A case study : exploring a DVD driven approach for teaching and learning mathematics, at secondary school level, with a framework of blended learning
- Authors: Padayachee, Pragashni
- Date: 2010
- Subjects: Matehmatics -- Study and Teaching (Secondary) -- Audio-visual aids , Matehmatics -- Study and Teaching (Secondary) -- South Africa -- Port Elizabeth
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10504 , http://hdl.handle.net/10948/1384 , Matehmatics -- Study and Teaching (Secondary) -- Audio-visual aids , Matehmatics -- Study and Teaching (Secondary) -- South Africa -- Port Elizabeth
- Description: Post-apartheid South Africa is witnessing an education crisis of significant proportions. The new outcomes-based education system has failed to deliver and universities are suffering the consequences of under-preparation of learners for tertiary studies especially in mathematics. The educator corps is lacking and it has become common practice for universities to deploy augmented programmes in mathematics for secondary school learners in the surrounding areas. This thesis describes a particular approach of blended learning, devised for the Incubator School Project (ISP), an initiative of the Nelson Mandela Metropolitan University (NMMU) in the Eastern Cape of South Africa. The defining feature of this blended approach is that it incorporates DVD technology, which offers an affordable and accessible option for the particular group of learners and the schools they attend. The thesis poses the research question: How did the use of the DVD approach within a blended learning environment support the learning of mathematics? This case study explores the particular blended approach and reports six fold on the approach – qualitatively based firstly on a questionnaire completed by learners and secondly on interviews of learners, thirdly on the facilitators reports, fourthly quantitatively on learner performance before and after the intervention. Fifthly six schools are used as a case study where the mathematics performance of the learners who participated in the ISP is compared to those who did not participate in the ISP. Finally the scope of blending of this model is evaluated by means of a radar chart, adapted from an existing radar measure. This research revealed that using the DVD approach within a blended learning environment did lead to an improvement in learners perceptions about mathematics, an improvement in the manner in which they learned mathematics, an extension in their mathematics knowledge and provided learners with a supportive environment in which to learn mathematics. The elements which supported learning in this approach are presented. The findings of the study suggest that this approach impacted favourably on the mathematics learning and enhanced the mathematics learning and performance of these learners. Recommendations are offered for practice, teachers and schools and for further research possibilities.
- Full Text:
- Date Issued: 2010
- Authors: Padayachee, Pragashni
- Date: 2010
- Subjects: Matehmatics -- Study and Teaching (Secondary) -- Audio-visual aids , Matehmatics -- Study and Teaching (Secondary) -- South Africa -- Port Elizabeth
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10504 , http://hdl.handle.net/10948/1384 , Matehmatics -- Study and Teaching (Secondary) -- Audio-visual aids , Matehmatics -- Study and Teaching (Secondary) -- South Africa -- Port Elizabeth
- Description: Post-apartheid South Africa is witnessing an education crisis of significant proportions. The new outcomes-based education system has failed to deliver and universities are suffering the consequences of under-preparation of learners for tertiary studies especially in mathematics. The educator corps is lacking and it has become common practice for universities to deploy augmented programmes in mathematics for secondary school learners in the surrounding areas. This thesis describes a particular approach of blended learning, devised for the Incubator School Project (ISP), an initiative of the Nelson Mandela Metropolitan University (NMMU) in the Eastern Cape of South Africa. The defining feature of this blended approach is that it incorporates DVD technology, which offers an affordable and accessible option for the particular group of learners and the schools they attend. The thesis poses the research question: How did the use of the DVD approach within a blended learning environment support the learning of mathematics? This case study explores the particular blended approach and reports six fold on the approach – qualitatively based firstly on a questionnaire completed by learners and secondly on interviews of learners, thirdly on the facilitators reports, fourthly quantitatively on learner performance before and after the intervention. Fifthly six schools are used as a case study where the mathematics performance of the learners who participated in the ISP is compared to those who did not participate in the ISP. Finally the scope of blending of this model is evaluated by means of a radar chart, adapted from an existing radar measure. This research revealed that using the DVD approach within a blended learning environment did lead to an improvement in learners perceptions about mathematics, an improvement in the manner in which they learned mathematics, an extension in their mathematics knowledge and provided learners with a supportive environment in which to learn mathematics. The elements which supported learning in this approach are presented. The findings of the study suggest that this approach impacted favourably on the mathematics learning and enhanced the mathematics learning and performance of these learners. Recommendations are offered for practice, teachers and schools and for further research possibilities.
- Full Text:
- Date Issued: 2010
A combination of platinum anticancer drugs and mangiferin causes increased efficacy in cancer cell lines
- Authors: Du Plessis-Stoman, Debbie
- Date: 2010
- Subjects: Cancer -- Chemotherapy , Antineoplastic agents , Platinum compounds -- Therapeutic use , Cancer cells
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10338 , http://hdl.handle.net/10948/d1016160
- Description: This thesis mainly deals with some biochemical aspects regarding the efficacy of novel platinum anticancer compounds alone and in combination with mangiferin, as part of a broader study in which both chemistry and biochemistry are involved. Various novel diamine and N-S donor chelate compounds of platinum II and IV have been developed in which factors such as stereochemistry, ligand exchange rate and biocompatibility were considered as additional parameters. In the first order testing, each of these compounds was tested with reference to their “killing” potential by comparing their rate of killing, over a period of 48 hours with those of cisplatin and oxaliplatin. Numerous novel compounds were tested in this way, using the MTT cell viability assay and the three cancer cell lines MCF7, HT29 and HeLa. Although only a few could be regarded as equal to or even better than cisplatin, CPA7 and oxaliplatin, the testing of these compounds on cancer cells provided useful knowledge for the further development of novel compounds. Three of the better compounds, namely Yol 25, Yol 29.1 and Mar 4.1.4 were selected for further studies, together with oxaliplatin and CPA7 as positive controls, to obtain more detailed knowledge of their anticancer action, both alone and when applied in combination with mangiferin. In addition to the above, resistant cells were produced for each of the three different cell lines tested and all the selected compounds, both in the presence and absence of mangiferin. The effects of these treatments on the activation of NFĸB when applied to normal and resistant cell lines were also investigated. All the compounds induced apoptosis in the cell lines tested as well as alter the DNA cycle at one or more phase. Additionally, combination of these compounds with mangiferin enhanced the above-mentioned effects. Mangiferin decreases the IC50 values of the platinum drugs by up to 3.4 times and, although mangiferin alone did not induce cell cycle arrest, the presence of mangiferin in combination with oxaliplatin and Yol 25 shows an earlier and greatly enhanced delay in the S-phase, while cells treated with CPA7, Yol 29.1 and Mar 4.1.4 in combination with mangiferin showed a later, but greatly enhanced delay in the S-phase. It was also found that mangiferin acts as an NFĸB inhibitor when applied in combination with these drugs, which, in turn, reduces the occurrence of resistance in the cell lines. Resistance to oxaliplatin was counteracted by the combination with mangiferin in HeLa and HT29, but not in MCF7 cells, while resistance to CPA7 was only counteracted in the MCF7 cell line. Yol 25 and Mar 4.1.4 did not seem to induce resistance in HeLa and MCF7 cells, but did in HT29 cells, whereas Yol 29.1 caused resistance in HeLa and HT29 cells, but not in MCF7 cells. Finally, an effort was made to evaluate the different compounds by comparing them with respect to their properties relating to anticancer action with and without the addition of mangiferin.
- Full Text:
- Date Issued: 2010
- Authors: Du Plessis-Stoman, Debbie
- Date: 2010
- Subjects: Cancer -- Chemotherapy , Antineoplastic agents , Platinum compounds -- Therapeutic use , Cancer cells
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10338 , http://hdl.handle.net/10948/d1016160
- Description: This thesis mainly deals with some biochemical aspects regarding the efficacy of novel platinum anticancer compounds alone and in combination with mangiferin, as part of a broader study in which both chemistry and biochemistry are involved. Various novel diamine and N-S donor chelate compounds of platinum II and IV have been developed in which factors such as stereochemistry, ligand exchange rate and biocompatibility were considered as additional parameters. In the first order testing, each of these compounds was tested with reference to their “killing” potential by comparing their rate of killing, over a period of 48 hours with those of cisplatin and oxaliplatin. Numerous novel compounds were tested in this way, using the MTT cell viability assay and the three cancer cell lines MCF7, HT29 and HeLa. Although only a few could be regarded as equal to or even better than cisplatin, CPA7 and oxaliplatin, the testing of these compounds on cancer cells provided useful knowledge for the further development of novel compounds. Three of the better compounds, namely Yol 25, Yol 29.1 and Mar 4.1.4 were selected for further studies, together with oxaliplatin and CPA7 as positive controls, to obtain more detailed knowledge of their anticancer action, both alone and when applied in combination with mangiferin. In addition to the above, resistant cells were produced for each of the three different cell lines tested and all the selected compounds, both in the presence and absence of mangiferin. The effects of these treatments on the activation of NFĸB when applied to normal and resistant cell lines were also investigated. All the compounds induced apoptosis in the cell lines tested as well as alter the DNA cycle at one or more phase. Additionally, combination of these compounds with mangiferin enhanced the above-mentioned effects. Mangiferin decreases the IC50 values of the platinum drugs by up to 3.4 times and, although mangiferin alone did not induce cell cycle arrest, the presence of mangiferin in combination with oxaliplatin and Yol 25 shows an earlier and greatly enhanced delay in the S-phase, while cells treated with CPA7, Yol 29.1 and Mar 4.1.4 in combination with mangiferin showed a later, but greatly enhanced delay in the S-phase. It was also found that mangiferin acts as an NFĸB inhibitor when applied in combination with these drugs, which, in turn, reduces the occurrence of resistance in the cell lines. Resistance to oxaliplatin was counteracted by the combination with mangiferin in HeLa and HT29, but not in MCF7 cells, while resistance to CPA7 was only counteracted in the MCF7 cell line. Yol 25 and Mar 4.1.4 did not seem to induce resistance in HeLa and MCF7 cells, but did in HT29 cells, whereas Yol 29.1 caused resistance in HeLa and HT29 cells, but not in MCF7 cells. Finally, an effort was made to evaluate the different compounds by comparing them with respect to their properties relating to anticancer action with and without the addition of mangiferin.
