Estudo de soroconversão com formulações da vacina Biken CAM-70 contra sarampo

OBJETIVO: Comparar a resposta sorológica induzida por formulações com diferentes concentrações de vírus da vacina contra sarampo da cepa Biken CAM-70. MÉTODOS: Crianças sadias de 9 a 18 meses de um centro de saúde do Rio de Janeiro, RJ, cujos responsáveis concordaram em participar, foram randomizadas em três grupos vacinados com concentrações de 5.000, 1.000 ou 200 CCID50 (50% Tissue Culture Infective Dose). Os participantes e o pessoal da pesquisa ignoravam o tipo de vacina administrado. A avaliação sorológica foi realizada pelo teste de redução em plaque de lise. Duas análises intermediárias dos dados foram programadas. RESULTADOS: Das 223 crianças recrutadas, 84% completaram todos os procedimentos; 79% tinham idade menor que 10 meses; e 93% não tinham anticorpos contra sarampo no soro pré-vacinal. As proporções de soroconversão (quadruplicação das concentrações pré-vacinais) foram 82%, 55% e 37% (p<0,0000), nos grupos vacinados com 5.000, 1.000 ou 200 CCID50, respectivamente. As diferenças nas concentrações médias de anticorpos pós-vacinais também foram substanciais e estatisticamente significativas (p<0,000). A soroconversão (independente da formulação da vacina) foi de 73% nas crianças com 10 ou mais meses de idade e 53% naquelas com menos de 10 meses. CONCLUSÕES: Formulações da vacina com concentrações inferiores a 5.000 CCID50 não induziram soroconversão satisfatória. O desempenho da vacina por faixas etárias foi compatível com o observado em outros estudos com a vacina Biken CAM-70 e indica que uma proporção apreciável de crianças vacinadas aos 9 meses pode não obter resposta imunológica plena.

Biological monitoring of aflatoxin (AFB1) in workers of swine and poultry production

Although a great body of literature exists concerning the ingestion of food contaminated with aflatoxin, there are still few studies regarding mycotoxin inhalation in occupational settings. Since mycotoxins are relatively non-volatile, inhalation exposure is cause by inhalation of airborne fungal particulates or fungi-contaminated substrates that contain aflatoxin. We intend to know if there is occupational exposure to aflatoxin in Portuguese poultry and swine production. A total of 19 individuals (11 swine; 8 poultry) agreed and provided blood samples during the course of this investigation. Measurement of AFB1 was performed by ELISA. The samples were treated with pronase (Merck), wash in a Column C18 and purification was made with immunoaffinity columns (R.biopharma), specific for AFB1. It was applied statistical test (Mann-Whitney) to verified statistical difference in AFB1 results between the two settings. Results varied with concentrations from

Environmental impact on the bacteriological quality of domestic water supplies in Lagos, Nigeria

OBJECTIVE: To assess the impact of town planning, infrastructure, sanitation and rainfall on the bacteriological quality of domestic water supplies. METHODS: Water samples obtained from deep and shallow wells, boreholes and public taps were cultured to determine the most probable number of Escherichia coli and total coliform using the multiple tube technique. Presence of enteric pathogens was detected using selective and differential media. Samples were collected during both periods of heavy and low rainfall and from municipalities that are unique with respect to infrastructure planning, town planning and sanitation. RESULTS: Contamination of treated and pipe distributed water was related with distance of the collection point from a utility station. Faults in pipelines increased the rate of contamination (p<0.5) and this occurred mostly in densely populated areas with dilapidated infrastructure. Wastewater from drains was the main source of contamination of pipe-borne water. Shallow wells were more contaminated than deep wells and boreholes and contamination was higher during period of heavy rainfall (p<0.05). E. coli and enteric pathogens were isolated from contaminated supplies. CONCLUSIONS: Poor town planning, dilapidated infrastructure and indiscriminate siting of wells and boreholes contributed to the low bacteriological quality of domestic water supplies. Rainfall accentuated the impact.

