This thesis uses one of Brain Computer Interface (BCI) products, NeuroSky headset, to design a prototype model to control window blind by using headset’s single channel electrode. Seven volunteers performed eight different exercises while the signal from the headset was recorded. The dataset was analyzed, and exercises with strongest power spectral density (PSD) were chosen to continue to work with. Matlabs spectrogram function was used to divide the signal in time segments, which were 0.25 seconds. One segment from each of these eight exercises was taken to form different combinations which were later classified.The classification result, while using two of proposed exercises (tasks) was successful with 97.0% accuracy computed by Nearest Neighbor classifier. Still, we continued to investigate if we could use three or four thoughts to create three or four commands. The result presented lower classification accuracy when using either 3 or 4 command thoughts with performance accuracy of 92% and 76% respectively.Thus, two or three exercises can be used for constructing two or three different commands.

Orientador: Vera Maria Barbosa de Moraes / Banca: Maria Cristina Thomaz / Banca: Alexandre Oba / Resumo: O objetivo do presente trabalho foi o de determinar a digestibilidade do extrato de leveduras, avaliação do desempenho, rendimento de carcaça, morfometria e ultra-estrutura da mucosa intestinal e resposta imune humoral de frangos de corte criados em diferentes temperaturas e que receberam, na fase pré-inicial, ração contendo ou não extrato de leveduras e/ou prebiótico. Foram utilizados 1440 pintainhos Cobb-500® machos de um dia de idade, criados sob diferentes temperaturas a partir do primeiro dia de vida. As rações acrescidas ou não com extrato de leveduras e/ou prebiótico foram oferecidas somente na fase pré- inicial (1-7 dias) e a partir do 8º dia todas as aves receberam a mesma ração. Conclui-se que o extrato de leveduras apresenta 92,49% de matéria seca, 48,07% de proteína bruta, 4.883 Kcal/kg de energia bruta, e apresenta, em média, um coeficiente de digestibilidade verdadeira dos aminoácidos de 99,42%, sendo rico em ácido glutâmico, leucina, ácido aspártico, serina, glicina. A alta temperatura ambiente prejudicou o desempenho, rendimento de carcaça, morfometria intestinal e resposta imune humoral. A inclusão de prebiótico na ração pré-inicial resultou em maior ganho de peso e melhor conversão alimentar nas aves criadas sob alta temperatura, ao final de 42 dias de idade, além de aumentar a viabilidade criatória até os 21 dias de idade. Observou-se também que a inclusão dos produtos nas temperaturas quente e fria produziu efeito benéfico sobre as vilosidades frente ao estresse ambiental e que os aditivos estudados não produziram efeito sobre a resposta imune humoral para VDN e VDG ao longo do ciclo produtivo do frango de corte. / Abstract: The aim of this experiment was to determine the digestibility of the yeast extract, estimate the performance, carcass yield, intestinal mucosa structure and ultra structure and humoral immune response of broiler reared in different temperatures and that received in started diet containing or not extract of yeasts or prebiotic. 1440 one-day old male Cobb-500® were reared in different temperatures from first day of life. The diets with or without yeast extract and/or prebiotic were provided only in starter diet (1 to 7 days), forward of 8 days all broiler were fed with same commercial diet. The yeast extract contain 92.49% of dry matter, 48.07% of crude protein, 4,883 kcal/kg of crude energy /kg, and, on average a coefficient of digestibility true of amino acids of 99.42%. The amino acids digestibilities are glutamic acid, leucine, aspartic acid, serine, glycine. The high temperature damages the performance, carcass yield, intestinal mucosa structure, villus densities and antibody titters. The prebiotic inclusion in started diet resulted in increase body weight and improved feed conversion in the birds reared in high temperature to the 42 days of age and besides increase the viability until the 21 days of age. It was also observed that the inclusion of the two products in the hot and cold temperatures produced beneficial effect on the villus front in environmental stress and that the yeast extract and prebiotic did not produce effect on immune humoral response along the productive cycle of broiler. / Mestre

