Este estudo teve por objetivo avaliar, in vitro, a viabilidade do uso de soluções de extratos naturais no tratamento restaurador da dentina, por meio do ensaio de resistência à microtração e análise qualitativa da superfície dentinária e interface adesiva em microscopia eletrônica de varredura (MEV). A amostra foi composta de 90 fragmentos de dentina bovina. Os fragmentos foram planificados, polidos e condicionados com ácido fosfórico 37% e, em seguida, divididos de acordo com o tratamento da dentina em 5 níveis: GI Sem tratamento (controle), GII Solução de extrato de Camellia sinensis (chá verde), GIII Solução de extrato de Punica granatum (romã), GIV Solução de extrato de Vitis vinifera (semente de uva), GV Solução de extrato de Lycium barbarum (goji berry). Os espécimes que foram utilizados na análise da resistência de união e interface adesiva tiveram as superfícies restauradas com adesivo (Adper Single Bond 2 - 3M) e resina composta (Filtek Z250 - 3M). Os 60 fragmentos utilizados para o teste de microtração (6 mm x 6 mm x 6 mm) foram seccionados em palitos de aproximadamente 1 mm&sup2, obtendo-se 4 palitos por dente (n=12). Após o teste, o padrão de falhas foi analisado em microscopia de varredura confocal a laser (MVCL). Para análise da interface adesiva, 15 fragmentos (6 mm x 3 mm x 3 mm) foram tratados com as soluções (n=3), restaurados e preparados para MEV. Os 15 fragmentos (3 mm x 3 mm x 3 mm) destinados à análise da superfície foram tratados com as soluções (n=3) e preparados para MEV. Os dados do teste de microtração (MPa) foram submetidos à ANOVA e teste de Tukey (p=0,05). Os maiores valores médios de resistência adesiva (p < 0,05) foram obtidos nos grupos controle (sem tratamento, GI - 25,69 ± 6,32), extrato de Camellia sinenis (chá verde, GII - 28,14 ± 7,66) e extrato de Vitis vinifera (semente de uva, GIV - 25,55 ± 4,00), sendo estes iguais entre si (p > 0,05). Os fragmentos tratados com o extrato de Punica granatum apresentaram os menores valores médios (p < 0,05) de resistência adesiva (Romã, GIII - 17,42 ± 4,82). Os fragmentos que receberam o extrato de Lycium barbarum apresentaram valores intermediários (Goji Berry, GV - 23,44 ± 6,78), ora similares aos dos grupos com maiores médias, ora iguais ao grupo de menor média de resistência adesiva (p > 0,05). Falhas adesivas foram predominantes nos grupos experimentais. Na análise em MEV da superfície dentinária, verificou-se superfície regular, sem camada de smear e com abertura dos túbulos dentinários em todos os grupos analisados. Na análise da interface adesiva, houve boa hibridização da embocadura dos canais, e presença de tags resinosos na maioria dos túbulos dentinários, sugerindo formação de camada hibrida. As soluções de Camellia sinensis, Vitis vinifera e Lycium barbarum não interferiram negativamente na resistência de união quando comparados ao grupo controle, sem tratamento. / This study aimed to evaluate, in vitro, the viability of using natural extracts solutions in dentin restorative treatment, by microtensile bond strength test and qualitative analysis of dentin morphology surface and adhesive interface under scanning electron microscopy (SEM). Sample was composed of 90 fragments of intracoronary bovine dentin. The fragments were flattened, etched with 37% phosphoric acid and then were divided according to the dentin treatments: GI - Untreated (control), GII - Camellia sinensis extract solution (green tea), GIII - Punica granatum extract solution (pomegranate), GIV - Vitis vinifera extract solution (grape seed), GV - Lycium barbarum extract solution (goji berry). Specimens that were used on the bond strength test and adhesive interface analysis were restored with adhesive (Single Bond 2 - 3M) and composite resin (Filtek Z250 - 3M). The 60 fragments used to microtensile test (MPa) (6mm x 6mm x 6mm) were sectioned at approximately 1mm2 sticks, 4 destined to the bonding interface analysis (n=12). After the test, the failure pattern was analyzed in confocal laser scanning microscopy (CLSM). For adhesive interface analysis, 15 pieces (6mm x 3mm x 3mm) were treated with solutions, restored and prepared for SEM (n=3). The 15 fragments (3mm x 3mm x 3mm) destined for surface analysis were treated with solutions and prepared for SEM (n=3). The microtensile test (MPa) data were submitted to ANOVA and Tukey test (p=0.05). The highest values of bond strength (p<0.05) were obtained in the control groups (untreated, GI - 25.69 ± 6.32), Camellia sinenis extract (green tea, GII - 28.14 ± 7. 66) and Vitis vinifera extract (grape seed, GIV - 25.55 ± 4.00), which are equal to each other (p>0.05). The fragments treated with Punica granatum extract (Pomegranate, GIII - 17.42 ± 4.82) showed lowest values of bond strength (p <0.05). Fragments treated with Lycium barbarum extract showed intermediate values (Goji Berry, GV - 23.44 ± 6.78), similar to those of groups with higher values and to the lower values group of bond strength (p>0.05). Adhesive failures were predominant in all groups. The SEM analysis of the dentin surface demonstrated that there was a regular surface, without smear layer and open dentinal tubules in all groups analyzed. The adhesive interface analysis, showed that there was good hybridization of the entrance of dentinal tubules, and presence of resin tags in most dentinal tubules, suggesting formation of hybrid layer. The extracts solutions of Camellia sinensis, Vitis vinifera and Lycium barbarum did not interfere negatively on the bond strength when compared to the control group without treatment.

