Darbo tikslas - uždaruose gruntuose integruotai auginamiems pomidorams parinkti optimalų tręšimo būdą, užtikrinantį gerą produkcijos kokybę ir derlių. Stebėjimai buvo atliekami 2009 metais Alytaus mieste, polietilenu dengtame privačiame šiltnamyje. Šiltnamio plotas 36 m2. Pomidorai – viena svarbiausių ir populiariausių šiltnamių daržovių, auginamų ne tik ūkininkų, bet ir daržininkų mėgėjų. Tyrimui parinktas Enza Zaden firmos pomidorų hibridas Buran H. Daigai pasodinti polietileniniuose 20 litrų talpos puoduose, užpildytuose dviejų skirtingų gruntų: mineralinio (1 bandymas) ir organinio (2 bandymas). Augimo ir derėjimo metu papildomai kartą per savaitę tręšiama mineralinėmis (1 bandymas) arba organinėmis (2 bandymas) trąšomis. Vertinant pomidoro savybes nustatoma vaisių rūgštingumas, vitaminas C, nitratai, sausos medžiagos ir skonio savybės. Skaičiuojamas vidutinė vaisiaus masė, pomidorų skaičius kekėje, derlingumas ir sutrūkusių vaisių skaičius. Tyrimo metu nustatyta, kad subalansuotame grunte, papildomai tręšiant mažomis normomis kaip N34P8K50 ir N103P24K151, vaisių kokybiniai rodikliai praktiškai nesiskiria nuo organiniame grunte ir papildomai tręštų raugintomis dilgėlėmis pomidorų kokybinių rodiklių. Didinimas mineralinių trąšų normos skatino greitesnį pomidorų brendimą, vaisių odelės trūkimą, vitamino C kiekio didėjimą ir nitratų kaupimąsi. Augalus auginant 20 litrų polietileniniuose puoduose užtikrino mažą drėgmę šiltnamyje, kas neleido plisti ligoms ir kenkėjams... [toliau žr. visą tekstą] / The investigation of soil substratum and fertilization impact on the properties of tomato was carried out at the private greenhouse in Alytus in 2009. Buran H breed was selected for the growing experiment in special isolated 20L plastic containers filled with two different kinds of soil substratum: mineral (1 trial) and organic (2 trial). The analyses of tomato acidity, nitrate quantity, vitamin C amount, dray matter harvest, level of technical maturity (evaluated by cracked tomato number) and comparative taste characteristics carried out. It was established that application of well-balanced complex fertilizers in quantities as small as N34P8K50 and N103P24K151 conditioned quality of tomato in mineral soil substratum identically to that of vegetables grown in the organic soil substratum. The increasing mineral fertilizer rates stimulated faster tomato maturing process and cracking of its skin. As a result of increasing fertilizer application the larger amounts of vitamin C and nitrate accumulated in the fruits. From the other hand, unfavorable conditions for tomato deceases and pest were tested in the experiment due to the vegetables growing in the special isolated 20L plastic containers. This treatment was highly effective in order to obtain all plants very healthy and therefore might be recommended for the farmers growing vegetables in light and poor soils of the Dzūkija region.

