Camptosemin, a tetrameric lectin of Camptosema ellipticum: structural and functional analysis

Lectins have been classified into a structurally diverse group of proteins that bind carbohydrates and glycoconjugates with high specificity. They are extremely useful molecules in the characterization of saccharides, as drug delivery mediators, and even as cellular surface makers. In this study, we present camptosemin, a new lectin from Camptosema ellipticum. It was characterized as an N-acetyl-d-galactosamine-binding homo-tetrameric lectin, with a molecular weight around 26 kDa/monomers. The monomers were stable over a wide range of pH values and exhibited pH-dependent oligomerization. Camptosemin promoted adhesion of breast cancer cells and hemagglutination, and both activities were inhibited by its binding of sugar. The stability and unfolding/folding behavior of this lectin was characterized using fluorescence and far-UV circular dichroism spectroscopies. The results indicate that chemical unfolding of camptosemin proceeds as a two-state monomer-tetramer process. In addition, small-angle X-ray scattering shows that camptosemin behaves as a soluble and stable homo-tetramer molecule in solution.

Molecular Basis of the Thermostability and Thermophilicity of Laminarinases: X-ray Structure of the Hyperthermostable Laminarinase from Rhodothermus marinus and Molecular Dynamics Simulations

Glycosyl hydrolases are enzymes capable of breaking the glycosidic linkage of polysaccharides and have considerable industrial and biotechnological applications. Driven by the later applications, it is frequently desirable that glycosyl hydrolases display stability and activity under extreme environment conditions, such as high temperatures and extreme pHs. Here, we present X-ray structure of the hyperthermophilic laminarinase from Rhodothermus marinus (RmLamR) determined at 1.95 angstrom resolution and molecular dynamics simulation studies aimed to comprehend the molecular basis, for the thermal stability of this class of enzymes. As most thermostable proteins, RmLamR contains a relatively large number of salt bridges, which are not randomly distributed on the structure. On the contrary, they form clusters interconnecting beta-sheets of the catalytic domain. Not all salt bridges, however, are beneficial for the protein thermostability: the existence of charge-charge interactions permeating the hydrophobic core of the enzymes actually contributes to destabilize the structure by facilitating water penetration into hydrophobic cavities, as can be seen in the case of mesophilic enzymes. Furthermore, we demonstrate that the mobility of the side-chains is perturbed differently in each class of enzymes. The side-chains of loop residues surrounding the catalytic cleft in the mesophilic laminarinase gain mobility and obstruct the active site at high temperature. By contrast, thermophilic laminarinases preserve their active site flexibility, and the active-site cleft remains accessible for recognition of polysaccharide substrates even at high temperatures. The present results provide structural insights into the role played by salt-bridges and active site flexibility on protein thermal stability and may be relevant for other classes of proteins, particularly glycosyl hydrolases.

Molecular characterization of Arabidopsis thaliana PUF proteins - binding specificity and target candidates

PUF proteins regulate both stability and translation through sequence-specific binding to the 3` UTR of target mRNA transcripts. Binding is mediated by a conserved PUF domain, which contains eight repeats of approximately 36 amino acids each. Found in all eukaryotes, they have been related to several developmental processes. Analysis of the 25 Arabidopsis Pumilio (APUM) proteins presenting PUF repeats reveals that 12 (APUM-1 to APUM-12) have a PUF domain with 50-75% similarity to the Drosophila PUF domain. Through three-hybrid assays, we show that APUM-1 to APUM-6 can bind specifically to the Nanos response element sequence recognized by Drosophila Pumilio. Using an Arabidopsis RNA library in a three-hybrid screening, we were able to identify an APUM-binding consensus sequence. Computational analysis allowed us to identify the APUM-binding element within the 3` UTR in many Arabidopsis transcripts, even in important mRNAs related to shoot stem cell maintenance. We demonstrate that APUM-1 to APUM-6 are able to bind specifically to APUM-binding elements in the 3` UTR of WUSCHEL, CLAVATA-1, PINHEAD/ZWILLE and FASCIATA-2 transcripts. The results obtained in the present study indicate that the APUM proteins may act as regulators in Arabidopsis through an evolutionarily conserved mechanism, which may open up a new approach for investigating mRNA regulation in plants.

