ABSTRACT One of the highlighted areas in the development of new materials is the generation of micro- and nanoparticles as drug carriers which allow the progress in formulations with the ability to release active agents in a controlled way. The proanthocyanidins (PAC) extracted from the bark of the Black Wattle have stood out for their biological activities. However, most polyflavonoids have some features which limit their application in the pharmaceutical field, such as light fastness, low bioavailability of active agents, and unpleasant taste. In this context, this study aims to present the synthesis and characterization of PAC-loaded lactic-co-glycolic acid (PLGA) microparticles obtained by the multiple emulsion method. The incorporation of PAC into PLGA was successfully achieved with PAC encapsulation efficiency around 73%. Spherical microparticles were obtained with a size distribution in the range of 0.6 to 2.4 μm. The presence of PAC modified the thermal properties of the PLGA matrix. The results of in vitro assays with Vero and T24 lineage celss showed that PLGA/PAC microparticles did not promote any effect on cell proliferation by MTT assay after 24 h. The novel Acacia mearnsii proanthocyanidin-loaded PLGA microparticles have potential for application in biological systems.

ABSTRACT Bacterial halo blight, caused by Pseudomonas syringae pv. garcae, is an important disease of coffee crop occurring in Brazil and other countries. In recent years, outbreaks of this disease have damaged several coffee crops in Brazil. Aggressiveness and genetic diversity of 25 strains of P. s. pv. garcae, obtained between the years 1958 and 2011, in 23 cities of São Paulo and Minas Gerais states, as well as three strains from Kenya were evaluated in this study. The strains were inoculated on coffee seedlings cultivar Mundo Novo, and their genetic diversity was evaluated by ERIC-PCR, REP-PCR, and their combination. All the strains were pathogenic to the coffee seedlings; the results of pathogenicity tests, in both experiments, could be divided in four aggressiviness classes (highly aggressive; aggressive; moderately aggressive and less aggressive). The Kenyan strains grouped separately from the Brazilian strains with ERIC-PCR and the combination of ERIC- and REP-PCR. The Brazilian strains could be grouped in two sub-clusters, the first including the older strains, obtained from 1958 to 1978, and the other comprising the remaining strains. With a few exceptions, strains isolated from 1997 to 2011, grouped mainly by their region of origin, were predominantly isolated from higher altitude regions, above 800 m. This probably occurred because the climatic conditions that prevail in these regions, characterized by milder temperatures and regular rainfall, are favorable for the coffee crop and for the production of high quality coffee beverage, but can be also favorable to bacterial halo blight.

RESUMO: Este estudo avaliou o status do cálcio sérico em 39 vacas com deslocamento de abomaso (DA), provenientes de 30 propriedades leiteiras selecionadas, na região de Campos Gerais no Paraná. O diagnóstico do deslocamento de abomaso foi realizado por percussão auscultatória, além dos sinais clínicos e informações da anamnese. Previamente ao procedimento cirúrgico, amostras de sangue foram coletadas para a mensuração dos níveis sanguíneos de cálcio, albumina, proteína e glicose. Como grupo controle, amostras de sangue foram coletadas de vacas hígidas que se encontravam em semelhante período de lactação. Das 39 vacas com deslocamento de abomaso, 35 apresentaram hipocalcemia e no grupo controle, apenas um animal. Concentrações sanguíneas de glicose e proteína foram inferiores nos animais com DA, quando comparados com animais do grupo controle.

ABSTRACT: This study evaluated the status of serum calcium in 39 Holstein cows with displacement of the abomasum (DA), from 30 dairy farms selected in the region of Campos Gerais, Paraná state, Brazil.. The diagnosis of abomasal displacement was performed by auscultation and percussion, besides the clinical signs and history information. Before surgery, blood samples were collected to measure blood levels of calcium, albumin, protein and glucose. As a control group, blood samples were collected from healthy cows that were in similar period of lactation. Of the 39 cows with abomasal displacement, 35 had hypocalcemia and in the control group only one cow. Blood glucose and protein concentrations were lower in the animals with DA when compared with the control group.