Controlled release systems aim at achieving a predictable and reproducible drug release profile over a desired time period. These controlled release formulations offer many advantages over conventional dosage forms. These advantages include: reduced dosing intervals, constant drug levels in the blood, increased patient compliance and decreased adverse effects. Complex controlled release formulations such as those with sustained release properties, often require additional steps during the production phase. The cost and economic impact associated with these complex controlled release dosage formulations often outweigh the short term benefits. Thus the development of an economic method to produce controlled release particles is of great importance especially in third world countries. In controlled release formulations the drug is often equally dispersed throughout a polymer matrix. In the presence of a thermodynamically compatible solvent, swelling occurs and the polymer releases its content to the surrounding medium. The rate of drug release can be controlled by interfering with the amount of swelling and rate of diffusion by manipulating the viscosity of the polymer matrix. Chitosan is an ideal candidate for controlled drug delivery through matrix release systems. It is a biodegradable polymer with absorption-enhancing properties. Cross-linking chitosan with different cross-linking agents allow the preparation of beads. Beads are frequently used in controlled release dosage forms as they are very flexible in dosage form development and show various advantages over single unit dosage forms. Because beads disperse freely in the gastrointestinal tract they maximize drug absorption, reduce fluctuation in peak plasma, and minimize potential side effects without lowering drug bio-availability. Chitosan beads and excipient containing chitosan beads were prepared and investigated as possible controlled release formulations. Pyrazinamide was chosen as the model drug. Chitosan beads and excipient containing chitosan beads were prepared by ionotropic gelation in tripolyphosphate. In this study chitosan/pyrazinamide beads containing pharmaceutical excipients (Ascorbic acid, Explotab and Ac-Di-Sol) were produced. The excipients were added individually and in combinations to the chitosadpyrazinamide dispersion and the beads were characterized on the basis of their morphology, solubility, fiability, drug loading capacity and swelling behaviour, as well as drug release (dissolution properties). The drug loading of the pyrazinarnide loaded chitosan beads, was 52.26 % 0.57%. It was noted that the inclusion of excipients in the beads resulted in an increase in drug loading with the combination of Ascorbic acid and Ac-Di-Sol giving the highest drug loading of 67.09 ± 0.22%. It was expected that the addition of the pharmaceutical excipients would lead to a sustained release of pyrazinamide. Dissolutions studies, however, revealed a burst release in both phosphate buffer solution (PBS) pH 5.60 and 7.40 over the first 15 minutes and the curve reached a plateau after 30 minutes. Thus, apparently the inclusion of the pharmaceutical excipients did not contribute to a sustained release of pyrazinamide over the tested period of six hours. In future studies the dissolution time can possibly be extended to a period of 24 hours. It might be possible for the remaining drug (approximately 40%) in the beads to be released over the extended period. Other polymers can also be investigated to control the release of pyrazinamide. Further studies are, however, necessary to investigate this possibility in the future. / Thesis (M.Sc. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2007.

We have investigated electropolymerisation for fabrication of a chemically modified working electrode for the determination of dopamine and catechol neurotransmitters in the presence of ascorbic acid. A variety of film compositions were investigated that would allow discrimination of the neurotransmitters through a combination of electrostatic barrier and the film porosity. The films investigated were based on different compositions of () poly-o-toluidine-co-aniline (POT-co-PA), () poly-o-toluidine-co-o-anisidine (POT-co-POA) and () polyacriflavine (PAF). The POT-co-PA and POT-co-POA gave the most promising result although the POT-co-PA was preferred because of higher current enhancement and better separation of dopamine and catechol neurotransmitters in the presence of ascorbic acid. The uses of electropolymerisation make the investigated films attractive candidates for the fabrication of a chemically modified microelectrode with application in capillary electrophoresis separation with electrochemical detection. The active area of nano particle (Au, Pt and Ag) screen printed electrodes was determined using cyclic voltammogram with ferro/ferricyanide couple. The active surface of the nano particle coated electrode was found surprisingly to be 5% - 65% lower than that geometrically calculated surface area for the electrode. This is ascribed to the limitation of the screen printing approach that was used. A low cost high replication approach that would allow development of a capillary electrophoresis microfluidic chip with electrochemical detection (CE-ECD) on a polymer substrate was investigated. A fluidic top layer was fabricated using hot embossing and an electrode bottom layer by metal patterning on a polymer substrate using metallisation and photolithography.