Novel immunoadjuvants based on cationic lipid: Preparation, characterization and activity in vivo

The interactions between three different protein antigens and dioctadecyldimethylammonium bromide (DODAB) dispersed in aqueous solutions from probe sonication or adsorbed its one bilayer onto particles was comparatively investigated. The three model proteins were bovine serum albumin (BSA), purified 18 kDa/14 kDa antigens from Taenia crassiceps (18/14-Tcra) and a recombinant, heat-shock protein hsp-18 kDa from Mycobacterium leprae. Protein-DODAB complexes in water solution were characterized by dynamic light scattering for sizing and zeta-potential analysis. Cationic complexes (80-100 nm of mean hydrodynamic diameter) displayed sizes similar to those of DODAB bilayer fragments (BF) in aqueous solution and good colloid stability over a range of DODAB and protein concentrations. The amount of cationic lipid required for attaining zero of zeta-potential at a given protein amount depended on protein nature being smaller for 18 kDa/14 kDa antigens than for BSA. Mean diameters for DODAB/protein complexes increased, whereas zeta-potentials decreased with NaCl or protein concentration. In mice, weak IgG production but significant cellular immune responses were induced by the complexes in comparison to antigens alone or carried by aluminum hydroxide as shown from IgG in serum determined by ELISA, delayed type hypersensitivity reaction from footpad swelling tests and cytokines analysis. The novel cationic adjuvant/protein complexes revealed good colloid stability and potential for vaccine design at a reduced DODAB concentration. (C) 2009 Elsevier Ltd. All rights reserved.

1,4-Dioxane enhances properties and biocompatibility of polyanionic collagen for tissue engineering applications

Polyanionic collagen obtained from bovine pericardial tissue submitted to alkaline hydrolysis is an acellular matrix with strong potential in tissue engineering. However, increasing the carboxyl content reduces fibril formation and thermal stability compared to the native tissues. In the present work, we propose a chemical protocol based on the association of alkaline hydrolysis with 1,4-dioxane treatment to either attenuate or revert the drastic structural modifications promoted by alkaline treatments. For the characterization of the polyanionic membranes treated with 1,4-dioxane, we found that (1) scanning electron microscopy (SEM) shows a stronger reorientation and aggregation of collagen microfibrils; (2) histological evaluation reveals recovering of the alignment of collagen fibers and reassociation with elastic fibers; (3) differential scanning calorimetry (DSC) shows an increase in thermal stability; and (4) in biocompatibility assays there is a normal attachment, morphology and proliferation associated with high survival of the mouse fibroblast cell line NIH3T3 in reconstituted membranes, which behave as native membranes. Our conclusions reinforce the ability of 1,4-dioxane to enhance the properties of negatively charged polyanionic collagen associated with its potential use as biomaterials for grafting, cationic drug- or cell-delivery systems and for the coating of cardiovascular devices.

Development of a new stir bar sorptive extraction coating and its application for the determination of six pesticides in sugarcane juice

In this article, a novel polydimethylsiloxane/activated carbon (PDMS-ACB) material is proposed as a new polymeric phase for stir bar sorptive extraction (SBSE). The PDMS-ACB stir bar, assembled using a simple Teflon (R)/glass capillary mold, demonstrated remarkable stability and resistance to organic solvents for more than 150 extractions. The SBSE bar has a diameter of 2.36 mm and a length of 2.2 cm and is prepared to contain 92 mu L of polymer coating. This new PDMS-ACB bar was evaluated for its ability to determine the quantity of pesticides in sugarcane juice samples by performing liquid desorption (LD) in 200 mu L of ethyl acetate and analyzing the solvent through gas chromatography coupled with mass spectrometry (GC-MS). A fractional factorial design was used to evaluate the main parameters involved in the extraction procedure. Then, a central composite design with a star configuration was used to optimize the significant extraction parameters. The method used demonstrated a limit of quantification (LOQ) of 0.5-40 mu g/L, depending on the analyte detected; the amount of recovery varied from 0.18 to 49.50%, and the intraday precision ranged from 0.072 to 8.40%. The method was used in the analysis of real sugarcane juice samples commercially available in local markets.

Product verification and defect repair - status and considerations

Product verifications have become a cost-intensive and time-consuming aspect of modern electronics production, but with the onset of an ever-increasing miniaturisation, these aspects will become even more cumbersome. One may also go as far as to point out that certain precision assembly, such as within the biomedical sector, is legally bound to have 0 defects within production. Since miniaturisation and precision assembly will soon become a part of almost any product, the verifications phases of assembly need to be optimised in both functionality and cost. Another aspect relates to the stability and robustness of processes, a pre-requisite for flexibility. Furthermore, as the re-engineering cycle becomes ever more important, all information gathered within the ongoing process becomes vital. In view of these points, product, or process verification may be assumed to be an important and integral part of precision assembly. In this paper, product verification is defined as the process of determining whether or not the products, at a given phase in the life-cycle, fulfil the established specifications. Since the product is given its final form and function in the assembly, the product verification normally takes place somewhere in the assembly line which is the focus for this paper.