Este estudo teve por objetivos avaliar diferentes níveis de suplementação de ?-ácidos do lúpulo como aditivos de rações para frangos de corte sobre o desempenho e a manutenção do equilíbrio da microbiota intestinal após desafio com Eimeria. No experimento 1, foram alojados 1440 pintos de corte da linhagem Cobb 500, no período de 1 a 42 dias de idade, com 6 tratamentos e 6 repetições. Os tratamentos utilizados foram: controle negativo, ração sem antimicrobiano; controle positivo, ração com 30 mg/kg de bacitracina de zinco e rações com 30, 60, 120 ou 240 mg/kg de ?-ácidos do lúpulo, compondo 4 tratamentos adicionais. Os ?-ácidos foram microencapsulados com 30%de extrato. As rações foram formuladas à base de milho e farelo de soja, com inclusão de 5% de farelo de trigo e 5% de farinha de penas e vísceras. Aos 7 dias de idade todas as aves foram vacinadas contra coccidiose. As aves e as rações foram pesadas semanalmente para cálculo de desempenho. No experimento 2, foram alojados 1440 pintos de corte da linhagem Ross 308, no período de 1 a 42 dias de idade, com 6 tratamentos e 6 repetições. Os tratamentos foram: controle negativo, dieta basal; controle positivo, ração com 30 mg/kg de bacitracina de zinco; controle negativo mais desafio; controle positivo mais desafio; controle negativo suplementado com 30 mg/kg de ?-ácidos mais desafio; controle negativo suplementado com 240 mg/kg de ?-ácidos mais desafio. As rações foram formuladas à base de milho e farelo de soja com inclusão de 5% de farinha de penas e vísceras. Aos 14 dias de idade, as aves dos tratamentos 3, 4, 5 e 6 foram desafiadas, via oral, com 2x105 e 5x104 oocistos de Eimeria acervulina e E. tenella, respectivamente. As aves e as rações foram pesadas semanalmente para obtenção dos dados de desempenho. Aos 21 e 35 dias de idade das aves, coletou-se o conteúdo do intestino delgado e cecos das aves para análise da microbiota intestinal, com auxílio de técnicas moleculares. No experimento 1, aos 21 dias de idade as aves os tratamentos com 30 ou 60 mg/kg de ?-ácidos apresentaram desempenho semelhante às do controle positivo. Aos 42 dias houve melhora na conversão alimentar das aves recebendo 30 e 240 mg/kg de ?-ácidos ou antimicrobiano. No segundo experimento, a coccidiose causou significativa redução no desempenho das aves e nenhum dos aditivos utilizados foi capaz de reverter esta situação. Houve aumento de bactérias do gênero Clostridium no intestino delgado aos 21 dias, em consequência do desafio, entretanto, o maior nível de ?-ácidos reduziu esta população. Aos 35 dias de idade a infecção de coccidiose não alterou a comunidade bacteriana do intestino delgado, mas nos cecos houve aumento do gênero Bacteroides.Os ?-ácidos possuem potencial para serem utilizados nas dietas de frangos de corte em situações de baixo desafio, e podem auxiliar no controle da proliferação de Clostridium, embora ineficazes contra a Eimeria. / The objective was to evaluate increasing level of hops ?-acids in the feed on performance of broiler chickens. A pen trial using 1440 one-day old chickens, from 1 to 42 days, with 6 treatments and 6 replicates was conducted (Experiment 1). The experimental treatments were: negative control, basal diet; positive control, basal diet supplemented with zinc bacitracin, 30 mg/kg; and basal diet supplemented with 30, 60, 120 or 240 mg/kg of hops ?-acids, for 4 additional treatments. The corn soybean meal basal diet was formulated with inclusion of 5% poultry by-product meal and 5% wheat bran. At 7 days of age all birds were vaccinated against coccidiosis. The chickens and the feed were weighted weekly to calculate the performance. In the second experiment, the objective was to evaluate the supplementation of hops ?-acids on performance and the balance of intestinal microbiota of broilers, following challenge with Eimeria acervulina and E. tenella. A pen trial using 1440 one-day-old chickens, from 1 to 42 days, with 6 treatments and 6 replicates was conducted. The experimental treatments were: negative control, basal diet; positive control, basal diet supplemented witn zinc bacitracin, 30 mg/kg; negative controle + challenge; Positive control + challenge; negative control supplemented with 30 mg/kg of hops ?-acids + challenge; negative control supplemented with 240 mg/kg of hops ?-acids + challenge. The corn soybean meal basal diet was formulated with inclusion of 5% poultry byproduct meal. At 14 days of age, the birds in treatments 3, 4, 5 and 6 were challenged with 2x105 and 5x104 oocists of Eimeria acervulina and E. tenella, respectively. The chickens and the feed were weighted weekly to calculate the performance. At 21 and 35 days of age, the small intestine and ceca content was collected to analyze the intestinal microbiota, using molecular techniques. In the first experiment, at 21 days of age the treatment with 30 or 60 mg/kg of ?-acids had the same performance of chickens in positive control. At 42 days, the treatments containing 30 or 240 mg/kg of ?-acids and positive control had improved feed conversion. In the second experiment, there was worse performance in broilers chickens challenged with coccidiosis and the additives were not able to oppose this situation. There was increase in the Clostridium population in the small intestine at 21 days, due to challenge, however, the highest level of ?-acids decreasead this genus. At 35 days, the coccidiosis did not alter the bacterial community in the small intestine, although the ceca had higher level of Bacteroides. The hops ?-acids have the potential to be used in the diets of broiler chickens, under low level of challenge, and to proliferation of Clostridium, although ineffective against Eimeria.