Os cães são afetados por doenças inflamatórias e neoplásicas que compartilham diversas similaridades com as desordens em humanos, assim seu estudo representa um importante modelo animal para as condições humanas. As células dendríticas (DCs) representam a população mais potente de células apresentadoras de antígenos. As DCs representam também um novo alvo promissor de imunoterapia em cães; no entanto o uso terapêutico de DC caninas é restrito, dentre outros fatores, devido a falta de padronização nas técnicas de isolamento e limitado numero de informações específicas da espécie a esse respeito. Este projeto tem por finalidade avaliar a geração de células dendríticas caninas geradas in vitro e ativadas por diferentes estímulos biológicos na presença e ausência de extrato tumoral de melanoma canino. Os resultados demonstraram que as DCs caninas geradas na presença de extrato tumoral em grandes concentrações apresentavam atividade funcional semelhante as DCs maduras / Dogs are affected by inflammatory and neoplastic diseases that share many similarities with the disorders in humans, so their study is an important animal model for the human condition. Dendritic cells (DCs) are the most potent population of antigen presenting cells. DCs also represent a promising new target for immunotherapy in dogs; However, the therapeutic use of canine DC is restricted among others factors due to lack of standardization in isolation techniques and limited number of species-specific information in this regard. This project aims to assess the generation of canine dendritic cells generated in vitro and activated by different biological stimuli in the presence and absence of tumor extract of canine melanoma. The results showed that the canine DCs generated in the presence of high concentrations tumor extract showed similar functional activity of mature DCs

Made available in DSpace on 2016-04-28T20:17:28Z (GMT). No. of bitstreams: 1 2008 - Alexander Santos.pdf: 2028526 bytes, checksum: 5876d3f1d1f305df3af598f0bef1531d (MD5) Previous issue date: 2008-04-15 / This work was developed in the Department of Microbiology and Immunology of the Institute of Veterinary Rural Federal University of Rio de Janeiro in Serop?dica-RJ. We evaluated the antifungal activity plant Cymbopogon citratus (lemon-grass), Peumus boldus (boldo) and Shinus terebinthifolia (aroeira), inhibition of growth of species of the genus Aspergillus (A. flavus, A. niger, A. ochraceus, A. Parasiticus and A. carbonarius. Using the method of minimum inhibitory concentration in agar, with the technique of dilution plate (Pour-Plate), were held dilutions of different extracts, resulting in the final concentrations of 5%, 2.5% and 1.25%. Testing of commercial sensitivity to antifungal - ketoconazole were made in a final concentration of 1933.18 ? g /mL, as recommended by the National Committee for Clinical Laboratory Standards (NCCLS). The results obtained in the experiments have shown that aqueous extracts of C. citratus and S. terebinthifolia, after 24 hours of incubation, were able to inhibit the growth of A. flavus and A. carbonarius, respectively. The other extracts being studied and tested in the concentrations were not able to inhibit the growth of mycelial species of the genus Aspergillus. / Este trabalho foi desenvolvido no Departamento de Microbiologia e Imunologia do Instituto de Veterin?ria da Universidade Federal Rural do Rio de Janeiro, em Serop?dica-RJ. Foram avaliadas a atividade antif?ngica das plantas Cymbopogon citratus (capim-lim?o), Peumus boldus (boldo) e Shinus terebinthifolia (aroeira), na inibi??o do crescimento de esp?cies do g?nero Aspergillus (A. flavus, A. niger, A. ochraceus, A. parasiticus e A. carbonarius). Utilizando o m?todo da concentra??o inibit?ria m?nima em ?gar, com a t?cnica de dilui??o em placa (Pour-Plate), foram realizadas dilui??es dos diferentes extratos, obtendo-se as concentra??es finais de 5%, 2,5% e 1,25%. Testes de sensibilidade ao antif?ngico comercial - cetoconazol foram realizados numa concentra??o final de 1933,18 ?g/mL, conforme recomendada pelo National Committee for Clinical Laboratory Standards (NCCLS). Os resultados obtidos nos experimentos demonstraram que os extratos aquosos de C. citratus e S. terebinthifolia, ap?s 24 horas de incuba??o, foram capazes de inibir o crescimento de A. flavus e A. carbonarius, respectivamente. Os demais extratos em estudo e nas concentra??es testadas, n?o foram capazes de inibir o crescimento micelial das esp?cies do g?nero Aspergillus.

