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Phytochemical analysis and bioactivity of Garcinia Kola (Heckel) seeds on selected bacterial pathogensSeanego, Christinah Tshephisho January 2012 (has links)
Garcinia kola is one of the plants used in folklore remedies for the treatment of microbial infections. Bacterial resistance to commonly used antibiotics has necessitated the search for newer and alternative compounds for the treatment of drug resistant microbial infections. This study focuses on the bioactivity of G. kola seeds on Streptococcus pyogenes (ATCC 49399), Staphylococcus aureus (NCTC 6571), Plesiomonas Shigelloides (ATCC 51903) and Salmonella typhimurium (ATCC 13311), organisms which can cause illnesses from mild to severe with potentially fatal outcomes. The crude ethyl acetate, ethanol, methanol, acetone and aqueous extracts were screened by agar-well diffusion method and the activities of the extract were further determined by Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays. The inhibition zones ranged from 0 - 24 mm, while MIC and MBC of the extract ranged between 0.04 - 1.25 mg/mL and 0.081 - 2.5 mg/mL respectively. Chloroform/ Ethyl Acetate/ Formic acid (CEF) solvent system separated more active compounds followed by Ethyl Acetate/ Methanol/ Water (EMW) and Benzene/ Ethanol/ Ammonium Hydroxide (BEA). The extracts were fractionated by Thin Layer Chromatography (TLC). Bioautography was used to assess the activity of the possible classes of compounds present in the more active extracts. Column chromatography was used to purify the active compounds from the mixture while Gas Chromatography-Mass Spectrometry (GC-MS) was used to identify the phyto components of the fractions. The MIC of the fractions ranged between 0.0006 - 2.5 mg/mL. CEF 3 (F3), CEF 11 (F11) and CEF 12 (F12) revealed the presence of high levels fatty acids Linoleic acid, 1, 2-Benzenedicarboxylic acid and 2, 3-Dihydro-3, 5-dihydroxy-6-methyl, respectively. The results obtained from this study justify the use of this plant in traditional medicine and provide leads which could be further exploited for the development of new and potent antimicrobials.
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Potencial antitumoral do composto 7-epi-clusianona em linhagens celulares de câncer de mama humano cultivadas como monocamadas e esferoides. / Antitumoral potential of 7-epi-clusianone in human breast cancer cell lines cultured in monolayer and as spheroids.Sales, Bianca Rocha 25 September 2015 (has links)
A biodiversidade de plantas brasileiras é uma fonte muito rica de moléculas bioativas, dentro da proposta da busca por novas drogas antitumorais, avaliamos neste estudo o potencial antiproliferativo do composto 7-epi-clusianona. Foram utilizadas duas linhagens celulares derivadas de tumor de mama humana, Hs 578T e MCF-7, cultivadas em monocamada e como esferoides. O IC50 após 48 horas de tratamento das células é de 20 μM para Hs 578T e 6 μM para MCF-7. A análise do ciclo celular mostrou que o composto é capaz de reter as células em fase G1/G0 em ambas as linhagens em 2D, mas não em 3D. O composto é capaz de induzir as células a senescência celular, como mostrado pelo ensaio de detecção de β-galactosidase. Esses dados indicam que o composto 7-epi-clusianona é uma molécula promissora, que demonstrou potencial antitumoral em células de tumor de mama. A cultura tridimensional se mostrou mais resistente ao tratamento com 7-epi-clusianona, portanto estudos mais abrangentes são necessários para melhor entendimento dos efeitos do composto sobre esse tipo de cultura. / Brazilian flora is considered one of the most diverse in the world and natural products are some of the important sources of new antitumoral compounds. The aim of this study was to evaluate the antiproliferative potential of 7-epi-clusianone. Two cell lines derived from human breast tumor were used, Hs 578T and MCF-7, cultured in monolayer and as spheroids. The IC50 after 48 hours of treatment is 20 μM to Hs 578T cells and 6 μM to MCF-7 cells. Cell cycle analysis showed induction of cell cycle arrest in G1/S phase in cells cultured in monolayers, but not in spheroids. The amount of cells in senescence after the treatment with 7-epi-clusianone is higher than the control group, as seen by the senescence β-galactosidase staining assay. These data suggest that 7-epi-clusianone is a promising molecule against breast cancer cells. We show that 3D culture was more resistant to treatment than 2D culture, therefore more comprehensive studies are needed to better understand the effects of 7-epi-clusianone on this kind of culture.