- Full Text:
- Date Issued: 2010
Aerial optical fibres in telecommunication systems : SOP and PMD monitoring, and tolerance of modulation formats
- Authors: Ireeta, Winston Tumps
- Date: 2010
- Subjects: Optical communications , Fiber optics
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10542 , http://hdl.handle.net/10948/1478 , http://hdl.handle.net/10948/d1012894 , Optical communications , Fiber optics
- Description: The topic of this thesis is aerial optical fibres in telecommunication systems: state of polarization (SOP) and polarization mode dispersion (PMD) monitoring and tolerance of modulation formats. Errors in optical fibre telecommunication systems are introduced when these polarization effects (SOP and PMD) change. These changes are so intense especially in aerial optical fibres. Part of the backbone of South Africa’s national grid includes long distances of aerial optical fibre between transmission exchange stations. The work in this thesis can be divided into three parts which all deal with the major aspects of PMD in deployed aerial optical fibres: characterization, environmental effects plus other perturbations, and tolerance of different modulation formats. In our work, SOP and PMD field measurements revealed that they both fluctuate more rapidly in deployed aerial optical fibres especially on windy and hot days. The SOP and PMD changes in the aerial optical fibres showed a significant correlation with these environmental parameters. SOP and PMD are stochastic in nature due to changes in the properties of the optical fibres and its positions because of both intrinsic and extrinsic perturbations. In our work, with only 184 PMD values measured and obtained by use of the FTB-5700 single-ended dispersion analyzer, the predicted theoretical Gaussian fit was obtained with a mean of 0.47 ps and standard deviation of 0.08 ps. This small standard deviation was justification for its robustness and accuracy. The statistical distributions for first-order polarization mode dispersion (FO-PMD) and second-order polarization mode dispersion (SO-PMD) for the first time were experimentally confirmed when measured using the FTB-5700 single-ended dispersion analyzer instrument for deployed aerial optical fibres. We were also able to determine the time scale over which to compensate FO-PMD in deployed aerial fibres using the directional time drift autocorrelation function method. It is slightly higher than 390 s for SOP measurements made on a particular windy and hot day. This is due to the fact that the changes of the PMD vector are known to be slower than the SOP changes. vi We also investigated the theoretical statistical distribution that corresponds to output SOP variations. The SOP variations can either be with wavelength (for buried fibre) or with time (for aerial fibre). Our results showed that the statistics of the relative SOP changes approached the distribution proposed by Foschini et al. (2000). Advanced optical modulation formats have become a key ingredient in the design of modern state-of-the-art wavelength-division-multiplexed (WDM) optical transmission systems. In our work, we investigated which of these advanced modulation formats is best suited for the South African network especially on systems that have links of aerial optical fibres. Keywords: aerial optical fibre, polarization mode dispersion (PMD), principal states of polarization (PSP), state of polarization (SOP), first-order PMD, second-order PMD.
- Full Text:
- Date Issued: 2010
- Authors: Ireeta, Winston Tumps
- Date: 2010
- Subjects: Optical communications , Fiber optics
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10542 , http://hdl.handle.net/10948/1478 , http://hdl.handle.net/10948/d1012894 , Optical communications , Fiber optics
- Description: The topic of this thesis is aerial optical fibres in telecommunication systems: state of polarization (SOP) and polarization mode dispersion (PMD) monitoring and tolerance of modulation formats. Errors in optical fibre telecommunication systems are introduced when these polarization effects (SOP and PMD) change. These changes are so intense especially in aerial optical fibres. Part of the backbone of South Africa’s national grid includes long distances of aerial optical fibre between transmission exchange stations. The work in this thesis can be divided into three parts which all deal with the major aspects of PMD in deployed aerial optical fibres: characterization, environmental effects plus other perturbations, and tolerance of different modulation formats. In our work, SOP and PMD field measurements revealed that they both fluctuate more rapidly in deployed aerial optical fibres especially on windy and hot days. The SOP and PMD changes in the aerial optical fibres showed a significant correlation with these environmental parameters. SOP and PMD are stochastic in nature due to changes in the properties of the optical fibres and its positions because of both intrinsic and extrinsic perturbations. In our work, with only 184 PMD values measured and obtained by use of the FTB-5700 single-ended dispersion analyzer, the predicted theoretical Gaussian fit was obtained with a mean of 0.47 ps and standard deviation of 0.08 ps. This small standard deviation was justification for its robustness and accuracy. The statistical distributions for first-order polarization mode dispersion (FO-PMD) and second-order polarization mode dispersion (SO-PMD) for the first time were experimentally confirmed when measured using the FTB-5700 single-ended dispersion analyzer instrument for deployed aerial optical fibres. We were also able to determine the time scale over which to compensate FO-PMD in deployed aerial fibres using the directional time drift autocorrelation function method. It is slightly higher than 390 s for SOP measurements made on a particular windy and hot day. This is due to the fact that the changes of the PMD vector are known to be slower than the SOP changes. vi We also investigated the theoretical statistical distribution that corresponds to output SOP variations. The SOP variations can either be with wavelength (for buried fibre) or with time (for aerial fibre). Our results showed that the statistics of the relative SOP changes approached the distribution proposed by Foschini et al. (2000). Advanced optical modulation formats have become a key ingredient in the design of modern state-of-the-art wavelength-division-multiplexed (WDM) optical transmission systems. In our work, we investigated which of these advanced modulation formats is best suited for the South African network especially on systems that have links of aerial optical fibres. Keywords: aerial optical fibre, polarization mode dispersion (PMD), principal states of polarization (PSP), state of polarization (SOP), first-order PMD, second-order PMD.
- Full Text:
- Date Issued: 2010
An investigation of IgE regulation by recombinant soluble IgE receptors and co-receptors in human cell culture models
- Authors: Bowles, Sandra Lyn
- Date: 2010
- Subjects: Immunoglobulin E , Allergy
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10321 , http://hdl.handle.net/10948/1231 , Immunoglobulin E , Allergy
- Description: Type I hypersensitivities are mediated by the IgE antibody. The effector functions and synthesis of IgE result from interactions with a network of proteins that include a high affinity (FcRIα) and a low affinity (CD23, FcRII) Fc receptor in conjunction with the B lymphocyte receptor, CD21. CD23 is a multifunctional type II transmembrane protein that binds its known ligands through its ectodomain either as a membrane-bound or soluble receptor generated in vivo by specific proteolytic cleavages. IgE production is primarily regulated by interactions between IgE, CD23 and CD21. Despite its importance for development of strategies to limit hypersensitivity, precise information about the molecular interactions remains limited. During this study, I engineered, expressed and purified from bacteria three soluble human CD23 fragments that are normally formed in vivo and shed from the cell surface (1) derCD23, amino acids 156-298 (2) sCD23, amino acids 150-321 and (3) the entire ectodomain, exCD23, amino acids 48-321 to examine the comparative binding of recombinant human CD21 SCR 1-2 and native human IgE to these fragments. Gel filtration HPLC revealed that derCD23 and sCD23 were monomeric whereas exCD23 assembled as a heterogeneous mixture that included trimers and monomers. At the concentrations utilized, CD23 fragments sCD23 and exCD23 bound CD21 with similar affinity, whereas interaction between derCD23 and CD21 was minimal when analyzed by surface plasmon resonance (SPR) spectroscopy. These findings suggest that penultimate “tail” amino acids between 298 and 321 stabilize CD21 attachment, although it cannot be ruled out, the region between Met 150 and Ser 156 may also play a role in binding CD21 SCR 1-2. In contrast, there is a progressive increment in the affinity of soluble fragments (exCD23>sCD23>derCD23) for IgE, upon increasing length of the proximal CD23 “stalk” domain. These findings highlight the differences in both the structural basis and affinity of the three physiological fragments of human CD23 for the ligands CD21 and IgE and underscore the complexity of CD23-mediated regulatory networks. It was found that B-cells only make up ~5% of the PBMC population, and that these cells were able to be activated, via STAT-6 phosphorylation, to enter class switch recombination (CSR) by the addition of switch factors (IL-4 and anti-CD40). Titration experiments dictated that 25 ng/mL of CD23 was the most efficient concentration to up-regulate IgE synthesis in PBMCs; furthermore, soluble CD23 proteins were incubated with PBMCs in the presence and absence of CD21 SCR 1-2 to investigate the effect that these recombinant proteins have on IgE synthesis. Results showed that the influence of recombinant proteins (both CD23 and CD21) on IgE synthesis was slight. It was shown that while derCD23 had no significant effect, monomeric sCD23 down-regulated, and the mixture of monomeric and oligomeric exCD23 up-regulated IgE synthesis. On addition of CD21 SCR 1-2 to the cells switched and treated with soluble CD23, it was found that in both cases for sCD23 and exCD23, IgE synthesis was increased, while for derCD23, there was no noticeable difference in IgE synthesis. This confirmed previous data showing the lack of binding between derCD23 and CD21 SCR 1-2. The exact binding site for CD21 on the CD23 molecule is unknown, and incompletely represented in the NMR and crystal structures. It is thought that CD21 binds to the C-terminal tail section, not present in derCD23. It is therefore likely that only a negative-feedback mechanism operates with derCD23 to regulate IgE synthesis. Further investigation of the binding of CD23 fragments to SCR 5-8 of CD21 and the effect of this on IgE synthesis may lead to a potential therapeutic role for derCD23 in the treatment of allergic disease. Data accumulated in this study suggests that investigating the modulation of oligomeric state and thus the activity of soluble CD23 fragments may be important in the construction of new regulators of IgE synthesis.
- Full Text:
- Date Issued: 2010
- Authors: Bowles, Sandra Lyn
- Date: 2010
- Subjects: Immunoglobulin E , Allergy
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10321 , http://hdl.handle.net/10948/1231 , Immunoglobulin E , Allergy
- Description: Type I hypersensitivities are mediated by the IgE antibody. The effector functions and synthesis of IgE result from interactions with a network of proteins that include a high affinity (FcRIα) and a low affinity (CD23, FcRII) Fc receptor in conjunction with the B lymphocyte receptor, CD21. CD23 is a multifunctional type II transmembrane protein that binds its known ligands through its ectodomain either as a membrane-bound or soluble receptor generated in vivo by specific proteolytic cleavages. IgE production is primarily regulated by interactions between IgE, CD23 and CD21. Despite its importance for development of strategies to limit hypersensitivity, precise information about the molecular interactions remains limited. During this study, I engineered, expressed and purified from bacteria three soluble human CD23 fragments that are normally formed in vivo and shed from the cell surface (1) derCD23, amino acids 156-298 (2) sCD23, amino acids 150-321 and (3) the entire ectodomain, exCD23, amino acids 48-321 to examine the comparative binding of recombinant human CD21 SCR 1-2 and native human IgE to these fragments. Gel filtration HPLC revealed that derCD23 and sCD23 were monomeric whereas exCD23 assembled as a heterogeneous mixture that included trimers and monomers. At the concentrations utilized, CD23 fragments sCD23 and exCD23 bound CD21 with similar affinity, whereas interaction between derCD23 and CD21 was minimal when analyzed by surface plasmon resonance (SPR) spectroscopy. These findings suggest that penultimate “tail” amino acids between 298 and 321 stabilize CD21 attachment, although it cannot be ruled out, the region between Met 150 and Ser 156 may also play a role in binding CD21 SCR 1-2. In contrast, there is a progressive increment in the affinity of soluble fragments (exCD23>sCD23>derCD23) for IgE, upon increasing length of the proximal CD23 “stalk” domain. These findings highlight the differences in both the structural basis and affinity of the three physiological fragments of human CD23 for the ligands CD21 and IgE and underscore the complexity of CD23-mediated regulatory networks. It was found that B-cells only make up ~5% of the PBMC population, and that these cells were able to be activated, via STAT-6 phosphorylation, to enter class switch recombination (CSR) by the addition of switch factors (IL-4 and anti-CD40). Titration experiments dictated that 25 ng/mL of CD23 was the most efficient concentration to up-regulate IgE synthesis in PBMCs; furthermore, soluble CD23 proteins were incubated with PBMCs in the presence and absence of CD21 SCR 1-2 to investigate the effect that these recombinant proteins have on IgE synthesis. Results showed that the influence of recombinant proteins (both CD23 and CD21) on IgE synthesis was slight. It was shown that while derCD23 had no significant effect, monomeric sCD23 down-regulated, and the mixture of monomeric and oligomeric exCD23 up-regulated IgE synthesis. On addition of CD21 SCR 1-2 to the cells switched and treated with soluble CD23, it was found that in both cases for sCD23 and exCD23, IgE synthesis was increased, while for derCD23, there was no noticeable difference in IgE synthesis. This confirmed previous data showing the lack of binding between derCD23 and CD21 SCR 1-2. The exact binding site for CD21 on the CD23 molecule is unknown, and incompletely represented in the NMR and crystal structures. It is thought that CD21 binds to the C-terminal tail section, not present in derCD23. It is therefore likely that only a negative-feedback mechanism operates with derCD23 to regulate IgE synthesis. Further investigation of the binding of CD23 fragments to SCR 5-8 of CD21 and the effect of this on IgE synthesis may lead to a potential therapeutic role for derCD23 in the treatment of allergic disease. Data accumulated in this study suggests that investigating the modulation of oligomeric state and thus the activity of soluble CD23 fragments may be important in the construction of new regulators of IgE synthesis.