Muitas drogas terapêuticas produzidas pela indústria farmacêutica são estruturas químicas isoladas de organismos encontrados na natureza ou moléculas baseadas nelas. Podem ser incluídas nesse grupo drogas isoladas de organismos marinhos, como corais, esponjas e algas marinhas, conhecidos como produtores de grandes quantidades de metabólitos secundários. Com base neste fato o presente estudo teve como objetivo realizar a prospecção de moléculas bioativas com propósito farmacológico, em extratos de algas marinhas vermelhas (Rhodophyta) e pardas (Heterokontophyta) coletadas no litoral brasileiro. A prospecção foi realizada por meio de avaliação de seus potenciais antioxidante, antibacteriano, antifúngico, anticancerígeno, e antiparasitário contra organismos causadores de leishmaniose e esquistossomose. Para as avaliações foram empregadas os extratos supercríticos de 5 espécies diferentes, sendo 2 pardas: Dictyota dichotoma e D. menstrualis e três vermelhas: Chondria littoralis, Spyridia hypnoides e Plocamium brasiliense. Os extratos foram avaliados quanto aos seus potenciais bioativos e os resultados mais promissores foram selecionados para as etapas seguintes do fracionamento. Em uma avaliação geral os extratos apresentaram bons resultados e representam uma potencial fonte de bioativos. Os extratos das espécies de D. dichotoma e D. menstrualis foram então submetidos a um procedimento de fracionamento bioguiado pela atividade esquistossomicida. Incorporou-se ainda um terceiro extrato de D. mertensii aos estudos e todas as etapas do fracionamento foram monitoradas por LC-MS. Comparando-se as massas detectadas em todas as frações que apresentaram atividade, para os 3 extratos, foi verificado que a substância de m/z 271,24 estava presente em todas elas, portanto os procedimentos de isolamento foram direcionados a esta molécula para a qual foi possível isolar 7 mg. Diferentemente do que era esperado a molécula quando avaliada isoladamente não apresentou atividade esquistossomicida, levando a hipótese de que a atividade seja decorrente de uma molécula diferente para cada espécie ou ainda que a mesma seja decorrente de uma interação com outras substâncias por um mecanismo de ação aditivo ou sinérgico. O trabalho avaliado de forma geral apresentou resultados promissores e representa um grande embasamento para servir como base para posteriores trabalho de fracionamento. / Several therapeutic drugs manufactured by the pharmaceutical industry are chemical structures isolated from organisms that are found in nature or molecules based on that. May be included at this group drugs isolated from marine organisms, like corals, sponges and seaweeds, known as great secondary metabolites producers. Based on this facts the objective of the present study is to perform a prospection study to achieve bioactive molecules with pharmaceutical purposes, on extracts made from red (Rhodophyta) and brown (Heterokontophyta) seaweed collected in the Brazilian shore. The prospection studies was performed by means of evaluation of the antioxidant, antibacterial, antifungal, anticancer and antiparasitic (against Leishmania and Schistosoma) potential. In the evaluation were tested the supercritical extracts of 5 different species, including 2 brown species: Dictyota dichotoma and D. menstrualis and 3 red species: Chondria littoralis, Spyridia hypnoides and Plocamium brasiliense. The extracts were evaluated by their potential bioactive compounds and the most promising results were selected for the following fractionation steps. Overall the extracts have shown good results and may be represent a potential source of bioactive molecules. The extracts of both D. dichotoma and D. menstrualis were submitted to a bioguided fractionation process by their antischistosomal activities. It was still included a third extract from D. mertensii to the studies and every step was monitored by LC-MS techniques. Comparing the detected mass for each active fraction, it was observed the presence of a substance with m/z 271,24 in all of the extracts, so the isolating procedures were directed to obtain that specific molecule, which was obtained in a biomass of 7 mg. Differently than expected the molecule when evaluated isolated do not show the antischistosomal activity, leading to the hypothesis that the activity was related to different molecules for each species or even the observed effect is resulted by an interaction mechanism with another substances by an additive or synergist mechanism. The overall evaluation of the whole work show some promising results and it represent a great support for future fractionation works.

