Reactivity of Rhenium (iii) and Rhenium (V) with multidentate NN-and no-donor ligands
- Yumata, Nonzaliseko Christine
- Authors: Yumata, Nonzaliseko Christine
- Date: 2010
- Subjects: Rhenium , Ligands
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10373 , http://hdl.handle.net/10948/1085 , Rhenium , Ligands
- Description: The reaction of the potentially tridentate Schiff-base chelate Hhaep [Haep = N’-(1- (2-hydroxyphenyl)ethylidene)benzohydrazide] with [ReCl3(benzil)(PPh3)] and trans-[ReOCl3(PPh3)2] produced the neutral oxorhenium(V) complexes cis- [ReOCl2(mep)] [Hmep = 2-(1-iminoethyl)phenol] and cis-[ReOCI2(meb)(PPh3)] [Hmeb = N’-(propan-2-ylidene)benzohydrazide] in ethanol and acetone respectively. In both reactions the Hhaep molecule cleaves to give different coordinated bidentate NO-donor chelates coordinated to the rhenium(V) centers. The X-ray studies reveal that mep is present as a bidentate, monoanionic Schiffbase coordinating through the neutral imino nitrogen and the deprotonated phenolate oxygen in cis-[ReOCl2(mep)]. The bond distances and angles in cis- [ReOCI2(meb)(PPh3)] confirm that meb coordinates to the metal in the enolate form. The distorted octahedral complex fac-[ReCl3(dpa)(PPh3)] was prepared by the reaction of trans-[ReCl3(MeCN)(PPh3)2] with a twofold molar excess of dpa in acetonitrile under a nitrogen atmosphere. The compound dpa.HCl.2H2O was obtained as a by-product in the reaction of dpa with trans-[ReCI3(MeCN)(PPh3)2] in acetonitrile. The reaction of trans-[ReCl3(MeCN)(PPh3)2] with a twofold molar excess of 6- amino-3-methyl-1-phenyl-4-azahept-2-ene-1-one (Hamp) in acetonitrile led to the isolation of cis-[ReCl2(bat)(PPh3)2]. On complexation to the metal center Hamp decomposed to give the coordinated benzoylacetone (bat). Bat is present as a monoanionic bidentate chelate. The complexes [ReVOCI(had)] and [ReIVCl(had)(PPh3)](ReO4) were prepared from the reaction of trans-[ReCl3(MeCN)(PPh3)2] with N,N-bis((2-hydroxybenzyl)-2- aminoethyl)dimethylamine (H2had) in ethanol under various reaction conditions. The treatment of [ReCl3(benzil)(PPh3)] with 2-[((2- pyridinylmethyl)amino)methyl]phenol (Hham) in a 2:1 molar ratio in acetonitrile led to the isolation of the hydrogen-bonded dimer [ReOCl2(ham)]2. The distorted octahedral complex [ReOCl(hap)] [H2hap = N,N-bis(2- hydroxybenzyl)aminomethylpyridine] was prepared from the reaction of trans- [ReCl3(MeCN)(PPh3)2] with a twofold molar excess of H2hap in acetonitrile. The X-ray crystal structure analysis shows that the chloride is coordinated trans to the tripodal tertiary amino nitrogen, with a phenolate oxygen trans to the oxo oxygen.
