No description available.

No description available.

No description available.

No description available.

Made available in DSpace on 2014-06-11T19:29:08Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-06-21Bitstream added on 2014-06-13T18:38:50Z : No. of bitstreams: 1 foguel_mv_me_araiq.pdf: 1424253 bytes, checksum: 63eeed20ff10405c2717879b8b49170a (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Muitas espécies de fungos pertencentes ao gênero Aspergillus sp. são capazes de contaminar e produzir micotoxinas em grãos e cereais, sendo responsáveis por doenças em animais e humanos. As espécies Aspergillus flavus e Aspergillus parasiticus são produtoras das aflatoxinas, que têm recebido grande atenção, devido a seus efeitos tóxicos no organismo como carcinogenicidade e mutagenicidade. Dentre os diversos tipos de aflatoxinas, destaca-se a aflatoxina B1, devido à alta potencialidade tóxica. Várias técnicas têm sido aplicadas na detecção desta toxina e o desenvolvimento de imunossensores eletroquímicos tem sido uma alternativa, devido à simplicidade, especificidade e alta sensibilidade. Superfícies metálicas de ouro, como CD-Rs de ouro (Au-CDtrodos), têm sido amplamente utilizadas como transdutores para a construção de imunossensores, devido à capacidade de imobilizar moléculas biológicas na presença ou ausência de monocamadas auto-organizadas (“selfassembled monolayers” - SAM). O presente trabalho relata o desenvolvimento de imunossensores, amperométrico e impedimétrico, para detecção de aflatoxina B1 em alimentos empregando CDtrodos de ouro. As etapas de construção dos imunossensores incluem as modificações do transdutor de ouro com p-aminotiofenol (p-ATP), proteína A ou ácido lipóico, seguido da imobilização do anticorpo anti-AFB1 e imunoensaios com o antígeno (AFB1). Empregando técnicas eletroquímicas (espectroscopia de impedância eletroquímica e voltametria cíclica) definiu-se que a SAM de ácido lipóico ativada com EDC/NHS foi o modificador que apresentou maior eficiência para a imobilização do anticorpo anti-AFB1. A partir de estudos quimiométricos, por meio do planejamento fatorial completo, definiram-se as melhores condições para o desenvolvimento do imunossensor:... / Many fungi species belonging to the genus Aspergillus sp. are able to infect and produce mycotoxins in grains and cereals, they are responsible for diseases in animals and humans. Aspergillus flavus and Aspergillus parasiticus are producers of aflatoxina, which have received great attention, due to toxic effects on the organism, such as carcinogenicity and mutagenicity. Among the different types of aflatoxins, aflatoxina B1 stands out due to high toxic potential. Several techniques have been applied in the detection of this toxin and the development of electrochemical immunosensors has been an option due to simplicity, specificity and high sensitivity. Metallic surfaces of gold, as gold CD-R (Au-CDtrodes), have been widely used as transducers for construction of immunosensors, due to the ability to immobilize biological molecules in presence or absence of self-assembled monolayers (SAM). This work describes the development of immunosensors, amperometric and impedimetric, for aflatoxin B1 detection in foods using gold CDtrodes. The construction steps of the immunosensors include modifications of the gold transducer with p-aminothiophenol (p-ATP), protein A or lipoic acid, followed by anti-AFB1 immobilization and immunoassays with antigen (AFB1). Employing electrochemical techniques (electrochemical impedance spectroscopy and cyclic voltammetry) was defined that the lipoic acid SAM activated with EDC/NHS was the modifier with the highest efficiency for the anti-AFB1 antibody immobilization. Using chemometric studies, through the full factorial design, was defined the best conditions for the immunosensor development: anti-AFB1 antibody at 1:2000 dilution and surface blocking with 0.5% bovine serum albumin, both with incubation 1 hour and antibody- AFB1 antigen immunoreaction for 30 minutes... (Complete abstract click electronic access below)

The main theme in this dissertation is to develop Scanning Electrochemical Microscopy (SECM) based electroanalytical techniques to study quantitative biology in real time. The multidrug resistance (MDR) phenomenon in live cancer cells was studied using mimic drug molecules such as menadione with the aid of SECM. Real time quantitative detection of thiodione (menadione-conjugate) pumped out of the cells was determined to be 140 μM due to exposure of 500 μM menadione to the cells. Selective blocking of these MDR pumps in live intact cells was also achieved by small molecules such as MK571 as well as by the MDR specific antibody. An approximately 50% drop in thiodione flux was observed in both cases of MDR pumps inhibition. This SECM technique was also extended to measure the permeability of a highly charged hydrophilic molecule passing through the membrane of a single living cell. The permeability was measured to be 6.5 ± 2.0 × 10-6 m/s. Real time monitoring of morphological changes in a live HeLa cell due to addition of varying concentration of surfactant such as Triton X-100 was also demonstrated by SECM. This electroanalytical technique was also expanded to study quantitative microbiology. Real time quantitative detection of pyocyanin produced by Pseudomonas aeruginosa (PA14 strain) biofilm locally was determined to be 2.5 μM after 6 h. Pyocyanin (PYO) was also observed to be reduced by PA14 biofilm, thus maintaining a reduced atmosphere above the biofilm even in presence of oxygen. Spatial mapping of this reduced PYO showed that this reduced zone was only formed up to 500 μm above the biofilm. The cells are also able to modulate the height of the reduced PYO zone in accordance to the availability of Fe(III/II) in the solution to scavenge iron from the surrounding environment. Real time spatial mapping hydrogen peroxide across polymicrobial biofilm (Sg and Aa) was also achieved with the aid of SECM. The local peroxide concentration produced by Sg was measured to be 1 mM, which is significantly higher than the bulk peroxide concentration. This study also showed that the local concentration across the microbial film is more important than the bulk concentration since bacteria communicate locally in real world. / text

