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Efeitos anti-neoplásicos da raiz de Pfaffia paniculata (Ginseng brasileiro) no modelo de hepatocarcinogênese murina e em cultura de células de hepatocarcinoma humano / Antineoplastic effects of the roots of Pfaffia paniculata (Brazilian ginseng) in a murine hepatocarcinogenesis model and in human hepatocarcinoma cellsTereza Cristina da Silva 17 March 2008 (has links)
A raiz pulverizada de Pfaffia paniculata e seu extrato butanólico apresentam propriedades antineoplásicas, quimiopreventivas e antiangiogênicas, onde muitos indícios conferem às saponinas triterpenóides presentes nestas raízes as propriedades antitumorais observadas. Sabe-se que saponinas de diversos tipos de plantas possuem capacidade de interferir diretamente no ciclo celular de células tumorais. Assim, considerando os efeitos inibitórios das raízes e extratos de P. paniculata sobre a proliferação celular, o objetivo deste trabalho foi compreender os mecanismos envolvidos na ação quimiopreventiva desta raiz, tanto na fase de iniciação da carcinogênese hepática, quanto sobre células tumorais já estabelecidas. Inicialmente, foram avaliados os efeitos de diferentes concentrações da raiz em pó de P. paniculata adicionada à ração em camundongos submetidos ao modelo de hepatocarcinogênese, pela análise da proliferação celular, indução da apoptose e a comunicação intercelular hepática. Seqüencialmente, foram avaliados os efeitos das frações purificadas do extrato butanólico destas raízes sobre linhagem de células de carcinoma hepatocelular humano. Neste experimento foram analisadas a influência do tratamento sobre a viabilidade celular, fases e proteínas do ciclo celular, proliferação e presença de morte celular. No modelo de hepatocarcinogênese o tratamento com a raiz reduziu a proliferação celular, aumentou a apoptose e desencadeou processo inflamatório crônico em intensidades dependentes da concentração testada e não afetou a comunicação intercelular. Estes resultados indicam que os efeitos quimiopreventivos da P. Paniculata são decorrentes do controle da proliferação celular e apoptose, e são diretamente influenciados pela concentração da raiz. No experimento in vitro o tratamento reduziu a concentração das células vivas sem aumentar a concentração de células mortas, diminuiu a porcentagem de células na fase G2 do ciclo celular, reduziu a expressão dos genes das proteínas ciclinas D1, E e CDK6 e aumentou a expressão de p27, e não induziu apoptose. Estes resultados mostraram que a redução na concentração de células observadas após o tratamento com a fração do extrato butanólico de P. paniculata não foi decorrente de indução de apoptose. O tratamento parou o ciclo celular das células tumorais HepG2 em G1, inibindo proteínas importantes para a progressão do ciclo e estimulando a expressão de p27 um conhecido gene inibidor de CDKs. Os efeitos antiproliferativos observados no experimento in vivo em camundongos, reproduziram-se in vitro em células tumorais humanas, o que pode indicar que as propriedades antineoplásicas anteriormente observadas não são espécie específica. Em linhas gerais, as raízes e/ou as frações do extrato butanólico obtido a partir das raízes de P. paniculata apresentam propriedades antineoplásicas por inibir a proliferação celular e induzir apoptose in vivo e por parar o ciclo celular in vitro. Estes resultados consagram as propriedades antineoplásicas de Pfaffia paniculata e motivam o desenvolvimento de mais estudos sobre as suas potencialidades. / The powdered roots of Pfaffia paniculata and their butanolic extract present antineoplastic, chemopreventive and antiangiogenic properties, and many evidences suggest that the triterpenoid saponins are the responsible for these properties. It is well known that saponins from several types of plants have the capacity to directly interfere on the tumor cell cycle. Therefore, considering the inhibitory effects of the roots and extracts of P. paniculata on cell proliferation, the aim of this study was to search for the mechanisms involved in the chemopreventive effects of this root, both in the initiation phase of the hepatocarcinogenesis and on tumor cell lineage. Initially, the effects of different concentrations of the powdered root of P. paniculata added to the mouse food were evaluated in mice submitted to hepatocarcinogenesis model. Cell proliferation, induction of apoptosis, and hepatic intercellular communication were evaluated. The effects of the purified fractions of the butanolic extract of these roots were then evaluated in human hepatocarcinoma cells. In this experiment, the influence of the treatment on cell viability, phases and proteins of cell cycle, cell proliferation and cell death were evaluated. In the hepatocarcinogenesis model, the treatment with the root decreased cell proliferation, increased apoptosis and induced a chronic inflammatory process dependent on the concentration tested, and did not affect cell communication. These results indicate that the chemopreventive effects of P. Paniculata are apparently dependent on the control of cell proliferation and apoptosis and are directly influenced by the concentration of the root. In the in vitro treatment, it has been observed a reduction in the concentration of live cells without increasing the concentration of dead cells, decreased the level of G2 phase cells, reduced the expression of proteins cyclins D1, E and CDK6, increased the expression of p27, and did not induce apoptosis. These results showed that the reduction in the concentration of cells after the treatment with the butanolic extract of P. paniculata was not due to induction of apoptosis. The treatment inhibited the progression of the cell cycle of HepG2 cells in phase G1, by the inhibition of the expression of proteins that are important to the progression of the cycle, and stimulating the expression of p27, a known inhibitor of CDKs. The antiproliferative effects observed in the in vivo experiments were repeated in the in vitro study with human tumor cells. This may indicate that the antineoplastic properties previously observed are not species-specific. In conclusion, the roots and/or butanolic extract obtained from P. paniculata present antineoplastic properties due to inhibition of cell proliferation and induction of apoptosis in vivo, and due to the cell cycle arrest in vitro. These results reinforce the antineoplastic properties of Pfaffia paniculata and motivate the development of more studies focusing on its antineoplastic potentials.