Muitas drogas terapêuticas produzidas pela indústria farmacêutica são estruturas químicas isoladas de organismos encontrados na natureza ou moléculas baseadas nelas. Podem ser incluídas nesse grupo drogas isoladas de organismos marinhos, como corais, esponjas e algas marinhas, conhecidos como produtores de grandes quantidades de metabólitos secundários. Com base neste fato o presente estudo teve como objetivo realizar a prospecção de moléculas bioativas com propósito farmacológico, em extratos de algas marinhas vermelhas (Rhodophyta) e pardas (Heterokontophyta) coletadas no litoral brasileiro. A prospecção foi realizada por meio de avaliação de seus potenciais antioxidante, antibacteriano, antifúngico, anticancerígeno, e antiparasitário contra organismos causadores de leishmaniose e esquistossomose. Para as avaliações foram empregadas os extratos supercríticos de 5 espécies diferentes, sendo 2 pardas: Dictyota dichotoma e D. menstrualis e três vermelhas: Chondria littoralis, Spyridia hypnoides e Plocamium brasiliense. Os extratos foram avaliados quanto aos seus potenciais bioativos e os resultados mais promissores foram selecionados para as etapas seguintes do fracionamento. Em uma avaliação geral os extratos apresentaram bons resultados e representam uma potencial fonte de bioativos. Os extratos das espécies de D. dichotoma e D. menstrualis foram então submetidos a um procedimento de fracionamento bioguiado pela atividade esquistossomicida. Incorporou-se ainda um terceiro extrato de D. mertensii aos estudos e todas as etapas do fracionamento foram monitoradas por LC-MS. Comparando-se as massas detectadas em todas as frações que apresentaram atividade, para os 3 extratos, foi verificado que a substância de m/z 271,24 estava presente em todas elas, portanto os procedimentos de isolamento foram direcionados a esta molécula para a qual foi possível isolar 7 mg. Diferentemente do que era esperado a molécula quando avaliada isoladamente não apresentou atividade esquistossomicida, levando a hipótese de que a atividade seja decorrente de uma molécula diferente para cada espécie ou ainda que a mesma seja decorrente de uma interação com outras substâncias por um mecanismo de ação aditivo ou sinérgico. O trabalho avaliado de forma geral apresentou resultados promissores e representa um grande embasamento para servir como base para posteriores trabalho de fracionamento. / Several therapeutic drugs manufactured by the pharmaceutical industry are chemical structures isolated from organisms that are found in nature or molecules based on that. May be included at this group drugs isolated from marine organisms, like corals, sponges and seaweeds, known as great secondary metabolites producers. Based on this facts the objective of the present study is to perform a prospection study to achieve bioactive molecules with pharmaceutical purposes, on extracts made from red (Rhodophyta) and brown (Heterokontophyta) seaweed collected in the Brazilian shore. The prospection studies was performed by means of evaluation of the antioxidant, antibacterial, antifungal, anticancer and antiparasitic (against Leishmania and Schistosoma) potential. In the evaluation were tested the supercritical extracts of 5 different species, including 2 brown species: Dictyota dichotoma and D. menstrualis and 3 red species: Chondria littoralis, Spyridia hypnoides and Plocamium brasiliense. The extracts were evaluated by their potential bioactive compounds and the most promising results were selected for the following fractionation steps. Overall the extracts have shown good results and may be represent a potential source of bioactive molecules. The extracts of both D. dichotoma and D. menstrualis were submitted to a bioguided fractionation process by their antischistosomal activities. It was still included a third extract from D. mertensii to the studies and every step was monitored by LC-MS techniques. Comparing the detected mass for each active fraction, it was observed the presence of a substance with m/z 271,24 in all of the extracts, so the isolating procedures were directed to obtain that specific molecule, which was obtained in a biomass of 7 mg. Differently than expected the molecule when evaluated isolated do not show the antischistosomal activity, leading to the hypothesis that the activity was related to different molecules for each species or even the observed effect is resulted by an interaction mechanism with another substances by an additive or synergist mechanism. The overall evaluation of the whole work show some promising results and it represent a great support for future fractionation works.

