structural adjustment (1990-2005); and the recent economic performance of both Latin America and Sub-Saharan Africaregions that truly embraced neoliberalismhas lagged well behind that of many Asian economies, which have instead pursued judicial and unorthodox combinations of state intervention and economic openness. As scholars and policy makers reconstruct alternatives to the Washington Consensus on development, it is important to underline that prudent and effective state intervention and selective integration with the global economy have been responsible for development success in the past; they are also likely to remain the recipes for upward mobility in the global economy in the future."

Assessing the dynamics of terms of trade in a model of cumulative causation and structural change

ABSTRACT The motivation for this paper stems from the steady decline in the share of consumer expenditures on goods produced in the global south, coupled with the (empirically ambiguous) Singer/Prebisch hypothesis that this can be explained by a secular decline in the southern terms of trade. Drawing on these sources of inspiration, the paper sets out to study the dynamics of the terms of trade using a multi-sector growth model based on the principle of cumulative causation. The upshot is a North-South model of growth and trade in which the evolution of the terms of trade depends on differential rates of productivity growth in different sectors of the economy - and in which terms of trade dynamics may not be the best guide as to whether or not there is an uneven development problem.