RESUMO: Há várias bactérias que causam problemas para o cafeeiro, incluindo Pseudomonas cichorii, P. syringae pv. garcae, P. syringae pv. tabaci, Burkholderia andropogonis e Xylella fastidiosa, todas elas já descritas no Brasil. Tentativas de diferenciar essas bactérias por testes serológicos de dupla difusão em ágar (dda), com antissoros produzidos contra células íntegras de P. s. pv. garcae, mostraram reações cruzadas, principalmente entre P. s. pv. garcae e P. s. pv. tabaci. Dessa forma, foram produzidos antissoros contra P. s. pv. garcae (linhagem patotipo IBSBF-248 - Coleção de Culturas de Fitobactérias do Instituto Biológico - IBSBF), obtidos por meio de imunizações de coelhos com antígenos de proteínas do complexo proteico da membrana (CPM). Esses antissoros foram testados por dupla difusão em agarose (dda) contra diversas formas de antígenos extraídos de P. cichorii, P. s. pv. garcae e P. s. pv. tabaci [células autoclavadas, células tratadas com formol, exopolissacarídeos (EPS), glicoproteínas (GP) da cápsula bacteriana, proteínas de membrana e suspensão bacteriana (SB) em NaCl 0,85%]. Os resultados mostraram que, dependendo do antígeno e do meio suporte da dupla difusão (com ou sem MgCl2 e/ou azul de tripano), os antissoros reagem somente com P. s. pv. garcae. Desse modo, esses antígenos podem ser usados para a rápida diagnose da mancha aureolada do cafeeiro nos testes de dda.

ABSTRACT: Some bacterial diseases have been described causing problems in coffee, whose causal agents are Pseudomonas cichorii, P. syringae pv. garcae, P. s. pv. tabaci and Burkholderia andropogonis, all of them also occurring in Brazil. Attempts to differentiate these bacteria by double diffusion agar (dda) technique with antisera produced against whole cells of P. s. pv. garcae showed cross-reactions, especially among P. s. pv. garcae and P. s. pv. tabaci. Thus, antisera were produced against P. s. pv. garcae (pathotype strain IBSBF-248 - Phytobacteria Culture Collection of Instituto Biológico - IBSBF), using rabbit immunizations with antigens of the protein complex of membranes. These antisera were tested against different kind of antigens (autoclaved cells, cells treated with formaldehyde, capsule polysaccharides, glycoproteins, bacterial membrane proteins and bacterial suspension in 0.85% NaCl) extracted from P. cichorii, P. s. pv. garcae and P. s. pv. tabaci. The results showed that depending on the kind of antigen and the medium used in the double diffusion test (with or without MgCl2 and/or trypan blue), the antiserum reacted only with P. s. pv. garcae. Thus, these antigens may be used as an auxiliary tool in the rapid diagnosis of bacterial halo blight of coffee in the double diffusion test.

Copaiba oil-resin has aromatic characteristics and pharmacological activities attributed to sesquiterpenes compounds. These purified compounds present application in perfume, pharmaceutical and cosmetic industries. Therefore, the objective of this work was to use molecular distillation to recover and purify sesquiterpenes of the copaiba oil-resin with purity high. The parameters evaporator temperature (TEV) from 45 to 100 ºC and feed flow rate (Q) between 15 and 30 ml min-1 were evaluated through design of experiments (22 with center points). The results showed that it was possible to recover sesquiterpenes with 98.12% purity in the distillate stream. Increased in TEV and reduction in Q were the effects observed experimentally and through the response surface. The best experimental condition was TEV of 100 °C and Q of 15 mL min-1. Collor differences were noticed between distillate streams (colorless) and residue (brown). These can be used as parameters for separation of sesquiterpenes and diterpenes. The molecular distillation proved to be an efficient processo to both recovery and purification of sesquiterpenes from the copaiba oil-resin.