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Levantamento da entomofauna de plantas medicinais, aromáticas e condimentares e desinfestação pelo processo de irradiação / Survey of insect fauna from plants medicinal, aromatic and seasoning and disinfestation the process of radiationFabrício Caldeira Reis 24 April 2013 (has links)
O presente trabalho teve como objetivo realizar o levantamento da entomofauna associada a plantas medicinais, aromáticas e condimentares desidratadas no comércio atacadista na cidade de São Paulo, empregar diferentes doses de radiação gama visando a desinfestação dos materiais e determina a dose letal de radiação gama de indivíduos adultos de Sphaericus gibboides. Foram coletados em 10 estabelecimentos no período de abril a maio de 2011 os materiais: Melissa officinalis L. (ervacidreira), Mentha piperita L. (hortelã), Ocimum basilicum L. (manjericão), Origanum vulgare L. (orégano), Rosmarinus officinalis L. (alecrim), Thymus vulgaris L. (tomilho), Senna alexandrina Mill (sene), Coriandrum sativum L. (coentro), Petroselinum crispum (Mill.) Fuss(salsa) e Pimpinella anisum L. (ervadoce), Baccharis trimera (Less.) DC. (carqueja), Matricaria chamomilla L. (= M. recutita L.) (camomila), Laurus nobilis L. (louro) (Lauraceae); Capsicum annuum L. (pápricadoce), Bixa orellana L. (colorífico) (Bixaceae); e Peumus boldus Molina (boldo). Na primeira triagem constatou-se que todos os materiais analisados não apresentaram presença de insetos adultos. Após o período de 45 dias foram encontrados 1838 insetos adultos, oriundos de ovos e larvas. Dos substratos analisados Matricaria recutita apresentou o maior índice de infestação, com 70,6 %. Pelmus boldus, Laurus nobilis, Matricaria recutita e Capsicum annuum, tiveram a maior diversidade de espécies. Baccharis strimera, Bixa orellana, Melissa officinalis, Origanum vulgares e Coriandrum sativum não apresentaram infestação. A espécie Lasioderma serricorne foi o inseto com maior número de indivíduos encontrados (936), maior porcentagem de infestação nos diferentes materiais (62,5 %) e maior ocorrência (68,75 %) nos materiais (M. piperita, S. alexandrina, P. anisum, M. chamomilla, P. crispum, L. nobilis, C. sativum, C. annuum, O. basilicum, P. boldus e T. vulgaris). Os seguintes materiais foram selecionados para os ensaios de desinfestação pelo processo de irradiação: Bixa orellana, Capsicum annuum, Cassia angustifolia, Coriandrum sativum, Mentha piperita, Petrosolium sativum, Matricaria recutita. Os materiais foram infestados com adultos de L. serricorne e Sphaericus gibboides, divididos em 5 parcelas de 30g com 20 indivíduos cada. Utilizando o Irradiador Multiprpósito cobalto-60 com taxa de dose no período dos ensaios de 6 kGy/h, as parcelas foram submetidas a doses crescentes de radiação gama de 0 (grupo controle); 0,25; 0,50; 0,75; 1,00; 1,25; 1,50; 1,75 e 2,00 kGy. A determinação da dose letal para adultos de Sphaericus gibboides, procedeu-se irradiando 4 repetições por tratamento contendo 25 indivíduos cada. As doses utilizadas foram: 2,4; 2,2; 2,0; 1,8; 1,6; 1,4; 1,2; 1,0; 0,8; 0,6 kGy. A menor dose para a não emergência de adultos de L. serricorne e S. gibboides em todos os materiais analisados foi a de 0,25 kGy. Não foi observada a presença de adultos nas demais doses crescentes. Foi observado que os materiais adquiridos no comércio apresentaram diferentes porcentagens de infestação justificando assim a aplicação de métodos de controle. A utilização do Irradiador Multipropósito Cobalto-60 com taxa de dose de 6 kGy/h foi eficiente no controle de L. serricorne e S. gibboides na dose de 0,25 kGy. A dose de 2,2 kGy atingiu 100% letalidade em indivíduos adultos de S. gibboides. / The present study aimed to survey the insect fauna associated with medicinal plants, aromatic dehydrated and seasoning trade in São Paulo city, using different doses of gamma radiation with the aim of disinfestation of the material and determine the lethal dose of gamma radiation on Sphaericus gibboides. From April to May 2011 were collected in 10 establishments the following sample materials: Melissa officinalis L. (Lemongrass), Mentha piperita L. (Mint), Ocimum basilicum L. (Basil), Origanum vulgare L. (Oregano), Rosmarinus officinalis L. (Rosemary), Thymus vulgaris L. (Thyme), Senna alexandrina Mill (senna), Coriandrum sativum L. (Coriander), Petroselinum crispum (Mill.) Fuss (salsa) and Pimpinella anisum L. (Fennel), Baccharis trimera (Less.) DC. (Gorse), Chamomilla recutita L. (= M. recutita L.) (chamomile), Laurus nobilis L. (Blonde) (Lauraceae); Capsicum annuum L. (Sweet paprika), Bixa orellana L. (Spice) (Bixaceae) and Peumus boldus Molina (Boldo). The first screening showed that all the tested materials did not show the presence of adult insects. After 45 days 940 adult insects were found and larvae from eggs. The substrates analyzed Chamomilla recutita showed the highest rate of infestation, with 70,6%. Pelmus boldus, Laurus nobilis, Chamomilla recutita and Capsicum annuum, had the highest species diversity. Baccharis trimera, Bixa orellana, Melissa officinalis, Origanum vulgare and Coriandrum sativum showed no infestation. The species was Lasioderma serricorne the insect with the largest number of individuals found (936), higher percentage of infestation in different materials (62.5%) and lots, and highest occurrence (68,75%) materials (M. piperita, S. alexandrian, P. anisum, Chamomilla recutita, P. crispum, L. nobilis, C. sativum, C. annuum, O. basilicum, P. boldus and T. vulgaris). The following materials were selected for testing disinfestation by irradiation process: Bixa orellana, Capsicum annuum, Cassia angustifolia, Coriandrum sativum, Mentha piperita, Petrosolium sativum, Chamomilla recutita. The materials were infested with adults of L. serricorne and S. gibboides, divided into 5 portions of 30g with 20 individuals each. Using Radiator Multiprpósito Cobalt-60 dose rate during the tests of 6 kGy/h, the plots were subjected to increasing doses of gamma radiation of 0 (control), 0,25; 0,50; 0,75; 1,00; 1,25; 1,50; 1,75 and 2,00 kGy. The determination of the lethal dose for adults S. gibboides, proceeded radiating 4 replicates per treatment each containing 25 individuals. The doses used were: 3,0; 2,8; 2,6; 2,4; 2,2; 2,0; 1,8; 1,6; 1,4; 1,2; 1,0; 0,8; 0,6 kGy. The lowest dose at no emergence of adult L. serricorne and S. gibboides in all materials tested was to 0,25 kGy. We did not observe the presence of other adults in increasing doses. It was observed that the materials acquired in trade showed different percentages of infestation thus justifying the application of control methods. Using Radiator Multiprpósito Cobalt-60 with dose rate of 6 kGy was effective in controlling L. serricorne and S. gibboides at a dose of 0,25 kGy. The dose of 2,2 kGy reached 100% mortality for adults of S. gibboides.