O experimento 1 teve o objetivo de avaliar o efeito de dietas suplementadas com sais de cálcio de óleo de palma contendo diferentes concentrações (3,8% e 7,0%) de extrato etéreo (EE) e suas interações com níveis crescentes de inclusão de lisolecitina (0, 0,5 e 1 g por unidade percentual do extrato etéreo da ração) para tourinhos da raça nelore em fase de terminação. Um total de 234 tourinhos, previamente adaptados a dietas de alta energia, com peso inicial médio de 297 kg, foram alojados por 106 dias em 30 baias. O delineamento experimental utilizado foi em blocos ao acaso, em arranjo fatorial 3 x 2 (três níveis de lisolecitina e dois níveis de EE). A ingestão de matéria seca, ganho médio diário e peso corporal final foram menores para a dieta de bovinos alimentados com maior concentração de EE (P <0,05). Houve interação (P <0,1) entre o nível dietético de EE e a lisolecitina. A lisolecitina aumentou o ganho médio diário e o peso corporal final de bovinos alimentados com dietas contendo maior concentração de EE, mas não de bovinos alimentados com dietas contendo menor concentração de EE. O objetivo do experimento 2 foi avaliar o efeito de dietas com diferentes fontes de gordura (sais de cálcio de óleo de soja, sais de cálcio de óleo de palma e óleo de soja degomado) e dois níveis de lisolecitina (0 e 7 g / cabeça / dia). sobre a fermentação ruminal e digestibilidade do trato total de nutrientes. Um total de 30 novilhos Nelore foram alojados em 30 baias individuais por 30 dias (23 dias de adaptação e 7 dias de coleta). O delineamento experimental utilizado foi em blocos ao acaso, em arranjo fatorial 3 x 2 (três fontes de gordura e dois níveis de lisolecitina). Houve interação entre fontes de gordura e lisolecitina para pH ruminal (P = 0.01), proporções molares no rúmen de acetato (P <0.01), propionato (P = 0.02), isobutirato (P <0.01), valerato (P <0.01), isovalerato (P <0,01) e concentração molar de AGV total (P = 0,09). De acordo com os dados, não houve efeito de interação (P> 0,05) entre a fonte de gordura e a lisolecitina para ingestão de matéria seca e a digestibilidade aparente dos nutrientes. A lisolecitina melhorou a fermentação ruminal de dietas com óleo de soja degomado e não teve efeito sobre a digestibilidade da EE independente da fonte de gordura, mas aumentou a digestibilidade aparente de fibra no trato total. / Experiment 1 was designed to evaluate the effect of diets supplemented with calcium salts of palm oil to contain different concentrations (3.8% and 7.0%) of ether extract (EE) and their interacions with increasing levels of inclusion of Lysolecithin (0, 0.5 and 1 g per percentage unit of ether extract of the diet) for finishing Nellore bulls. A total of 234 Nelore bulls, previously adapted to high energy diets, with a mean initial body weight of 297 kg were housed for 106 days in 30 pens. The experimental design was randomized blocks in a 3 x 2 factorial arrangement (three levels of lysolecithin and two EE levels). Intakes of DM, average daily gain en final body weight were lower for cattle fed hihg EE diets (P<0.05). There was an interaction (P<0.1) between dietary EE level and Lysolecithin. Lysolecithin increased cattle average daily gain and final body weight of cattle fed hihg EE diets, but not of cattle fed low EE diets. The objective of experiment 2 was to evaluate the effect of diets with different fat sources (calcium salts of soybean oil, calcium salts of palm oil and degummed soybean oil) and two levels of inclusion of Lysolecithin (0 and 7 g/head/day) on rumen fermentation and total tract digestibility of nutrients. A total of 30 fininshing Nellore steers were housed in 30 individual pens for 30 days (23 days adptation and 7 days collection period). The experimental design was randomized blocks in a 3 x 2 factorial arrangement (three sources of fat and two levels of Lysolecithin). There was interaction between fat sources and Lysolecithin for rumen pH (P=0.01), for rumen molar proportions of acetate (P<0.01), propionate (P=0.02), isobutyrate (P<0.01), valerate (P<0.01), isovalerate (P<0.01), and molar concentration of total VFA (P=0.09). According to the data, there was no interaction effect (P>0.05) between fat source and Lysolecithin for DMI and nutrient digestibility. Lysolecithin improved rumen fermentation of diets with degummed soybean oil and had no effect on EE digestibility independent of fat source, but it increased total tract fiber digestibility.