- Full Text:
- Date Issued: 2010
Beta-N-methylamino-L-alanine in South African fresh water cyanobacteria : incidence, prevalence, ecotoxicological considerations and human exposure risk
- Authors: Esterhuizen-Londt, Maranda
- Date: 2010
- Subjects: Cyanobacteria , Bioaccumulation , Chromatographic analysis , Neurotoxic agents
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10306 , http://hdl.handle.net/10948/1473 , Cyanobacteria , Bioaccumulation , Chromatographic analysis , Neurotoxic agents
- Description: β-N-methylamino-L-alanine (BMAA) is a non-proteinogenic amino acid associated with human neurodegenerative disease. Due to the cosmopolitan nature of cyanobacteria, detection of BMAA in cyanobacteria has caused concerns about human exposure risk. This study was therefore based on the hypothesis that BMAA poses a health risk to humans either by direct ingestion or by indirect exposure to BMAA from a cyanobacterial source via a freshwater food chain. A validated gas chromatography-mass spectrometry (GC-MS) BMAA analysis method and a confirmatory liquid chromatography-mass spectrometry (LC-MS) method, with improved sensitivity, were developed in addition to a LC-MS/MS method for analyte confirmation. These methods were used to quantify BMAA in South African cyanobacteria, isolated from various potable water reservoirs. The majority of the isolates tested, contained BMAA. Possible human exposure by direct consumption of BMAA released from cyanobacterial blooms was investigated by the development of a robust solid phase extraction (SPE) method used for BMAA concentration and quantification in raw and treated tap water. Despite the use of the SPE method that facilitated the concentration of BMAA from large quantities of water, no free dissolved BMAA was detected in raw or processed fresh water. The fate of exogenous BMAA was therefore investigated firstly by evaluating the efficacy of standard water treatment processes employed in South Africa and secondly by investigating the possibility of BMAA bioaccumulation and biomagnification in aquatic food chains. Standard water treatment processes proved highly efficient at removing free dissolved BMAA, explaining the absence of BMAA in treated tap water. However, the cause of the BMAA absence in raw potable water remained unknown. Uptake of BMAA by model aquatic organisms was investigated in controlled experiments. BMAA uptake was documented in both Ceratophyllum demersum and Daphnia magna, however, BMAA-protein association and biomagnification were not observed in D. magna. BMAA had an inhibitory effect on the oxidative stress enzyme acitivties of both organisms tested (as well as human S9 extracts), resulting in accumulation of detrimental reactive oxygen species (ROS) in the cells. Exposure of crop plants to BMAA in controlled experiments resulted in BMAA uptake, protein association, and subsequent inhibition of the antioxidative enzyme activities. However, BMAA was detected in neither free nor protein-associated form in natural crop plants irrigated with known BMAA-containing bloom water. Post-mortem liver samples of Clarias gariepinus (Catfish) and Crocodylus niloticus (Crocodile), from a natural fresh water ecosystem that experienced frequent cyanobacterial blooms, contained both free and protein-associated BMAA. Higher BMAA concentrations were found in crocodile liver samples compared to fish liver samples, strongly suggesting biomagnification from one trophic level to the next. BMAA concentrations corresponded to crocodile age. This is the first report of bioaccumulation and biomagnification in two trophic levels in a fresh water ecosystem. These findings strongly suggest possible human exposure via aquatic food chains of cyanobacterial origin. Direct BMAA exposure via drinking water is not plausible due to the efficiency of standard water treatment processes to remove BMAA. The use of raw water for agricultural and recreational use, however, remains a problem. The development of management strategies as well as daily tolerable levels for BMAA is urgently required.
- Full Text:
- Date Issued: 2010
- Authors: Esterhuizen-Londt, Maranda
- Date: 2010
- Subjects: Cyanobacteria , Bioaccumulation , Chromatographic analysis , Neurotoxic agents
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10306 , http://hdl.handle.net/10948/1473 , Cyanobacteria , Bioaccumulation , Chromatographic analysis , Neurotoxic agents
- Description: β-N-methylamino-L-alanine (BMAA) is a non-proteinogenic amino acid associated with human neurodegenerative disease. Due to the cosmopolitan nature of cyanobacteria, detection of BMAA in cyanobacteria has caused concerns about human exposure risk. This study was therefore based on the hypothesis that BMAA poses a health risk to humans either by direct ingestion or by indirect exposure to BMAA from a cyanobacterial source via a freshwater food chain. A validated gas chromatography-mass spectrometry (GC-MS) BMAA analysis method and a confirmatory liquid chromatography-mass spectrometry (LC-MS) method, with improved sensitivity, were developed in addition to a LC-MS/MS method for analyte confirmation. These methods were used to quantify BMAA in South African cyanobacteria, isolated from various potable water reservoirs. The majority of the isolates tested, contained BMAA. Possible human exposure by direct consumption of BMAA released from cyanobacterial blooms was investigated by the development of a robust solid phase extraction (SPE) method used for BMAA concentration and quantification in raw and treated tap water. Despite the use of the SPE method that facilitated the concentration of BMAA from large quantities of water, no free dissolved BMAA was detected in raw or processed fresh water. The fate of exogenous BMAA was therefore investigated firstly by evaluating the efficacy of standard water treatment processes employed in South Africa and secondly by investigating the possibility of BMAA bioaccumulation and biomagnification in aquatic food chains. Standard water treatment processes proved highly efficient at removing free dissolved BMAA, explaining the absence of BMAA in treated tap water. However, the cause of the BMAA absence in raw potable water remained unknown. Uptake of BMAA by model aquatic organisms was investigated in controlled experiments. BMAA uptake was documented in both Ceratophyllum demersum and Daphnia magna, however, BMAA-protein association and biomagnification were not observed in D. magna. BMAA had an inhibitory effect on the oxidative stress enzyme acitivties of both organisms tested (as well as human S9 extracts), resulting in accumulation of detrimental reactive oxygen species (ROS) in the cells. Exposure of crop plants to BMAA in controlled experiments resulted in BMAA uptake, protein association, and subsequent inhibition of the antioxidative enzyme activities. However, BMAA was detected in neither free nor protein-associated form in natural crop plants irrigated with known BMAA-containing bloom water. Post-mortem liver samples of Clarias gariepinus (Catfish) and Crocodylus niloticus (Crocodile), from a natural fresh water ecosystem that experienced frequent cyanobacterial blooms, contained both free and protein-associated BMAA. Higher BMAA concentrations were found in crocodile liver samples compared to fish liver samples, strongly suggesting biomagnification from one trophic level to the next. BMAA concentrations corresponded to crocodile age. This is the first report of bioaccumulation and biomagnification in two trophic levels in a fresh water ecosystem. These findings strongly suggest possible human exposure via aquatic food chains of cyanobacterial origin. Direct BMAA exposure via drinking water is not plausible due to the efficiency of standard water treatment processes to remove BMAA. The use of raw water for agricultural and recreational use, however, remains a problem. The development of management strategies as well as daily tolerable levels for BMAA is urgently required.
- Full Text:
- Date Issued: 2010
Implementation of novel flow cytometric methods to assess the in vitro antidiabetic mechanism of a Sutherlandia Frutescens extract
- Authors: Elliot, Gayle Pamela
- Date: 2010
- Subjects: Insulin resistance -- South Africa , Insulin -- Therapeutic use -- South Africa , Medicinal plants -- South Africa , Non-insulin-dependent diabetes -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10304 , http://hdl.handle.net/10948/1439 , Insulin resistance -- South Africa , Insulin -- Therapeutic use -- South Africa , Medicinal plants -- South Africa , Non-insulin-dependent diabetes -- South Africa
- Description: The ability of insulin to stimulate glucose uptake into muscle and adipose tissue is central to the maintenance of whole-body glucose homeostasis. Deregulation of insulin action manifests itself as insulin resistance, a key component of type 2 diabetes. Insulin resistance is also observed in HIV patients receiving protease inhibitors. An agent that can reversibly induce an insulin-resistant state would be a very useful tool in developing model systems that mimic the pathogenesis of type 2 diabetes. Insulin resistance can arise from defects in insulin signal transduction, changes in the expression of proteins or genes that are targets of insulin action, cross talk from other hormonal systems or metabolic abnormalities. Deterioration of the insulin-receptor-signalling pathway at different levels leading to decreased levels of signalling pathway intermediates and/or decreased activation through phosphorylation accounts for the evolution from an insulin-resistant state to type 2 diabetes. In addition, defects in GLUT4 glucose transporter translocation are observed, further fuelling impairments in skeletal muscle glucose uptake. Levels of insulin-induced GLUT4 translocation in the skeletal muscle of type 2 diabetic patients are typically reduced by 90%. Many cellular pathways & their intermediates are in some way or another linked to insulin signalling. This study focused on three of these namely the PI3-kinase/Akt pathway, the Mitogen Activated Protein Kinase (MAPK) cascade and the AMP Kinase pathway, with successful monitoring of the PI3-K pathway. Investigations involved observing and evaluating the effects of various compounds as well as an indigenous medicinal plant, Sutherlandia frutescens on the activities of key insulin signalling pathway intermediates within the three fore mentioned pathways including Akt, AMPK and MEK1/2 as well as membrane surface GLUT4 levels. Scientific research has in the past leant heavily on Western blotting as the method of choice for gaining vital information relating to signal transduction pathways, however for research into cellular mechanisms the negatives of this method outweigh the positives. The drawbacks include a need for large amount of cells, multiple washing steps which may be disadvantageous to any weak and transient interactions as well as lysing of cells which may interfere with the maintenance of the subcellular localisation of a specific signalling event. Based on these, the need for a better method in terms of speed & reliability to monitor phosphorylation states of signal transduction pathway intermediates & GLUT4 translocation was evident and was one VII of the main aims & successes of this study. The method created used the mouse muscle cell line C2C12 in conjunction with the quick, sensitive method of flow cytometry which allowed us to monitor these processes in these cells through immune-labelling. Adherent cell cultures such as the C2C12 cell line pose the problem of possible damage to plasma membrane receptors (including insulin receptors) during harvesting to obtain a cell suspension for flow cytometry. We however used C2C12 mouse myocytes to optimize a method yielding insulin responsive cells in suspension that were successfully used for flow cytometry after immunelabelling of insulin signalling intermediates. Insulin (0.1μM) significantly raised the levels of both P-Akt and GLUT4 above basal levels. This effect was shown to be dose dependent. At a concentration of 50μg/ml, Sutherlandia frutescens was able to act as an insulin-mimetic in terms of its ability to increase P-Akt levels, GLUT4 translocation and glucose utilisation in an acute manner. These increases could be reduced with the addition of wortmannin, a PI3-K inhibitor. Therefore, these results suggest the mechanism of the plant extract’s insulin-like activity may be in part due to the activation of the insulin signalling pathway leading to GLUT4 translocation, which involves the phosphorylation of insulin receptor- and subsequent PI3-K activity, leading to P-Akt activity. These results provide further evidence of this plant extract’s anti-diabetic potential. The effect of Sutherlandia frutescens on insulin secretion, calcium signalling and proliferation in INS-1 rat pancreatic cells was also investigated and it was found to increase the activities of all of these processes. However no change in the levels of GLUT2 glucose transporter was seen. Ritonavir is prescribed by the South African Department of Health in co-formulation with other protease inhibitors within its second regime in the treatment of HIV and AIDS. Using C2C12 cells, ritonavir decreased glucose uptake acutely and had no effect on GLUT4 translocation however surprisingly increased P-Akt levels. In conclusion, it was found that Sutherlandia frutescens has antidiabetic benefits, diverse in nature depending on tissue type as well as length of time administered. The establishment of novel flow cytometry techniques to assess antidiabetic properties using in vitro cell culture was achieved. These methods will be useful in the future for the assessment of insulin sensitivity and in the identification of novel compounds that stimulate the insulin signalling pathways.