Bioconversion of D-glucose into D-glucosone by Glucose 2-oxidase from Coriolus versicolor at Moderate Pressures

Glucose 2-oxidase (pyranose oxidase, pyranose: oxygen-2-oxidoreductase, EC 1.1.3.10) from Coriolus versicolor catalyses the oxidation of D-glucose at carbon 2 in the presence of molecular O(2) producing D-glucosone (2-keto-glucose and D-arabino-2-hexosulose) and H(2)O(2). It was used to convert D-glucose into D-glucosone at moderate pressures (i.e. up to 150 bar) with compressed air in a modified commercial batch reactor. Several parameters affecting biocatalysis at moderate pressures were investigated as follows: pressure, [enzyme], [glucose], pH, temperature, nature of fluid and the presence of catalase. Glucose 2-oxidase was purified by immobilized metal affinity chromatography on epoxy-activated Sepharose 6B-IDA-Cu(II) column at pH 6.0. The rate of bioconversion of D-glucose increased with the pressure since an increase in the pressure with compressed air resulted in higher rates of conversion. On the other hand, the presence of catalase increased the rate of reaction which strongly suggests that H(2)O(2) acted as inhibitor for this reaction. The rate of bioconversion of D-glucose by glucose 2-oxidase in the presence of either nitrogen or supercritical CO(2) at 110 bar was very low compared with the use of compressed air at the same pressure. The optimum temperature (55 degrees C) and pH (5.0) of D-glucose bioconversion as well as kinetic parameters for this enzyme were determined under moderate pressure. The activation energy (E(a)) was 32.08 kJmol(-1) and kinetic parameters (V(max), K(m), K(cat) and K(cat)/K(m)) for this bioconversion were 8.8 Umg(-1) protein, 2.95 mM, 30.81 s(-1) and 10,444.06 s(-1)M(-1), respectively. The biomass of C. versicolor as well as the cell-free extract containing glucose 2-oxidase activity were also useful for bioconversion of D-glucose at moderate pressures. The enzyme was apparently stable at moderate pressures since such pressures did not affect significantly the enzyme activity.

Bioconversion of D-glucose into D-glucosone by immobilized glucose 2-oxidase from Coriolus versicolorat moderate pressures

The immobilized glucose 2-oxidase (pyranose oxidase, pyranose:oxygen-2-oxidoreductase, EC 1.1.3.10) from Coriolus versicolor was used to convert D-glucose into D-glucosone at moderate pressures, up to 150 bar, with compressed air in a modified commercial batch reactor. Several parameters affecting biocatalysis at moderate pressures were investigated as follows: pressure, different forms of immobilized biocatalysts, glucose concentration, pH, temperature and the presence of catalase. Glucose 2-oxidase (GOX2) was purified by immobilized metal affinity chromatography on epoxy-activated Sepharose 6B-IDA-Cu(II) column at pH 6.0. Purified enzyme and catalase were immobilized into a polyethersulfone (PES) membrane in the presence of glutaraldehyde and gelatin. Enhancement of the bioconversion of D-glucose was done by the pressure since an increase in the pressure with compressed air increases the conversion rates. The optimum temperature and pH for bioconversion of D-glucose were found to be 62 degrees C and pH 6.0, respectively and the activation energy (E(a)) was 28.01 kJ mol(-1). The apparent kinetic constants (V(max)' K(m)', K(cat)' and K(cat)/K(m)') for this bioconversion were 2.27 U mg(-1) protein, 11.15 mM, 8.33 s(-1) and 747.38 s(-1) M(-1), respectively. The immobilized biomass of C. versicolor as well as crude extract containing GOX2 activity were also useful for bioconversion of D-glucose at 65 bar with a yield of 69.9 +/- 3.8% and 91.3 +/- 1.2%, respectively. The immobilized enzyme was apparently stable for several months without any significant loss of enzyme activity. On the other hand, this immobilized enzyme was also stable at moderate pressures, since such pressures did not affect significantly the enzyme activity. (C) 2010 Elsevier Ltd. All rights reserved.