Bifurcations and bistability in cavity-assisted photoassociation of Bose-Einstein-condensed molecules

We study the photoassociation of Bose-Einstein condensed atoms into molecules using an optical cavity field. The driven cavity field introduces a dynamical degree of freedom into the photoassociation process, whose role in determining the stationary behavior has not previously been considered. The semiclassical stationary solutions for the atom and molecules as well as the intracavity field are found and their stability and scaling properties are determined in terms of experimentally controllable parameters including driving amplitude of the cavity and the nonlinear interactions between atoms and molecules. For weak cavity driving, we find a bifurcation in the atom and molecule number occurs that signals a transition from a stable steady state to nonlinear Rabi oscillations. For a strongly driven cavity, there exists bistability in the atom and molecule number.

Extension And Testing Of A 2D Hydrodynamic Model For Direct Rainfall Runoff Simulation

While the simulation of flood risks originating from the overtopping of river banks is well covered within continuously evaluated programs to improve flood protection measures, flash flooding is not. Flash floods are triggered by short, local thunderstorm cells with high precipitation intensities. Small catchments have short response times and flow paths and convective thunder cells may result in potential flooding of endangered settlements. Assessing local flooding and pathways of flood requires a detailed hydraulic simulation of the surface runoff. Hydrological models usually do not incorporate surface runoff at this detailedness but rather empirical equations are applied for runoff detention. In return 2D hydrodynamic models usually do not allow distributed rainfall as input nor are any types of soil/surface interaction implemented as in hydrological models. Considering several cases of local flash flooding during the last years the issue emerged for practical reasons but as well as research topics to closing the model gap between distributed rainfall and distributed runoff formation. Therefore, a 2D hydrodynamic model, depth-averaged flow equations using the finite volume discretization, was extended to accept direct rainfall enabling to simulate the associated runoff formation. The model itself is used as numerical engine, rainfall is introduced via the modification of waterlevels at fixed time intervals. The paper not only deals with the general application of the software, but intends to test the numerical stability and reliability of simulation results. The performed tests are made using different artificial as well as measured rainfall series as input. Key parameters of the simulation such as losses, roughness or time intervals for water level manipulations are tested regarding their impact on the stability.

Empirical evidence on real convergence across brazilian states

This paper examines the real convergence hypothesis across Brazilian states. In order to test for the existence of income convergence the or- der of integration of real Gross State Product (GSP) per capita series is examined as well as their di¤erences with respect to the São Paulo state which is used as a benchmark state. Both parametric and semiparametric methods are used and the results show that convergence is achieved in the cases of Alagoas, Amazonas, Bahia, Goiás, Mato Grosso, Minas Gerais, Pernambuco, Piauí, Rio Grande do Sul, Rio de Janeiro and Santa Cata- rina and convergence is weakly achieved in the cases of Ceará, Maranhao, Pará, Paraná and Sergipe .The states of Espírito Santo, Paraíba and Rio Grande do Norte show no convergence. O artigo examina a hipótese de convergência real entre os estados brasileiros. Para testar a existência ou não da convergência da renda a ordem da integração da série do produto real bruto do estado per capita é examinada assim como suas diferenças com respeito ao estado de São Paulo que é usado como base. Foram utilizados métodos paramétricos e semiparametric e os resultados mostram que ocorre convergência nos estados: Alagoas, Amazonas, Baía, Goiás, Mato Grosso, Minas Gerais, Pernambuco, Piauí, Rio Grande do Sul, Rio de Janeiro e Santa Catarina e ocorre convergência fraca nos estados: Ceará, de Maranhão, Pará, Paraná e Sergipe. Nos estado

Differences in the perception of interest representation: a comparision of Brasília and Brussels lobbying activity