- Full Text:
- Date Issued: 2010
- Authors: Elliot, Gayle Pamela
- Date: 2010
- Subjects: Insulin resistance -- South Africa , Insulin -- Therapeutic use -- South Africa , Medicinal plants -- South Africa , Non-insulin-dependent diabetes -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10304 , http://hdl.handle.net/10948/1439 , Insulin resistance -- South Africa , Insulin -- Therapeutic use -- South Africa , Medicinal plants -- South Africa , Non-insulin-dependent diabetes -- South Africa
- Description: The ability of insulin to stimulate glucose uptake into muscle and adipose tissue is central to the maintenance of whole-body glucose homeostasis. Deregulation of insulin action manifests itself as insulin resistance, a key component of type 2 diabetes. Insulin resistance is also observed in HIV patients receiving protease inhibitors. An agent that can reversibly induce an insulin-resistant state would be a very useful tool in developing model systems that mimic the pathogenesis of type 2 diabetes. Insulin resistance can arise from defects in insulin signal transduction, changes in the expression of proteins or genes that are targets of insulin action, cross talk from other hormonal systems or metabolic abnormalities. Deterioration of the insulin-receptor-signalling pathway at different levels leading to decreased levels of signalling pathway intermediates and/or decreased activation through phosphorylation accounts for the evolution from an insulin-resistant state to type 2 diabetes. In addition, defects in GLUT4 glucose transporter translocation are observed, further fuelling impairments in skeletal muscle glucose uptake. Levels of insulin-induced GLUT4 translocation in the skeletal muscle of type 2 diabetic patients are typically reduced by 90%. Many cellular pathways & their intermediates are in some way or another linked to insulin signalling. This study focused on three of these namely the PI3-kinase/Akt pathway, the Mitogen Activated Protein Kinase (MAPK) cascade and the AMP Kinase pathway, with successful monitoring of the PI3-K pathway. Investigations involved observing and evaluating the effects of various compounds as well as an indigenous medicinal plant, Sutherlandia frutescens on the activities of key insulin signalling pathway intermediates within the three fore mentioned pathways including Akt, AMPK and MEK1/2 as well as membrane surface GLUT4 levels. Scientific research has in the past leant heavily on Western blotting as the method of choice for gaining vital information relating to signal transduction pathways, however for research into cellular mechanisms the negatives of this method outweigh the positives. The drawbacks include a need for large amount of cells, multiple washing steps which may be disadvantageous to any weak and transient interactions as well as lysing of cells which may interfere with the maintenance of the subcellular localisation of a specific signalling event. Based on these, the need for a better method in terms of speed & reliability to monitor phosphorylation states of signal transduction pathway intermediates & GLUT4 translocation was evident and was one VII of the main aims & successes of this study. The method created used the mouse muscle cell line C2C12 in conjunction with the quick, sensitive method of flow cytometry which allowed us to monitor these processes in these cells through immune-labelling. Adherent cell cultures such as the C2C12 cell line pose the problem of possible damage to plasma membrane receptors (including insulin receptors) during harvesting to obtain a cell suspension for flow cytometry. We however used C2C12 mouse myocytes to optimize a method yielding insulin responsive cells in suspension that were successfully used for flow cytometry after immunelabelling of insulin signalling intermediates. Insulin (0.1μM) significantly raised the levels of both P-Akt and GLUT4 above basal levels. This effect was shown to be dose dependent. At a concentration of 50μg/ml, Sutherlandia frutescens was able to act as an insulin-mimetic in terms of its ability to increase P-Akt levels, GLUT4 translocation and glucose utilisation in an acute manner. These increases could be reduced with the addition of wortmannin, a PI3-K inhibitor. Therefore, these results suggest the mechanism of the plant extract’s insulin-like activity may be in part due to the activation of the insulin signalling pathway leading to GLUT4 translocation, which involves the phosphorylation of insulin receptor- and subsequent PI3-K activity, leading to P-Akt activity. These results provide further evidence of this plant extract’s anti-diabetic potential. The effect of Sutherlandia frutescens on insulin secretion, calcium signalling and proliferation in INS-1 rat pancreatic cells was also investigated and it was found to increase the activities of all of these processes. However no change in the levels of GLUT2 glucose transporter was seen. Ritonavir is prescribed by the South African Department of Health in co-formulation with other protease inhibitors within its second regime in the treatment of HIV and AIDS. Using C2C12 cells, ritonavir decreased glucose uptake acutely and had no effect on GLUT4 translocation however surprisingly increased P-Akt levels. In conclusion, it was found that Sutherlandia frutescens has antidiabetic benefits, diverse in nature depending on tissue type as well as length of time administered. The establishment of novel flow cytometry techniques to assess antidiabetic properties using in vitro cell culture was achieved. These methods will be useful in the future for the assessment of insulin sensitivity and in the identification of novel compounds that stimulate the insulin signalling pathways.
- Full Text:
- Date Issued: 2010
Leonotis leonurus: the anticoagulant and antidiabetic activity of Leonotis leonurus
- Authors: Mnonopi, Nandipha
- Date: 2010
- Subjects: Medicinal plants -- South Africa , Materia medica, Vegetable -- South Africa , Diabetes -- Alternative treatment -- South Africa , Plant bioactive compounds , Leonotis leonurus -- Physiological aspects
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10323 , http://hdl.handle.net/10948/1194 , Medicinal plants -- South Africa , Materia medica, Vegetable -- South Africa , Diabetes -- Alternative treatment -- South Africa , Plant bioactive compounds , Leonotis leonurus -- Physiological aspects
- Description: Commercial marrubiin, aqueous and organic extracts of Leonotis leonurus were tested in vitro for their anticoagulant and antiplatelet activities. The aqueous extract inhibited platelet aggregation by 69.5 percent (100 μg/mL), while the organic extract (100 μg/mL) and marrubiin (5 μg/mL) showed 92.5 percent and 91.6 percent inhibition, respectively, by inhibiting the binding of fibrinogen to glycoprotein IIb/IIIa receptor in a concentration dependent manner. The extracts significantly prolonged activated partial thromboplastin time compared to untreated plasma controls. Fibrin and D-Dimer formation were drastically decreased. The extracts and marrubiin concentration-dependently inhibited calcium mobilization induced by collagen and thrombin. The formation of thromboxane A2 was also significantly reduced by both the extracts and marrubiin. Protein secretion and platelet adhesion were significantly reduced by both the extracts and marrubiin. The organic extract and marrubiin showed a more pronounced effect than the aqueous extracts in all the in vitro assays. The ex-vivo animal model confirmed the results obtained in vitro. Similar to the in vitro studies, activated partial thromboplastin time clotting time was prolonged by marrubiin and the number of aggregated platelets were significantly reduced relative to aspirin. The findings reflect that marrubiin largely contributes to the organic extract's anticoagulant and antiplatelet effect in vitro. INS-1 cells were cultured under normo- and hyperglycaemic conditions. Marrubiin and the two Leonotis leonurus extracts were screened for anti-diabetic activity in vitro. The stimulatory index of INS-1 cells cultured under hyperglycaemic conditions was significantly increased by 60 percent and 61 percent (p<0.01; n=5) in cells exposed to the organic extract (10 μg/mL) and marrubiin (500 ng/mL), respectively, relative to the normoglycaemic conditions. The gene expression of insulin was significantly increased by 76.5 and 71 percent, and of glucose transporter-2 by 93 and 92.5 percent for marrubiin and the organic extract, respectively, under the same conditions stipulated above (p<0.01; n=4). The extract and marrubiin similarly showed an increase in respiratory rate under hyperglycaemic conditions. Marrubiin increased insulin secretion, HDL-cholesterol, while it decreased total cholesterol, LDL-cholesterol and the atherogenic index in the in vivo rat model.
- Full Text:
- Date Issued: 2010
- Authors: Mnonopi, Nandipha
- Date: 2010
- Subjects: Medicinal plants -- South Africa , Materia medica, Vegetable -- South Africa , Diabetes -- Alternative treatment -- South Africa , Plant bioactive compounds , Leonotis leonurus -- Physiological aspects
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10323 , http://hdl.handle.net/10948/1194 , Medicinal plants -- South Africa , Materia medica, Vegetable -- South Africa , Diabetes -- Alternative treatment -- South Africa , Plant bioactive compounds , Leonotis leonurus -- Physiological aspects
- Description: Commercial marrubiin, aqueous and organic extracts of Leonotis leonurus were tested in vitro for their anticoagulant and antiplatelet activities. The aqueous extract inhibited platelet aggregation by 69.5 percent (100 μg/mL), while the organic extract (100 μg/mL) and marrubiin (5 μg/mL) showed 92.5 percent and 91.6 percent inhibition, respectively, by inhibiting the binding of fibrinogen to glycoprotein IIb/IIIa receptor in a concentration dependent manner. The extracts significantly prolonged activated partial thromboplastin time compared to untreated plasma controls. Fibrin and D-Dimer formation were drastically decreased. The extracts and marrubiin concentration-dependently inhibited calcium mobilization induced by collagen and thrombin. The formation of thromboxane A2 was also significantly reduced by both the extracts and marrubiin. Protein secretion and platelet adhesion were significantly reduced by both the extracts and marrubiin. The organic extract and marrubiin showed a more pronounced effect than the aqueous extracts in all the in vitro assays. The ex-vivo animal model confirmed the results obtained in vitro. Similar to the in vitro studies, activated partial thromboplastin time clotting time was prolonged by marrubiin and the number of aggregated platelets were significantly reduced relative to aspirin. The findings reflect that marrubiin largely contributes to the organic extract's anticoagulant and antiplatelet effect in vitro. INS-1 cells were cultured under normo- and hyperglycaemic conditions. Marrubiin and the two Leonotis leonurus extracts were screened for anti-diabetic activity in vitro. The stimulatory index of INS-1 cells cultured under hyperglycaemic conditions was significantly increased by 60 percent and 61 percent (p<0.01; n=5) in cells exposed to the organic extract (10 μg/mL) and marrubiin (500 ng/mL), respectively, relative to the normoglycaemic conditions. The gene expression of insulin was significantly increased by 76.5 and 71 percent, and of glucose transporter-2 by 93 and 92.5 percent for marrubiin and the organic extract, respectively, under the same conditions stipulated above (p<0.01; n=4). The extract and marrubiin similarly showed an increase in respiratory rate under hyperglycaemic conditions. Marrubiin increased insulin secretion, HDL-cholesterol, while it decreased total cholesterol, LDL-cholesterol and the atherogenic index in the in vivo rat model.