Thesis (M.Sc. (Biochemistry) -- University of Limpopo, 2018 / Dicerocaryum senecioides is a plant widely used as a nutritional source. It is used also for treatment of measles, wounds and to facilitate birth in domestic animal and humans in many parts of southern Africa (Mampuru et al., 2012). Findings in our laboratory have shown that a dichloromethane fraction of D. senecioides possesses antiinflammatory properties in human t-lymphocytes (Madiga, 2009), while the methanol crude extract possesses anti-proliferative and proapoptotic properties against Jurkat T cancer cells (Mphahlele, 2008). In this study, the probable anti-cancer effect of D. senecioides crude methanol leaf extract was investigated on cervical HeLa cancer cells. Dried powdered leaves of D. senecioides were extracted with absolute methanol to obtain a crude extract. To assess the cytotoxicity effect of the extract, KMST-6 and HeLa cell cultures were exposed to various extract concentrations (0 to 600 µg/ml) for 24 and 48 hours and subjected to the MTT assay. The results showed the extract to have no significant increase in the viability inhibition of HeLa cells at all tested concentrations after 24 hours of treatment. However, treatment with 400, 500 and 600 µg/ml of the extract for 48 hours revealed significantly increased HeLa cell viability inhibition. Furthermore, the extract showed to have no effect on the viability of normal human fibroblast KMST-6 cells at concentrations below 600 µg/ml, after 24 and 48 hours of treatment, thus showing selective cytotoxicity of the extract. To determine the mode of cell death associated with the increase in HeLa cell viability inhibition, the Hoechst 33258 nuclear staining assay and inverted light microscopy were employed. The data proposed apoptosis as the mode of cell death associated with the inhibition of HeLa cell viability. This was evidenced by changes in cell morphology such as the loss of HeLa cell radial extensions, cell shrinkage, as well as nuclear morphological features such as chromatin condensation. Apoptosis induction was further confirmed by the annexin-V/PI and multicaspase assays, using flow cytometry. The results showed an increase in the percentage of cells stained with annexin-V/PI, as well as increased caspase activity in extract-treated HeLa cells. To elucidate proapoptotic mechanisms of the extract, Western blotting analysis as well as the human apoptosis antibody array kit were used. This was to measure the expression profile of a number of apoptosis regulatory proteins. The results demonstrated modulation of some anti- and pro-apoptotic proteins, as well as the release of mitochondrial proteins required xiii for initiation of apoptosis, in the cytoplasm. The D. senecioides extract showed to have no effect on the cell division cycle of HeLa cells as determined by the PI staining assay. In conclusion, D. senecioides crude methanol leaf extract induced some degree of apoptosis in cervical HeLa cancer cells via the intrinsic apoptosis pathway. This was by modulating some of the members of the Bcl-2 family of proteins, which, facilitated the release of cytochrome C and activation of a caspase cascade. / South African Medical Research Council (SAMRC)

Declines in some fish populations in the Upper Danube River, Germany, have been reported during the past decades despite extensive stocking efforts. Many theories exist for why such declines have occurred including habitat change, dams, invasive species, disease and pollution. One of the factors of concern in the Upper Danube River is pollution because a number of studies have shown that sediments collected from this area were acutely and/or chronically toxic to fish. Although it can be difficult to link bioassay results to direct effects on the population level, bioassays can give us insight into the potential of exposure of wildlife including fish to sediment. In combination with other researchers a large battery of sediment testing on the Upper Danube River is being performed. Testing includes sediment testing of estrogen receptor mediated processes, dioxin-like responses and genotoxic effects. In this study, four sediment extracts from the Upper Danube River in Germany were used with a novel fractionation technique to characterize the sediment extracts and fractions for their ability to disrupt steroidogenesis, for their mutagenic activities and their teratogenic effects. Fractionation of each of the four sediment samples was performed by separating compounds according to their polarity, planarity, and the size of the aromatic ring system in an on-line fractionation procedure on coupled high performance liquid chromatography columns.<p> Mutagenic activity was measured in the raw sediment extracts and all 18 fractions using the Ames fluctuation assay and the Danio rerio embryo assay was used to assess lethal endpoints. Furthermore, disruptions of steroidogenesis were assessed by first establishing methods and a proof of concept of the H295R assay by exposing H295R cells to 7 model chemicals and measuring changes from a control in estradiol, testosterone and aromatase activity. Once methods were established all sediments and their fractions were analyzed using the Assay.