- Full Text:
- Date Issued: 2010
- Authors: Yumata, Nonzaliseko Christine
- Date: 2010
- Subjects: Rhenium , Ligands
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10373 , http://hdl.handle.net/10948/1085 , Rhenium , Ligands
- Description: The reaction of the potentially tridentate Schiff-base chelate Hhaep [Haep = N’-(1- (2-hydroxyphenyl)ethylidene)benzohydrazide] with [ReCl3(benzil)(PPh3)] and trans-[ReOCl3(PPh3)2] produced the neutral oxorhenium(V) complexes cis- [ReOCl2(mep)] [Hmep = 2-(1-iminoethyl)phenol] and cis-[ReOCI2(meb)(PPh3)] [Hmeb = N’-(propan-2-ylidene)benzohydrazide] in ethanol and acetone respectively. In both reactions the Hhaep molecule cleaves to give different coordinated bidentate NO-donor chelates coordinated to the rhenium(V) centers. The X-ray studies reveal that mep is present as a bidentate, monoanionic Schiffbase coordinating through the neutral imino nitrogen and the deprotonated phenolate oxygen in cis-[ReOCl2(mep)]. The bond distances and angles in cis- [ReOCI2(meb)(PPh3)] confirm that meb coordinates to the metal in the enolate form. The distorted octahedral complex fac-[ReCl3(dpa)(PPh3)] was prepared by the reaction of trans-[ReCl3(MeCN)(PPh3)2] with a twofold molar excess of dpa in acetonitrile under a nitrogen atmosphere. The compound dpa.HCl.2H2O was obtained as a by-product in the reaction of dpa with trans-[ReCI3(MeCN)(PPh3)2] in acetonitrile. The reaction of trans-[ReCl3(MeCN)(PPh3)2] with a twofold molar excess of 6- amino-3-methyl-1-phenyl-4-azahept-2-ene-1-one (Hamp) in acetonitrile led to the isolation of cis-[ReCl2(bat)(PPh3)2]. On complexation to the metal center Hamp decomposed to give the coordinated benzoylacetone (bat). Bat is present as a monoanionic bidentate chelate. The complexes [ReVOCI(had)] and [ReIVCl(had)(PPh3)](ReO4) were prepared from the reaction of trans-[ReCl3(MeCN)(PPh3)2] with N,N-bis((2-hydroxybenzyl)-2- aminoethyl)dimethylamine (H2had) in ethanol under various reaction conditions. The treatment of [ReCl3(benzil)(PPh3)] with 2-[((2- pyridinylmethyl)amino)methyl]phenol (Hham) in a 2:1 molar ratio in acetonitrile led to the isolation of the hydrogen-bonded dimer [ReOCl2(ham)]2. The distorted octahedral complex [ReOCl(hap)] [H2hap = N,N-bis(2- hydroxybenzyl)aminomethylpyridine] was prepared from the reaction of trans- [ReCl3(MeCN)(PPh3)2] with a twofold molar excess of H2hap in acetonitrile. The X-ray crystal structure analysis shows that the chloride is coordinated trans to the tripodal tertiary amino nitrogen, with a phenolate oxygen trans to the oxo oxygen.
- Full Text:
- Date Issued: 2010
Quantification of greenhouse gas fluxes from soil in agricultural fields
- Authors: Nkongolo, Nsalambi Vakanda
- Date: 2010
- Subjects: Greenhouse gases -- Environmental aspects
- Language: English
- Type: Thesis , Doctoral , DPhil
- Identifier: vital:10645 , http://hdl.handle.net/10948/1474 , Greenhouse gases -- Environmental aspects
- Description: Field studies were conducted at Lincoln University of Missouri (USA) and Hokkaido University (Japan) to: (i) study the relationships between greenhouse gases emissions and soil properties, (ii) assess the influence of agricultural practices on greenhouse gas fluxes and soil properties and (iii) improve the quantification of greenhouse gases from soil in agricultural fields using geospatial technologies. Results showed that besides soil temperature (T), soil thermal properties such as thermal conductivity (K), resistivity (R) and diffusivity (D) and soil pore spaces indices such as the pore tortuosity factor and the relative gas diffusion coefficient (Ds/Do) are controlling factors for greenhouse gases emissions. Soil thermal properties correlated with greenhouse gases emissions when soil temperature could not. The study has found that predicted Ds/Do and correlate with greenhouse gas fluxes even when the air-filled porosity and the total porosity from which they are predicted did not. We have also showed that Ds/Do and can be predicted quickly from routine measurements of soil water and air and existing diffusivity models found in the literature. Agricultural practices do seriously impact greenhouse gases emissions as showed by the effect of mechanized tillage operations on soil physical properties and greenhouse gas fluxes in a corn and soybean fields. In fact, our results showed that tractor compaction increased soil resistance to penetration, water, bulk density and pore tortuosity while reducing air-filled porosity, total pore space and the soil gas diffusion coefficient. Changes in soil properties resulted in increased CO2, NO and N2O emissions. Finally, our results also confirmed that greenhouse gas fluxes vary tremendously in space and time. As estimates of greenhouse gas emissions are influenced by the data processing approach, differences between the different calculation approaches leads to uncertainty. Thus, techniques for developing better estimates are needed. We have showed that Geographic Information Systems (GIS), Global Positioning System (GPS), computer mapping and geo-statistics are technologies that can be used to better understand systems containing large amounts of spatial and temporal variability. Our GIS-based approach for quantifying CO2, CH4 and N2O fluxes from soil in agricultural fields showed that estimating (extrapolating) total greenhouse gas fluxes using the “standard” approach – multiplying the average flux value by the total field area – results in biased predictions of field total greenhouse gases emissions. In contrast, the GIS-based approach we developed produces an interpolated map portraying the spatial distribution of gas fluxes across the field from point measurements and later process the interpolated map produced to determine flux zones. Furthermore, processing, classification and modeling enables the computation of field total fluxes as the sum of fluxes in different zones, therefore taking into account the spatial variability of greenhouse gas fluxes.