The research topic of this paper is focused on the analysis of how trade associations perceive lobbying in Brussels and in Brasília. The analysis will be centered on business associations located in Brasília and Brussels as the two core centers of decision-making and as an attraction for the lobbying practice. The underlying principles behind the comparison between Brussels and Brasilia are two. Firstof all because the European Union and Brazil have maintained diplomatic relations since 1960. Through these relations they have built up close historical, cultural, economic and political ties. Their bilateral political relations culminated in 2007 with the establishment of a Strategic Partnership (EEAS website,n.d.). Over the years, Brazil has become a key interlocutor for the EU and it is the most important market for the EU in Latin America (European Commission, 2007). Taking into account the relations between EU and Brazil, this research could contribute to the reciprocal knowledge about the perception of lobby in the respective systems and the importance of the non-market strategy when conducting business. Second both EU and Brazilian systems have a multi-level governance structure: 28 Member States in the EU and 26 Member States in Brazil; in both systems there are three main institutions targeted by lobbying practice. The objective is to compare how differences in the institutional environments affect the perception and practice of lobbying, where institutions are defined as ‘‘regulative, normative, and cognitive structures and activities that provide stability and meaning to social behavior’’ (Peng et al., 2009). Brussels, the self-proclaimed "Capital of Europe”, is the headquarters of the European Union and has one of the highest concentrations of political power in the world. Four of the seven Institutions of the European Union are based in Brussels: the European Parliament, the European Council, the Council and the European Commission (EU website, n.d.). As the power of the EU institutions has grown, Brussels has become a magnet for lobbyists, with the latest estimates ranging from between 15,000 and 30,000 professionals representing companies, industry sectors, farmers, civil society groups, unions etc. (Burson Marsteller, 2013). Brasília is the capital of Brazil and the seat of government of the Federal District and the three branches of the federal government of Brazilian legislative, executive and judiciary. The 4 city also hosts 124 foreign embassies. The presence of the formal representations of companies and trade associations in Brasília is very limited, but the governmental interests remain there and the professionals dealing with government affairs commute there. In the European Union, Brussels has established a Transparency Register that allows the interactions between the European institutions and citizen’s associations, NGOs, businesses, trade and professional organizations, trade unions and think tanks. The register provides citizens with a direct and single access to information about who is engaged in This process is important for the quality of democracy, and for its capacity to deliver adequate policies, matching activities aimed at influencing the EU decision-making process, which interests are being pursued and what level of resources are invested in these activities (Celgene, n.d). It offers a single code of conduct, binding all organizations and self-employed individuals who accept to “play by the rules” in full respect of ethical principles (EC website, n.d). A complaints and sanctions mechanism ensures the enforcement of the rules and addresses suspected breaches of the code. In Brazil, there is no specific legislation regulating lobbying. The National Congress is currently discussing dozens of bills that address regulation of lobbying and the action of interest groups (De Aragão, 2012), but none of them has been enacted for the moment. This work will focus on class lobbying (Oliveira, 2004), which refers to the performance of the federation of national labour or industrial unions, like CNI (National Industry Confederation) in Brazil and the European Banking Federation (EBF) in Brussels. Their performance aims to influence the Executive and Legislative branches in order to defend the interests of their affiliates. When representing unions and federations, class entities cover a wide range of different and, more often than not, conflicting interests. That is why they are limited to defending the consensual and majority interest of their affiliates (Oliveira, 2004). The basic assumption of this work is that institutions matter (Peng et al, 2009) and that the trade associations and their affiliates, when doing business, have to take into account the institutional and regulatory framework where they do business.

Franchisor-franchisee relationship and performance: influence of personality traits, entrepreneurial drive, and time of relationship

A literatura em franchising tem virtualmente ignorado o papel de aspectos psicologicos nos resultados interorganizacionais das empresas, a despeito de sua influencia nos resultados das organizações e da qualidade de relacionamento. Este estudo, portanto, tem por objetivo analisar a influência da personalidade e do potencial empreendedor na qualidade de relacionamento e desempenho financeiro na relação franqueador-franqueado, ao longo do tempo, sob a perspectiva dos franqueados. Este estudo analisa também o papel do tempo de relacionamento sobre a qualidade de relacionamento e o desempenho financeiro. Foi utilizado neste estudo um questionário de auto-preenchimento, enviado por e-mail, com o objetivo de recolher dados de uma amostra de 342 franqueados de 3 redes de franquias. A personalidade foi mensurada por meio dos “Cinco Grandes” traços de personalidade (escalas IPIP-B5): extroversão, agradabilidade, consciencia, estabilidade emocional e imaginação. O potencial empreendedor foi mensurado por meio do índice CEI (Carland Entrepreneurship Index). A qualidade do relacionamento foi estruturada como um constructo de segunda ordem, composto por 23 itens (incorporando confiança, comprometimento e satisfação com o relacionamento), e o desempenho financeiro foi representado por meio de uma escala de mensuração de crescimento de vendas e de rentabilidade. O tempo de relacionamento foi medido por meio dos meses de relacionamento entre franqueado e franqueador. As hipoteses foram testadas por meio de modelagem por equações estruturais, com a utilização do método de mínimos quadrados parciais (PLS), análise de regressão e análise de médias. Três das cinco dimensões da personalidade apresentaram o efeito previsto sobre as variáveis qualidade do relacionamento – agradabilidade (positivamente), estabilidade emocional (positivamente), e imaginação (positivamente). O desempenho financeiro foi influenciado, como previsto por consciência (positivamente), estabilidade emocional (positivamente), e imaginação (positivamente). Como esperado, a qualidade do relacionamento apresentou efeito positivo e significativo em relação ao desempenho financeiro. O potencial empreendedor apresentou o efeito positivo previsto apenas sobre desempenho. O tempo de relacionamento teve o efeito positivo esperado sobre o relacionamento franqueador-franqueado, em relação à qualidade do relacionamento e o desempenho financeiro, mas as diferenças entre as fases de relacionamento propostas foram apenas parcialmente confirmadas, uma vez que em somente duas fases (rotina e estabilização) a análise de médias mostrou diferenças significativas. Os resultados indicam que a personalidade influencia a qualidade de relacionamento e o desempenho, mas a meneira pela qual isso ocorre é diferente no contexto brasileiro, onde esta pesquisa foi realizada, dos achados da pesquisa conduzida na Austrália, sugerindo que fatores como cultura e estabilidade de mercado podem ter influencia sobre a relação entre traços de personalidade e qualidade de relacionamento, e traços de personalidade e desempenho financeiro. O potencial empreendedor parece influenciar positivamente o desempenho do franqueado, mas a sua influência não foi significativa em relação à qualidade do relacionamento. Os resultados também indicam a importância do tempo no desenvolvimento da qualidade de relacionamento e desempenho. Além disso, os relacionamentos de longo prazo estão relacionados a melhores avaliações de qualidade de relacionamento e desempenho financeiros por parte dos franqueados. As limitações do trabalho e sugestões para estudos futuros também são discutidos.