<p> Specifically, in the <i>Danio rerio</i> assay, two raw sediment extracts killed 100% of <i>Danio rerio</i> embryos at a concentration of 33.3 mg sediment equivalents (SEQ)/ml, but none of the 18 fractions of these samples produced any measured toxicity at a concentration of 100 mg SEQ/ml. In the Ames fluctuation assay, significant mutagenic activity was measured in raw sediment extracts and in the fractions. Fraction 10 produced a significant mutagenic response in all sediment samples measured only in S9 bio-activated samples. Furthermore, fraction 15 produced a significant mutagenic response in all sediment samples measured only in non bio-activated samples.<p> All raw extracts tested in the H295R assay caused an increase in estradiol production up to 4-fold from controls. Testosterone production increased slightly from controls in only two of the raw extract samples. Of the 18 fractions, fractions 7, 10 and 15 increased estradiol in at least three of the samples studied (Sigmaringen2006, Opfingen2006, Lauchert2006 and Lauchert2004). Furthermore, fraction 7 significantly decreased testosterone production compared to controls in three of the four sediment samples.<p> Taken as a whole, these results show the value of using multiple bioassays and fractionation to characterize sediments that covers a variety of different biological endpoints. This study also demonstrates the usefulness of the H295R assay when combined with a new fraction technique to assess endocrine disrupting chemicals in sediment samples.

Declines in some fish populations in the Upper Danube River, Germany, have been reported during the past decades despite extensive stocking efforts. Many theories exist for why such declines have occurred including habitat change, dams, invasive species, disease and pollution. One of the factors of concern in the Upper Danube River is pollution because a number of studies have shown that sediments collected from this area were acutely and/or chronically toxic to fish. Although it can be difficult to link bioassay results to direct effects on the population level, bioassays can give us insight into the potential of exposure of wildlife including fish to sediment. In combination with other researchers a large battery of sediment testing on the Upper Danube River is being performed. Testing includes sediment testing of estrogen receptor mediated processes, dioxin-like responses and genotoxic effects. In this study, four sediment extracts from the Upper Danube River in Germany were used with a novel fractionation technique to characterize the sediment extracts and fractions for their ability to disrupt steroidogenesis, for their mutagenic activities and their teratogenic effects. Fractionation of each of the four sediment samples was performed by separating compounds according to their polarity, planarity, and the size of the aromatic ring system in an on-line fractionation procedure on coupled high performance liquid chromatography columns.<p> Mutagenic activity was measured in the raw sediment extracts and all 18 fractions using the Ames fluctuation assay and the Danio rerio embryo assay was used to assess lethal endpoints. Furthermore, disruptions of steroidogenesis were assessed by first establishing methods and a proof of concept of the H295R assay by exposing H295R cells to 7 model chemicals and measuring changes from a control in estradiol, testosterone and aromatase activity. Once methods were established all sediments and their fractions were analyzed using the Assay.<p> Specifically, in the <i>Danio rerio</i> assay, two raw sediment extracts killed 100% of <i>Danio rerio</i> embryos at a concentration of 33.3 mg sediment equivalents (SEQ)/ml, but none of the 18 fractions of these samples produced any measured toxicity at a concentration of 100 mg SEQ/ml. In the Ames fluctuation assay, significant mutagenic activity was measured in raw sediment extracts and in the fractions. Fraction 10 produced a significant mutagenic response in all sediment samples measured only in S9 bio-activated samples. Furthermore, fraction 15 produced a significant mutagenic response in all sediment samples measured only in non bio-activated samples.<p> All raw extracts tested in the H295R assay caused an increase in estradiol production up to 4-fold from controls. Testosterone production increased slightly from controls in only two of the raw extract samples. Of the 18 fractions, fractions 7, 10 and 15 increased estradiol in at least three of the samples studied (Sigmaringen2006, Opfingen2006, Lauchert2006 and Lauchert2004). Furthermore, fraction 7 significantly decreased testosterone production compared to controls in three of the four sediment samples.<p> Taken as a whole, these results show the value of using multiple bioassays and fractionation to characterize sediments that covers a variety of different biological endpoints. This study also demonstrates the usefulness of the H295R assay when combined with a new fraction technique to assess endocrine disrupting chemicals in sediment samples.