- Full Text:
- Date Issued: 2010
Metabolic energy relations in the Eastern Cape Angulate Tortoise (Chersina Anguluta)
- Authors: Setlalekgomo, Mpho Rinah
- Date: 2010
- Subjects: Eastern Cape angulate Tortoise , Oxygen consumption (Physiology)
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10690 , http://hdl.handle.net/10948/1400 , Eastern Cape angulate Tortoise , Oxygen consumption (Physiology)
- Description: The daily oxygen consumption (VO2) pattern, the effects of varying ambient temperatures, season and mass on the resting oxygen consumption (RVO2) of Chersina angulata of the Eastern Cape were investigated. The RVO2 was measured using flow-through respirometry and specific resting oxygen consumption (sRVO2) calculated. To determine the daily pattern in the VO2 of C. angulata, the tortoises were acclimated in an environmentally controlled room (ECR) to an ambient temperature of 26 ± 1°C and a light regime of 14 hours of light and 10 hours of darkness (14L:10D) for at least a week prior to the RVO2 measurements. The RVO2 was measured at a constant temperature of 26 ± 1°C, and at three different light regimes, namely: 14L:10D, constant darkness (DD) and constant light (LL). There were no significant effects of mass or gender on the sRVO2 of the tortoises used. Rhythms in the sRVO2 were detected under all three light regimes. The amplitudes of the rhythm were largest at 14L:10D, followed by DD and smallest at LL regime. The persistence of the rhythmic pattern under constant conditions suggests the existence of an endogenous circadian rhythm in the sRVO2 for adult C. angulata. To test for the effect of ambient temperature on the sRVO2 of adult C. angulata, the tortoises were acclimated to 22 ± 1°C and a 14L:10D light regime prior to the RVO2 measurements. RVO2 was measured at eight experimental temperatures; 14°C, 18°C, 22°C, 26°C, 30°C, 35°C, 38°C and 40°C. The sRVO2 was not influenced by gender and increased with experimental temperatures, but this did not happen consistently over the whole range of temperatures tested. A plateau, possibly a thermal preferendum zone, was detected within the temperature range of 26 - 38°C. Determination of seasonal effect on the sRVO2 of adult C. angulata was accomplished by acclimating tortoises to standard summer and winter conditions. Seasonal effects were tested in the appropriate seasons. Winter experiments were conducted in winter and summer experiments conducted in summer. The RVO2 was measured at experimental temperature 14°C, 18°C and 22°C. In addition RVO2 of iv winter-acclimated tortoises was also measured at 10°C. The sRVO2 increased significantly with increasing temperature within the temperature range tested. No distinct pattern was observed in the seasonal acclimation of adult C. angulata. The metabolic rate-temperature curves of the summer and the winter-acclimated tortoises cross each other. Season and temperature had no significant effects on the mass-scaling exponent of the sRVO2. The exponent ranged from 0.48 to 0.73 within the temperature range of 22 - 38°C. Below and above this temperature range, the exponent ranged from 1.47 to 1.67. An inverse relationship was observed between sRVO2 and body mass over the temperature range of 22 - 38°C. At 14°C and 18°C, sRVO2 increased with body mass, while at 10°C and at 40°C the slope was 1.01.
- Full Text:
- Date Issued: 2010
- Authors: Setlalekgomo, Mpho Rinah
- Date: 2010
- Subjects: Eastern Cape angulate Tortoise , Oxygen consumption (Physiology)
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10690 , http://hdl.handle.net/10948/1400 , Eastern Cape angulate Tortoise , Oxygen consumption (Physiology)
- Description: The daily oxygen consumption (VO2) pattern, the effects of varying ambient temperatures, season and mass on the resting oxygen consumption (RVO2) of Chersina angulata of the Eastern Cape were investigated. The RVO2 was measured using flow-through respirometry and specific resting oxygen consumption (sRVO2) calculated. To determine the daily pattern in the VO2 of C. angulata, the tortoises were acclimated in an environmentally controlled room (ECR) to an ambient temperature of 26 ± 1°C and a light regime of 14 hours of light and 10 hours of darkness (14L:10D) for at least a week prior to the RVO2 measurements. The RVO2 was measured at a constant temperature of 26 ± 1°C, and at three different light regimes, namely: 14L:10D, constant darkness (DD) and constant light (LL). There were no significant effects of mass or gender on the sRVO2 of the tortoises used. Rhythms in the sRVO2 were detected under all three light regimes. The amplitudes of the rhythm were largest at 14L:10D, followed by DD and smallest at LL regime. The persistence of the rhythmic pattern under constant conditions suggests the existence of an endogenous circadian rhythm in the sRVO2 for adult C. angulata. To test for the effect of ambient temperature on the sRVO2 of adult C. angulata, the tortoises were acclimated to 22 ± 1°C and a 14L:10D light regime prior to the RVO2 measurements. RVO2 was measured at eight experimental temperatures; 14°C, 18°C, 22°C, 26°C, 30°C, 35°C, 38°C and 40°C. The sRVO2 was not influenced by gender and increased with experimental temperatures, but this did not happen consistently over the whole range of temperatures tested. A plateau, possibly a thermal preferendum zone, was detected within the temperature range of 26 - 38°C. Determination of seasonal effect on the sRVO2 of adult C. angulata was accomplished by acclimating tortoises to standard summer and winter conditions. Seasonal effects were tested in the appropriate seasons. Winter experiments were conducted in winter and summer experiments conducted in summer. The RVO2 was measured at experimental temperature 14°C, 18°C and 22°C. In addition RVO2 of iv winter-acclimated tortoises was also measured at 10°C. The sRVO2 increased significantly with increasing temperature within the temperature range tested. No distinct pattern was observed in the seasonal acclimation of adult C. angulata. The metabolic rate-temperature curves of the summer and the winter-acclimated tortoises cross each other. Season and temperature had no significant effects on the mass-scaling exponent of the sRVO2. The exponent ranged from 0.48 to 0.73 within the temperature range of 22 - 38°C. Below and above this temperature range, the exponent ranged from 1.47 to 1.67. An inverse relationship was observed between sRVO2 and body mass over the temperature range of 22 - 38°C. At 14°C and 18°C, sRVO2 increased with body mass, while at 10°C and at 40°C the slope was 1.01.
- Full Text:
- Date Issued: 2010
Molecular and cellular analysis of the interaction between soluble CD23 and CD11/CD18 integrins
- Authors: Daniels, Brodie Belinda
- Date: 2010
- Subjects: CD23 antigen , Immune response -- Regulation
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10303 , http://hdl.handle.net/10948/1217 , CD23 antigen , Immune response -- Regulation
- Description: The low affinity IgE receptor, CD23, is expressed by a wide variety of cells and cleaved from its original 45 kDa size to several smaller soluble CD23 proteins. Soluble CD23 function depends on the form of the protein and its interaction with various ligands. CD23 is believed to play an important role in regulating allergic responses and in inflammation, amongst others. β2 integrins are important in a variety of cell-adhesion reactions during immune-inflammatory mechanisms and the binding of their natural ligands generates outside-in cellular signalling, leading to cell activation. Although the binding of CD23 to β2 integrins contributes to this signalling in monocytes, the interaction site for CD23 is unknown. This study focused on the interaction of three soluble CD23 proteins with the β2 integrins CD11b/CD18 and CD11c/CD18. Differentiated HL60, THP1 and U937 monocytic cells were used to demonstrate the binding of three recombinant CD23 constructs (corresponding to 16, 25 and 33 kDa human soluble CD23) to upregulated CD11b/CD18 and CD11c/CD18. This binding was partially blocked by an antibody specific for the CD11b/CD18 αI domain, demonstrating that αI domains are involved in binding to CD23. Recombinant αI domain proteins of CD11b and CD11c were demonstrated to bind CD23 using ELISA and in surface plasmon resonance spectroscopy. The dissociation constants for CD23-CD11b/CD18 and CD23-CD11c/CD18 are comparable to other integrin ligands. This study has shown that CD23 interacts directly with the αI domains of β2 integrins and that the interaction surface likely spans the lectin domain as well as either the stalk and/or C-terminal tail of CD23. This study also looked at the effect that soluble CD23 proteins had on monocyte biology. It appears that iv sCD23 proteins have little effect on the phagocytic or chemotactic ability of monocytes, while an increase in oxidative burst was shown with the 16 kDa and 25 kDa CD23 proteins. Signalling pathways for the production of reactive oxygen species were investigated and it appears that the CD23 proteins signal mainly through the phosphoinositide-3 kinase pathway, although the mitogen activated protein kinase and Src kinase pathways may also play a role. These data suggest that sCD23 proteins induce outside-in signalling of β2 integrins and are able to change the activation state of CD11b/CD11c by stimulating oxidative burst. This needs to be further investigated by determining how the three sCD23 proteins are binding the CD11 proteins and investigating further leukocyte function and inflammatory responses by the cells.
