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Análise da atividade antimicrobiana de extratos e frações purificadas da planta arrabidaea chica verlMota, Milena Rodrigues Soares 25 March 2011 (has links)
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Previous issue date: 2011-03-25 / Não Informada / This study describes the therapeutic potential of extracts and standardized fractions
of Arrabidaea chica leaves. A. chica is a Bignoniaceae popularly known as “crajiru”.
The genus Arrabidaea occurs in Tropical America, from the Southern Mexico to the
Southern Brazil. The red color of its dried leaves is attributed to two flavonoidal
pigments: carajurina (main pigment) and carajurona. Chemical studies described the
isolation of saponins and flavonoids from the plant leaves; purified 3-
desoxyanthocyanidins were reported as anti-inflammatory. The infusion or decoction
of the plant leaves is used in the folk medicine to treat anemia, inflammation and in
skin wound-healing. In this work, the antimicrobial activity of the standardized
extracts and fractions of A. chica cultivated at Embrapa Amazônia Ocidental,
Manaus, were evaluated against fungal and bacterial microorganisms grown either
from local domestic dogs and cats, or from human samples supplied by the
Microorganism Collection of FIOCRUZ, in Manaus, Brazil. The plant dried leaves
were extracted with increasing polarity solvents and progressively purified in
preparative thin layer chromatography (TLC) and silica-gel column; the semi-purified
extracts were standardized in TLC. The agar diffusion method; bioautography;
minimum inhibitory concentrations tested against Staphylococcus epidermidis
(CBAM 293), Staphylococcus aureus (CBAM 324), Pseudomonas aeruginosa
(CBAM 232), Escherichia coli (CBAM 002), Trichophyton mentagrophytes (CFAM
1288), Microsporum canis (CFAM 1289), Malassezia pachydermatis (CFAM 1290) e
Candida albicans (CFAM 1285). The standardized fractions were effective against all
these microorganisms, but more intensively against Microsporum canis and
Staphylococcus epidermidis. The results might favour the use of the standardized
sub-fractions of A. chica as topic phytotherapic agent to treat canine external otitis.
So far oleanolic and ursolic acids were identified as the main compounds in the
active semi-purified fraction but other compounds of the leaves extract were not
discarded. Later studies will consider the veterinarian use of the standardized extract,
of the active pure entities and the convenience of the natural active antibiotic mix. / Este estudo analisou o potencial terapêutico de extratos e frações purificadas da
planta amazônica Arrabidaea chica visando seu uso tópico como medicamento e
eficácia comprovada em doenças cutâneas. A. chica Verl., é uma Bignoniaceae
conhecida popularmente como crajiru. O gênero Arrabidaea ocorre na América
tropical, do sul do México ao sul do Brasil. A cor avermelhada da folha seca e sua
propriedade tintorial são devidas a dois pigmentos flavonoídicos: a carajurina, que é
o pigmento principal e a carajurona. Dela foram isolados saponinas e flavonóides;
As 3-desoxiantocianidinas, descritas na planta parecem possuir atividade
antiinflamatória. A medicina popular utiliza o decocto ou a infusão das folhas para
tratar anemia, inflamações e na cicatrização da pele. Neste trabalho, a atividade
antimicrobiana de extratos e frações padronizadas da A. chica cultivada na Embrapa
Amazônia Ocidental, em Manaus/AM, foi avaliada contra fungos e bactérias de
amostras clínicas coletadas de animais domésticos e contra amostras humanas
depositadas na coleção de Microrganimos da FIOCRUZ, Manaus/AM. Para isso, as
folhas secas da planta foram extraídas com solventes de polaridade crescente, as
frações foram progressivamente purificadas em cromatografia de placa ou coluna de
sílica-gel, os extratos semi-purificados foram padronizados em cromatografia líquida
de alta eficiência acoplada à espectrometria de massas. Os testes de difusão em
ágar, bioautografia e concentração inibitória mínima foram usados para avaliar a
atividade antimicrobiana das subfrações padronizadas frente aos microrganismos
Staphylococcus epidermidis (CBAM 293), Staphylococcus aureus (CBAM 324),
Pseudomonas aeruginosa (CBAM 232), Escherichia coli (CBAM 002), Trichophyton
mentagrophytes (CFAM 1288), Microsporum canis (CFAM 1289), Malassezia
pachydermatis (CFAM 1290) e Candida albicans (CFAM 1285). As frações
padronizadas foram ativas contra todos esses microrganismos, com melhores
resultados contra M. pachydermatis e S. epidermidis. Os resultados foram favoráveis
à utilização das subfrações padronizadas na formulação de um produto fitoterápico
para uso tópico em otite canina. Nas frações ativas foram identificados os ácidos
oleanólico e ursólico. Estudos posteriores deverão avaliar a possibilidade de uso
humano das frações purificadas ou dos compostos identificados.