New business dynamics and agressiveness between competitors

Pesquisas recentes tem se concentrado em analisar se as vantagens competitivas podem ser sustentáveis em um contexto hipercompetitivo. Literatura existente que tem analisado economias emergentes descobriu que o desenvolvimento do contexto institucional contribui para a criação de condições hipercompetitivas. Reconhecendo a importância dessas pesquisas no campo da gestão estratégica, mas ao invés de concentrar a atenção em países emergentes, este trabalho considera crucial ter um espectro mais amplo como objeto de estudo com países que diferem em suas características. Nesta pesquisa foi usada uma metodologia diferente dos estudos que foram tomados como referência para o trabalho, como a feita por Hermelo e Vassolo (2010) que centraram a sua atenção nos países em desenvolvimento analisando o efeito das instituições e os ambientes hipercompetitivos na sustentabilidade das vantagens competitivas. As pesquisas anteriores foram complementadas incluindo dados de empresas em países em desenvolvimento e desenvolvidos, isto para determinar a diferença de desempenho econômico superior persistente ao longo do tempo entre eles com a ideia de mostrar como o meio ambiente não é apenas um fator-chave, mas também um componente principal no desenvolvimento de vantagens competitivas. Usando o test Kolmogorov Smirnov e o Propensity Score Matching para reduzir viés, as questões de vantagem competitiva sustentável e os efeitos do contexto institucional foram analisados, tomando cuidado com a comparação uma vez que as suas diferenças podem trazer resultados incorretos. Os resultados empíricos sustentam a tese de que há uma diferença significativa no desempenho econômico superior e sua sustentabilidade entre as empresas em países com essas diferenças. Considerando-se que o foco de estudo está nas instituições e reconhecendo a importância que elas têm na sociedade e crescimento econômico, estudá-las pode nos ajudar a determinar como o desempenho economico superior e o desempenho economico superior persistente poderiam ser alcançados com a estabilidade das instituições e fatores macroeconômicos favoráveis. Uma contribuição importante deste trabalho é que ele usa outras teorias relacionadas com as vantagens competitivas e as relaciona com o entorno macroeconomico e institucional com o proposito de comparar países com realidades muito diferentes através de testes estatísticos que dão uma resposta à hipótese formulada.

Cyclicality of SME Lending and Government Involvement in Banks

Recent regulatory efforts aim at lowering the cyclicality of bank lending because of its potential detrimental effects on financial stability and the real economy. We investigate the cyclicality of SME lending by local banks with vs. without a public mandate, controlling for location, size, loan maturity, funding structure, liquidity, profitability, and credit demand-side factors. The public mandate is set by local governments and stipulates a deviation from strict profit maximization and a sustainable provision of financial services to local customers. We find that banks with a public mandate are 25 percent less cyclical than other local banks. The result is credit supply-side driven and especially strong for savings banks with high liquidity and stable deposit funding. Our findings have implications for the banking structure, financial stability and the finance-growth nexus in a local context.