Chromatographic behaviour of monoclonal antibodies against wild-type amidase from Pseudomonasaeruginosa on immobilized metal chelates

The aim of this work was to devise a one-step purification procedure for monoclonal antibodies (MAbs) of IgG class by immobilized metal affinity chromatography (IMAC). Therefore, several stationary phases were prepared containing immobilized metal chelates in order to study the chromatographic behaviour of MAbs against wild-type amidase from Pseudomonas aeruginosa. Such MAbs adsorbed to Cu(II), Ni(II), Zn(II) and Co(II)-IDA agarose columns. The increase in ligand concentration and the use of longer spacer arms and higher pH values resulted in higher adsorption of MAbs into immobilized metal chelates. The dynamic binding capacity and the maximum binding capacity were 1.33 +/- 0.015 and 3.214 +/- 0.021 mg IgG/mL of sedimented commercial matrix, respectively. A K(D) of 4.53 x 10(-7) M was obtained from batch isotherm measurements. The combination of tailor-made stationary phases of IMAC and the correct selection of adsorption conditions permitted a one-step purification procedure to be devised for MAbs of IgG class. Culture supernatants containing MAbs were purified by IMAC on commercial-Zn(II) and EPI-30-IDA-Zn(II) Sepharose 6B columns and by affinity chromatography on Protein A-Sepharose CL-4B. This MAb preparation revealed on SDS-PAGE two protein bands with M(r) of 50 and 22 kDa corresponding to the heavy and light chains, respectively. Copyright (C) 2011 John Wiley & Sons, Ltd.

Aedes albopictus may not be vector of dengue virus in human epidemics in Brazil

Over 60,500 dengue cases were reported in the state of Espírito Santo (ES), Brazil, between 1995 and 1998. The study's purpose was to identify whether Aedes albopictus was transmitting the dengue virus during an epidemic in the locality of Vila Bethânia (Viana County),Vitória, ES. From April 3 to 9, 1998, blood and serum samples were collected daily for virus isolation and serological testing. Four autochthonous cases were confirmed through DEN 1 virus isolation and two autochthonous cases through MAC ELISA testing. Of 37 Ae. aegypti and 200 Ae. albopictus adult mosquitoes collected and inoculated, DEN1 virus was isolated only from a pool of two Ae. aegypti female mosquitoes. The study results suggest that Ae. albopictus still cannot be considered an inter-human vector in dengue epidemics in Brazil.

A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations

The conjugation of antigens with ligands of pattern recognition receptors (PRR) is emerging as a promising strategy for the modulation of specific immunity. Here, we describe a new Escherichia coli system for the cloning and expression of heterologous antigens in fusion with the OprI lipoprotein, a TLR ligand from the Pseudomonas aeruginosa outer membrane (OM). Analysis of the OprI expressed by this system reveals a triacylated lipid moiety mainly composed by palmitic acid residues. By offering a tight regulation of expression and allowing for antigen purification by metal affinity chromatography, the new system circumvents the major drawbacks of former versions. In addition, the anchoring of OprI to the OM of the host cell is further explored for the production of novel recombinant bacterial cell wall-derived formulations (OM fragments and OM vesicles) with distinct potential for PRR activation. As an example, the African swine fever virus ORF A104R was cloned and the recombinant antigen was obtained in the three formulations. Overall, our results validate a new system suitable for the production of immunogenic formulations that can be used for the development of experimental vaccines and for studies on the modulation of acquired immunity.

Nanofiltration for the treatment of coke plant ammoniacal wastewaters

This work addresses the treatment by nanofiltration (NF) of solutions containing NaCN and NH(4)Cl at various pH values. The NF experiments are carried out in a Lab-Unit equipped with NF-270 membranes for model solutions that are surrogates of industrial ammoniacal wastewaters generated in the coke-making processes. The applied pressure is 30 bar. The main objective is the separation of the compounds NaCN and NH(4)Cl and the optimization of this separation as a function of the pH. Membrane performance is highly dependent on solution composition and characteristics, namely on the pH. In fact, the rejection coefficients for the binary model solution containing sodium cyanide are always higher than the rejections coefficients for the ammonium chloride model solution. For ternary solutions (cyanide/ammonium/water) it was observed that for pH values lower than 9 the rejection coefficients to ammonium are well above the ones observed for the cyanides, but for pH values higher than 9.5 there is a drastic decrease in the ammonium rejection coefficients with the increase of the pH. These results take into account the changes that occur in solution, namely, the solute species that are predominant, with the increase of the pH. The fluxes of the model solutions decreased with increased pH. (C) 2010 Elsevier B.V. All rights reserved.

