Agar extraction from integrated multitrophic aquacultured Gracilaria vermiculophylla: Evaluation of a microwave-assisted process using response surface methodology

Microwave-assisted extraction (MAE) of agar from Gracilaria vermiculophylla, produced in an integrated multitrophic aquaculture (IMTA) system, from Ria de Aveiro (northwestern Portugal), was tested and optimized using response surface methodology. The influence of the MAE operational parameters (extraction time, temperature, solvent volume and stirring speed) on the physical and chemical properties of agar (yield, gel strength, gelling and melting temperatures, as well as, sulphate and 3,6-anhydro-Lgalactose contents) was evaluated in a 2^4 orthogonal composite design. The quality of the extracted agar compared favorably with the attained using traditional extraction (2 h at 85ºC) while reducing drastically extraction time, solvent consumption and waste disposal requirements. Agar MAE optimum results were: an yield of 14.4 ± 0.4%, a gel strength of 1331 ± 51 g/cm2, 40.7 ± 0.2 _C gelling temperature, 93.1 ± 0.5ºC melting temperature, 1.73 ± 0.13% sulfate content and 39.4 ± 0.3% 3,6-anhydro-L-galactose content. Furthermore, this study suggests the feasibility of the exploitation of G. vermiculophylla grew in IMTA systems for agar production.

Clay body wrap with microcurrent: effects in central adiposity

Introduction Increased fat mass is becoming more prevalent in women and its accumulation in the abdominal region can lead to numerous health risks such as diabetes mellitus. The clay body wrap using compounds such as green clay, green tea and magnesium sulfate, in addition to microcurrent, may reduce abdominal fat mass and minimize or prevent numerous health problems. Objective This study aims at measuring the influence of the clay body wrap with microcurrent and aerobic exercise on abdominal fat. Methods Nineteen female patients, randomized into intervention (n = 10) and control (n = 9) groups, were evaluated using ultrasound for visceral and subcutaneous abdominal fat, calipers and abdominal region perimeter for subcutaneous fat and bioimpedance for weight, fat mass percentage and muscular mass. During 10 sessions (5 weeks, twice a week) both groups performed aerobic exercise in a cycloergometer and a clay body wrap with microcurrent was applied to the intervention group. Results When comparing both groups after 5 weeks of protocol, there was a significant decrease in the subcutaneous fat around left anterior superior iliac spine in the intervention group (ρ = 0.026 for a confidence interval 95%). When comparing initial and final abdominal fat in the intervention group, measured by ultrasound (subcutaneous and visceral fat) and by skinfold (subcutaneous fat), we detected a significant abdominal fat reduction. Conclusion This study demonstrated that the clay body wrap used with microcurrent and aerobic exercise can have a positive effect on central fat reduction.

The isolation and characterization of cytochrome c nitrite reductase subunits (NrfA and NrfH) from Desulfovibrio desulfuricans ATCC 27774

Eur. J. Biochem. 270, 3904–3915 (2003)

Kinetic behavior of Desulfovibrio gigas aldehyde oxidoreductase encapsulated in reverse micelles

Biochemical and Biophysical Research Communications 308 (2003) 73–78

Legal and health variations in drug litigation injunctions granted in Minas Gerais

OBJECTIVE To investigate the factors related to the granting of preliminary court orders [injunctions] in drug litigations. METHODS A retrospective descriptive study of drug lawsuits in the State of Minas Gerais, Southeastern Brazil, was conducted from October 1999 to 2009. The database consists of 6,112 lawsuits, out of which 6,044 had motions for injunctions and 5,167 included the requisition of drugs. Those with more than one beneficiary were excluded, which totaled 5,072 examined suits. The variables for complete, partial, and suppressed motions were treated as dependent and assessed in relation to those that were independent – lawsuits (year, type, legal representation, defendant, court in which it was filed, adjudication time), drugs (level five of the anatomical therapeutic chemical classification), and diseases (chapter of the International Classification of Diseases). Statistical analyses were performed using the Chi-square test. RESULTS Out of the 5,072 lawsuits with injunctions, 4,184 (82.5%) had the injunctions granted. Granting varied from 95.8% of the total lawsuits in 2004 to 76.9% in 2008. Where there was legal representation, granting exceeded 80.0% and in lawsuits without representation, it did not exceed 66.9%. In public civil actions (89.1%), granting was higher relative to ordinary lawsuits (82.8%) and injunctions (80.1%). Federal courts granted only 68.6% of the injunctions, while the state courts granted 84.8%. Diseases of the digestive system and neoplasms received up to 87.0% in granting, while diseases of the nervous system, mental and behavioral disorders, and diseases of the skin and subcutaneous tissue received granting below 78.6% and showed a high proportion of suspended injunctions (10.9%). Injunctions involving paroxetine, somatropin, and ferrous sulfate drugs were all granted, while less than 54.0% of those involving escitalopram, sodium diclofenac, and nortriptyline were granted. CONCLUSIONS There are significant differences in the granting of injunctions, depending on the procedural and clinical variances. Important trends in the pattern of judicial action were observed, particularly, in the reduced granting [of injunctions] over the period.