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Development of Sensitive In Vitro Assays to Assess the Ocular Toxicity Potential of Chemicals and Ophthalmic ProductsMcCanna, David January 2009 (has links)
The utilization of in vitro tests with a tiered testing strategy for detection of mild ocular irritants can reduce the use of animals for testing, provide mechanistic data on toxic effects, and reduce the uncertainty associated with dose selection for clinical trials. The first section of this thesis describes how in vitro methods can be used to improve the prediction of the toxicity of chemicals and ophthalmic products. The proper utilization of in vitro methods can accurately predict toxic threshold levels and reduce animal use in product development. Sections two, three and four describe the development of new sensitive in vitro methods for predicting ocular toxicity. Maintaining the barrier function of the cornea is critical for the prevention of the penetration of infections microorganisms and irritating chemicals into the eye. Chapter 2 describes the development of a method for assessing the effects of chemicals on tight junctions using a human corneal epithelial and canine kidney epithelial cell line. In Chapter 3 a method that uses a primary organ culture for assessing single instillation and multiple instillation toxic effects is described. The ScanTox system was shown to be an ideal system to monitor the toxic effects over time as multiple readings can be taken of treated bovine lenses using the nondestructive method of assessing for the lens optical quality. Confirmations of toxic effects were made with the utilization of the viability dye alamarBlue. Chapter 4 describes the development of sensitive in vitro assays for detecting ocular toxicity by measuring the effects of chemicals on the mitochondrial integrity of bovine cornea, bovine lens epithelium and corneal epithelial cells, using fluorescent dyes.
The goal of this research was to develop an in vitro test battery that can be used to accurately predict the ocular toxicity of new chemicals and ophthalmic formulations. By comparing the toxicity seen in vivo animals and humans with the toxicity response in these new in vitro methods, it was demonstrated that these in vitro methods can be utilized in a tiered testing strategy in the development of new chemicals and ophthalmic formulations.
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Development of Sensitive In Vitro Assays to Assess the Ocular Toxicity Potential of Chemicals and Ophthalmic ProductsMcCanna, David January 2009 (has links)
The utilization of in vitro tests with a tiered testing strategy for detection of mild ocular irritants can reduce the use of animals for testing, provide mechanistic data on toxic effects, and reduce the uncertainty associated with dose selection for clinical trials. The first section of this thesis describes how in vitro methods can be used to improve the prediction of the toxicity of chemicals and ophthalmic products. The proper utilization of in vitro methods can accurately predict toxic threshold levels and reduce animal use in product development. Sections two, three and four describe the development of new sensitive in vitro methods for predicting ocular toxicity. Maintaining the barrier function of the cornea is critical for the prevention of the penetration of infections microorganisms and irritating chemicals into the eye. Chapter 2 describes the development of a method for assessing the effects of chemicals on tight junctions using a human corneal epithelial and canine kidney epithelial cell line. In Chapter 3 a method that uses a primary organ culture for assessing single instillation and multiple instillation toxic effects is described. The ScanTox system was shown to be an ideal system to monitor the toxic effects over time as multiple readings can be taken of treated bovine lenses using the nondestructive method of assessing for the lens optical quality. Confirmations of toxic effects were made with the utilization of the viability dye alamarBlue. Chapter 4 describes the development of sensitive in vitro assays for detecting ocular toxicity by measuring the effects of chemicals on the mitochondrial integrity of bovine cornea, bovine lens epithelium and corneal epithelial cells, using fluorescent dyes.
The goal of this research was to develop an in vitro test battery that can be used to accurately predict the ocular toxicity of new chemicals and ophthalmic formulations. By comparing the toxicity seen in vivo animals and humans with the toxicity response in these new in vitro methods, it was demonstrated that these in vitro methods can be utilized in a tiered testing strategy in the development of new chemicals and ophthalmic formulations.
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