Estudos de absorção de CO2 com soluções aminadas

A absorção química de dióxido de carbono (CO2) através de soluções aquosas de aminas tem sido estudada nos últimos anos devido à preocupação ambiental face ao aquecimento global. Nestes estudos, foram tidos como principais objectivos a realização de ensaios preliminares de absorção e desabsorção de CO2 em soluções aquosas de aminas bem como a construção de uma instalação piloto para a mesma finalidade. Inicialmente, a nível laboratorial, procedeu-se à absorção de CO2 através de soluções aquosas de aminas. As aminas utilizadas nestes estudos foram a monoetanolamina (MEA), etilenodiamina (EDA), 1,6- hexanodiamina (HDA) e piperazina anidra (PZ). A absorção de CO2 através destas aminas foi realizada experimentalmente às condições normais de pressão e temperatura. A concentração das soluções aquosas foi de 20% em massa de cada amina. Foram também realizados estudos de regeneração das soluções aquosas de aminas saturadas de CO2 em banho de glicerina, para determinar as condições de regeneração. Para além disso, observou-se o estado físico das aminas no estado puro até saturação com CO2 para garantir a não ocorrência de danos a nível de entupimento numa posterior utilização na instalação piloto. Por fim, voltaram-se a repetir todos estes ensaios experimentais utilizando-se, em vez da água destilada, um solvente polar aprótico, dimetilsulfóxido (DMSO). Numa segunda fase destes estudos, a absorção de CO2 através de soluções aquosas de aminas foi investigada experimentalmente numa instalação piloto. O objectivo era utilizar nesta fase do estudo as mesmas aminas utilizadas nos ensaios preliminares mas uma vez que não se dispunha das quantidades necessárias de aminas e para a sua aquisição teria que se despender bastante tempo e dinheiro, utilizaram-se duas soluções aquosas de alcanolaminas. As alcanolaminas utilizadas no presente estudo foram a amina secundária dietanolamina (DEA) e a amina terciária N-metildietanolamina (MDEA), duas aminas amplamente utilizadas nas indústrias químicas e petroquímicas para a purificação dos gases de combustão. A absorção de CO2 através destas duas alcanolaminas foi realizada experimentalmente às condições normais de pressão e temperatura. As concentrações das soluções aquosas foram de (10, 20 e 30) % em massa de MDEA e de DEA. O processo de adição de cloreto de bário (BaCl2.2H2O) às alcanolaminas ajuda à formação de carbonato de bário, quando o CO2 passa através da solução de alcanolamina. A quantidade de carbonato de bário formado foi utilizado para determinar a solubilidade do CO2 (mol CO2/mol alcanolamina). O principal desafio na captura de CO2 dos gases de combustão é o de reduzir o consumo de energia necessária para a regeneração do solvente. Deste modo, foram também realizados estudos de regeneração das soluções de alcanolaminas saturadas, para determinar as condições de regeneração. Os resultados obtidos, a nível laboratorial, revelaram que uma amina secundária (PZ) e uma amina primária de cadeia longa (HDA) são mais favoráveis ao processo de absorção e regeneração de CO2. No entanto, e devido a essa mesma estrutura molecular, necessitam de maiores valores de temperaturas para desabsorver o CO2. Garantiu-se poder trabalhar com as quatro aminas no estado puro em estudos futuros, na instalação piloto, garantindo que não ocorrerão danos a nível de entupimento. Relativamente ao solvente utilizado concluiu-se que um solvente polar aprótico não é um solvente favorável para estes estudos. Os resultados obtidos, na instalação piloto, revelaram que a amina terciária, MDEA, consegue absorver maiores quantidades de CO2 do que a amina secundária, EDA, bem como é a mais fácil de regenerar com menor perda de capacidade de absorção do que a EDA.