- Full Text:
- Date Issued: 2010
- Authors: Nkongolo, Nsalambi Vakanda
- Date: 2010
- Subjects: Greenhouse gases -- Environmental aspects
- Language: English
- Type: Thesis , Doctoral , DPhil
- Identifier: vital:10645 , http://hdl.handle.net/10948/1474 , Greenhouse gases -- Environmental aspects
- Description: Field studies were conducted at Lincoln University of Missouri (USA) and Hokkaido University (Japan) to: (i) study the relationships between greenhouse gases emissions and soil properties, (ii) assess the influence of agricultural practices on greenhouse gas fluxes and soil properties and (iii) improve the quantification of greenhouse gases from soil in agricultural fields using geospatial technologies. Results showed that besides soil temperature (T), soil thermal properties such as thermal conductivity (K), resistivity (R) and diffusivity (D) and soil pore spaces indices such as the pore tortuosity factor and the relative gas diffusion coefficient (Ds/Do) are controlling factors for greenhouse gases emissions. Soil thermal properties correlated with greenhouse gases emissions when soil temperature could not. The study has found that predicted Ds/Do and correlate with greenhouse gas fluxes even when the air-filled porosity and the total porosity from which they are predicted did not. We have also showed that Ds/Do and can be predicted quickly from routine measurements of soil water and air and existing diffusivity models found in the literature. Agricultural practices do seriously impact greenhouse gases emissions as showed by the effect of mechanized tillage operations on soil physical properties and greenhouse gas fluxes in a corn and soybean fields. In fact, our results showed that tractor compaction increased soil resistance to penetration, water, bulk density and pore tortuosity while reducing air-filled porosity, total pore space and the soil gas diffusion coefficient. Changes in soil properties resulted in increased CO2, NO and N2O emissions. Finally, our results also confirmed that greenhouse gas fluxes vary tremendously in space and time. As estimates of greenhouse gas emissions are influenced by the data processing approach, differences between the different calculation approaches leads to uncertainty. Thus, techniques for developing better estimates are needed. We have showed that Geographic Information Systems (GIS), Global Positioning System (GPS), computer mapping and geo-statistics are technologies that can be used to better understand systems containing large amounts of spatial and temporal variability. Our GIS-based approach for quantifying CO2, CH4 and N2O fluxes from soil in agricultural fields showed that estimating (extrapolating) total greenhouse gas fluxes using the “standard” approach – multiplying the average flux value by the total field area – results in biased predictions of field total greenhouse gases emissions. In contrast, the GIS-based approach we developed produces an interpolated map portraying the spatial distribution of gas fluxes across the field from point measurements and later process the interpolated map produced to determine flux zones. Furthermore, processing, classification and modeling enables the computation of field total fluxes as the sum of fluxes in different zones, therefore taking into account the spatial variability of greenhouse gas fluxes.
- 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
Integer optimisation for the selection of a fantasy league cricket team
- Authors: Brettenny, Warren James
- Date: 2010
- Subjects: Sports -- Research -- Methodology , Teamwork (Sports) , Cricket players
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10565 , http://hdl.handle.net/10948/1230 , Sports -- Research -- Methodology , Teamwork (Sports) , Cricket players
- Description: Sports fans often scrutinise the team selection strategies employed by their favourite team's coach or selection panel. Many of these fans believe that they can perform the selection process far better than those tasked with the responsibility. Fantasy leagues, provide a platform for fans to test their hand at this selection procedure. Twenty20 cricket is a new and exciting form of cricket and has become very popular in recent years. This research focuses on bringing these concepts together by proposing a binary integer program to determine a team selection strategy for fantasy league cricket. This is done in a Twenty20 setting. The approach used in this study focuses on evaluating the effectiveness of previously developed performance measures in a fantasy league setting. Adjustments to these measures are made and new measures are proposed. These measures are then used to select a fantasy league team using a prospective approach. This is done to provide fantasy league participants with a mathematical procedure for fantasy league team selection.