Abstract The removal of water content present in fuels such as biodiesel and diesel is quite important to adequate the fuel to standards for commercialization and to avoid corrosion of storage tanks and injection equipment in diesel engines. In this study, hydrophilic hydrogels were employed to remove the water content in biodiesel and diesel fuel samples. The results showed that the hydrogels were capable of decreasing the free water content and also the soluble water content present in the samples. The highest decrease of total water content in samples of biodiesel was 53.3% wt and for diesel samples the reduction of water content was 32.0%, starting with samples that had 2160 ppm and 240 ppm of water, respectively. The highest decrease of total water content (free and soluble water) for diesel samples was 80.4% wt, from a diesel sample initially containing 348 ppm of total water content.

RESUMO Introdução: Avanços no campo da biologia molecular têm proporcionado a diferenciação dos subtipos moleculares das neoplasias mamárias, fornecendo melhor prognóstico e ferramentas importantes para a terapêutica de pacientes com câncer de mama. Entre esses subtipos, as alterações ocorridas no gene receptor tipo 2 do fator de crescimento epidérmico humano (HER2/neu) amplificam o seu número de cópias e geram o aumento da proteína HER2. Estudos mostram que pacientes portadoras de câncer de mama HER2/neu amplificado tendem a ter recaída mais cedo e tempo de sobrevida menor, sendo o anticorpo monoclonal Trastuzumab a terapia indicada. A elegibilidade das pacientes para a terapia é feita inicialmente pela técnica de imuno-histoquímica (IHQ), que avalia o nível de expressão da proteína HER2. Após essa avaliação, os casos que apresentam diagnósticos equívocos (escore 2+) são encaminhados para uma técnica mais precisa, a hibridização cromógena in situ (CISH). Objetivo: Analisar a sensibilidade e a especificidade do anticorpo SP3, além de determinar o seu nível de concordância com a técnica de CISH. Material e métodos: Estudo retrospectivo no banco de dados de um laboratório anatomopatológico, em laudos de exames de CISH para HER2/neu. Conclusão: Os resultados revelaram que o clone SP3 apresentou 100% de especificidade e 92% de sensibilidade. A IHQ revela variabilidade em seus resultados, porém é sabido que a técnica é uma importante ferramenta na rotina diária dos laboratórios, contribuindo na triagem inicial das pacientes portadoras de câncer de mama, que, posteriormente, mostram resultados satisfatórios quando comparados com a técnica de CISH.

ABSTRACT Introduction: Advances in the field of molecular biology have provided the differentiation of molecular subtypes of breast tumors, providing better prognosis and important tools for the treatment of patients with breast cancer. Among these subtypes, the changes in the human epidermal growth factor receptor 2 gene (HER2/neu), increase its copy number and generating HER2 protein amplification. Studies show that patients with breast cancer HER2/neu amplified tend to relapse earlier and have shorter survival time, the monoclonal antibody Trastuzumab is the therapy indicated. The eligibility of patients for therapy is initially made by the immunohistochemistry (IHC) technique, which evaluates the expression level of the HER2 protein. After this evaluation, the cases with equivocal diagnosis (score 2+), are referred to a more accurate technique, the chromogenic in situ hybridization (CISH). Objective: To analyze the sensitivity and specificity of the antibody SP3, and determine their level of agreement with the CISH technique. Material and methods: Retrospective study in the database of the anatomy-pathology laboratory, in CISH tests reports for HER2/neu. Conclusion: The results revealed that clone SP3 showed 100% specificity and 92% sensitivity. IHC reveals variability in its results; however, it is known that the technique is an important tool in the daily routine of laboratories, contributing to the initial screening of patients with breast cancer, which later showed satisfactory results when compared with the CISH technique.