- Full Text:
- Date Issued: 2010
- Authors: Daniels, Brodie Belinda
- Date: 2010
- Subjects: CD23 antigen , Immune response -- Regulation
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10303 , http://hdl.handle.net/10948/1217 , CD23 antigen , Immune response -- Regulation
- Description: The low affinity IgE receptor, CD23, is expressed by a wide variety of cells and cleaved from its original 45 kDa size to several smaller soluble CD23 proteins. Soluble CD23 function depends on the form of the protein and its interaction with various ligands. CD23 is believed to play an important role in regulating allergic responses and in inflammation, amongst others. β2 integrins are important in a variety of cell-adhesion reactions during immune-inflammatory mechanisms and the binding of their natural ligands generates outside-in cellular signalling, leading to cell activation. Although the binding of CD23 to β2 integrins contributes to this signalling in monocytes, the interaction site for CD23 is unknown. This study focused on the interaction of three soluble CD23 proteins with the β2 integrins CD11b/CD18 and CD11c/CD18. Differentiated HL60, THP1 and U937 monocytic cells were used to demonstrate the binding of three recombinant CD23 constructs (corresponding to 16, 25 and 33 kDa human soluble CD23) to upregulated CD11b/CD18 and CD11c/CD18. This binding was partially blocked by an antibody specific for the CD11b/CD18 αI domain, demonstrating that αI domains are involved in binding to CD23. Recombinant αI domain proteins of CD11b and CD11c were demonstrated to bind CD23 using ELISA and in surface plasmon resonance spectroscopy. The dissociation constants for CD23-CD11b/CD18 and CD23-CD11c/CD18 are comparable to other integrin ligands. This study has shown that CD23 interacts directly with the αI domains of β2 integrins and that the interaction surface likely spans the lectin domain as well as either the stalk and/or C-terminal tail of CD23. This study also looked at the effect that soluble CD23 proteins had on monocyte biology. It appears that iv sCD23 proteins have little effect on the phagocytic or chemotactic ability of monocytes, while an increase in oxidative burst was shown with the 16 kDa and 25 kDa CD23 proteins. Signalling pathways for the production of reactive oxygen species were investigated and it appears that the CD23 proteins signal mainly through the phosphoinositide-3 kinase pathway, although the mitogen activated protein kinase and Src kinase pathways may also play a role. These data suggest that sCD23 proteins induce outside-in signalling of β2 integrins and are able to change the activation state of CD11b/CD11c by stimulating oxidative burst. This needs to be further investigated by determining how the three sCD23 proteins are binding the CD11 proteins and investigating further leukocyte function and inflammatory responses by the cells.
- Full Text:
- Date Issued: 2010
Photoluminescence study of ZnO doped with nitrogen and arsenic
- Authors: Dangbegnon, Julien Kouadio
- Date: 2010
- Subjects: Photoluminescence , Zinc oxide , Nitrogen , Arsenic
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10518 , http://hdl.handle.net/10948/1216 , Photoluminescence , Zinc oxide , Nitrogen , Arsenic
- Description: In this work, the optical properties of ZnO doped with arsenic and nitrogen were studied. The ZnO samples were grown by Metalorganic Chemical Vapor Deposition (MOCVD). The solubility of nitrogen in the ZnO films, as well as its activation upon annealing, was also investigated. Hydrogen is known as a major source for passivation of the acceptors in ZnO:N. Therefore, it is crucial to dissociate the complex(es) formed by nitrogen and hydrogen and diffuse out the hydrogen in order to prevent the reformation of such complexes. High temperatures (≥ 600 C) are required for these purposes. In order to effectively remove the hydrogen impurities from the sample, it is important to know the optical fingerprints of hydrogen and its thermal stability. Therefore, the effects of annealing and hydrogen plasma treatment on bulk ZnO (hydrothermally grown) were first studied. The use of bulk material for this purpose was motivated by the well-resolved photoluminescence (PL) lines observed for bulk ZnO, which allow the identification of the different lines related to hydrogen after plasma treatment. Annealing at 850 C was effective for the removal of most of the hydrogen related transitions in the near-band-edge emission. Also, additional transitions at ~3.364 eV and ~3.361 eV were observed after hydrogen plasma treatment, which were ascribed to hydrogen-Zn vacancy complexes. In this work, a comparative study of the annealing ambient and temperature on ZnO films grown on GaAs substrate, using diethyl zinc (DEZn) and tertiary butanol (TBOH), showed that arsenic diffuses in the ZnO films and gives a shallow level in the band gap, which is involved in an acceptor-bound exciton line at 3.35 eV. This shallow level is visible when annealing is performed in oxygen, but not when annealing is performed in nitrogen, and indeed only for annealing temperatures around 550 C. However, annealing in either ambient also causes zinc to diffuse from the ZnO films into the GaAs substrate, rendering the electrical properties deduced from Hall measurements ambiguous. For ZnO:N, NO was used as both oxygen and nitrogen sources. Monitoring the concentration of nitrogen, carbon and hydrogen in the ZnO films, the formation of different complexes from these impurities were deduced. Furthermore, an investigation of the effect of annealing on the concentrations of impurities showed that their out- diffusion was strongly dependent on the crystalline quality of the ZnO films. For porous ZnO films, obtained at low growth temperatures (≤310 C), the out-diffusion of impurities was efficient, whereas for films grown at higher temperatures, which have improved crystalline quality, the out-diffusion was practically nonexistent. The out-diffusion of unwanted impurities may activate the nitrogen dopant in the ZnO films, as was confirmed by the PL measurements on the different samples grown at different temperatures. PL transitions at ~3.24 eV and ~3.17 eV were related to substitutional NO. These transitions were more dominant in the spectra of samples grown at low temperatures. An additional transition at ~3.1 eV was assigned to a donor-acceptor pair transition involving VZn, instead of NO, as previously reported.
- Full Text:
- Date Issued: 2010
- Authors: Dangbegnon, Julien Kouadio
- Date: 2010
- Subjects: Photoluminescence , Zinc oxide , Nitrogen , Arsenic
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10518 , http://hdl.handle.net/10948/1216 , Photoluminescence , Zinc oxide , Nitrogen , Arsenic
- Description: In this work, the optical properties of ZnO doped with arsenic and nitrogen were studied. The ZnO samples were grown by Metalorganic Chemical Vapor Deposition (MOCVD). The solubility of nitrogen in the ZnO films, as well as its activation upon annealing, was also investigated. Hydrogen is known as a major source for passivation of the acceptors in ZnO:N. Therefore, it is crucial to dissociate the complex(es) formed by nitrogen and hydrogen and diffuse out the hydrogen in order to prevent the reformation of such complexes. High temperatures (≥ 600 C) are required for these purposes. In order to effectively remove the hydrogen impurities from the sample, it is important to know the optical fingerprints of hydrogen and its thermal stability. Therefore, the effects of annealing and hydrogen plasma treatment on bulk ZnO (hydrothermally grown) were first studied. The use of bulk material for this purpose was motivated by the well-resolved photoluminescence (PL) lines observed for bulk ZnO, which allow the identification of the different lines related to hydrogen after plasma treatment. Annealing at 850 C was effective for the removal of most of the hydrogen related transitions in the near-band-edge emission. Also, additional transitions at ~3.364 eV and ~3.361 eV were observed after hydrogen plasma treatment, which were ascribed to hydrogen-Zn vacancy complexes. In this work, a comparative study of the annealing ambient and temperature on ZnO films grown on GaAs substrate, using diethyl zinc (DEZn) and tertiary butanol (TBOH), showed that arsenic diffuses in the ZnO films and gives a shallow level in the band gap, which is involved in an acceptor-bound exciton line at 3.35 eV. This shallow level is visible when annealing is performed in oxygen, but not when annealing is performed in nitrogen, and indeed only for annealing temperatures around 550 C. However, annealing in either ambient also causes zinc to diffuse from the ZnO films into the GaAs substrate, rendering the electrical properties deduced from Hall measurements ambiguous. For ZnO:N, NO was used as both oxygen and nitrogen sources. Monitoring the concentration of nitrogen, carbon and hydrogen in the ZnO films, the formation of different complexes from these impurities were deduced. Furthermore, an investigation of the effect of annealing on the concentrations of impurities showed that their out- diffusion was strongly dependent on the crystalline quality of the ZnO films. For porous ZnO films, obtained at low growth temperatures (≤310 C), the out-diffusion of impurities was efficient, whereas for films grown at higher temperatures, which have improved crystalline quality, the out-diffusion was practically nonexistent. The out-diffusion of unwanted impurities may activate the nitrogen dopant in the ZnO films, as was confirmed by the PL measurements on the different samples grown at different temperatures. PL transitions at ~3.24 eV and ~3.17 eV were related to substitutional NO. These transitions were more dominant in the spectra of samples grown at low temperatures. An additional transition at ~3.1 eV was assigned to a donor-acceptor pair transition involving VZn, instead of NO, as previously reported.
- Full Text:
- Date Issued: 2010
Spatially resolved opto-electric measurements of photovoltaic materials and devices
- Authors: Thantsha, Nicolas Matome
- Date: 2010
- Subjects: Photovoltaic cells , Photovoltaic power systems , Photovoltaic power generation
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10520 , http://hdl.handle.net/10948/1123 , Photovoltaic cells , Photovoltaic power systems , Photovoltaic power generation
- Description: The objective of this study is to characterize and analyse defects in solar cell devices. Materials used to fabricate solar cells are not defects free and therefore, there is a need to investigate defects in cells. To investigate this, a topographical technique was developed and employed which uses a non-destructive methodology to analyse solar cells. A system was built which uses a technique based on a laser beam induced current (LBIC). LBIC technique involves focusing light on to a surface of a solar cell device in order to create a photo-generated current that can be measured in the external circuit for analyses. The advantage of this technique is that it allows parameter extraction. Parameters that can be extracted include short-circuit current, carrier lifetime and also the external and internal quantum efficiency of a solar cell. In this thesis, LBIC measurements in the form of picture maps are used to indicate the distribution of the localized beam induced current within solar cells. Areas with low minority carrier lifetime in solar cells are made visible by LBIC mapping. Surface reflection intensity measurements of cells can also be mapped using the LBIC system developed in this study. The system is also capable of mapping photo-generated current of a cell below and above room temperature. This thesis also presents an assessment procedure capable of assessing the device and performance parameters with reference to I-V measurements. The dark and illuminated I-V characteristics of solar cells were investigated. The illuminated I-V characteristics of solar cells were obtained using a defocused laser beam. Dark I-V measurements were performed by applying voltage across the cell in the dark and measuring a current through it. The device parameters which describe the behaviour of I-V characteristic were extracted from the I-V data using Particle Swarm Optimization (PSO) method based on a one-and two-diode solar cell models. Solar cells of different technologies were analysed, namely, single-crystalline (c-Si) and multicrystalline (mc-Si) silicon, Edge-defined Film-fed Growth Si (EFG-Si) and Cu(In,Ga)(Se,S)2 (CIGSS) thin film based cells. The LBIC results illustrated the effect of surface reflection features and material defects in the solar cell investigated. IQE at a wavelength of 660 nm were measured on these cells and the results in general emphasised the importance of correcting optical losses, i.e. reflection loss, when characterizing different types of defects. The agreement between the IQE measurements and I-V characteristics of a cell showed that the differences in crystal grains influence the performance of a mc-Si cell. The temperature-dependence of I-V characteristics of a CIGSS solar cell was investigated. The results showed that, for this material, the photo response is reduced at elevated temperatures. In addition to LBIC using a laser beam, solar spectral radiation was employed to obtained device performance parameters. The results emphasised the effect of grain boundaries as a recombination centres for photo-generated hole-pairs. Lastly, mesa diode characterizations of solar cells were investigated. Mesa diodes are achieved by etching down a solar cell so that the plateau regions are formed. Mesa diodes expose the p-n junction, and therefore mesa diode analysis provides a better way of determining and revealing the fundamental current conduction mechanism at the junction. Mesa diodes avoid possible edge effects. This study showed that mesa diodes can be used to characterize spatial non-uniformities in solar cells. The results obtained in this study indicate that LBIC is a useful tool for defect characterization in solar cells. Also LBIC complements other characterization techniques such as I-V characterization.