Thesis MTech (Food Technology))--Cape Peninsula University of Technology, 2017. / Bambara groundnut (Vigna subterranea) an indigenous legume cultivated in Sub-Saharan Africa has been proclaimed to have medicinal properties from communities and in rural areas. However, there is not enough scientific information to validate these claims. Therefore, this study aimed to identify possible medicinal properties of Bambara groundnut (BGN), by analysing the phytochemical and antimicrobial properties of BGN seed and product extracts from Mpumalanga province within South Africa. The BGN extracts (70% methanol, 70% ethanol, milli-Q water) from seeds and products (milk and yoghurt) were screened for the presence of alkaloids, flavonoids, phenols, riboflavin and thiamine using analytical laboratory methods for basic screening, high-performance liquid chromatography (HPLC) and gas chromatography (GC) for quantification. The antimicrobial activity involved direct bioautography and minimum inhibitory concentration (MIC) against six antibiotic-resistant microorganisms, Acinetobacter baumannii ATCC 19606T, Enterococcus faecalis ATCC 29212, Klebsiella pneumoniae subsp. pneumoniae ATCC 700603, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus subsp. aureus ATCC 33591 and Candida albicans ATCC 24433. For the seed extracts, flavonoids and phenols were highly concentrated in the red and brown hulls of BGN compared to whole and dehulled BGN. Organic solvents in comparison to water yielded the highest concentration of flavonoids, whilst water yielded the highest concentration for phenols. Flavonoid compounds that were detected at the highest concentrations were rutin (24.458 ± 0.234 mg.g-1, brown hull extracted with 70% methanol), quercetin (0.070 ± 0.043 mg.g-1, red hull extracted with 70% methanol), kaempferol (0.391 ± 0.161 mg.g-1; brown hull extracted with 70% ethanol) and myricetin (1.800 ± 0.771 mg.g-1; red hull extracted with 70% methanol). For phenol compounds, gallic acid (0.009 ± 0.004 mg.g-1; brown hull extracted with milli-Q water), catechin (0.026 ± 0.041 mg.g-1; brown hull extracted with milli-Q water), methyl gallate (0.008 ± 0.013 mg.g-1; brown whole extracted with milli-Q water), chlorogenic acid (0.115 ± 0.199 mg.g-1; brown hull extracted with milli-Q water) and ellagic acid (0.105 ± 0.082 mg.g-1; red hull extracted with milli-Q water) were detected. Vitamins B1 and B2 (riboflavin and thiamine) were mostly present in milli-Q water extracts. Black-eye hull had the highest concentration of thiamine (vitamin B1) and riboflavin (vitamin B2) consisting of 0.072 mg.g-1 (extracted with milli-Q water) and 0.002 mg.g-1 (extracted with 70% ethanol and 70% methanol). Red and brown hull extracts from organic solvents (70% ethanol and 70% methanol) showed the highest antimicrobial activity, whereas the whole, dehulled and hulls (black-eye and brown-eye) extracts had no antimicrobial activity. As for BGN products extracts, flavonoid compounds that were detected at the highest concentrations were rutin (5.694 mg.g-1, whole BGN milk, milli-Q water), quercetin (0.703 mg.g-1, whole BGN yoghurt, milli-Q water) and myricetin (0.987 mg.g-1, whole BGN yoghurt, 70% ethanol).

Submitted by admin tede (tede@pucgoias.edu.br) on 2017-06-29T13:18:42Z No. of bitstreams: 1 MARIA JOSÉ BATISTA DE SOUSA.pdf: 2843064 bytes, checksum: f400a0df10a20e086f4f74a5b6cf9480 (MD5) / Made available in DSpace on 2017-06-29T13:18:42Z (GMT). No. of bitstreams: 1 MARIA JOSÉ BATISTA DE SOUSA.pdf: 2843064 bytes, checksum: f400a0df10a20e086f4f74a5b6cf9480 (MD5) Previous issue date: 2017-03-28 / The species Brosimum gaudichaudii (family Moraceae) and Caesalpinia ferrea (family Fabaceae) are widely distributed throughout Brazil and are considered medicinal plants. The extract of Brosimum gaudichaudii bark has been indicated for the treatment of skin blemishes and vitiligo. On the other hand, the extract of Caesalpinia ferrea fruit has been used due to its therapeutic properties as antibacterial, anti-inflammatory and analgesic action. Much of the medicinal plant extracts constituents are unknown and may be toxic to human and animal health, so it is necessary to study the qualitative phytochemical of secondary metabolites and to evaluate the cytotoxic, genotoxic and mutagenic potential of the extracts of these species. In this study, in order to evaluate the mutagenic and / or genotoxic effects, different concentrations of the extractive solutions of B. gaudichaudii and C. ferrea were evaluated in vivo in Astyanax sp and Allium cepa, and ex vivo, by the micronucleus test in T lymphocytes humans. Data were submitted to Kruskall-Wallis a non-parametric test and then to simple linear regression with a significance level of 5%. The Allium cepa test, micronucleus test for human T lymphocytes and erythrocytes of Astyanax sp did not indicate mutagenic and / or genotoxic potential of phytochemicals (p> 0.05) when compared to the non-exposed controls, except the concentration of 5 mg/L of B. gaudichaudii that showed cytotoxicity. On the other hand, the comet assay revealed genotoxic action for all concentrations evaluated for the tail length parameter of the comet. For the moment parameter of Olive's tail only the 20mg /L concentration of Caesalpinia ferrea extract was genotoxic. Therefore, apical meristematic cells from the roots of Allium cepa and human T lymphocytes did not present genotoxic and / or mutagenic changes induced by exposure to both plant extracts detectable by micronuclei tests or mitotic index reduction. Genotoxic effect was evidenced by the tail length and tail moment parameter of Olive in the Comet Assay only for C. ferrea extract in the erythrocytes of Astyanax sp. In order to understand the genotoxic and mutagenic activities of B. gaudichaudii and C. ferrea it is important to increase the number of studies to establish safer doses for human consumption. / As espécies Brosimum gaudichaudii (família Moraceae) e Caesalpinia ferrea da família Fabaceae são amplamente distribuídas pelo território brasileiro e são consideradas plantas medicinais. O extrato das cascas de Brosimum gaudichaudii tem sido indicado para tratamento de mancha de pele e vitiligo. Por outro lado, o extrato dos frutos de Caesalpinia ferrea tem sido usado devido suas propriedades terapêuticas como ação antibacteriana, antiinflamatória e analgésica. A maioria dos fitoquímicos presentes nos extratos de plantas medicinais ainda não foram completamente estudados e podem ser tóxicos para a saúde humana e animal. Nesse sentido, é necessário estudos fitoquímicos qualitativos de metabólitos secundários e avaliação do potencial citotóxico, genotóxico e mutagênico dos extratos destas espécies. Nesse estudo, visando avaliar os efeitos mutagênico e/ou genotóxico, diferentes concentrações das soluções extrativas de B. gaudichaudii e C. ferrea foram avaliadas in vivo em Astyanax sp e em Allium cepa, e em ex vivo, pelo teste de micronúcleos em linfócitos T humanos. Os resultados observados das análises foram submetidos ao teste não paramétrico Kruskall-Wallis e posteriormente a regressão linear simples com nível de significância 5%. O teste em Allium cepa, teste de micronúcleo em linfócitos T humanos e em eritrócitos de Astyanax sp não indicaram potencial mutagênico e/ou genotóxico dos fitoconstituintes (p>0,05) quando comparado aos controles não expostos, exceto a concentração de 5g/L de B. gaudichaudii que apresentou citotoxicidade (p=0,038). Por outro lado, o ensaio cometa, revelou ação genotóxica para todas as concentrações avaliadas no parâmetro comprimento da cauda do cometa, para o parâmetro momento da cauda de Olive, apenas a concentração de 20mg/L do extrato de Caesalpinia ferrea mostrou-se genotóxica. Nenhum dos parâmetros avaliados evidenciou danos genéticos resultantes da exposição aos extratos das cascas do caule de B. gaudichaudii. Portanto, as células meristemáticas apicais das raízes de Allium cepa e os linfócitos T humanos não apresentaram alterações genotóxicas e/ou mutagênicas induzidas pela exposição a ambos extratos vegetais que pudesse ser detectadas pelos testes do micronúcleos ou redução do índice mitóticos. Enquanto, nos eritrócitos de Astyanax sp foi evidenciado ação genotóxica pelo parâmetro comprimento da cauda do cometa e momento de da cauda de Olive somente para o extrato de C. ferrea. Diante do exposto, há necessidade de ampliar os estudos para melhor compreensão das atividades genotóxicas e/ou mutagênicas dos extratos de B. gaudichaudii e C. ferrea visando o estabelecimento de doses mais seguras para o consumo humano.