FABRICATION OF NANOPOROUS GOLD AND BIOLOGICAL APPLICATIONS By Badharinadh Uppalapati A Dissertation submitted in partial fulfillment of the requirements for the degree of Master of Science at Virginia Commonwealth University. Virginia Commonwealth University, 2014 Major Director: Maryanne M. Collinson, Professor, Department of Chemistry Fabrication of nanoporous gold electrodes by dealloying Au:Ag alloys has attracted much attention in sensing applications. In the first part of this work, the electrochemical response of the redox active molecule, potassium ferricyanide, in a solution of bovine serum albumin in buffer, serum or blood was studied using nanoporous gold and comparisons made to planar gold. Nanoporous gold electrodes with different surface areas and porosity were prepared by dealloying Au:Ag alloy in nitric acid for different dealloying times, specifically, 7.5, 10, 12.5, 20 minutes. Characterization was done using scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), energy dispersive X-ray spectroscopy (EDX), and cyclic voltammetry (CV). Using cyclic voltammetry, planar gold electrodes exposed to bovine serum albumin in buffer showed a decrease in Faradaic peak current and an increase in peak splitting for potassium ferricyanide. The time required for the peak Faradaic current to drop to one-half of its original value was 3 minutes. At nanoporous gold electrodes, however, no significant reduction in Faradaic peak current or increase in peak splitting was observed. Nanoporous gold electrodes having the smallest pore size and largest surface area showed ideal results to biofouling. These electrodes are believed to impede the mass transport of large biomolecules while allowing small redox molecules to exchange electrons effectively with the electrode. In the second part of this work, the open circuit potential (OCP) of biologic solutions (e.g., blood) was measured using nanoporous gold electrodes. Historically, the measurement of blood redox potential has been hindered due to significant fouling and surface passivation of the metal electrodes. As nanoporous gold electrodes retained electrochemical activity of redox probes like potassium ferricyanide in human serum and rabbit blood, they were used to measure the OCP of blood and plasma from various animals like pig, rabbit, rat, monkey and humans. Comparisons were made to planar gold electrodes. The OCP values at both the planar gold and nanoporous gold electrodes were different from each other and there was variability due to different constituents present in blood and plasma. The OCP of rabbit blood and crashed rabbit blood was measured and the values were found to be different from each other indicating that ORP helps in measuring the animal condition. Ascorbic acid was added to rabbit and sheep blood and OCP measured at the nanoporous electrodes. Addition of reducing agent to blood at different intervals and different concentrations showed a change in potential with concentration.

Based upon experimental evidence from the 1970’s we proposed that a reduced form of hydroxocobalamin should be capable of producing carbon dioxide (CO2) from carbon monoxide (CO) in blood, and that this conversion should be detectable. Using resonance raman spectroscopy we demonstrated that a mixture of hydroxocobalamin and ascorbic acid could create the reduced form of hydroxocobalamin. We used a closed-loop circulation system with a hollow-fiber membrane oxygenator to produce carboxyhemoglobin. Using sensitive gas monitoring equipment to the gas-out port of the oxygenator we analyzed the CO and CO2 concentrations coming from the oxygenator. The mixture of hydroxocobalamin and ascorbic acid caused a 5-fold increase in the CO2 concentration of the gas-out flow, in comparison to baseline and negative controls. These findings offer initial support for the potential use of a mixture of hydroxocobalamin and ascorbic acid as an injectable antidote for carbon monoxide poisoning.