- Full Text:
- Date Issued: 2010
- Authors: Brettenny, Warren James
- Date: 2010
- Subjects: Sports -- Research -- Methodology , Teamwork (Sports) , Cricket players
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10565 , http://hdl.handle.net/10948/1230 , Sports -- Research -- Methodology , Teamwork (Sports) , Cricket players
- Description: Sports fans often scrutinise the team selection strategies employed by their favourite team's coach or selection panel. Many of these fans believe that they can perform the selection process far better than those tasked with the responsibility. Fantasy leagues, provide a platform for fans to test their hand at this selection procedure. Twenty20 cricket is a new and exciting form of cricket and has become very popular in recent years. This research focuses on bringing these concepts together by proposing a binary integer program to determine a team selection strategy for fantasy league cricket. This is done in a Twenty20 setting. The approach used in this study focuses on evaluating the effectiveness of previously developed performance measures in a fantasy league setting. Adjustments to these measures are made and new measures are proposed. These measures are then used to select a fantasy league team using a prospective approach. This is done to provide fantasy league participants with a mathematical procedure for fantasy league team selection.
- Full Text:
- Date Issued: 2010
The importance of estuarine head waters for fishes in selected Eastern Cape systems, with particular emphasis on the influence of freshwater inflow, migration barriers and non-native predators on the juvenile and small fish component
- Authors: Wasserman, Ryan
- Date: 2010
- Subjects: Estuaries -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10685 , http://hdl.handle.net/10948/1457 , Estuaries -- South Africa -- Eastern Cape
- Description: The utilisation of estuary headwater environments by young estuary- and marine-spawned fish species was investigated together with the effects of riverflow alteration, in-stream barrier effects and non-native ichthyofauna on the nursery function of these habitats. The distribution and abundance of young estuary- and marine-spawned fish were sampled using seine and fyke nets in the headwater environments of four permanently open Eastern Cape systems, namely the Great Fish, Kowie, Kariega and Sundays Estuaries. Within the suite of study systems, the first of two case studies focussed on barrier effects of in-stream structures on fish migration. This was undertaken in the Sundays River. In the second case study, predation and competition dynamics of the non-native piscivorous Micropterus salmoides on estuary-dependent fish was investigated in the estuary headwater regions of the Kowie River system. In all four estuaries, young estuary-spawned fish species dominated the ichthyofaunal community followed by marine-spawned species, despite varied freshwater inflow resulting in headwaters varying in salinity from fresh to hypersaline. Fish community structure however, differed largely between estuaries, with both freshwater abstraction and unnatural elevation of freshwater into estuaries, as a result of inter-basin transfers, affecting these communities. In-stream structures were found to effect upstream movement of fish in two ways, dependent on the type of barrier. Partial (size-dependent) and complete (species-dependent) restriction to upstream migration of fish by causeway-type instream structures were observed. Weir-type in-stream structures acted as a complete barrier to most species, regardless of fish size. Predation of estuary- and marine-spawned fish species by large sized M. salmoides was recorded, although these fish did not contribute significantly to their diet during this study. However, the main dietary components found in smaller sized M. salmoides stomachs overlap with those of juvenile estuary- and marinespawned fish species, suggesting feeding competition between the juveniles of indigenous and non-native fish species.