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Mulheres camponesas do Oeste Catarinense: organiza??o, trabalho e cuidado / Peasant women of west of Santa Catarina: Organization, work and carebarbosa, Yamira Rodrigues de Souza 16 March 2016 (has links)
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Previous issue date: 2016-03-16 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / This following work aimed to study the trajectory of the Peasant Women's Movement (MMC) in the west of Santa Catarina. The study was conducted based on the intersection of oral and written sources, as well as the participant observation, which was systematized in the field journal. The Movement of Agricultural Women (MMA) emerged in the eighties due to the need for guaranteeing social rights such as retirement and paid maternity leave for women farmers, this being one of the first fights of the movement. In its trajectory, the MMA joined the National Articulation of Rural Women Workers (ANMTR) in the mid-nineties, whose emergence was an embryo for the consolidation of the MMC, created in 2004. With the MMC, which brings together different organizations of rural women workers in Brazil, among them the MMA, emerged a peasant political identity and a defense of an ecological and feminist peasant agricultural project. It is perceived that the discussions regarding feminism gained prominence, especially through the dialogue with Agroecology, and have also been consolidated with the emergence of a new political category within the MMC / SC, the young peasants. Finally, it is considered that the construction of peasant feminism by the MMC / SC is the core element to guide the creation of more egalitarian relationships between men and women, constituting as part of a new rurality / O presente trabalho teve como objetivo geral analisar a trajet?ria do Movimento de Mulheres Camponesas (MMC) no Oeste de Santa Catarina. A pesquisa foi realizada com base no cruzamento de fontes orais e escritas al?m da observa??o participante que foi sistematizada no di?rio de campo. O Movimento de Mulheres Agricultoras (MMA) surgiu nos anos 1980 na busca pela garantia de direitos sociais como aposentadoria e sal?rio maternidade ?s agricultoras, o que exigiu o reconhecimento profissional das mulheres agricultoras, sendo esta uma das primeiras bandeiras de luta do movimento. Em sua trajet?ria, o MMA passou a integrar a Articula??o Nacional de Mulheres Trabalhadoras Rurais (ANMTR) em meados dos anos 1990, cujo surgimento foi um embri?o para a consolida??o do MMC, que foi criado em 2004. Com o MMC, que re?ne diversas organiza??es de mulheres trabalhadoras rurais do Brasil, dentre elas o MMA, emerge a identidade pol?tica camponesa e a defesa de um projeto de agricultura camponesa ecol?gica e feminista. Percebe-se que as discuss?es sobre feminismo ganharam destaque, sobretudo, a partir do di?logo com a Agroecologia e v?m se consolidando tamb?m com a emerg?ncia de uma nova categoria pol?tica no MMC/SC, a de jovens camponesas. Por fim, percebe-se que a constru??o de um feminismo campon?s pelo MMC/SC ? central para orientar a cria??o de rela??es mais igualit?rias entre homens e mulheres, constituindo-se como elemento de uma nova ruralidade
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Avaliação in vitro das atividades antioxidante, antimicrobiana e citotóxica de extratos orgânicos das cascas da do fruto jabuticaba (Myrciaria cauliflora Berg) / In vitro evaluation of antioxidant, antimicrobial and cytotoxic activities from organic extracts of the jabuticaba fruit’s peel (Myrciaria cauliflora Berg)Coelho, Jéssica Aparecida 08 February 2017 (has links)
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Previous issue date: 2017-02-08 / Currently, research related to natural products, mainly those of plant origin, has been strongly encouraged by the World Health Organization (WHO), once the use of plants with proven or unproven medicinal properties are of great influence on public health. Among the high medicinal potential plants, the species Myrciaria cauliflora Berg, which belongs to the Myrtaceae family, popularly known as `Jabuticaba`, a typical Brazilian fruit, is used in popular medicine to treat angina pectoris, dysentery, inflammation and asthma. Aiming to highlight the importance of the use of plants for the treatment of various diseases, the aim of this work was to identify the bioactive compounds present in Jabuticaba peel, and to perform in vitro tests of antioxidant action, antimicrobial potential and cytotoxic effect against red blood cells. The tests were carried out with the aim of obtaining results that could prove the possible pharmacological activities of the organic extracts, for further development of formulation with the use of pharmaceutical technology. Bioprospecting studies from the Brazilian flora represent concrete possibilities in the discovery of new antimicrobial agents, as well as the improvement and the discovery of new technologies in the most diverse industrial branches, with emphasis in the health sector. / Atualmente as pesquisas relacionadas aos produtos naturais, principalmente os de origem vegetal, têm sido incentivadas fortemente pela Organização Mundial da Saúde (OMS), visto que o uso de plantas com propriedades medicinais, comprovadas ou não, são de grande influência na saúde pública. Dentre as plantas com um alto potencial medicinal, destaca-se a espécie vegetal Myrciaria cauliflora Berg, pertencente à família Myrtaceae, conhecida popularmente como jabuticaba, fruta tipicamente brasileira, utilizada na medicina popular para tratar anginas do peito, disenterias, inflamações e asma. Visando ressaltar a importância do uso de plantas para o tratamento de diversos males, o intuito desse trabalho foi realizar a identificação dos compostos bioativos presentes nas cascas da jabuticaba, além, de realizar testes in vitro de ação antioxidante, potencial antimicrobiano e efeito citotóxico frente a hemácias humanas. Os testes foram realizados com o intuito de obter resultados que comprovassem as possíveis atividades farmacológicas dos extratos orgânicos, para posterior desenvolvimento de formulação com o uso de tecnologia farmacêutica. Estudos de bioprospecção a partir da flora brasileira, representam possibilidades concretas na descoberta de novos agentes antimicrobianos, assim como o aprimoramente e descobrimento de novas tecnologias nos mais diversos ramos industriais, com ênfase no setor da saúde.