Nas últimas décadas, a produção de carne de frango vem se intensificando por expressivos avanços tecnológicos. Nutricionalmente, os promotores de crescimento antimicrobianos (antibióticos e quimioterápicos) foram essenciais, beneficiando o desempenho e a eficiência alimentar, quando utilizados como aditivos nas dietas, em doses subterapêuticas. Apesar da comprovada contribuição no desempenho das aves, os antibióticos promotores de crescimento passaram a ser vistos como fatores de risco para a saúde humana, devido ao potencial desenvolvimento da resistência bacteriana cruzada em humanos. Recentemente, têm sido desenvolvidas diversas alternativas aos antibióticos promotores de crescimento, incluindo probióticos, prebióticos, ácidos orgânicos, enzimas e extratos vegetais. Seguindo essa tendência, este estudo teve o objetivo de avaliar a eficácia do uso de extratos vegetais como alternativas aos antimicrobianos promotores de crescimento em dietas de frangos de corte. Foram realizados dois experimentos com frangos de corte para avaliar os efeitos de diferentes extratos vegetais no desempenho, na energia metabolizável da dieta e na morfometria dos órgãos. O experimento de desempenho envolveu 1200 frangos de corte machos criados em galpão experimental no período de 1 a 42 dias de idade, separados em grupos de 40 aves por boxe. O delineamento experimental foi em blocos casualizados, sendo seis tratamentos e cinco repetições. O ensaio de metabolismo contou com 96 frangos machos em crescimento alojados em gaiolas metabólicas para coleta total de excretas, foram seis tratamentos e quatro repetições em delineamento interiramente casualizado. Ambos os experimentos receberam os mesmos tratamentos: dieta controle (DC); DC + 10 ppm de Avilamicina; DC + 1000 ppm de extrato de orégano; DC + 1000 ppm de extrato de cravo; DC + 1000 ppm de extrato de canela e DC + 1000 ppm de extrato de pimenta vermelha. Os produtos à base de extratos vegetais consistiram de microencapsulados contendo 20% do óleo essencial. As variáveis determinadas foram: peso vivo, ganho de peso, consumo de ração, conversão alimentar, mortalidade, energia metabolizável aparente (EMAn) das dietas e peso relativo dos órgãos. Os resultados foram submetidos à análise de variância e as médias comparadas pelo teste de Tukey. Não foi possível observar diferenças significativas (P>0.05) entre os tratamentos propostos para as variáveis de desempenho analisadas em todo o período de criação das aves. Da mesma maneira, a suplementação dos extratos nas dietas não interferiu (P>0.05) nos valores de EMAn das mesmas. A morfometria dos órgãos, em geral, também não foi alterada pela utilização dos extratos vegetais. Pôde ser observado que, os animais que receberam ração controle apresentaram maior peso relativo do fígado em relação aos demais tratamentos, diferindo significativamente (P<0.05) das aves alimentadas com a dieta suplementada com extrato de pimenta, que foi o menor valor observado. / Broiler meat production has experienced expressive technological improvements in the last decades. Nutritionally, the antimicrobial growth promoters (antibiotics and chemotherapeutics) contributed to that, when utilized as feed additives. These antimicrobials are very effective in performance improvement, but there are human health risks associated to their use because of the possibility of development of bacterial cross resistance. In recent years, several alternatives to the antibiotic growth promoters have been proposed including probiotics, prebiotics, organic acids, enzymes and plant extracts. In this context, the objective of the present study was to evaluate the efficacy of plant extracts as alternative growth promoters to the antibiotics as feed additives in chicken diets. Two experiments were conducted to determine the effects of different plant on the metabolizable energy of the diet and performance and organ morphometry of broiler chickens. The performance trial involved 1,200 male chicks raised from 1 to 42 days of age, in groups of 40 birds per pen, in a experimental poultry house. A randomized complete block design, with 6 treatments and 5 blocks, was used. In the metabolism trial, 96 chickens in the grower period were allotted to battery cages and total excreta collection was conducted with 6 treatments and 4 replicates in a completely randomized design. The treatments for both experiments were: Corn-soybean meal control diet (C); C + 10 ppm Avilamycin; C + 1,000 ppm oregano extract; C + 1,000 ppm clove extract; C + 1,000 ppm cinnamon extract; C + 1,000 ppm red pepper extract. The plant extract products added consisted of microencapsulated preparations containing 20% essential oil. The variables evaluated were liveweight, weight gain, feed intake, feed conversion, mortality, apparent metabolizable energy (AMEn) of the diets and organ weights. The results were statistically analyzed by ANOVA and means compared using Tukey&#39;s test. It was not detected any significant (P>0.05) difference among the treatments for the performance variables of the chickens along the growth period. Likewise, the plant extracts supplementation did not affect the AMEn of the diets (P>0.05). Organ morphometry was also, in general, not influenced by the additives. The only significant effect was a higher liver relative weight of the control birds (P<0.05) compared to the red pepper fed birds.