L – Ascorbic acid is a compound of great importance as it is vital to immune response and wound healing but it cannot be synthesised by some mammals, including humans. L- Ascorbic acid was also found to play an important role in the body as a free – radical scavenger, which may help to prevent free – radical induced diseases such as cancer and Parkinson’s disease. In foods, it is found in many fruits, especially in citrus fruits and berries, in vegetables and in beverages, especially those derived from fruit juices. Despite its wide presence in vegetables, many people do not acquire much Vitamin C this way. So very important is vitamin C. PI–50-1 model potentiostat, connected to Epsilon EC-2000-XP ( Ver.1,50,69_XP) programmer, and computer. The experiments were performed in a three-electrode cell, containing glassy carbon working electrode and counter-electrode (A = 0,07 cm2). Saturated Ag/AgCl reference electrode. All potential values reported refer to this reference electrode. The electrochemical oxidation of ascorbic acid at poly(N-methylaniline) modified electrode has been studied. The proposed mechanism explains the ability of poly(N-methylaniline) to electro-catalyze the anodic oxidation of ascorbic acid. It has been shown that poly(N-methylaniline) modified electrodes can be used for ascorbic acid assay within a pH range of 4,41 – 6,70, operating potential window of 0,25 – 0,4 V vs. Ag/AgCl, showing a linear range of response up to 1 mM, and a lower detection... [to full text]

No description available.

No description available.

Orientador: Hideko Yamanaka / Coorientador: Antonio Aparecido Pupim Ferreira / Banca: Zeki Naal / Banca: Lauro Tatsuo Kubota / Resumo: Muitas espécies de fungos pertencentes ao gênero Aspergillus sp. são capazes de contaminar e produzir micotoxinas em grãos e cereais, sendo responsáveis por doenças em animais e humanos. As espécies Aspergillus flavus e Aspergillus parasiticus são produtoras das aflatoxinas, que têm recebido grande atenção, devido a seus efeitos tóxicos no organismo como carcinogenicidade e mutagenicidade. Dentre os diversos tipos de aflatoxinas, destaca-se a aflatoxina B1, devido à alta potencialidade tóxica. Várias técnicas têm sido aplicadas na detecção desta toxina e o desenvolvimento de imunossensores eletroquímicos tem sido uma alternativa, devido à simplicidade, especificidade e alta sensibilidade. Superfícies metálicas de ouro, como CD-Rs de ouro (Au-CDtrodos), têm sido amplamente utilizadas como transdutores para a construção de imunossensores, devido à capacidade de imobilizar moléculas biológicas na presença ou ausência de monocamadas auto-organizadas ("selfassembled monolayers" - SAM). O presente trabalho relata o desenvolvimento de imunossensores, amperométrico e impedimétrico, para detecção de aflatoxina B1 em alimentos empregando CDtrodos de ouro. As etapas de construção dos imunossensores incluem as modificações do transdutor de ouro com p-aminotiofenol (p-ATP), proteína A ou ácido lipóico, seguido da imobilização do anticorpo anti-AFB1 e imunoensaios com o antígeno (AFB1). Empregando técnicas eletroquímicas (espectroscopia de impedância eletroquímica e voltametria cíclica) definiu-se que a SAM de ácido lipóico ativada com EDC/NHS foi o modificador que apresentou maior eficiência para a imobilização do anticorpo anti-AFB1. A partir de estudos quimiométricos, por meio do planejamento fatorial completo, definiram-se as melhores condições para o desenvolvimento do imunossensor:... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Many fungi species belonging to the genus Aspergillus sp. are able to infect and produce mycotoxins in grains and cereals, they are responsible for diseases in animals and humans. Aspergillus flavus and Aspergillus parasiticus are producers of aflatoxina, which have received great attention, due to toxic effects on the organism, such as carcinogenicity and mutagenicity. Among the different types of aflatoxins, aflatoxina B1 stands out due to high toxic potential. Several techniques have been applied in the detection of this toxin and the development of electrochemical immunosensors has been an option due to simplicity, specificity and high sensitivity. Metallic surfaces of gold, as gold CD-R (Au-CDtrodes), have been widely used as transducers for construction of immunosensors, due to the ability to immobilize biological molecules in presence or absence of self-assembled monolayers (SAM). This work describes the development of immunosensors, amperometric and impedimetric, for aflatoxin B1 detection in foods using gold CDtrodes. The construction steps of the immunosensors include modifications of the gold transducer with p-aminothiophenol (p-ATP), protein A or lipoic acid, followed by anti-AFB1 immobilization and immunoassays with antigen (AFB1). Employing electrochemical techniques (electrochemical impedance spectroscopy and cyclic voltammetry) was defined that the lipoic acid SAM activated with EDC/NHS was the modifier with the highest efficiency for the anti-AFB1 antibody immobilization. Using chemometric studies, through the full factorial design, was defined the best conditions for the immunosensor development: anti-AFB1 antibody at 1:2000 dilution and surface blocking with 0.5% bovine serum albumin, both with incubation 1 hour and antibody- AFB1 antigen immunoreaction for 30 minutes... (Complete abstract click electronic access below) / Mestre