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Potencial antitumoral do composto 7-epi-clusianona em linhagens celulares de câncer de mama humano cultivadas como monocamadas e esferoides. / Antitumoral potential of 7-epi-clusianone in human breast cancer cell lines cultured in monolayer and as spheroids.Bianca Rocha Sales 25 September 2015 (has links)
A biodiversidade de plantas brasileiras é uma fonte muito rica de moléculas bioativas, dentro da proposta da busca por novas drogas antitumorais, avaliamos neste estudo o potencial antiproliferativo do composto 7-epi-clusianona. Foram utilizadas duas linhagens celulares derivadas de tumor de mama humana, Hs 578T e MCF-7, cultivadas em monocamada e como esferoides. O IC50 após 48 horas de tratamento das células é de 20 μM para Hs 578T e 6 μM para MCF-7. A análise do ciclo celular mostrou que o composto é capaz de reter as células em fase G1/G0 em ambas as linhagens em 2D, mas não em 3D. O composto é capaz de induzir as células a senescência celular, como mostrado pelo ensaio de detecção de β-galactosidase. Esses dados indicam que o composto 7-epi-clusianona é uma molécula promissora, que demonstrou potencial antitumoral em células de tumor de mama. A cultura tridimensional se mostrou mais resistente ao tratamento com 7-epi-clusianona, portanto estudos mais abrangentes são necessários para melhor entendimento dos efeitos do composto sobre esse tipo de cultura. / Brazilian flora is considered one of the most diverse in the world and natural products are some of the important sources of new antitumoral compounds. The aim of this study was to evaluate the antiproliferative potential of 7-epi-clusianone. Two cell lines derived from human breast tumor were used, Hs 578T and MCF-7, cultured in monolayer and as spheroids. The IC50 after 48 hours of treatment is 20 μM to Hs 578T cells and 6 μM to MCF-7 cells. Cell cycle analysis showed induction of cell cycle arrest in G1/S phase in cells cultured in monolayers, but not in spheroids. The amount of cells in senescence after the treatment with 7-epi-clusianone is higher than the control group, as seen by the senescence β-galactosidase staining assay. These data suggest that 7-epi-clusianone is a promising molecule against breast cancer cells. We show that 3D culture was more resistant to treatment than 2D culture, therefore more comprehensive studies are needed to better understand the effects of 7-epi-clusianone on this kind of culture.
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Effects of dietary Garcinia kola supplementation and oxidative stress in isolated perfused rat heartsNyepetsi, Naledi Gape January 2014 (has links)
Thesis submitted in fulfilment of the requirements for the degree of
Master of Technology: Biomedical Technology
In the Faculty of Health and Wellness Sciences
At the Cape Peninsula University of Technology
Supervisors: Prof. Adriaan J Esterhuyse
Dr Dirk J Bester
Bellville
January 2014 / Background: Oxidative stress and chronic inflammation contributes significantly to the pathogenesis of several ischaemic heart diseases, including atherosclerotic plaque rupture and myocardial infarction. It is widely demonstrated that ischaemia, followed by reperfusion, results in alterations of the mitochondrial and endothelial function through uncontrolled cascades of events characterized by free radical release and inflammation. Recent experimental evidence shows that modulation of inflammatory and antioxidant signaling mediators may determine the host outcome following myocardial ischaemia-reperfusion injury.
Investigations from the past decade indicate that food supplements may play an important role in the prevention and management of chronic inflammatory diseases. Garcinia kola seeds are flavonoid rich nut from a tropical flowering, non-timber plant of the Guttiferae family. This plant is highly valued in several African cultures for its use in herbal medicine. Recently, the majority of experimental research has linked phytochemicals found in Garcinia kola nut, to its proposed beneficial effects in treatment and management of oxidative stress related-chronic diseases. Research performed in our laboratory demonstrated that kolaviron, a prominent Garcinia kola flavonoid extract, reduces myocardial apoptosis during ischaemia-reperfusion injury. Therefore, the aim of our current study was to determine the effects of Garcinia kola supplementation on cardiac inflammatory and antioxidant signaling pathways during ischaemia-reperfusion using a Wistar rat heart model.
Materials and Methods: Male wistar rats were randomly divided into two groups: a control group receiving 2ml/kg corn oil and the experimental group receiving 100mg/kg Garcinia kola dissolved in corn oil, daily for 4 weeks. After the feeding period, blood samples were collected and lymphocytic DNA damage was analyzed using the alkaline comet assay. Furthermore, rat hearts were isolated and perfused with Krebs-Henseleit buffer on a working heart perfusion apparatus to measure myocardial functional parameters. Myocardial functional recovery was measured after 15 minutes global ischaemia followed by 25 minutes reperfusion. Hearts were freeze clamped at three different time points for myocardial cytokine concentration determinations using multiplex electrochemilunescent immunoassay. Nuclear factor
kappa beta (NF- kβ), p38 mitogen activated protein kinases (p38 MAPK), protein kinase B/Akt (PKB/Akt), nitro-tyrosine, inducible nitric oxide (iNOS), cyclooxygenase-2 (COX-2), poly (adenosine-di-phosphate) ribose polymerase-1 (PARP-1) and caspase-3 expression and their phosphorylated forms (where applicable) were analyzed using the Western blot technique.
Results: Dietary Garcinia kola supplementation significantly improved functional recovery when compared to the control group as reflected by the improved aortic output recovery (68.47 ± 6.16% versus 44.96 ± 7.00%; p<0.05). Our biochemical results supports the hypothesis that, dietary Garcinia kola supplementation modulates different cardiac proteins in terms of expression and activation at different time points when compared to the control group. We show that, before induction of ischaemia, Garcinia kola supplementation attenuates expression of inflammatory mediators and pro-apoptotic proteins when compared to the control group. The improved functional recovery was associated with a prompt inflammatory response, activation of PKB/Akt and attenuation of protein nitrosylation after 10 minutes of reperfusion. Modulation of NF-kβ and the p38 MAPK family proteins expression could have also played a significant role in myocardial functional recovery.
Conclusion: We have shown that a 4 week period of dietary Garcinia kola supplementation at 100mg/kg daily improves cardiac functional recovery following ischaemia-reperfusion injury. We propose that dietary Garcinia kola supplementation protects cardiac myocytes from ischaemia-reperfusion induced oxidative stress through the induction of a prompt inflammatory response and controlled expression and/or activation of the, NF-kβ, PKB/Akt and p38 MAPK protein signaling pathways PARP-1 and caspase. Finally, we demonstrated that dietary Garcinia kola supplementation did not induce rat lymphocytic DNA damage when compared to the control group.
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