Ampelopsis grossedentata is a wildly used herb in South China as sleep aid beverage for many years. Yet the active ingredients and mechanisms of this herb were unknown. In the present study, extract from Ampelopsis grossedentata which we named ECBRC-AG, and one of its active ingredient myricetin were proved having significant hypnotic/sedative effects in multiple animal models. ECBRC-AG shortened sleep latency, increase NREM sleep and decrease locomotor activity when treated before the onset of light period in rats. ECBRC-AG could decrease active awake and increase REM sleep in the late part of light period. ECBRC-AG also decreased the caffeine induced hyperactivity in rats. Among the three suspected active ingredients from ECBRC-AG, myricetin showed similar active profile with ECBRC-AG. Myricetin increased NREM and REM sleep, decreased sleep latency, decreased locomotor activity and also active awake. All the above evidences have implicated that myricetin is the most important active ingredient of ECBRC-AG ECBRC-AG and myricetin did not show any obvious side effects on rats. / Based on these findings, we propose that myricetin facilitates GABA function on PVN neurons through a T-type calcium channel and CaM-KII mechanism. The hypnotic/sedative effects of ECBRC-AG and myricetin are mediated by PVN. ECBRC-AG treatment decreased corticosterone levels in rats, which also indicated that PVN/HPA axis was the target of these herbal derivates. PVN has broad interactions with GABAergic, hypocretinergic, cytokine and NPY system and all these systems are proved to be deeply involved in sleep regulation. / In conclusion, the present study has identified that myricetin is the most important active ingredient of the herbal extract ECBRC-AG. We confirmed the hypnotic/sedative effects of ECBRC-AG and myricetin on rats, and also revealed the different action profiles of these herbal derivates compared with zolpidem. T-type calcium channels and the HPA axis were shown to be involved in the mechanisms of ECBRC-AG and myricetin, indicating that they may be the new targets for insomnia treatment with these herbal derivates. / Insomnia is the most common sleep disorder and affects about one third of the general population. Insomnia is always combined with physical and mental illness, as either a consequence or a contributing factor. Insomnia produces sleepiness, impairment in psychomotor performance, absenteeism, frequent accidents, memory impairment and a high risk of depression. Pharmacologic therapies are the most important interventions for insomnia. However, the currently available hypnotics are associated with residual effects and risks of abuse and dependence. More efficient and safe hypnotics are needed. / The DNA array and RT-PCR studies revealed that GABA, hypocretin, cytokine and NPY systems were involved in the mechanisms of ECBRC-AG and myricetin. In calcium imaging study, we found that myricetin induced a transient Ca 2+ influx in the primary culture of rat hypothalamus neurons. This Ca2+ influx could be blocked by T-type channel blocker mibefradil. RT-PCR study also showed that ECBRC-AG and myricetin treatment changed the mRNA expression level of T-type calcium channel al G subunit in rat hypothalamus. The present results are consistent with our previous study showing that myricetin enhanced GABA function in the neurons of rat hypothalamic paraventricular nucleus (PVN), and that blocking CaM-KII pathway eliminated this effect. / Zhang, Xiaohu. / "March 2008." / Adviser: Chan Hsiao Chang. / Source: Dissertation Abstracts International, Volume: 70-03, Section: B, page: 1516. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (p. 156-174). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Orientador: Luciane Dias de Oliveira / Banca: Antonio Olavo Cardoso Jorge / Banca: Marcos Roberto Furlan / Resumo: Visando o potencial terapêutico do extrato glicólico de B. pendula em tratamentos de infecções bacterianas e fúngicas e de doenças inflamatórias, foram avaliadas suas atividades antimicrobiana, antiinflamatória, citotóxica e genotóxica. A atividade antimicrobiana do extrato foi analisada em Candida albicans, C. dubliniensis, C. glabrata, C. guilliermondii, C. krusei, C. tropicalis, Staphylococcus aureus, Streptococcus mutans, Escherichia coli e Pseudomonas aeruginosa com a determinação das Concentrações Inibitória Mínima e Microbicida Mínima (CIM e CMM) em culturas planctônicas e posteriormente testado sobre biofilmes monotípicos. A atividade citotóxica foi avaliada em cultura de macrófagos de camundongo (RAW 264.7) e fibroblastos gengivais humanos (FMM-1) após exposição de 5 min e 24 h ao extrato, com o teste MTT e teste imunoenzimático (ELISA) que quantificou a produção das citocinas IL-1β e TNF-α produzidas por RAW 264.7. A genotoxicidade do extrato foi avaliada pelo teste de micronúcleos. A atividade antiinflamatória foi avaliada nos sobrenadantes coletados de culturas de RAW 264.7 estimuladas com LPS de E. coli e expostas aos extratos (5 min e 24 h) pela quantificação de citocinas IL-1β, TNF-α e IL-10 por ELISA e óxido nítrico (ON) pelo método de Griess. A análise estatística foi realizada por ANOVA e teste de Tukey, com significância de 5%. O extrato teve ação antimicrobiana em todos biofilmes, com redução acima de 39,5% em 5 min e acima de 78% em 24 h. A viabilidade celular foi satisfatória em concentrações abaixo de 50 mg/mL em 5 min tanto para FMM-1, quanto para RAW 264.7, contudo em 24 h