L'acide ascorbic acid (AA) a été considéré, pendant longtempss, comme une molecule devantêtre absorbée dans la nutrition, et prévenant le scorbut. Notre hypothèse, fondé sur desrésulats de notre groupe, suggèrent de nouvelles fonctions.Parmi celles-ci, nous nous sommes posé la question de l'AA molècule de signalling, durantl'embryogenèse et chez l'adulte, commme l'acide rétinoique (principe actif de la vitamine A)l'est. A cet effet, nous avons utilisé deux modèles cellulaires : des cellules souchesembryonnaires murines et des lignées de cellules souches/progénétrices adultes. Nous avonsainsi montré que l'AA stimule la différentiation de ces cellules en cellules musculairessquelettiques et en osteoblastes et inhibe l'adipogenèse et la neurogenèse. Cet effet passe parle transporteur de l'AA SVCT2 et implique la voie p38/MAPK. D'autre part, nous avonsdemontré que l'AA agit en compétition avec le RA, sur la neurogenèse et la myogenèse.Enfin, dans des cellules mésenchymateuses adultes, nous avons montré que l'AA inhibel'adipogenèse et stimule l'ostéogenèse. Cette action, comme chez l'embryon implique SVCT2et une modulation du pool du cAMP.En conclusion, l'AA pousse les cellules à se différencier en cellule musculaire squelettique eten ostéoblste et inhibie l'adipogenèse et la neurogenèse. / AA has been considered for a long time as a molecule involved in nutrition, to prevent scurvy. Our hypothesis is that AA could also be involved in development during embryogenesis, as well as in cell differentiation in adults. The aim of this study is to evaluate the potential implication of AA in cell differentiation, especially of mesenchyme cells, and to propose potential pathways that could be involved in these processes. Using murine ESCs we observed that AA markedly enhance the differentiation of ESCs toward muscle cells. Furthermore, we demonstrated that induction of myocytes by AA involves p38MAPK pathway and p-CREB. Moreover, we demonstrated that AA acts in mirror with retinoic acid. ESCs treated with RA mainly differentiate into neuronal cells, but AA compete, in a dosage dependent way to this differentiation. AA induces differentiation of ESCs into cardiac myocytes and could probably acts through p38MAPK pathway. Regarding adipocyte we revealed that SVCT2 expression significantly decreased as preadipocytes cells differentiate to adipocytes. This data suggests that mature adipocytes could not receive signals from AA. In addition, our results show that the expression of SVCT2 is increased in cells treated with AA and without IBMX. Moreover, we demonstrated that AA evolves in decreasing of cells containing lipids. Finally, we demonstrated that AA is not only involved in muscle differentiation of mesenchyme but is also involved in adipose tissue as a negative inducer. In conclusion, AA drives differentiation of ESCs toward muscle cells and osteoblast, incompetition with RA, and has a negative effect on adipogenesis and neurogenesis differentiation.

A neuropatia autonômica que se manifesta em animais portadores de diabetes mellitus (DM) compromete vários órgãos. A nível gastrointestinal pode acarretar diarréia, constipação, estase e dilatação gástrica, levando a anorexia, perda de peso e vômito. Embora os mecanismos que desencadeiam esses distúrbios não estejam totalmente esclarecidos, parece que alterações do plexo mioentérico estão envolvidas. O estresse oxidativo juntamente com o aumento do sorbitol resultante das alterações metabólicas presentes no DM resultam em edema e lesão neuronal e, entre as conseqüências desta lesão, tem sido descrita alterações nos neurônios mioentéricos. O ácido ascórbico (AA) além da ação antioxidante pode reduzir a concentração de sorbitol atuando, desse modo, como neuroprotetor. O presente trabalho teve como objetivos investigar possíveis alterações no número e no tamanho dos neurônios mioentéricos NADH-diaforase reativos (NADH-dr) do estômago de ratos diabéticos e o efeito da suplementação com AA (1g/L de água) nos referidos parâmetros. Para tanto, 15 ratos (Rattus norvegicus) foram separados em três grupos (n = 5): controle (C); diabético (D); e diabético suplementado com AA (DS). O DM foi induzido por estreptozootocina (35 mg/kg de peso corporal). Após 120 dias de experimento, os animais foram anestesiados, sacrificados e seus estômagos retirados. Os neurônios mioentéricos foram evidenciados pelo método