- Full Text:
- Date Issued: 2010
- Authors: Thantsha, Nicolas Matome
- Date: 2010
- Subjects: Photovoltaic cells , Photovoltaic power systems , Photovoltaic power generation
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10520 , http://hdl.handle.net/10948/1123 , Photovoltaic cells , Photovoltaic power systems , Photovoltaic power generation
- Description: The objective of this study is to characterize and analyse defects in solar cell devices. Materials used to fabricate solar cells are not defects free and therefore, there is a need to investigate defects in cells. To investigate this, a topographical technique was developed and employed which uses a non-destructive methodology to analyse solar cells. A system was built which uses a technique based on a laser beam induced current (LBIC). LBIC technique involves focusing light on to a surface of a solar cell device in order to create a photo-generated current that can be measured in the external circuit for analyses. The advantage of this technique is that it allows parameter extraction. Parameters that can be extracted include short-circuit current, carrier lifetime and also the external and internal quantum efficiency of a solar cell. In this thesis, LBIC measurements in the form of picture maps are used to indicate the distribution of the localized beam induced current within solar cells. Areas with low minority carrier lifetime in solar cells are made visible by LBIC mapping. Surface reflection intensity measurements of cells can also be mapped using the LBIC system developed in this study. The system is also capable of mapping photo-generated current of a cell below and above room temperature. This thesis also presents an assessment procedure capable of assessing the device and performance parameters with reference to I-V measurements. The dark and illuminated I-V characteristics of solar cells were investigated. The illuminated I-V characteristics of solar cells were obtained using a defocused laser beam. Dark I-V measurements were performed by applying voltage across the cell in the dark and measuring a current through it. The device parameters which describe the behaviour of I-V characteristic were extracted from the I-V data using Particle Swarm Optimization (PSO) method based on a one-and two-diode solar cell models. Solar cells of different technologies were analysed, namely, single-crystalline (c-Si) and multicrystalline (mc-Si) silicon, Edge-defined Film-fed Growth Si (EFG-Si) and Cu(In,Ga)(Se,S)2 (CIGSS) thin film based cells. The LBIC results illustrated the effect of surface reflection features and material defects in the solar cell investigated. IQE at a wavelength of 660 nm were measured on these cells and the results in general emphasised the importance of correcting optical losses, i.e. reflection loss, when characterizing different types of defects. The agreement between the IQE measurements and I-V characteristics of a cell showed that the differences in crystal grains influence the performance of a mc-Si cell. The temperature-dependence of I-V characteristics of a CIGSS solar cell was investigated. The results showed that, for this material, the photo response is reduced at elevated temperatures. In addition to LBIC using a laser beam, solar spectral radiation was employed to obtained device performance parameters. The results emphasised the effect of grain boundaries as a recombination centres for photo-generated hole-pairs. Lastly, mesa diode characterizations of solar cells were investigated. Mesa diodes are achieved by etching down a solar cell so that the plateau regions are formed. Mesa diodes expose the p-n junction, and therefore mesa diode analysis provides a better way of determining and revealing the fundamental current conduction mechanism at the junction. Mesa diodes avoid possible edge effects. This study showed that mesa diodes can be used to characterize spatial non-uniformities in solar cells. The results obtained in this study indicate that LBIC is a useful tool for defect characterization in solar cells. Also LBIC complements other characterization techniques such as I-V characterization.
- Full Text:
- Date Issued: 2010
Synthesis, characterization and reactions of novel ferrocenylimidazoles as donor ligands
- Authors: Onyancha, Douglas Okerio
- Date: 2010
- Subjects: Ferrocene
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10390 , http://hdl.handle.net/10948/1150 , Ferrocene
- Description: In this project a series of ferrocenylimidazole derivatives were successfully prepared and characterized using an array of analytical techniques. Reactions of ferrocenylcarbinols with N,N′-thiocarbonyldiimidazole under both solvent-free conditions and in dichloromethane are described. The reaction of ferrocenylmethanol afforded S,S-bis(ferrocenylmethyl)dithiocarbonate, the reaction of α-substituted ferrocenylcarbinols provided ferrocenylalkylimidazolides, while ω-ferrocenylcarbinols yielded ferrocenylimidazolecarbothioates. The reactions were carried out under solvent-free conditions, consistent with the principles of Green Chemistry. Ferrocenyl benzyl ethers were successfully prepared by reacting 4-ferrocenylbenzyl-1Hcarboxylate with a series of primary alcohols under catalytic condition and under a catalyst– free environment. Refluxing a mixture of alcohol-water and 4-ferrocenylbenzyl-1Hcarboxylate provided the corresponding ethers in modest yields. The same ethers were obtained at room temperature by employing potassium tetrachloroplatinate or hydrochloric acid as catalyst. Ferrocenyl-1H-imidazole, 4-ferrocenylphenyl-1H-imidazole and their corresponding ferrocenyl-3-subsitituted imidazolium salts have been successfully synthesized. The compounds were characterized and the electrochemical properties of selected imidazolium salts were examined by cyclic voltammetry. Additionally, X-ray structures of two of the imidazolium salts were determined. The imidazolium salts were found to be good catalysts for the Heck and Suzuki cross-coupling reactions.
- Full Text:
- Date Issued: 2010
- Authors: Onyancha, Douglas Okerio
- Date: 2010
- Subjects: Ferrocene
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10390 , http://hdl.handle.net/10948/1150 , Ferrocene
- Description: In this project a series of ferrocenylimidazole derivatives were successfully prepared and characterized using an array of analytical techniques. Reactions of ferrocenylcarbinols with N,N′-thiocarbonyldiimidazole under both solvent-free conditions and in dichloromethane are described. The reaction of ferrocenylmethanol afforded S,S-bis(ferrocenylmethyl)dithiocarbonate, the reaction of α-substituted ferrocenylcarbinols provided ferrocenylalkylimidazolides, while ω-ferrocenylcarbinols yielded ferrocenylimidazolecarbothioates. The reactions were carried out under solvent-free conditions, consistent with the principles of Green Chemistry. Ferrocenyl benzyl ethers were successfully prepared by reacting 4-ferrocenylbenzyl-1Hcarboxylate with a series of primary alcohols under catalytic condition and under a catalyst– free environment. Refluxing a mixture of alcohol-water and 4-ferrocenylbenzyl-1Hcarboxylate provided the corresponding ethers in modest yields. The same ethers were obtained at room temperature by employing potassium tetrachloroplatinate or hydrochloric acid as catalyst. Ferrocenyl-1H-imidazole, 4-ferrocenylphenyl-1H-imidazole and their corresponding ferrocenyl-3-subsitituted imidazolium salts have been successfully synthesized. The compounds were characterized and the electrochemical properties of selected imidazolium salts were examined by cyclic voltammetry. Additionally, X-ray structures of two of the imidazolium salts were determined. The imidazolium salts were found to be good catalysts for the Heck and Suzuki cross-coupling reactions.
- Full Text:
- Date Issued: 2010
The in vitro biological activities of three Hypoxis species and their active compounds
- Authors: Boukes, Gerhardt Johannes
- Date: 2010
- Subjects: Potatoes -- Africa , Potatoes -- Therapeutic use , Medicinal plants
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10322 , http://hdl.handle.net/10948/1228 , Potatoes -- Africa , Potatoes -- Therapeutic use , Medicinal plants
- Description: The African potato is used as an African traditional medicine for its nutritional and medicinal properties. Most research has been carried out on H. hemerocallidea, with very little or nothing on other Hypoxis spp. The main aim of this project was to provide scientific data on the anticancer, anti-inflammatory and antioxidant properties of H. hemerocallidea, H. stellipilis and H. sobolifera chloroform extracts and their active compounds. The hypoxoside and phytosterol contents of the three Hypoxis spp. were determined using TLC, HPLC and GC. H. hemerocallidea and H. sobolifera chloroform extracts contained the highest amounts of hypoxoside and β-sitosterol, respectively. For the anticancer properties, cytotoxicity of the Hypoxis extracts and its purified compounds were determined against the HeLa, HT-29 and MCF-7 cancer cell lines (using MTT), and PBMCs (using CellTiter-Blue®). H. sobolifera had the best cytotoxicity against the three cancer cell lines, whereas H. stellipilis stimulated HeLa and HT-29 cancer cell growth. IC50 values of hypoxoside and rooperol were determined. DNA cell cycle arrest (using PI staining) occurred in the late G1/early S (confirmed by increased p21Waf1/Cip1 expression) and G2/M phases after 15 and 48 hrs, respectively, when treated with Hypoxis extracts and rooperol. H. sobolifera and rooperol activated caspase-3 and -7 (using fluorescently labelled antibodies) in HeLa and HT-29 cancer cells, and caspase-7 in MCF-7 cancer cells after 48 hrs. Annexin V binding to phosphatidylserines in rooperol treated U937 cells confirmed early apoptosis after 15 hrs. The TUNEL assay showed DNA fragmentation in the three cancer cell lines when treated with H. sobolifera and rooperol for 48 hrs. A shift pass the G2/M phase has led to the investigation of endoreduplication, which was confirmed by cell/nucleus size, and anti-apoptotic proteins (Akt, phospho-Akt, phospho-Bcl-2 and p21Waf1/Cip1). U937 cell differentiation to monocyte-macrophages was optimized using PMA and 1,25(OH)2D3, which was confirmed by morphological and biochemical changes. For the anti-inflammatory properties, Hypoxis extracts and rooperol significantly increased NO production in monocyte-macrophages (pre-loaded with DAF-2 DA) and phagocytosis of pHrodoTM E. coli BioParticles®. The treatments had no effect on COX-2 expression in monocyte-macrophages. The phytosterols significantly increased IL-1β and IL-6 secretion xv (using the FlowCytomix Multiplex human Th1/Th2 10plex Kit I) in the PBMCs of one donor. For the antioxidant properties, Hypoxis extracts and rooperol significantly increased ROS production in undifferentiated and differentiated U937 cells, which were pre-loaded with DCFH-DA. Hypoxis extracts and purified compounds had ferric reducing activities, but only rooperol had ferric reducing activities significantly greater than ascorbic acid. β-sitosterol, campesterol and cholesterol significantly increased SOD activity in Chang liver cells, while H. stellipilis, H. sobolifera and rooperol decreased SOD activity. Anticancer, anti-inflammatory and antioxidant properties of the Hypoxis extracts may be attributed to the β-sitosterol content, because Hypoxis chloroform extracts contained very little or no hypoxoside. Unidentified compounds, and synergistic and additive effects of the compounds may have contributed to the biological effects. This study confirms previous reports that rooperol is the active compound. Results provide scientific data on the medicinal properties of one of the most frequently used medicinal plants in South Africa.