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Efeitos do chá de Bauhinia forficata (subsp. pruinosa) sobre o dano oxidativo induzido por altas concentrações de glicose em eritrócitos humanos e por estreptozotocina em camundongosSalgueiro, Andréia Caroline Fernandes 15 January 2013 (has links)
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Previous issue date: 2013-01-15 / O Diabetes mellitus (DM) é uma desordem metabólica caracterizada pela deficiência absoluta ou relativa na produção de insulina pelas células β do pâncreas, ou ainda por uma incapacidade de utilização da insulina produzida. Acredita-se que a hiperglicemia crônica no DM esteja envolvida na gênese de diversos eventos bioquímicos, os quais resultam em um desequilíbrio entre a produção de espécies reativas de oxigênio e a capacidade dos sistemas de defesa antioxidante em neutralizá-las, levando a um estado de estresse oxidativo. Popularmente, muitas plantas conhecidas como medicinais, são utilizadas para tratar os sintomas do DM. Dentre estas plantas, estão as do gênero Bauhinia (Fabaceae, Leguminosae), cuja atividade antioxidante tem sido comprovada em modelos utilizando diferentes formas de extração de compostos químicos. Porém, pouco se sabe sobre os efeitos do chá das folhas de Bauhinia, tal como a mesma é popularmente utilizada. Com base no exposto, este trabalho teve como 13 objetivos investigar a atividade antioxidante da infusão das folhas de Bauhinia forficata (subsp. pruinosa) (BF), popularmente conhecida como “Pata de vaca” e largamente utilizada sob a forma de chá pela população, sobre o dano oxidativo induzido por altas concentrações de glicose em eritrócitos humanos (Manuscrito 1) e pela estreptozotocina (ETZ) em camundongos (Manuscrito 2). Os resultados do Manuscrito 1 mostraram que o chá de BF foi efetivo em reduzir o dano lipídico e em manter, ou mesmo aumentar, os níveis de antioxidantes em eritrócitos humanos incubados com elevadas concentrações de glicose (0 a 500mM). Acreditamos que estes resultados estejam diretamente relacionados aos elevados níveis de polifenóis e flavonoides presentes no chá de BF, os quais podem explicar a sua significativa atividade quelante de ferro e de neutralização de radicais hidroxilas (OH•), seja via neutralização do radical DPPH• seja via neutralização da reação de Fenton. Além de reduzir o dano oxidativo determinado por elevadas concentrações de glicose em eritrócitos humanos in vitro, observamos que o efeito antioxidante do chá de BF foi mantido em um modelo ex vivo de intoxicação por ETZ em camundongos (Manuscrito 2). Os resultados do Manuscrito 2 mostraram que o chá de BF foi capaz de minimizar as alterações oxidativas e morfológicas em diferentes órgãos, e também em reduzir a perda de peso e a hiperglicemia provocada pela ETZ em camundongos. O chá das folhas de BF foi bem aceito pelos animais e não teve efeitos per se sobre os achados bioquímicos ou histológicos deste trabalho. Acreditamos que os resultados observados estejam diretamente relacionados aos níveis de polifenóis e flavonoides previamente relatados. Em resumo, os efeitos antioxidantes do chá de BF foram observados tanto in vitro quanto ex vivo, e estes resultados contribuem para ressaltar e justificar os benefícios do uso do chá das folhas desta planta medicinal, tal como a mesma é popularmente 14 utilizada, em minimizar as alterações oxidativas de processos patológicos com incidência crescente na sociedade contemporânea, a exemplo do DM. / Diabetes mellitus (DM) is a metabolic disorder characterized by an absolute or relative deficiency in insulin production by pancreatic β cells, or by an inability to use the insulin produced. It is believed that the chronic hyperglycemia in DM is involved in the genesis of several biochemical events that result in an imbalance between the production of reactive oxygen species and the antioxidant defense systems capacity to neutralize them, leading to an oxidative stress state. Many plants known as medicinal are used to alleviate the symptoms of DM. Among these are the plants of the Bauhinia genus (Fabaceae, Leguminosae), which antioxidant activity was already proven in models of different extractions of chemical compounds. However, little is known about the effects of Bauhinia leaves tea, as it is popularly used. Based on the above, this study aimed to investigate the antioxidant activity of Bauhinia forficata (subsp. pruinosa) 16 (BF) leaves infusion, popularly known as “Paw cow” and used as tea in folk medicine, on the oxidative damage induced by high glucose concentrations in human erythrocytes (Manuscript 1) and by streptozotocin (STZ) in mice (Manuscript 2). The results of Manuscript 1 showed that the BF tea was effective in reducing lipid damage and maintain, or even increase, the antioxidant levels in human erythrocytes incubated with high glucose concentrations. We believe that these results are directly related to high levels of polyphenols and flavonoids present in BF tea, which can explain its significant iron chelating activity and hydroxyl radical (OH •) neutralization, either via DPPH• radical scavenger or via Fenton reaction neutralization. More than the oxidative damage reduction determined by high glucose concentrations in human erythrocytes in vitro, we found that the antioxidant potential of BF tea was kept in an ex vivo model of STZ intoxication in mice (Manuscript 2). The results of Manuscript 2 showed that BF tea was able to minimize the oxidative and morphological damage in various organs, and also to reduce the weight loss and hyperglycemia caused by STZ in mice. BF leaves tea was well accepted by the mice and had no per se effect on biochemical or histological findings of this study. We believe that the observed results are directly related with the polyphenols and flavonoids levels previously reported. In summary, the antioxidant effects of BF tea were observed both in vitro and ex vivo, and these results help to highlight and explain the benefits of using leaves tea of this medicinal plant, as it is popularly used, to minimize oxidative changes of pathological processes with increasing incidence in contemporary society, such as the DM.