Comparative studies of Yersinia pestis outer membrane isolation techniques and their potential use in plague epidemiology

In the present study three techniques for obtaining outer membrane enriched fractions from Yersinia pestis were evaluated. The techniques analysed were: differential solubilization of the cytoplasmic membrane with Sarkosyl or Triton X-100, and centrifugation in sucrose density gradients. The sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of outer membrane isolated by the different methods resulted in similar protein patterns. The measurement of NADH-dehydrogenase and succinate dehydrogenase (inner membrane enzymes) indicated that the outer membrane preparations obtained by the three methods were pure enough for analytical studies. In addition, preliminary evidences on the potential use of outer membrane proteins for the identification of geographic variants of Y. pestis wild isolates are presented.

Thermodynamic and kinetic modelling of the redox properties of tetrahaem cytochromes C3

Dissertação apresentada para obtenção do grau de Doutor em Bioquímica,especialidade Bioquímica-Física, pela Universidade Nova de Lisboa, Faculdade de Cincias e Tecnologia

Occurence of Giardia lamblia in children of municipal day-care centers from Botucatu, São Paulo State, Brazil

Considering that the number of day-care centers for pre-school-age children has expanded rapidly in developing countries, and that these institutions presenting conditions that facilitate the transmission of many enteric agents, a parasitological survey was carried out in three municipal day-cares from Botucatu: two in the urban area (one in downtown area and the other one in the city periphery area) and the third in the rural area. Three separate stool specimens were collected from 147 children ranging from 0 to 72 months old and 20 staff members. Each stool specimen was processed by Lutz and zinc sulfate flotation methods. The frequency of giardiasis observed among children of downtown, periphery and rural day-cares was 69.6%, 52.7% and 69.6%, respectively. Only one employee was positive for G. lamblia. The examination of three stool specimens increased the positivity for G. lamblia: from the ninety three final positive examinations, 24 (25.5%) and 8 (8.5%) were positives only after examination of the second and third samples, respectively. Others intestinal organisms like Ascaris lumbricoides (20.4%), Trichuris trichiura (19.0%). Hymenolepis nana (8.8%), Entamoeba coli (22.4%) and Blastocystis hominis (32.0%) were frequently found in the children. There was no significant association among localization of the day-cares, sex of the children and the levels of G. lamblia infection. According to the age, G. lamblia was found mainly in children between 12 to 47 months old.

Effect of b-Propiolactone treatment on the complement activation mediated by equine antisera

Reduction of complement activation through an alteration of the Fc fragment of immunoglobulins by b-propiolactone treatment was carried out in equine antisera raised against rabies virus, Bothrops venoms and diphtherial toxin. Results were evaluated by means of an anaphylactic test performed on guinea-pigs, and compared to the ones obtained with the same sera purified by saline precipitation (ammonium sulfate), followed or not by enzymatic digestion with pepsin. Protein purity levels for antibothropic serum were 184.5 mg/g and 488.5 mg/g in b-propiolactone treated and pepsin-digested sera, respectively. The recovery of specific activity was 100% and 62.5% when using antibothropic serum treated by b-propiolactone and pepsin digestion, respectively. The antidiphtherial and anti-rabies sera treated with b-propiolactone and pepsin presented protein purity levels of 5,698 and 7,179 Lf/g, 16,233 and 6,784 IU/g, respectively. The recovery of specific activity for these antisera were 88.8%, 77.7%, 100% and 36,5%, respectively. b-propiolactone treatment induced a reduction in complement activation, tested "in vivo", without significant loss of biological activity. This treatment can be used in the preparation of heterologous immunoglobulins for human use.