Mapping is a basic capability for mobile robots. Most applications demand some level of knowledge about the environment to be accomplished. Most mapping approaches in the literature are designed to perform in small structured (indoor) environments. This paper addresses the problems of localization and mapping in large urban (outdoor) environments. Due to their complexity, lack of structure and dimensions, urban environments presents several difficulties for the mapping task. Our approach has been extensively tested and validated in realistic situations. Our experimental results include maps of several city blocks and a performance analysis of the algorithms proposed.

In this paper, a novel approach for establishing the route for process intensification through the application of two developed softwares to characterize reactive mixtures is presented. A robust algorithm was developed to build up reactive phase diagrams and to predict the existence and the location of reactive azeotropes. The proposed algorithm does not depend on initial estimates and is able to compute all reactive azeotropes present in the mixture. It also allows verifying if there are no azeotropes, which are the major troubles in this kind of programming. An additional software was developed in order to calculate reactive residue curve maps. Results obtained with the developed program were compared with the published in the literature for several mixtures, showing the efficiency and robustness of the developed softwares.

The Acacia genus has presented various secondary metabolites, such as tannins, flavonoids, alkaloids and gums. Preparations from different species have been applied for diabetes, gastrointestinal disorders and inflammatory diseases in the traditional medicine and have demonstrated cytotoxic, antimicrobial and antiparasitic activities. Acacia podalyriifolia A. Cunn. ex G. Don (Fabaceae) is a small wood, indigenous to Australia and cultivated worldwide for its ornamental feature. This work aimed to characterize the anatomy of the phyllode and stem, in order to contribute to the species identification. The botanical material was fixed, sectioned and prepared according to usual light and scanning microtechniques. The epidermal cells, in surface view, are polygonal and coated with striate and thick cuticle, and filaments of epicuticular wax. Paracytic stomata and unicellular non-glandular trichomes are seen. Palisade and ground parenchymas, and minor collateral bundles with xylem directed alternately to upper and lower sides occur in the blade. The midrib shows two collateral bundles facing each other. The stem, in incipient secondary growth, exhibits epidermis, annular collenchyma, sclerenchymatic sheath and collateral vascular organization. Cells containing phenolic compounds and prisms of calcium oxalate are observed.

Objetivos: comparar as taxas de colonização e a resistência antimicrobiana de pneumococos em nasofaringe de crianças sadias e com pneumonia. Métodos: estudo de tipo transversal. As crianças sadias foram recrutadas em centros de vacinação (CV) e creches públicas (CP), selecionados aleatoriamente, e aquelas com pneumonia em emergências pediátricas. Foram utilizados swabs flexíveis de alginato para a colheita do material de nasofaringe. O isolamento e a identificação dos pneumococos seguiram procedimentos padronizados. As concentrações inibitórias mínimas (CIM) foram determinadas por microdiluição em placas. Resultados: foram estudadas 911 crianças, 429 sadias (60% portadoras de pneumococo, sendo 72% destas recrutadas em CP e 49% em CV) e 482 com pneumonia (50% de portadoras) (p=0,002). De 441 isolados com CIM determinadas, 198 (45%) apresentavam resistência intermediária, e 16 (4%) apresentavam resistência plena à penicilina. As taxas de resistência dos pneumococos isolados de portadores sadios e com pneumonia, respectivamente, foram: penicilina 48% (CV 37% e CP 55%) e 50% (p>0,05); eritromicina: 28% e 19% (p=0,05); cotrimoxazol 81% e 76% (p>0,05); cloranfenicol 6% e 7% (p>0,05), rifampicina 5% e 3% (p>0,05), ceftriaxone 2 e 4% (p>0,05) e vancomicina 0%, para ambos grupos. Foi constatada associação entre as resistências do pneumococo à penicilina, eritromicina e ao cotrimoxazol. Conclusões: a taxa de portador de pneumococos foi maior em crianças sadias do que naquelas com pneumonia. As resistências dos pneumococos à penicilina e ao cotrimoxazol foram elevadas, sobretudo entre os usuários de creches públicas.