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The antimicrobial and immunomodulatory effects of Cotyledon Orbiculata extractsTyavambiza, Caroline January 2018 (has links)
Thesis (MSc (Biomedical Technology))--Cape Peninsula University of Technology, 2018. / The challenge of antimicrobial resistance has increased drastically over the years as more microorganisms are becoming resistant to the available conventional treatments. The burden of antimicrobial resistant infections is intensified by the increase in immunocompromising conditions such as HIV/AIDS and cancer. Due to this challenge, pharmaceutical companies, health sectors and researches are in search of new antimicrobial agents that can solve the problem at hand. Medicinal plants are a reliable source for drug discovery as it is estimated that 25% of modern medicine originated from plants. They have also been used traditionally as sources of medicine in the treatment of many human ailments. Plants can also be applied in the field of nanotechnology. Nanotechnology is a promising field in medicine as it has the potential to offer improved methods for disease diagnostics and therapeutics. The use of plants in nanotechnology brings about biologically friendly nanomaterials. Cotyledon orbiculata is one of the well-known and common plants of South Africa that is used in traditional medicinal practices. The nanotechnology applications as well as the antimicrobial and immunomodulatory effects of this plant were evaluated. The ability of C. orbiculata to synthesize silver nanoparticles was determined. Optimisation of silver nanoparticle synthesis using water extract of C. orbiculata was done at different conditions. The conditions evaluated include, reaction temperature (25 and 70°C), silver nitrate concentration (1 and 3mM), plant extract concentration (1.5, 3 and 6mg/ml) and reaction time. The synthesis of silver nanoparticles using this plant was successful. The optimal conditions for the synthesis of silver nanoparticles using C. orbiculata were 3mg/ml of the C. orbiculata extract, 3mM silver nitrate at a reaction temperature of 70°C for 2 hours. Under these conditions, spherical, crystalline nanoparticles with sizes of 20-40nm were produced. The antimicrobial and immunomodulatory properties of C. orbiculata extracts and silver nanoparticles were evaluated. Antimicrobial activity was evaluated against Staphylococcus aureus, Staphylococcus epidermidis, Methicillin resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa and Candida albicans, using the micro-dilution assay to determine the minimum inhibitory concentration (MIC). The results obtained revealed that all extracts of C. orbiculata have antimicrobial properties against all the microorganisms tested. The MICs of the extracts ranged from 3.13 to 50mg/ml and the MBC/MFC from 6.25 to >100mg/ml. The methanol extract exhibited better antimicrobial activity in comparison to the others extracts whereas the water extract had better antifungal properties. The chloroform extract showed the lowest activity in both antibacterial and antifungal studies. Silver nanoparticles also exhibited antimicrobial activity against all the microorganisms tested. It’s MICs against these microorganisms ranged from 5–80μg/ml and MBC/MFC from 20-160μg/ml. The silver nanoparticles were highly active than the water extract against both the bacteria and the fungi. Immunomodulatory effects of the plant extracts and silver nanoparticles were determined by evaluating cytokine production using the enzyme linked immunoassay (ELISA) assay. All the extracts and silver nanoparticles of C. orbiculata were found to have anti-inflammatory properties. The water extracts showed more anti-inflammatory activity against the cytokines than the other extracts. However the silver nanoparticles were more active than the water extract. The findings from this study confirmed that C. orbiculata have antimicrobial and immunomodulatory effects. This provided scientific evidence of the traditional use of this plant in the treatment of skin infections and inflammatory conditions.