da NADH-diaforase. Por meio de microscópio de luz foram contados os neurônios NADH-dr e, pelo programa computadorizado para análise de imagens, foi mensurado o perfil do corpo celular (PCC) desses neurônios. As regiões aglandular e glandular do estômago foram analisadas separadamente. O número de neurônios NADH-dr foi maior (P<0,05) na região glandular do grupo DS quando comparado com o grupo D. A média dos PCCs foi maior (P<0,05) para os neurônios dos grupos D e DS do que para o grupo C. Ocorreu aumento na incidência de neurônios com PCC superior a 200 &micro;m2 no grupo D quando comparada aos grupos DS e C. Os resultados sugerem que a suplementação com AA teve efeito neuroprotetor sobre os neurônios mioentéricos NADH-dr representado pelo aumento no número de neurônios e diminuição de neurônios grandes na região glandular do grupo DS. / The autonomous neuropathy present in animals with diabetes mellitus (DM) compromises several organic systems. On the gastrointestinal tract it may provoke diarrhea, constipation, estase and gastric dilation, leading to anorexia, weight loss and vomit. Although the triggering mechanisms of these disturbances are not completely clear, it seems that alterations of the myenteric plexus may be involved. The oxidizing stress - together with the sorbitol increase resulting from the metabolic alterations due to dm - results in an edema and in a neuronal lesion. Among the consequences of this lesion, alterations in the myenteric neurons have been described. The ascorbic acid (AA), besides its antioxidant effect, may also reduce the sorbitol concentration working as a neuroprotector. The aims of the present work were to investigate the possible alterations in the number and size of nadh-dr stained myenteric neurons of the stomach of diabetic rats and the effect of the aa-supplementation (1g/l of water). In order to do so, 15 rats (rattus norvegicus) were divided into three groups (n = 5): controls (C); diabetic (D); and diabetic supplemented with AA (DS). DM was induced by streptozotocin (35 mg/kg of body weight). 120 days later, the animals were sacrificed for obtaining their stomachs. The myentric neurons were stained employing the nadh-diaforase method. We counted the nadh-dr neurons employing the light microscope and we measured the cellular body profile (CBP) using an image analysis software. The aglandular and glandular areas of the stomach were analyzed separately. The number of neurons nadh-dr was larger (p <0,05) in the glandular area of group ds when compared to group d. The average of cbps was larger (p <0,05) for neurons of groups d and ds than for group c. There was an increase in the incidence of neurons with cbps over 200 mm2 in group D when compared to groups DS and C. The results suggest that the AA supplementation had a neuroprotetor effect on the nadh-dr myenteric neurons represented by the increase in the number of neurons and the decrease of big neurons in the glandular area of group DS.

Este trabalho consta de uma avaliação da qualidade intrínseca e da qualidade percebida por potenciais consumidores espanhóis de polpa de camu-camu [Myrciaria dubia (H.B.K.) McVaugh] submetida a quatro diferentes tratamentos: congelamento, pasteurização, pressurização (alta pressão hidrostática) e liofilização, armazenada congelada por quatro meses. A avaliação da qualidade intrínseca foi realizada por meio de análises dos parâmetros pH, sólidos solúveis, acidez, coloração, teores de ácido ascórbico, de compostos fenólicos e de antocianinas, e a avaliação da qualidade percebida por meio de testes sensoriais para os parâmetros aparência, cor, aroma e sabor. Este estudo justifica-se pelo reduzido consumo de camu-camu, uma fruta nativa da Amazônia com o mais elevado teor de vitamina C observado em estudos científicos (entre 1.721 e 2.900 mg de ácido ascórbico.100 g-1), e que contém outros importantes compostos com atividade antioxidante como compostos fenólicos e carotenoides. Uma das razões do baixo consumo da fruta fresca é sua elevada acidez (entre 2,31 e 3,08 g de ácido cítrico.100 g-1), portanto, é importante a apresentação de alternativas de consumo que preservem sua principal característica nutricional (elevado teor de vitamina C) e demais atividades biológicas, ainda pouco exploradas, enquanto enriquece seus valores sensoriais. Neste trabalho os valores dos parâmetros físicos e químicos encontrados