concentrações acima de 3,13 e 12,5 mg/mL foram citotóxicas para RAW 264.7 e FMM-1, respectivamente. Não houve indícios de ação genotóxica do extrato. A atividade anti-inflamatória foi evidenciada pela redução significativa na produção de TNF-a e ON nos grupos tratados . Conclui-se que o... / Abstract: In order to verify the therapeutic potential of glicolic extract of B. pendula on the threatment of bacterial and fungical infection and on inflammatory diseases, its antimicrobial, anti-inflammatory actions, cytotoxic and genotoxic effects were evaluated. The antimicrobial activity of the extracts was tested on Candida albicans, C. dubliniensis, C. glabrata, C. guilliermondii, C. krusei, C. tropicalis, Staphylococcus aureus, Streptococcus mutans, Escherichia coli and Pseudomonas aeruginosa with determination of Minimum Inhibitory and Minimum Microbicide Concentrations (MIC and MMC) in planktonic growth, following of test in monotypic biofilms. The cytotoxic activity was evaluated in mouse macrophages (RAW 264.7) and human gingival fibroblast (FMM-1) after exposure of 5 min and 24 h to extract, through the MTT test and by Enzyme Linked Immunosorbent Assay (ELISA) to quantify the production of the cytokines IL-1β and TNF-α by RAW 264.7. The genotoxicity of the extracts was evaluated by micronucleus test. The anti-inflammatory activity was evaluated in RAW 264.7 stimulated with LPS from E. coli, after the period of exposure to the extract (5 min and 24 h) the supernatant was removed to quantify pro-inflammatory (IL-1β and TNF-α.) and antiinflammatory (IL-10) cytokines by ELISA and nitric oxide (NO) by Griess method. Statistical analysis was performed by ANOVA and Tukey's test, with significance level of 5%. The extract has shown antimicrobial activity with reduction above of 39.5% of biofilm in 5 min and more than 78% in 24 h. The cell viability was satisfactory at concentrations below 50 mg/mL in 5 min to RAW 264.7 and FMM-1, however in 24 h concentrations above 3.13 and 12.5 mg/mL were cytotoxic to RAW 264.7 and FMM-1, respectively. There was no evidence of genotoxic action of the extract. The anti-inflammatory activity was evidenced by the significant reduction in the production o... / Mestre

Orientador: Luciane Dias de Oliveira / Banca: Luis Felipe das Chagas e Silva de Carvalho / Banca: Jônatas Rafael de Oliveira / Resumo: As plantas medicinais e os fitoterápicos têm sido utilizados como coadjuvantes e alternativos no combate a diversas doenças com crescente frequência, no entanto, na Odontologia seu uso ainda é bastante limitado. Com isso, o objetivo deste estudo é avaliar a ação antimicrobiana dos extratos aquoso e glicólico de própolis verde sobre micro-organismos anaeróbios de interesse odontológico, bem como sua citotoxicidade, a fim de introduzir e incentivar o uso efetivo e sistemático desse fitoterápico em produtos como dentifrícios e enxaguatórios bucais no combate a cáries e doenças periodontais. Os extratos comerciais aquoso e glicólico de própolis foram obtidos das empresas Apis Flora e Mapric, respectivamente. Para avaliação da atividade antimicrobiana foram utilizadas cepas-padrão (ATCC) de Fusobacterium nucleatum, Parvimonas micra, Porphyromonas endodontalis Porphyromonas gingivalis e Prevotella intermedia em cultura planctônica, verificando a concentração inibitória mínima e concentração microbicida mínima (CIM e CMM), segundo Clinical and Laboratory Standards Institute. Para biofilmes monotípicos, suspensões padronizadas (107 céls/mL) foram adicionadas em poços de microplacas e após 48 h em anaerobiose foram tratados com 3 concentrações do extrato de própolis (n=12) por 5 min. Foi incluído um controle positivo (solução fisiológica) e um controle negativo (clorexidina). O biofilme foi mensurado pelos testes MTT e Cristal Violeta. Para análise de citotoxicidade, queratinócitos hu... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Medicinal plants and herbal medicines have been used as adjuvants and alternatives in fighting various diseases with increasing frequency. However, in dentistry its use is still quite limited. Therefore, the objective of this study is to evaluate the antimicrobial action of the aqueous and glycolic extracts of green propolis on anaerobic microorganisms of dental interest, in order to introduce and encourage the effective and systematic use of this herbal medicine in products such as dentifrices and mouthwashes in the fight against tooth decay and periodontal diseases. Aqueous and glycolic commercial extracts of propolis were obtained from Apis Flora and Mapric companies, respectively.To evaluate the antimicrobial activity, standard strains (ATCC) of Fusobacterium nucleatum, Parvimonas micra, Porphyromonas endodontalis, Porphyromonas gingivalis, and Prevotella intermedia in planktonic culture was used, verifying the minimum inhibitory concentration and minimum microbicide concentration (MIC and CMM), according to Clinical and Laboratory Standards Institute. For monotypic biofilms, standardized suspensions (107 cells / mL) was added to microplate wells and after 48 h in anaerobiosis was treated with 3 concentrations of propolis extracts (n = 12) for 5 min. A positive control (saline solution) and a negative control (chlorhexidine) was included. The biofilm was measured by the MTT and Violet Crystal tests. For cytotoxicity analysis, human keratinocytes (HaCaT) were cultured and ... (Complete abstract click electronic access below) / Mestre