Objectives: to compare colonization rates and antimicrobial resistance of nasopharyngeal pneumococci in healthy carriers and children with pneumonia. Methods: a cross-sectional study. Healthy subjects of this study were selected from randomly chosen immunization centers and day-care centers, and those with pneumonia were selected in pediatric emergency rooms. Flexible pernasal alginate swabs were employed to collect nasopharyngeal pneumococci specimens. Isolation and identification were performed according to standard procedures. Minimum Inhibitory Concentrations were assessed by microdilution techniques. Results: we studied 911 children, 429 healthy controls (60% of carriers, 72% attending day care centers and 49% recruited in immunization centers) and 482 children with pneumonia (50% of carriers) (P=0.02). The Minimum Inhibitory Concentration of penicillin to 441 isolates detected 198 (45%) of intermediate and 16 (4%) fully resistant pneumococci. Antimicrobial resistance rates of isolates from healthy carriers and children with pneumonia were, respectively: penicillin 48% (37% for immunization centers and 55% for day-care centers) and 50% (P>0.05), erythromycin 28% and 19% (P=0.05); cotrimoxazole 81% and 76% (P>0.05), chloramphenicol 6% and 7% (P>0.05), rifampin 5 and 3% (P>0.05) ceftriaxone 2 and 4% (P>0.05) and vancomycin 0% in both groups. An association among pneumococcal resistance to penicillin, erythromycin and cotrimoxazole was detected. Conclusions: pneumococcal carriage rate was higher in healthy children than in children with pneumonia. Penicillin and cotrimoxazole resistance rates were high, especially among those attending day-care centers.

Foram analisados os resultados de teste tuberculínico, efetuado três meses após vacinação com BCG (amostra Moreau-Rio de Janeiro), pela via intradérmica. A casuística correspondeu a 7.790 pessoas, incluindo as de grupo pertinente ao primeiro trimestre de vida e merecedor, atualmente, de especial atenção. Ficou constatada, globalmente, a percentagem de positividade de 69%. Houve apreciação, especifica e detalhada, referente a indivíduos de idades diversas, e a abordagem do tema tornou-se oportuna em face ao interesse que esse tipo de imunização vem despertando no Brasil e, também, em virtude da necessidade de coletar informes aptos a apoiar trabalhos profiláticos.

Results of tuberculin skin tests performed three months after intradermal vaccination with the Moreau-Rio de Janeiro strain of BCG were analyzed. The tests were made upon 7,790 persons, including a group of infants in their first trimester of life, a delicate and obviously necessary object of attention today. The percentage of positive reactors in the whole population group tested was 69%. A detailed analysis of the reactiveness in the different age groups was carried out. Moreover, that this study is opportune has been stressed not only because of the rising interest in Brazil in this type of immunization but also because of the need to compile information supporting prophylactic action.

Foram analisados os resultados de teste tuberculínico, efetuado três meses após vacinação com BCG (amostra Moreau-Rio de Janeiro), pela via intradérmica. A casuística correspondeu a 7.790 pessoas, incluindo as de grupo pertinente ao primeiro trimestre de vida e merecedor, atualmente, de especial atenção. Ficou constatada, globalmente, a percentagem de positividade de 69%. Houve apreciação, especifica e detalhada, referente a indivíduos de idades diversas, e a abordagem do tema tornou-se oportuna em face ao interesse que esse tipo de imunização vem despertando no Brasil e, também, em virtude da necessidade de coletar informes aptos a apoiar trabalhos profiláticos.

Results of tuberculin skin tests performed three months after intradermal vaccination with the Moreau-Rio de Janeiro strain of BCG were analyzed. The tests were made upon 7,790 persons, including a group of infants in their first trimester of life, a delicate and obviously necessary object of attention today. The percentage of positive reactors in the whole population group tested was 69%. A detailed analysis of the reactiveness in the different age groups was carried out. Moreover, that this study is opportune has been stressed not only because of the rising interest in Brazil in this type of immunization but also because of the need to compile information supporting prophylactic action.