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Antidiabetic and profertility mechanisms of aqueous extract of Basella alba in male Wistar ratsArokoyo, Dennis Seyi January 2017 (has links)
Thesis (PhD (Biomedical Sciences))--Cape Peninsula University of Technology, 2017. / The use of medicinal plants in the management of various health problems date back to the ancient times. However, only in recent years, researchers are starting to focus on the use of natural plant products as alternative treatment in disease control. Basella alba (Ba), commonly called Ceylon or Indian spinach is one of such medicinal plants, wildly cultivated and consumed mostly as vegetable. Studies have established many beneficial effects of Ba, including androgenic effects as well as antidiabetic effects which have been described in rats following oral administration of the leave extract. However, the actual mechanisms underlying the antihyperglyceamic effect of Ba have not been reported in any study and little or no research details are yet available on the potential beneficial effects of Ba in reproductive dysfunction resulting from diabetes mellitus. This study was aimed at investigating the mechanisms underlying the antidiabetic effect of Ba and the possibility of a role for the plant in correcting diabetic-induced reproductive dysfunctions in male Wistar rats. The first part of the study involved comparing of three different solvent extracts of Ba leaves namely ethyl acetate, methanolic and aqueous extracts for their antioxidant potentials, after which the aqueous extract was selected for further use in the experiments. Animal experimentation involved male rats (n=40) aged 8-10 weeks, randomly divided into four equal groups as follows: Healthy Control, Diabetic Control, Healthy Treatment and Diabetic Treatment. Diabetes was induced via a single intraperitoneal injection of streptozotocin (55mg/kg) and all animals subsequently received treatment via gavage (Rats in Control groups received 0.5ml/100g normal saline daily and treatment groups received 200mg/kg plant extract daily) for a period of four weeks. Fasting blood sugar and body weights were recorded weekly throughout the study. Animals were sacrificed upon completion of the treatment and blood samples and tissues collected for further analysis which included computer aided sperm analysis, Luminex® technology and enzyme-linked immunosorbent hormonal assays, inflammatory cytokine assays, analysis of oxidative stress markers and Histopathological analysis. The single intraperitoneal injection of a high streptozotocin dose resulted in hyperglycaemia, weight loss, subnormal sperm parameters, negative balance of inflammatory cytokines and endogenous antioxidants and degenerative changes in the pancreas, testes and epididymis as observed in the diabetic control rats. Oral administration with the aqueous extract of Ba for four weeks in diabetic treatment rats led to a significant reduction in blood sugar and improvement of sperm parameters by modulating the production of gonadal hormones, in vivo antioxidants and inflammatory cytokines. There was also significant recovery of normal islet histology and reduction in testicular and epididymal degeneration in the diabetic treatment rats when compared to their diabetic control counterparts. It was concluded from the findings of this study that the antidiabetic and profertility effects of Ba are largely dependent on the modulation of in vivo production of antioxidants, gonadal hormones and inflammatory cytokines, probably stimulated by one or more phytochemical component(s) that can be isolated in the aqueous extract of the plant
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Molecular authentication of Chinese herbs derived from Aristolochia.January 2008 (has links)
Lam, Hilary. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 182-191). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.I / ABSTRACT --- p.III / TABLE OF CONTENTS --- p.V / LIST OF FIGURES --- p.X / LIST OF TABLES --- p.XIX / LIST OF ABBREVIATIONS --- p.XXII / Chapter Chapter 1: --- LITERATURE REVIEW --- p.1 / Chapter 1. --- Aristolochia --- p.1 / Chapter 1.1 --- "Aristolochia, as a plant" --- p.1 / Chapter 1.2 --- The chemicals in Aristolochia --- p.1 / Chapter 1.3 --- "Aristolochia, as herbal remedies" --- p.3 / Chapter 1.4 --- The Aristolochia poisoning cases --- p.4 / Chapter 1.5 --- The mechanism of AAs --- p.6 / Chapter 1.6 --- Renaming CHN to AAN --- p.6 / Chapter 1.7 --- Banning Aristolochia herbs --- p.7 / Chapter 1.8 --- The possible cause of ANN --- p.8 / Chapter 1.8.1 --- Misuse of Chinese Medicine --- p.8 / Chapter 1.8.2 --- Substitution --- p.9 / Chapter 1.8.3 --- The complexities of the herbal nomenclature --- p.9 / Chapter 1.8.4 --- Adulteration --- p.11 / Chapter 1.9 --- Methods for authenication --- p.12 / Chapter 1.9.1 --- Traditional methods for authentication --- p.12 / Chapter 1.9.2 --- The advantage of using molecular methods --- p.13 / Chapter 1.9.2.1 --- DNA fingerprinting --- p.13 / Chapter 1.9.2.2 --- DNA sequencing --- p.15 / Chapter 1.10 --- Method selection rationale --- p.15 / Chapter 1.11 --- The need for molecular authentication of six medicinal herbs --- p.17 / Chapter 1.11.1 --- The herb Mutong --- p.17 / Chapter 1.11.1.1 --- The poisoning cases reported --- p.19 / Chapter 1.11.1.2 --- Other authentication studies of Mutong --- p.19 / Chapter 1.11.2 --- The herb Muxiang --- p.20 / Chapter 1.11.2.1 --- Chemical profile --- p.21 / Chapter 1.11.2.2 --- Other authentication studies of Muxiang --- p.21 / Chapter 1.11.3 --- The herb Baiying --- p.22 / Chapter 1.11.3.1 --- The poisoning cases reported --- p.23 / Chapter 1.11.3.2 --- Other authentication studies of Baiying --- p.24 / Chapter 1.11.4 --- The herb Fangj --- p.i 24 / Chapter 1.11.4.1 --- Chemical profile --- p.25 / Chapter 1.11.4.2 --- The poisoning cases reported --- p.26 / Chapter 1.11.5 --- The herb Madouling --- p.26 / Chapter 1.11.6 --- The herb Zhushalian --- p.27 / Chapter 1.12 --- Aristolochia specific markers --- p.28 / Chapter 1.13 --- Significance of the research --- p.29 / Chapter Chapter 2: --- OBJECTIVE --- p.30 / Chapter Chapter 3: --- MATERIALS AND METHODS --- p.31 / Chapter 3.1 --- Samples source --- p.31 / Chapter 3.2 --- Total DNA extraction --- p.39 / Chapter 3.2.1 --- Cetyltriethylammonium bromide extraction --- p.39 / Chapter 3.2.2 --- Commercial kit extraction --- p.40 / Chapter 3.3 --- DNA amplification --- p.42 / Chapter 3.4 --- DNA fingerprinting --- p.43 / Chapter 3.4.1 --- DNA concentration determination --- p.43 / Chapter 3.4.2 --- ISSR fingerprinting --- p.44 / Chapter 3.5 --- Agarose gel electrophoresis --- p.45 / Chapter 3.6 --- Purification of PCR product --- p.46 / Chapter 3.7 --- Cloning of PCR product --- p.47 / Chapter 3.7.1 --- Ligation --- p.47 / Chapter 3.7.2 --- Transformation --- p.48 / Chapter 3.7.3 --- Cell cultivation --- p.48 / Chapter 3.7.4 --- Plasmid extraction --- p.49 / Chapter 3.7.5 --- Insert confirmation --- p.49 / Chapter 3.8 --- DNA sequencing --- p.50 / Chapter 3.8.1 --- Cycle sequencing --- p.50 / Chapter 3.8.2 --- Purification of cycle sequencing product --- p.51 / Chapter 3.8.3 --- DNA analysis --- p.51 / Chapter 3.9 --- Sequence analysis --- p.52 / Chapter Chapter 4: --- AUTHENICATION OF MUTONG --- p.53 / Chapter 4.1 --- Results --- p.53 / Chapter 4.1.1 --- Sequence alignment --- p.54 / Chapter 4.1.1.1 --- trnL-trnF sequences --- p.54 / Chapter 4.1.1.2 --- psbA-trnH sequences --- p.55 / Chapter 4.1.2 --- Percentage similarity analysis --- p.64 / Chapter 4.1.3 --- Dendrogram analysis --- p.67 / Chapter 4.2 --- Discussion --- p.73 / Chapter 4.2.1 --- Evaluation of chloroplast trnL-trnF region in differentiation of Mutong --- p.73 / Chapter 4.2.2 --- Evaluation of chloroplast psbA-trnH region in differentiation of Mutong --- p.74 / Chapter 4.2.3 --- Evaluation of using DNA sequencing in differentiation of Mutong --- p.75 / Chapter 4.3 --- Conclusion --- p.77 / Chapter Chapter 5: --- AUTHENICATION OF MUXIANG --- p.78 / Chapter 5.1 --- Results --- p.78 / Chapter 5.1.1 --- Sequence alignment --- p.79 / Chapter 5.1.1.1 --- trnL-trnF sequences --- p.79 / Chapter 5.1.1.2 --- psbA-trnH sequences --- p.80 / Chapter 5.1.2 --- Percentage similarity analysis --- p.88 / Chapter 5.1.3 --- Dendrogram study --- p.91 / Chapter 5.2 --- Discussion --- p.97 / Chapter 5.2.1 --- Evaluation of chloroplast trnL-trnF region in differentiation of Muxiang --- p.97 / Chapter 5.2.2 --- Evaluation of chloroplast psbA-trnH region in differentiation of Muxiang --- p.99 / Chapter 5.3 --- Conclusion --- p.100 / Chapter Chapter 6: --- AUTHENICATION OF BAIYING --- p.102 / Chapter 6.1 --- Results --- p.102 / Chapter 6.1.1 --- Sequence alignment --- p.103 / Chapter 6.1.2 --- Percentage similarity analysis --- p.107 / Chapter 6.1.3 --- Dendrogram analysis --- p.107 / Chapter 6.2 --- Discussion --- p.109 / Chapter 6.2.1 --- Evaluation of chloroplast psbA-trnH region in differentiation of Solarium and Aristolochia --- p.109 / Chapter 6.2.2 --- Molecular authentication of Baiying --- p.112 / Chapter 6.3 --- Conclusion --- p.113 / Chapter Chapter 7: --- AUTHENICATION OF FANGJI --- p.114 / Chapter 7.1 --- Results --- p.114 / Chapter 7.1.1 --- Sequence alignment --- p.115 / Chapter 7.1.1.1 --- trnL-trnF sequence --- p.115 / Chapter 7.1.1.2 --- psbA-trnH sequence --- p.116 / Chapter 7.1.2 --- Percentage similarity analysis --- p.123 / Chapter 7.1.3 --- Dendrogram study --- p.126 / Chapter 7.2 --- Discussion --- p.132 / Chapter 7.2.1 --- Evaluation of chloroplast trnL-trnF region in differentiation of Fangji --- p.132 / Chapter 7.2.2 --- Evaluation of chloroplast psbA-trnH region in differentiation of Fangji --- p.133 / Chapter 7.3 --- Conclusion --- p.133 / Chapter Chapter 8: --- AUTHENICATION OF MADOULING --- p.135 / Chapter 8.1 --- Results --- p.135 / Chapter 8.1.1 --- Sequence alignment --- p.136 / Chapter 8.1.1.1 --- trnL-trnF sequence --- p.136 / Chapter 8.1.1.2 --- psbA-trnH sequence --- p.136 / Chapter 8.1.2 --- Percentage similarity analysis --- p.143 / Chapter 8.1.3 --- Dendrogram study --- p.146 / Chapter 8.2 --- Discussion --- p.152 / Chapter 8.2.1 --- Evaluation of chloroplast trnL-trnF region in differentiation of Madouling --- p.152 / Chapter 8.2.2 --- Evaluation of chloroplast psbA-trnH region in differentiation of Madouling --- p.153 / Chapter 8.3 --- Conclusion --- p.153 / Chapter Chapter 9: --- AUTHENICATION OF ZHUSHALIAN --- p.155 / Chapter 9.1 --- Results --- p.155 / Chapter 9.1.1 --- Sequence alignment --- p.156 / Chapter 9.1.1.1 --- trnL-trnF sequence --- p.156 / Chapter 9.1.1.2 --- psbA-trnH sequence --- p.157 / Chapter 9.1.2 --- Percentage similarity analysis --- p.157 / Chapter 9.1.3 --- Dendrogram study --- p.162 / Chapter 9.2 --- Discussion --- p.166 / Chapter 9.2.1 --- Evaluation of chloroplast trnL-trnF region in differentiation of Zhushalian --- p.166 / Chapter 9.2.2 --- Evaluation of chloroplast psbA-trnH region in differentiation of Zhushalian --- p.171 / Chapter 9.3 --- Conclusion --- p.171 / Chapter Chapter 10: --- ARISTOLOCHIA SPECIFIC MARKER --- p.172 / Chapter 10.1 --- ISSR fingerprinting --- p.172 / Chapter 10.2 --- Discussion --- p.178 / Chapter Chapter 11: --- CONCLUSION --- p.180 / BIBLIOGRAPHY --- p.182 / APPENDIX - MATERIALS PREPARATION --- p.192
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Molecular authentication of baihuasheshecao and icefish.January 2012 (has links)
Yu, Jing. / "November 2011." / Thesis (M.Phil.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 161-172). / Abstracts in English and Chinese. / Abstract --- p.I / 摘要 --- p.III / Acknowledgements --- p.V / Contents --- p.VI / List of Figures --- p.VIII / List of Tables --- p.X / Abbreviations and Symbols --- p.XII / Chapter CHAPTER 1 --- Introduction --- p.1 / Chapter 1.1 --- Phylogenetic study of Hedyotis --- p.2 / Chapter 1.1.1 --- Rubiaceae --- p.2 / Chapter 1.1.2 --- Controversial taxonomic issues --- p.9 / Chapter 1.2 --- Traditional Chinese medicine (TCM) --- p.19 / Chapter 1.2.1 --- Introduction --- p.19 / Chapter 1.2.2 --- Baihuasheshecao --- p.20 / Chapter 1.2.3 --- Authentication of Baihuasheshecao --- p.23 / Chapter 1.3 --- Icefishes in the Hong Kong market --- p.23 / Chapter 1.3.1 --- Introduction --- p.23 / Chapter 1.4 --- Molecular approach --- p.25 / Chapter 1.4.1 --- Introduction to molecular phylogeny --- p.25 / Chapter 1.4.2 --- FINS (Forensically Informative Nucleotide Sequencing) for species identification --- p.27 / Chapter 1.4.3 --- DNA sequence markers --- p.28 / Chapter 1.5 --- Objectives --- p.38 / Chapter CHAPTER 2 --- MATERIALS AND METHODOLOGY --- p.41 / Chapter 2.1 --- Materials --- p.42 / Chapter 2.2 --- DNA extraction --- p.50 / Chapter 2.3 --- Polymerase chain reaction (PCR) method --- p.51 / Chapter 2.4 --- Gel electrophoresis --- p.54 / Chapter 2.5 --- PCR production purification --- p.54 / Chapter 2.6 --- Ligation and transformation and transformation of PCR product --- p.56 / Chapter 2.7 --- DNA sequencing and sequence analyses --- p.58 / Chapter CHAPTER 3 --- USING FORENSICALLY INFORMATIVE NUCLEOTIDE SEQUENCING (FINS) TECHNOLOGY FOR SPECIES IDENTIFICATION --- p.64 / Chapter 3.1 --- Authentication of Baihuasheshecao by FINS Analysis --- p.65 / Chapter 3.1.1 --- Authentication using FINS technology --- p.65 / Chapter 3.1.2 --- Relative effectiveness of DNA regions for FINS analysis --- p.70 / Chapter 3.1.3 --- Phylogenetic interpretation --- p.72 / Chapter 3.2 --- Authentication of Salangids (Icefishes) by FINS Analysis --- p.74 / Chapter 3.2.1 --- Analysis based on mitochondrial ribosome DNA region --- p.74 / Chapter 3.2.2 --- Analysis based on mitochondrial 16S rRNA --- p.77 / Chapter 3.2.3 --- Analysis based on combined regions --- p.79 / Chapter 3.2.4 --- Phylogenetic analysis --- p.81 / Chapter 3.2.5 --- Discussion --- p.85 / Chapter 3.3 --- Conclusions --- p.88 / Chapter CHAPTER 4 --- PHYLOGENTIC STUDY OF HEDYOTIS IN CHINA AND THEIR POSITION IN SPERMACOCEAE --- p.89 / Chapter 4.1 --- Phylogentic study of Hedyotis species in Chinese --- p.90 / Chapter 4.1.1 --- Nuclear ITS region --- p.90 / Chapter 4.1.2 --- Plastid trnL intron and trnL-V intergenic spacer region --- p.94 / Chapter 4.1.3 --- Plastid trnH-psbA intergenic spacer region --- p.98 / Chapter 4.1.4 --- Plastid rbcL region --- p.102 / Chapter 4.1.5 --- Plastid matK region --- p.106 / Chapter 4.1.7 --- Combined analysis --- p.114 / Chapter 4.2 --- The phylogenetic position of Hedyotis (species in China) in the tribe of Spermacoceae s.1 --- p.121 / Chapter 4.2.1 --- Plastid trnL - F intergenic spacer region --- p.121 / Chapter 4.2.2 --- Plastid rbcL region --- p.133 / Chapter 4.2.3 --- Plastid rps16 region --- p.141 / Chapter 4.3 --- Discussion --- p.153 / Chapter 4.3.1 --- Comparison of phylogenetic utility of the six DNA regions --- p.153 / Chapter 4.3.2 --- Diplophragma section --- p.154 / Chapter 4.3.3 --- "Hedyotis, Dimetia, Euoldendandia and Gonotheca sections" --- p.156 / Chapter 4.3.4 --- The position of Hedyotis (species in China) in Spermacoceae --- p.158 / Chapter 4.4 --- Conclusions --- p.160 / REFERENCES --- p.161 / APPENDIX --- p.173
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Investigation of the probable anti-cancer effects of the crude methanol extract of dicerocaryum senecioides, (Klotzch) J. Abels, leaves on cervical HeLa cancer cellMalemela, Kholofelo Mmanoko January 2018 (has links)
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
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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)
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