Sun Huimin. / Thesis submitted in: December 2002. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 77-89). / Abstracts in English and Chinese. / TITLE PAGE --- p.i / ACKNOWLEDGEMENTS --- p.ii / ABSTRACT --- p.iii / ABSTRACT IN CHINESE --- p.v / LIST OF PUBLICATIONS --- p.vii / ABBREVIATIONS --- p.viii / TABLE OF CONTENTS --- p.ix / Chapter CHAPTER 1. --- General Introduction --- p.1 / Chapter 1.1 --- Current Status of Herbal Product Use --- p.1 / Chapter 1.2 --- Herb-drug interactions --- p.2 / Chapter 1.2.1. --- Mechanisms of herb-drug interaction --- p.3 / Chapter 1.2.2. --- Pharmacodynamic interaction --- p.3 / Chapter 1.2.3. --- Pharmacokinetic interaction --- p.4 / Chapter 1.2.4. --- Herb-drug interaction involving drug metabolizing enzymes --- p.5 / Chapter 1.3 --- Methodologies for studying herb-drug interactions involving CYP enzymes --- p.7 / Chapter 1.3.1. --- Animal studies (Ex vivo approach) --- p.7 / Chapter 1.3.2. --- In vitro inhibition/induction studies --- p.8 / Chapter 1.3.3. --- Clinical studies --- p.9 / Chapter CHAPTER 2. --- Effect of flavonoid-containing herbs on CYP 450 enzyme activities: a screening study in rat --- p.11 / Chapter 2.1 --- Introduction --- p.11 / Chapter 2.2 --- Materials and Methods --- p.12 / Chapter 2.2.1. --- Chemicals --- p.12 / Chapter 2.2.2. --- Herbs --- p.12 / Chapter 2.2.3. --- Preparation of herbal extracts --- p.13 / Chapter 2.2.3.1. --- Preparation of Green Tea extract --- p.13 / Chapter 2.2.3.2. --- Preparation of Decaffeinated Green Tea (DGT) and its extracts --- p.13 / Chapter 2.2.3.3. --- "Preparation of extracts of Huang Qin, Ge Gen and Huai Mi" --- p.14 / Chapter 2.2.3.4. --- Preparation of Ginkgo biloba extract suspension --- p.14 / Chapter 2.2.4. --- Animal treatment --- p.14 / Chapter 2.2.5. --- Preparation of rat liver microsomes --- p.15 / Chapter 2.2.6. --- Determination of protein content of liver microsomes --- p.16 / Chapter 2.2.7. --- Determination of microsomal CYP content --- p.17 / Chapter 2.2.8. --- Statistical analysis --- p.19 / Chapter 2.3 --- Results --- p.19 / Chapter 2.4 --- Discussion --- p.24 / Chapter 2.5 --- Conclusion --- p.25 / Chapter CHAPTER 3 --- Rationale of the clinical study --- p.26 / Chapter CHAPTER 4 --- Development of HPLC methods for simultaneous determination of multiple probe drugs and their metabolites in human plasma or urine --- p.30 / Chapter 4.1 --- Introduction --- p.30 / Chapter 4.2 --- Materials and Methods --- p.33 / Chapter 4.2.1. --- Chemicals and reagents --- p.33 / Chapter 4.2.2. --- Preparation of stock and working solutions --- p.33 / Chapter 4.2.3. --- Equipment and chromatographic conditions --- p.34 / Chapter 4.2.4. --- Treatment of plasma and urine samples with β-glucuronidase --- p.35 / Chapter 4.2.5. --- Extraction procedures --- p.36 / Chapter 4.2.6. --- Preparation of working solutions for calibration curve --- p.37 / Chapter 4.3 --- Results --- p.39 / Chapter 4.3.1. --- Separation of the analytes --- p.39 / Chapter 4.3.2. --- Calibration and linearity --- p.39 / Chapter 4.3.3. --- Sensitivity --- p.39 / Chapter 4.3.4 --- Accuracy and precision --- p.50 / Chapter 4.4 --- Discussion --- p.52 / Chapter 4.5 --- Conclusions --- p.53 / Chapter CHAPTER 5 --- Stability study of probe drugs --- p.54 / Chapter 5.1 --- Introduction --- p.54 / Chapter 5.2 --- Materials and Methods --- p.54 / Chapter 5.2.1. --- Preparation of standard solutions of probe drugs --- p.54 / Chapter 5.2.2. --- Preparation of stability study mediums --- p.54 / Chapter 5.2.2.1. --- Gastric juice (pH=1.2) --- p.54 / Chapter 5.2.2.2. --- Intestine fluid (pH=6.8) --- p.54 / Chapter 5.2.2.3. --- Human plasma (pH=7.4) --- p.55 / Chapter 5.2.2.4. --- Phosphate buffer (pH=7.4) --- p.55 / Chapter 5.2.3. --- Incubation --- p.55 / Chapter 5.2.4. --- Determination of probe drug concentrations in incubation samples --- p.56 / Chapter 5.3 --- Results --- p.57 / Chapter 5.4 --- Discussion --- p.59 / Chapter 5.5 --- Conclusion --- p.59 / Chapter CHAPTER 6 --- Effect of the extract of Ginkgo biloba leaf (761) on CYP isozymes in human subjects --- p.60 / Chapter 6.1 --- Introduction --- p.60 / Chapter 6.2 --- Materials and Methods --- p.60 / Chapter 6.2.1. --- Drugs --- p.60 / Chapter 6.2.2. --- Subjects --- p.61 / Chapter 6.2.3. --- Study design --- p.62 / Chapter 6.2.4. --- Determination of probe drugs/metabolites in the plasma and urine --- p.63 / Chapter 6.2.5. --- Data analysis --- p.65 / Chapter 6.2.6. --- Statistical analysis --- p.66 / Chapter 6.3 --- Results --- p.66 / Chapter 6.3.1. --- Effect of EGb761on CYP1A2 activity --- p.66 / Chapter 6.3.2. --- Effect of EGb761on CYP2E1 activity --- p.67 / Chapter 6.3.3. --- Effect of EGb761 on CYP450 3A activity --- p.68 / Chapter 6.3.4. --- Effect of EGb761 on NAT2 activity --- p.69 / Chapter 6.3.5. --- Effect of EGb761 on CYP2D6 activity --- p.70 / Chapter 6.3.6. --- Effects of EGb761 on CYP2C19 activity --- p.71 / Chapter 6.4 --- Discussion --- p.72 / Chapter 6.5 --- Conclusion --- p.76 / References --- p.77 / Appendix --- p.90