para as polpas de camu-camu ao longo do armazenamento permaneceram estáveis. Foram observados diferentes efeitos dos tratamentos sobre os teores de ácido ascórbico, compostos fenólicos e antocianinas, sendo que a pasteurização foi o tratamento que causou maior impacto negativo. Sensorialmente a polpa reconstituída de liofilizado de camu-camu apresentou resultados inferiores às polpas submetidas aos demais tratamentos. Os teores de ácido ascórbico das polpas de camu-camu após 125 dias de armazenamento estiveram entre 1.200 e 1.796 mg por 100 g de polpa, valores relativamente inferiores aos apresentados em publicações sobre camu-camu nos últimos anos, entretanto, comparáveis aos apresentados para acerola (entre 1.191 e 1.921 mg.100 g-1) e kakadu (1.240 mg.100 g-1) e superiores ao apresentado para rosehip (1.141 mg.100 g-1) outras fontes destacadas deste nutriente. Os néctares de camu-camu são uma alternativa para o consumo, visto que a fruta e suas polpas por serem destacadamente ácidas sofrem rejeição por potenciais consumidores / The purpose of this work is to present an assessment of the intrinsic quality and the quality perceived by potential Spanish consumers of camu-camu [Myrciaria dubia (H.B.K.) McVaugh] pulp submitted to four different treatments: freezing, pasteurization, pressurization (high hydrostatic pressure) and lyophilization, stored frozen for four months. The intrinsic quality assessment was performed by analysis of pH, soluble solids, acidity, color, contents of ascorbic acid, phenolic compounds and anthocyanins, and assessment of the quality perceived by sensory tests for the parameters of appearance, color, aroma and flavor. This study is justified by the low consumption of camu-camu, an Amazon native fruit with the highest vitamin C content observed in scientific studies (between 1,721 and 2,900 mg of ascorbic acid .100 g-1), and contains other important compounds with antioxidant activity such as phenolics compounds and carotenoids. One reason for the low consumption of fresh fruit is its high acidity (between 2.31 and 3.08 g of citric acid.100 g-1), so it is important to introduce consumption alternatives that preserve their main nutritional characteristic (high vitamin C content) and other biological activities, yet little explored, while enriching their sensory values. In this study, it was detected that the values of physical and chemical parameters found for the camu-camu pulp throughout the storage period remained stable. It was observed that different effects of treatments on ascorbic acid, phenolic compounds and anthocyanins; and that pasteurization was the treatment that caused major negative impact. Sensorially reconstituted freeze-dried camu-camu pulp showed results lower than those subjected to other treatments. The ascorbic acid content of camu-camu pulp after 125 days of storage was between 1,200 and 1,796 mg per 100 g of pulp, relatively lower than the values presented in publications on camu-camu in recent years; however, they were closely comparable to those for acerola fruit (between 1,191 and 1,921 mg.100 g-1) and kakadu fruit (1,240 g.100 g-1) and superior to that presented to rosehip fruit (1,141 mg.100 g-1) other highlighted sources of this nutrient. The camu-camu nectars are an alternative way for the consumer, since the fruit and its pulp, by being prominently acidic, suffer rejection by potential consumers

Objetivou-se realizar a caracterização física, química e funcional do fruto de camu-camu classificados nos estádios 2 (verde-avermelhado) e 4 (roxo), utilizando-se distintas doses de radiação gama (0; 0,5; 1 kGy) e acompanhar a atividade respiratória dos frutos, monitorados por um período de 25 dias em dois níveis de temperatura (5 e 25 oC). Os frutos foram avaliados quanto à cor instrumental da casca, firmeza, sólidos solúveis, acidez, ratio, pH, antocianinas, compostos fenólicos, atividade antioxidante, atividade respiratória e teor de ácido ascórbico. A irradiação com dose de 0,5 kGy teve influência positiva nos frutos no estádio 2 armazenados em temperatura ambiente, prolongando seu tempo de vida útil em dois dias, totalizando 9 dias. Os frutos tratados com 1 kGy tiveram seu tempo de vida útil reduzido em 6 dias. Em relação à firmeza, o ponto de maturação teve grande influência nas respostas; comportamento semelhante aconteceu com as antocianinas, onde os frutos no estádio 4 