Synthesis, characterization and antiproliferative activity on cancer cell lines of new ionic liquids from ampicillin

Ionic Liquids (ILs) are ionic compounds that possess melting temperature below 100ºC and they have been a topic of great interest since the mid-1990s due to their unique properties. The range of IL uses has been broadened, due to a significant increase in the variety of physical, chemical and biological ILs properties. They are now used as Active Pharmaceutical Ingredients (APIs) and recent interests are focused on their application as innovative solutions in new medical treatment and delivery options.1 In this work, our principal objective was the synthesis and investigation of physicochemical and medical properties of ionic liquids (ILs) and organic salts from ampicillin. This approach is of huge interest in pharmaceutical industry as cation and anion composition of ILs and organic salts can greatly alter their desired properties, namely the melting temperature and even synergistic effects can be obtained.2,3 For the synthesis of these compounds we used a recently developed method proposed by Ohno et al.4 for the preparation of quaternary ammonium and phosphonium hydroxides, that were neutralized by ampicillin. After purification we obtained pure ILs and salts in good yields. These ILs shows good antimicrobial and antifungal activities. As it is well known that some ionic liquids containing phosphonium and ammonium cation also shows anti-cancer activity1,5 we also decided to study these compounds against some cancer cell lines.

Isolamento e caracterização de uma proteína tóxica produzida por um isolado açoriano de Bacillus weihenstephanensis

Dissertação de Mestrado, Biotecnologia em Controlo Biológico, 27 de Junho de 2013, Universidade dos Açores.

Assessment of saccharide fractionation by ultrafiltration and nanofiltration

This paper addresses the investigation of the fractionation of saccharide mixtures and saccharide mixtures with calcium using ultrafiltration (UF) and nanofiltration (NF). A set of cellulose acetate membranes covered a wide range of molecular weight cut-off (MWCO) ranging from 250 to 46,000 Da and the total feed concentration of saccharides mixtures varied from 1550 to 4700 ppm with the ratio of the two saccharides-solutes (glucose to raffinose) being kept constant at the value of 1.8. The evolution pattern of the saccharide concentration ratio in the UF/NF permeate streams displayed a dependence on the membrane MWCO, on the total sugar concentration and on the presence of calcium ions. For the highest total sugar content, the membranes with MWCO from 2000 to 7000 Da showed saccharide fractionation capability that was enhanced in the presence of calcium. The Steric Pore Flow Model was used to predict individual solute permeation behaviours and to assess the deviations to steric hindered transport of the solutes in multi-component saccharide solutions. (C) 2008 Elsevier B.V. All rights reserved.

Production of hydroxamic acids by immobilized Pseudomonas aeruginosa cells Kinetic analysis in reverse micelles

Intact cells from Pseudomonas aeruginosa strain L10 containing amidase were used as biocatalysts both free and immobilized in a reverse micellar system. The apparent kinetic constants for the transamidation reaction in hydroxamic acids synthesis, were determined using substrates such as aliphatic, amino acid and aromatic amides and esters, in both media. In reverse micelles, K-m values decreased 2-7 fold relatively to the free biocatalyst using as substrates acetamide, acrylamide, propionamide and glycinamide ethyl ester. We have concluded that overall the affinity of the biocatalyst to each substrate increases when reactions are performed in the reversed micellar system as opposed to the buffer system. The immobilized biocatalyst in general, exhibits higher stability and faster rates of reactions at lower substrates concentration relatively to the free form, which is advantageous. Additionally, the immobilization revealed to be suitable for obtaining the highest yields of hydroxamic acids derivatives, in some cases higher than 80%. (C) 2013 Elsevier B.V. All rights reserved.

Pesquisa de péptidos em Macroalgas dos Açores com potencial atividade inibitória da Enzima Conversora da Angiotensina I (ECA): Extração, purificação, caracterização e avaliação da sua atividade anti-hipertensiva

Jornadas "Ciência nos Açores – que futuro? Tema Ciências Naturais e Ambiente", Ponta Delgada, 7-8 de Junho de 2013.