Clay body wrap with microcurrent: Effects in central adiposity

Introduction: Increased fat mass is becoming more prevalent in women and its accumulation in the abdominal region can lead to numerous health risks such as diabetes mellitus. The clay body wrap using compounds such as green clay, green tea and magnesium sulfate, in addition to microcurrent, may reduce abdominal fat mass and minimize or prevent numerous health problems. Objective: This study aims at measuring the influence of the clay body wrap with microcurrent and aerobic exercise on abdominal fat. Methods: Nineteen female patients, randomized into intervention (n = 10) and control (n = 9) groups, were evaluated using ultrasound for visceral and subcutaneous abdominal fat, calipers and abdominal region perimeter for subcutaneous fat and bioimpedance for weight, fat mass percentage and muscular mass. During 10 sessions (5 weeks, twice a week) both groups performed aerobic exercise in a cycloergometer and a clay body wrap with microcurrent was applied to the intervention group. Results: When comparing both groups after 5 weeks of protocol, there was a significant decrease in the subcutane- ous fat around left anterior superior iliac spine in the intervention group (ρ = 0.026 for a confidence interval 95%). When comparing initial and final abdominal fat in the intervention group, measured by ultrasound (subcutaneous and visceral fat) and by skinfold (subcutaneous fat), we detected a significant abdominal fat reduction. Conclusion: This study demonstrated that the clay body wrap used with microcurrent and aerobic exercise can have a positive effect on central fat reduction.

Crystallographic and biochemical studies on dissimilatory sulfite reductases

Thesis dissertation presented to obtain a PhD degree in Biochemistry at Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa

Desenvolvimento de coagulante/ floculante para tratamento de águas residuais numa indústria de adesivos

Esta dissertação teve como objetivo clarificar a qualidade das águas residuais de uma indústria de fabricação de adesivos. Para esta clarificação optou-se por um tratamento de coagulação/floculação a ser efetuado com a adição de apenas um produto no estado sólido. Para este estudo, selecionou-se substâncias comercialmente disponíveis destacando-se para esta formulação um coagulante, sulfato de alumínio (Al2 (SO4).3H2O, dois adjuvantes, hidróxido de cálcio, Ca(OH)2 (adjuvante H) e Montmorillonite, Al2O3.4SiO2.H2O (adjuvante M) e um floculante, uma poliacrilamida aniónica (C3H5NO)n. O efluente bruto da indústria de adesivos foi caraterizado e verificou-se que dos parâmetros analisados, apenas o pH obedece aos valores limite de emissão para descarga no coletor municipal. Determinaram-se as quantidades ótimas das substâncias selecionadas, a quantidade mínima de produto formulado e foi feita a otimização da composição do produto formulado, para três níveis de pH e duas condições de agitação Numa primeira fase, a realização de ensaios preliminares sobre a quantidade de cada uma das substâncias conduziu aos valores 5 e 30 g de coagulante, 2 e 10 g de adjuvante M; 3 g de adjuvante H e 0,01 g de floculante por 500 mL de efluente. Na otimização da quantidade mínima de produto formulado obteve-se o valor de 5g de produto por 500 mL de efluente. Relativamente à formulação do produto foi definido estudar o efeito da variação das razões, %(m/m), dos adjuvantes (M e H) em função do coagulante, para diferentes valores de pH inicial (6,2; 8,1 e 10) e para duas condições de agitação (A e B), sobre os parâmetros analisados (CQO, SST, turvação, pH final e IVL). Na condição A manteve-se uma agitação de 150 rpm durante 2 minutos, reduzindo-se para 60 rpm durante 15 minutos e, por fim, para 30 rpm durante 5 minutos. Na condição B foi usada uma agitação constante de 90 rpm durante 22 minutos. O estudo do efeito das variações das razões % (m/m) dos adjuvantes M e H foi dividido em três grupos com três subgrupos: 1º grupo - 0,6 a razão mássica adjuvante M/coagulante e 0,4, 1,2 e 2 a razão mássica adjuvante H/ coagulante; 2º grupo - 0,2 a razão mássica adjuvante M/coagulante e 0,1, 0,4 e 0,6 a razão mássica adjuvante H/ coagulante; 3º grupo - 0,1 a razão mássica adjuvante M/coagulante e 0,1, 0,2 e 0,3 a razão mássica adjuvante H/ coagulante. Após este estudo obteve-se para cada condição de agitação (A e B), as razões %(m/m) e o pH inicial mais adequados, com base nos melhores resultados obtidos no efeito sobre a CQO. Para a condição de agitação A o melhor resultado obteve-se para pH inicial igual a 8,1 com a razão mássica de adjuvante M/ coagulante igual a 0,1 e a razão mássica de adjuvante H/coagulante igual a 0,1 (percentagens de remoção de 98,01%, 99,60% e 99,43% para a CQO, SST e turvação, respetivamente). Para a condição de agitação B o melhor resultado obteve-se para pH inicial igual a 8,1 com a razão mássica de adjuvante M/ coagulante igual a 0,1 e a razão mássica de adjuvante H/coagulante igual a 0,1 (percentagens de remoção de 98,67%, 99,77% e 98,40% para a CQO, SST e turvação, respetivamente). Nestas condições é necessário efetuar a correção do pH após o tratamento. Apesar das elevadas percentagens de remoção obtidas, o índice volumétrico de lamas indica fraca qualidade de sedimentação.