tiveram valores até 10 vezes superiores aos frutos verde-avermelhado (estádio 2), mas, de forma geral, as antocianinas se mantiveram estáveis em relação às doses testadas e ao tempo de armazenamento. A dose de 1 kGy influenciou nos compostos antioxidantes dos frutos refrigerados, diminuindo 83,4 e 67,46% do teor para os frutos nos estádios 2 e 4, respectivamente. O ácido ascórbico no camu-camu se mostrou altamente estável, mesmo quando o fruto já estava impróprio para o consumo, apresentando valores acima de 1000 mg de ácido ascórbico 100 g-1 de fruto. Exceção foi para os frutos refrigerados tratados com 1 kGy, que apresentaram valores de 263,67 mg 100 g-1 para os frutos no estádio 2 e 626,1 mg 100 g-1 para os frutos no estádio 4. De forma geral, a refrigeração teve maior influência na conservação do camu-camu, aumentando em média 18 dias o seu armazenamento, a radiação gama nas doses estudadas não se mostrou um tratamento vantajoso para aumentar a vida útil do camu-camu refrigerado. O fruto do camu-camu apresentou baixos valores de ratio (índice de palatabilidade), o que faz do fruto não palatável para o consumo in natura, elevados teores de antocianinas, compostos fenólicos, atividade antioxidante e ácido ascórbico / The objective of this work was to characterize physical, chemical and functional of the camu-camu fruit in its 2 stage (green- reddish) and 4 (purple), using different doses of gamma radiation (0, 0.5, 1 kGy) and to keep up with the respiratory activity over a period of 25 days in two temperature levels (5 and 25 ºC). The fruits were evaluated when the instrumental color of skin, firmness, soluble solids, acidity, ratio, pH, anthocyanins, phenolic compounds, antioxidant, respiratory activity and ascorbic acid content. Irradiation with 0,5 kGy had a positive influence on the 2 stage fruits stored at room temperature, extending its lifetime two days, totaling nine days. The fruit treated with 1 kGy had its lifetime reduced by 6 days. About the firmness, the maturation point had great influence on the answers; responses similar happened with anthocyanins, where the purple fruits had values up to 10 times higher than the green-reddish fruits, but, in general, anthocyanins were stable in relation to the doses tested and the storage time. There was a decrease of antioxidants in 83,4 and 67,46% for 2 and 4stages fruits, respectively. Ascorbic acid in camu-camu was highly stable even when the fruit was already unfit for consumption, with values above 1000 mg ascorbic acid 100 g-1 fruit. There was an exception with the refrigerated fruits treated with 1 kGy, which showed values of 263,67 mg 100 g-1 for the 4 stage fruits and 626.1 mg 100 g-1 for the 2 stage fruits. In general, the cooling had a greater influence on the conservation of camu-camu, increasing on average 18 days your storage, the gamma radiation doses studied did not prove advantageous treatment to extend the lifetime of the camu-camu in refrigeration. The camu-camu fruit showed low values of ratio (index of palatability), which makes the fruit unsuitable for fresh consumption, high levels of anthocyanins, phenolic compounds, antioxidant activity and ascorbic acid

Nesse trabalho são apresentados resultados sobre o desenvolvimento de microeletrodos por desgaste eletroquímico de microfibras de dimensões da ordem de 100 µm de diferentes materiais (platina, ouro e fibra de carbono) visando à utilização em micro-ambientes. Na grande maioria dos casos esse processo foi realizado utilizando transformadores AC. Arranjo de microeletrodos construídos a partir de técnicas litográficas também foram construídos. A aplicabilidade de microeletrodos de Pt construídos por desgaste eletroquímico foi avaliada para monitorar a concentração \"in-situ\" de ácido ascórbico em diferentes frutas. O sensor foi capaz de avaliar a distribuição espacial da concentração de ácido ascórbico \"in-situ\" em laranjas. A partir desses resultados, mapas para a concentração de ácido ascórbico foram construídos. A correlação entre o estado de maturação e a concentração de ácido ascorbico também foi observada a partir das medições realizadas e concentrações maiores foram encontradas em frutas mais maduras. O arranjo de microeletrodos foi utilizado para avaliar a concentração de iodato em amostra de sal de cozinha utilizado pequenos volumes de amostra (500 µL). Verificada a possibilidade de fabricação