- Full Text:
- Date Issued: 2010
- Authors: Boukes, Gerhardt Johannes
- Date: 2010
- Subjects: Potatoes -- Africa , Potatoes -- Therapeutic use , Medicinal plants
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10322 , http://hdl.handle.net/10948/1228 , Potatoes -- Africa , Potatoes -- Therapeutic use , Medicinal plants
- Description: The African potato is used as an African traditional medicine for its nutritional and medicinal properties. Most research has been carried out on H. hemerocallidea, with very little or nothing on other Hypoxis spp. The main aim of this project was to provide scientific data on the anticancer, anti-inflammatory and antioxidant properties of H. hemerocallidea, H. stellipilis and H. sobolifera chloroform extracts and their active compounds. The hypoxoside and phytosterol contents of the three Hypoxis spp. were determined using TLC, HPLC and GC. H. hemerocallidea and H. sobolifera chloroform extracts contained the highest amounts of hypoxoside and β-sitosterol, respectively. For the anticancer properties, cytotoxicity of the Hypoxis extracts and its purified compounds were determined against the HeLa, HT-29 and MCF-7 cancer cell lines (using MTT), and PBMCs (using CellTiter-Blue®). H. sobolifera had the best cytotoxicity against the three cancer cell lines, whereas H. stellipilis stimulated HeLa and HT-29 cancer cell growth. IC50 values of hypoxoside and rooperol were determined. DNA cell cycle arrest (using PI staining) occurred in the late G1/early S (confirmed by increased p21Waf1/Cip1 expression) and G2/M phases after 15 and 48 hrs, respectively, when treated with Hypoxis extracts and rooperol. H. sobolifera and rooperol activated caspase-3 and -7 (using fluorescently labelled antibodies) in HeLa and HT-29 cancer cells, and caspase-7 in MCF-7 cancer cells after 48 hrs. Annexin V binding to phosphatidylserines in rooperol treated U937 cells confirmed early apoptosis after 15 hrs. The TUNEL assay showed DNA fragmentation in the three cancer cell lines when treated with H. sobolifera and rooperol for 48 hrs. A shift pass the G2/M phase has led to the investigation of endoreduplication, which was confirmed by cell/nucleus size, and anti-apoptotic proteins (Akt, phospho-Akt, phospho-Bcl-2 and p21Waf1/Cip1). U937 cell differentiation to monocyte-macrophages was optimized using PMA and 1,25(OH)2D3, which was confirmed by morphological and biochemical changes. For the anti-inflammatory properties, Hypoxis extracts and rooperol significantly increased NO production in monocyte-macrophages (pre-loaded with DAF-2 DA) and phagocytosis of pHrodoTM E. coli BioParticles®. The treatments had no effect on COX-2 expression in monocyte-macrophages. The phytosterols significantly increased IL-1β and IL-6 secretion xv (using the FlowCytomix Multiplex human Th1/Th2 10plex Kit I) in the PBMCs of one donor. For the antioxidant properties, Hypoxis extracts and rooperol significantly increased ROS production in undifferentiated and differentiated U937 cells, which were pre-loaded with DCFH-DA. Hypoxis extracts and purified compounds had ferric reducing activities, but only rooperol had ferric reducing activities significantly greater than ascorbic acid. β-sitosterol, campesterol and cholesterol significantly increased SOD activity in Chang liver cells, while H. stellipilis, H. sobolifera and rooperol decreased SOD activity. Anticancer, anti-inflammatory and antioxidant properties of the Hypoxis extracts may be attributed to the β-sitosterol content, because Hypoxis chloroform extracts contained very little or no hypoxoside. Unidentified compounds, and synergistic and additive effects of the compounds may have contributed to the biological effects. This study confirms previous reports that rooperol is the active compound. Results provide scientific data on the medicinal properties of one of the most frequently used medicinal plants in South Africa.
- Full Text:
- Date Issued: 2010
Towards social impact assessment of copper-nickel mining in Botswana
- Authors: Mengwe, Moses Seargent
- Date: 2010
- Subjects: Copper mines and mining -- Environmental aspects -- Botswana , Nickel mines and mining -- Environmental aspects -- Botswana , Mineral industry -- Botswana , Mines and mineral resources -- Botswana
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10644 , http://hdl.handle.net/10948/1443 , Copper mines and mining -- Environmental aspects -- Botswana , Nickel mines and mining -- Environmental aspects -- Botswana , Mineral industry -- Botswana , Mines and mineral resources -- Botswana
- Description: This research study is more of an initiative towards Social Impact Assessment of copper-nickel mining in Botswana. The specific objectives of the study were centred on the assessment of the social impacts of copper-nickel mining in Botswana from the initial mining stage of exploration, surveying and mine site development to mine closure. The study was carried out under the broad hypotheses that mining influences population movement that impact on areas of mining; mining activities have both economic benefits and deleterious social impacts on the local communities found in the areas where mining is taking place; and mine closure has far reaching socio-economic, investment and developmental implications over and above the obvious interests of project owners. To achieve the broad aim as summarised above, the research study used a multi-disciplinary methodology and approach that required several kinds of expertise and sources of information. Hence it used both primary and secondary sources centred on interactive informative interviews, site visits and observations, questionnaires, census data records, mining companies’ publications, published textbooks and journal articles. The research study comprised of three different mines operated by three different mining companies in three varied socio-cultural and ethnic regions of Botswana. First was a detailed Social Impact Assessment of the initial phase of exploration, surveying and mine site development represented by Mowana mine project operated by African Copper in the rural areas of Dugwi and Mosetse. This case study yielded results showing that the social impacts of mining in the area are diverse and extensive. The findings suggest that the impacts relate not only to the possible economic benefits of foreign exchange, employment, the optimal use of available mineral resources and the possible development of Dugwi and Mosetse villages, but extends to the deleterious social impacts. The results also indicated that the social impacts have just begun in the two communities. Hence they point towards a possible disruption within the socio-cultural system of the local people if serious mitigation measures are not put in place; thus suggesting that the early stages of exploration and mine site development results in the most conflict between the mine and the local people. Second was a comprehensive Social Impact Assessment of Tati-Nickel Phoenix mining project in the peri-urban areas of Matshelagabedi and Matsiloje areas representing the mining stage of mine production and expansion. The results from this case study suggest that during vi mine production and expansion, many people were relocated. However, the overriding impression gained from the case study was Tati-Nickel Mining Company’s elaborate corporate policies that suggested good corporate governance and best practices that promote sustainable development. A notable milestone on good corporate governance and best practice that the other two case studies (mining company) could benchmark on is Tati-Nickel’s corporate social responsibility programme that has been designed to ensure that the communities within a fifty kilometre mine radius benefit from the mine. The results from the case study also distinguished the mining stage of production and expansion from the other two because it is associated with the deep entrenchment of the social impacts into the communities near to mining areas. Third was a detailed Social Impact Assessment on Bamangwato Concession Limited mine in the industrial town of Selebi-Phikwe. The case study represented the stage of mine closure. Through the findings of this case study, it became apparent that the economic dependence of Selebi-Phikwe on mining has seen the town developing into a mining town, increasing its vulnerability at mine closure. The results from the case study further suggest that mine closure will degrade the socio-economic sector of the town with ever far reaching socio-economic implications as many people lose their gainful employment, hence suggesting that a possible complete mine closure will be the most traumatic phase leading to major social conflict within the area. Thus the results suggest that at mine closure, the deleterious social impacts will overspill to other areas in Botswana with disastrous effects for the economy of the country. The results yielded through this study established in clear and passionate language that copper-nickel mining in Botswana influences population movements that lead to positive and negative impacts on the communities found in mining areas. Another major finding of the study is that copper-nickel mining activities have both economic benefits and deleterious social impacts on the local communities, hence the recommendation that the copper-nickel mining companies should embrace the concept of sustainable mining for sustainable development to avoid most of the negative impacts of their operations on the local communities.
- Full Text:
- Date Issued: 2010
- Authors: Mengwe, Moses Seargent
- Date: 2010
- Subjects: Copper mines and mining -- Environmental aspects -- Botswana , Nickel mines and mining -- Environmental aspects -- Botswana , Mineral industry -- Botswana , Mines and mineral resources -- Botswana
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10644 , http://hdl.handle.net/10948/1443 , Copper mines and mining -- Environmental aspects -- Botswana , Nickel mines and mining -- Environmental aspects -- Botswana , Mineral industry -- Botswana , Mines and mineral resources -- Botswana
- Description: This research study is more of an initiative towards Social Impact Assessment of copper-nickel mining in Botswana. The specific objectives of the study were centred on the assessment of the social impacts of copper-nickel mining in Botswana from the initial mining stage of exploration, surveying and mine site development to mine closure. The study was carried out under the broad hypotheses that mining influences population movement that impact on areas of mining; mining activities have both economic benefits and deleterious social impacts on the local communities found in the areas where mining is taking place; and mine closure has far reaching socio-economic, investment and developmental implications over and above the obvious interests of project owners. To achieve the broad aim as summarised above, the research study used a multi-disciplinary methodology and approach that required several kinds of expertise and sources of information. Hence it used both primary and secondary sources centred on interactive informative interviews, site visits and observations, questionnaires, census data records, mining companies’ publications, published textbooks and journal articles. The research study comprised of three different mines operated by three different mining companies in three varied socio-cultural and ethnic regions of Botswana. First was a detailed Social Impact Assessment of the initial phase of exploration, surveying and mine site development represented by Mowana mine project operated by African Copper in the rural areas of Dugwi and Mosetse. This case study yielded results showing that the social impacts of mining in the area are diverse and extensive. The findings suggest that the impacts relate not only to the possible economic benefits of foreign exchange, employment, the optimal use of available mineral resources and the possible development of Dugwi and Mosetse villages, but extends to the deleterious social impacts. The results also indicated that the social impacts have just begun in the two communities. Hence they point towards a possible disruption within the socio-cultural system of the local people if serious mitigation measures are not put in place; thus suggesting that the early stages of exploration and mine site development results in the most conflict between the mine and the local people. Second was a comprehensive Social Impact Assessment of Tati-Nickel Phoenix mining project in the peri-urban areas of Matshelagabedi and Matsiloje areas representing the mining stage of mine production and expansion. The results from this case study suggest that during vi mine production and expansion, many people were relocated. However, the overriding impression gained from the case study was Tati-Nickel Mining Company’s elaborate corporate policies that suggested good corporate governance and best practices that promote sustainable development. A notable milestone on good corporate governance and best practice that the other two case studies (mining company) could benchmark on is Tati-Nickel’s corporate social responsibility programme that has been designed to ensure that the communities within a fifty kilometre mine radius benefit from the mine. The results from the case study also distinguished the mining stage of production and expansion from the other two because it is associated with the deep entrenchment of the social impacts into the communities near to mining areas. Third was a detailed Social Impact Assessment on Bamangwato Concession Limited mine in the industrial town of Selebi-Phikwe. The case study represented the stage of mine closure. Through the findings of this case study, it became apparent that the economic dependence of Selebi-Phikwe on mining has seen the town developing into a mining town, increasing its vulnerability at mine closure. The results from the case study further suggest that mine closure will degrade the socio-economic sector of the town with ever far reaching socio-economic implications as many people lose their gainful employment, hence suggesting that a possible complete mine closure will be the most traumatic phase leading to major social conflict within the area. Thus the results suggest that at mine closure, the deleterious social impacts will overspill to other areas in Botswana with disastrous effects for the economy of the country. The results yielded through this study established in clear and passionate language that copper-nickel mining in Botswana influences population movements that lead to positive and negative impacts on the communities found in mining areas. Another major finding of the study is that copper-nickel mining activities have both economic benefits and deleterious social impacts on the local communities, hence the recommendation that the copper-nickel mining companies should embrace the concept of sustainable mining for sustainable development to avoid most of the negative impacts of their operations on the local communities.
- Full Text:
- Date Issued: 2010
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