Chlorella sp. coagulation‐flocculation by inducing a modification on the pH broth medium

Dissertação apresentada à Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para a obtenção do grau de Mestre em Bioenergia

Toxocara spp. eggs in public squares of Sorocaba, São Paulo State, Brazil

The visceral larva migrans (VLM) is a syndrome observed in human infection with helminth larval eggs such as the Toxocara spp. that usually infects dogs and cats. Among the risk factors involved in the occurrence of VLM, particularly important is the size of these animal populations. Sorocaba is a city with a dog population twice as large as that recommended by the World Health Organization. This fact has led to a survey of the presence of Toxocara spp. eggs in public square soils of this city. Thirty squares were selected, fifteen located in the outskirts of the city and fifteen downtown. Soil samples were collected from five distinct sites in the same area. The material was homogenized and drained and 100 g was mixed with a saturated solution of magnesium sulfate and 5% potassium iodine. The floating material was analyzed under the light microscope. Toxocara spp. eggs were found in 16 squares, nine of which were located in the outskirts of the city and seven downtown. It was concluded that Sorocaba squares present a high rate of contamination with Toxocara spp. eggs. The squares in the outskirts of the city presented a higher occurrence of these eggs in comparison with those downtown, although the difference was not statistically significant (p > 0.05).

Desenvolvimento e aplicação de um biossensor para monitorização de creatinina, um biomarcador associado a doenças emergentes

Este trabalho descreve o desenvolvimento de um material sensor para creatinina por impressão molecular em estrutura polimérica (MIP) e a sua aplicação no desenvolvimento de um dispositivo de natureza potenciométrica para a determinação da molécula alvo em fluidos biológicos. A creatinina é um dos biomarcadores mais utilizados no acompanhamento da doença renal, já que é um bom indicador da taxa de filtração glomerular (TFG). Os materiais biomiméticos desenhados para interação com a creatinina foram obtidos por polimerização radicalar, recorrendo a monómeros de ácido metacríclico ou de vinilpiridina e a um agente de reticulação apropriado. De modo a aferir o efeito da impressão da creatinina na resposta dos materiais MIP à sua presença, foram também preparados e avaliados materiais de controlo, obtidos sem impressão molecular (NIP). O controlo da constituição química destes materiais, incluindo a extração da molécula impressa, foi realizado por Espectroscopia de Raman e de Infravermelho com Transformada de Fourrier. A afinidade de ligação entre estes materiais e a creatinina foi também avaliada com base em estudos cinéticos. Todos os materiais descritos foram integrados em membranas selectivas de elétrodos seletivos de ião, preparadas sem ou com aditivo iónico lipófilo, de carga negativa ou positiva. A avaliação das características gerais de funcionamento destes elétrodos, em meios de composição e pH diferentes, indicaram que as membranas com materiais impressos e aditivo aniónico eram as únicas com utilidade analítica. Os melhores resultados foram obtidos em solução tampão Piperazine-N,N′-bis(2- ethanesulfonic acid), PIPES, de pH 2,8, condição que permitiu obter uma resposta quasi-Nernstiana, a partir de 1,6×10-5 mol L-1. Estes elétrodos demonstraram ainda uma boa selectividade ao apresentaram uma resposta preferencial para a creatinina quando na presença de ureia, carnitina, glucose, ácido ascórbico, albumina, cloreto de cálcio, cloreto de potássio, cloreto de sódio e sulfato de magnésio. Os elétrodos foram ainda aplicados com sucesso na análise de amostras sintéticas de urina, quando os materiais sensores eram baseados em ácido metacrilico, e soro, quando os materiais sensores utilizados eram baseados em vinilpiridina.