de microeletrodos de diferentes geometrias e dimensões, estudos visando ao monitoramento e à detecção de DNA e ácido ascórbico foram efetuados. Superfícies eletródicas modificadas com filmes de óxido de rutênio hexacianoferrato (RuOHCF) foram utilizadas para a detecção amperométrica de 2\'-deoxiguanosina (dG) e ácido ascórbico por análise por injeção em fluxo (FIA). O método para detecção de dG apresentou um resposta linear de 3,8 a 252 µmol L-1, com um limite de detecção de 94 nmol L-1. Aplicações em amostras de DNA também foram realizadas, e os resultados para determinação da dG concordaram com os obtidos pelo método padrão (HPLC). Estudos sobre o processo eletrocatalítico de oxidação da dG em meio ácido em filmes de RuOHCF foram investigados utilizando eletrodo rotativo. Sobre esse mesma superfície modificada, o processo eletrocatalítico permitiu a determinação de ácido ascórbico em pH = 6,9 a 0,0 V vs Ag/AgCl em FIA, com um limite de detecção de 2,2 µmol L-1. Estudos sobre o processo eletrocatalítico de oxidação do ácido ascórbico também foram avaliados com eletrodo rotativo. O método foi aplicado para a determinação de ácido ascórbico em urina, obtendo-se bons valores de recuperação (96 - 104 %). Microeletrodos contendo filmes eletrodepositados de RuOHCF foram utilizados para monitorar o consumo de ácido ascórbico em 3 diferentes tipos de células neuronais (SH-SY5Y, células transfectadas com SOD humana e mutante típica da doença de Esclerose Lateral Amiotrófica (ALS)). Os resultados do consumo de ácido ascórbico para essas células estão de acordo com os níveis de estresse oxidativo induzido pela SOD mutante. Experimentos visando a aplicabilidade de microeletrodos inseridos em células também foram efetuados. / Results on the development of microelectrodes fabricated by electrochemical etching using fibers of different materials (platinum, gold and carbon fiber) and dimensions (starting from 100 µm) for the application in micronenvironments are presented. In almost all cases this procedure was carried out using AC transformer. Array of microelectrodes were also fabricated by using litographic techniques. The applicability of Pt microelectrodes fabricated by electrochemical etching was evaluated in the in-situ monitoring of the ascorbic acid concentration in different fruits. The sensor allowed spatial distribution of ascorbic acid concentration in oranges to be found and concentration maps were constructed. A correlation between the ripening stage and the ascorbic acid concentration was also observed from electrochemical measurements, the ascorbic acid content being higher in mature fruits. Studies on the detection of species involved in the oxidative stress process, such as DNA and ascorbic acid, were also performed. Ruthenium oxide hexacyanoferrate (RuOHCF) modified electrode surfaces were used as amperometric detectors for 2\'-deoxyguanosine (dG) and ascorbic acid determinations in FIA apparatus. The method exhibited a linear response range to 2\'-deoxyguanosine from 3.8 to 252 µmol L-1 dG with detection limit of 94 nmol L-1. Applications in DNA samples were examined, and the results for determination of 2\'-deoxyguanosine were in good agreement with those obtained by HPLC analysis. The dG electrocatalytic oxidation at a RuOHCF glassy carbon (GC) modified electrode was investigated in acid medium by using rotating disc electrode (RDE) voltammetry. On this modified surface, the electrocatalytic process allowed the determination of ascorbic acid to be performed at 0.0 V and pH = 6.9 with a limit of detection of 2.2 µmol L-1 in a flow injection configuration. The usefulness of the method was demonstrated by an addition-recovery experiment with urine samples and the recovered values were in the 96 to 104 % range. Investigations on the mechanism of the electrocatalytic oxidation of ascorbic acid was also investigated at pH = 6.9 by using RDE voltammetry. The RuOHCF carbon fiber modified microelectrode was used to monitor the ascorbate uptake by control SH-SY5Y cells, and cells transfected with wild-type Cu,Zn-Superoxide Dismutase (SOD) or with a mutant SOD (SOD G93A) typical of familial Amyotrophic Lateral Sclerosis (ALS). Data on the rate of ascorbate uptake by these cells were in agreement with the level of oxidative stress induced by the mutant SOD. Attemps to use the microelectrodes inside single cells were also performed