Yim Wan Sze. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 95-109). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / 槪論 --- p.iv / Table of contents --- p.v / List of abbreviations --- p.xii / List of Figures --- p.xv / List of Tables --- p.xviii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Oxidation stress --- p.1 / Chapter 1.1.1 --- Free radicals --- p.2 / Chapter 1.1.1.1 --- Hydrogen peroxide --- p.3 / Chapter 1.1.1.2 --- Menadione --- p.4 / Chapter 1.1.2 --- Diseases related to oxidative stress --- p.5 / Chapter 1.1.3 --- Liver Injury --- p.5 / Chapter 1.1.4 --- Antioxidants --- p.7 / Chapter 1.1.4.1 --- Importance of antioxidant --- p.7 / Chapter 1.1.4.2 --- Examples of antioxidant --- p.7 / Chapter 1.2 --- Traditional Chinese Medicinal (TCM) formula Wu-zi-yan-zong-wan (WZ) --- p.8 / Chapter 1.2.1 --- The WZ medicinal formula --- p.8 / Chapter 1.2.2 --- Pharmacological actions of WZ --- p.9 / Chapter 1.2.3 --- Pharmacological actions of individual herbs --- p.10 / Chapter 1.2.3.1 --- Fructus Lycii --- p.10 / Chapter 1.2.3.2 --- Semen Cuscuta --- p.11 / Chapter 1.2.3.3 --- Fructus Rubi --- p.12 / Chapter 1.2.3.4 --- Semen Plantaginis --- p.12 / Chapter 1.2.3.5 --- Fructus Schisandrae --- p.13 / Chapter 1.3 --- The relationship between the liver and kidney --- p.14 / Chapter 1.4 --- Objectives of study --- p.15 / Chapter Chapter 2 --- Preparation of Aqueous Extraction of Wu-zi-yan-zong-wan --- p.17 / Chapter 2.1 --- Introduction --- p.17 / Chapter 2.2 --- Materials and Methods --- p.18 / Chapter 2.2.1 --- Preparation of Wu-zi-yan-zong-wan (WZ) --- p.18 / Chapter 2.2.1.1 --- WZ extracts from raw materials (WZ-e) --- p.18 / Chapter 2.2.1.2 --- WZ extracts from commercial available ready-to-use powders (WZ-p) --- p.20 / Chapter 2.3 --- Results --- p.22 / Chapter 2.3.1 --- Extraction Yield for WZ-e --- p.22 / Chapter Chapter 3 --- In vitro Total Antioxidant Capacity of Aqueous Extracts of Wu-zi-yan-zong-wan and its Components --- p.24 / Chapter 3.1 --- Introduction --- p.24 / Chapter 3.1.1 --- Total Antioxidants: Trolox Equavalent Antioxidant Capacity (TEAC) --- p.24 / Chapter 3.1.2 --- Objectives --- p.25 / Chapter 3.2 --- Materials and methods --- p.26 / Chapter 3.2.1 --- Materials --- p.26 / Chapter 3.2.1.1 --- Reagents --- p.26 / Chapter 3.2.1.2 --- Instruments --- p.26 / Chapter 3.2.2 --- Methods --- p.26 / Chapter 3.2.2.1 --- Total Antioxidnats: Trolox Equavalent Antioxidnat Capacity (TEAC) --- p.26 / Chapter 3.2.3 --- Statistical analysis --- p.27 / Chapter 3.3 --- Results --- p.28 / Chapter 3.3.1 --- Antioxidnat Capacity of Trolox --- p.28 / Chapter 3.3.2 --- Antioxidant capacity of WZ-e formula --- p.28 / Chapter 3.3.3 --- Antioxidant capacity of WZ-p formula --- p.28 / Chapter 3.3.4 --- The total antioxidant capacity of WZ-e and its simplified formulae --- p.32 / Chapter 3.3.5 --- The total antioxidant capacity of WZ-p and its simplified formulae --- p.32 / Chapter 3.3.6 --- Synergetic effect of WZ-e and its simplified formulae --- p.34 / Chapter 3.3.7 --- Orthogonal analysis of WZ-e and its simplified formulae on TEAC assay --- p.40 / Chapter 3.4 --- Discussion --- p.42 / Chapter Chapter 4 --- Antioxidant Activity of Aqueous Extracts of Simplified Formulae of Wu-zi-yan-zong-wan in vitro --- p.44 / Chapter 4.1 --- Introduction --- p.44 / Chapter 4.1.1 --- In vitro antioxidant --- p.44 / Chapter 4.1.2 --- Antioxidant effect of Catechins --- p.44 / Chapter 4.1.3 --- MTT assay --- p.45 / Chapter 4.1.4 --- Objectives --- p.46 / Chapter 4.2 --- Materials and methods --- p.47 / Chapter 4.2.1 --- Materials --- p.47 / Chapter 4.2.1.1 --- Cell Culture --- p.47 / Chapter 4.2.1.2 --- Reagents --- p.47 / Chapter 4.2.2 --- Methods --- p.47 / Chapter 4.2.2.1 --- Cell Culture --- p.47 / Chapter 4.2.2.2 --- MTT Cytotoxicity Assay --- p.48 / Chapter 4.2.3 --- Statistical analysis --- p.48 / Chapter 4.3 --- Results --- p.49 / Chapter 4.3.1 --- The effect of WZ-e formula on HepG2 cells --- p.49 / Chapter 4.3.2 --- The effect of hydrogen peroxide on HepG2 cells --- p.49 / Chapter 4.3.3 --- The effect of menadione on HepG2 cells --- p.52 / Chapter 4.3.4 --- The effect of catechin on HepG2 cells --- p.52 / Chapter 4.3.5 --- The effect of WZ-e and its simplified formulae against hydrogen peroxide-induced cytotoxicty on HepG2 cells --- p.55 / Chapter 4.3.6 --- The effect of WZ-e and its simplified formulae against menadione-induced cytotoxicity on HepG2 cells --- p.60 / Chapter 4.3.7 --- The effect of WZ-p on HepG2 cells --- p.65 / Chapter 4.3.8 --- The effect of WZ-p and its simplified formulae against hydrogen peroxide-induced cytotoxicity on HepG2 cells --- p.67 / Chapter 4.3.9 --- The effect of WZ-p and its simplified formulae against menadione-induced cytotoxicity on HepG2 cells --- p.72 / Chapter 4.4 --- Discussion --- p.77 / Chapter Chapter 5 --- Effect of Aqueous Extract of Wu-zi-yan-zong-wan on Menadione-induced Oxidative Damage in Mouse Liver --- p.79 / Chapter 5.1 --- Introduction --- p.79 / Chapter 5.2 --- Materials and methods --- p.81 / Chapter 5.2.1 --- Materials --- p.81 / Chapter 5.2.1.1 --- Animals --- p.81 / Chapter 5.2.1.2 --- Reagents --- p.81 / Chapter 5.2.1.3 --- Apparatus --- p.81 / Chapter 5.2.2 --- Methods --- p.82 / Chapter 5.2.2.1 --- Animal treatments --- p.82 / Chapter 5.2.2.2 --- Collection of samples --- p.82 / Chapter 5.2.2.3 --- Marker enzyme measurement (ALT) --- p.83 / Chapter 5.2.3 --- Statistical analysis --- p.83 / Chapter 5.3 --- Results --- p.84 / Chapter 5.3.1 --- Dose-dependent effect of WZ-e on menadione hepatotoxicity --- p.84 / Chapter 5.3.2 --- Dose-dependent effect of WZ-e on menadione hepatotoxicity as illustrated by histopathological observations --- p.86 / Chapter 5.3.3 --- Analyzing he effect of WZ-e and its simplified formulae on menadione-induced hepatotoxicity by Orthogonal Analysis89 --- p.89 / Chapter 5.4 --- Discussion --- p.91 / Conclusion --- p.93 / References --- p.95