- Full Text:
- Date Issued: 2010
- Authors: Wasserman, Ryan
- Date: 2010
- Subjects: Estuaries -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10685 , http://hdl.handle.net/10948/1457 , Estuaries -- South Africa -- Eastern Cape
- Description: The utilisation of estuary headwater environments by young estuary- and marine-spawned fish species was investigated together with the effects of riverflow alteration, in-stream barrier effects and non-native ichthyofauna on the nursery function of these habitats. The distribution and abundance of young estuary- and marine-spawned fish were sampled using seine and fyke nets in the headwater environments of four permanently open Eastern Cape systems, namely the Great Fish, Kowie, Kariega and Sundays Estuaries. Within the suite of study systems, the first of two case studies focussed on barrier effects of in-stream structures on fish migration. This was undertaken in the Sundays River. In the second case study, predation and competition dynamics of the non-native piscivorous Micropterus salmoides on estuary-dependent fish was investigated in the estuary headwater regions of the Kowie River system. In all four estuaries, young estuary-spawned fish species dominated the ichthyofaunal community followed by marine-spawned species, despite varied freshwater inflow resulting in headwaters varying in salinity from fresh to hypersaline. Fish community structure however, differed largely between estuaries, with both freshwater abstraction and unnatural elevation of freshwater into estuaries, as a result of inter-basin transfers, affecting these communities. In-stream structures were found to effect upstream movement of fish in two ways, dependent on the type of barrier. Partial (size-dependent) and complete (species-dependent) restriction to upstream migration of fish by causeway-type instream structures were observed. Weir-type in-stream structures acted as a complete barrier to most species, regardless of fish size. Predation of estuary- and marine-spawned fish species by large sized M. salmoides was recorded, although these fish did not contribute significantly to their diet during this study. However, the main dietary components found in smaller sized M. salmoides stomachs overlap with those of juvenile estuary- and marinespawned fish species, suggesting feeding competition between the juveniles of indigenous and non-native fish species.
- Full Text:
- Date Issued: 2010
A chemo-enzymatic process for the production of beta-thymidine, a key intermediate in antiretrovirol manufacture
- Gordon, Gregory Ernest Robert
- Authors: Gordon, Gregory Ernest Robert
- Date: 2010
- Subjects: HIV infections -- Treatment -- South Africa , HIV infections -- South Africa -- Prevention , Antiretroviral agents
- Language: English
- Type: Thesis , Doctoral , DTech
- Identifier: vital:10423 , http://hdl.handle.net/10948/d1016217
- Description: The socio-economic impact of HIV/AIDS on South Africa has resulted in lower gross domestic product, loss of skills in key sectors such as education, and increased health-care costs in providing access to treatment. Currently active pharmaceutical ingredients (API’s) such as stavudine (d4T) and azidothymidine (AZT) are imported from India and China, while formulation is conducted locally. A strategy was initiated between CSIR Biosciences and LIFElab under the auspices of Arvir Technologies to investigate the feasibility of local antiretroviral manufacture (d4T and AZT) or the manufacture of a key intermediate such as β- thymidine (dT). Several advantages associated with successful implementation of this strategy include ensuring a local supply of API’s, thus reducing reliance on procurement from foreign sources and reducing the effect of foreign exchange rate fluctuations on providing cost effective access to treatment. A local supply source would also reduce the imports and thus aid the balance of payments deficit, and in addition to this, provide stimulus in the local pharmaceutical manufacturing industry (which has been in decline for several decades), resulting in increased skills and employment opportunities. This thesis describes the development of a superior chemo-enzymatic process for the production of β-thymidine (72 percent yield, prior to isolation), a key intermediate in the preparation of anti-retrovirals. Alternative processes based purely on chemical or bioprocess transformations to prepare either 5-methyluridine (5-MU) or dT suffer from several disadvantages: lengthy transformations due to protection/deprotection strategies, low selectivties and product yields (30 percent in the chemical process) and isolation of the product from dilute process streams requiring the use of large uneconomical reactors (bioprocesss). This contributes significantly to the cost of d4T and AZT manufacture. Our novel chemoenzymatic process comprises of a biocatalytic reaction for the production of 5-MU, with subsequent chemical transformation into dT (3 steps) negating and circumventing the limitations of the chemical or bioprocess routes. During the course of this project development, the β-thymidine selling price declined from 175 $/kg (2005) to 100 $/kg (2008). However, the process described in this work is still competitive based on the current β- thymidine selling price of 100 $/kg. The process economics show that with further optimization and increasing the isolated dT yield from 70 percent to 90 percent, the variable cost decreases from 136 $/kg to 110 $/kg. The increase in isolated yield is highly probable, based on solubility data of β-thymidine. The decrease in β-thymidine selling price and technological improvement in dT manufacture should translate into lower API manufacture costs and more cost effective access to treatment. Our novel biocatalytic process producing 5-MU uses a coupled enzyme system employing PNP, Purine Nucleoside Phosphorylase and PyNP, Pyrimidine Nucleoside Phosphorylase. The overall transglycosylation reaction may be decoupled into the phosphorolysis reaction (PNP) and synthesis reaction (PyNP). During the phosphorolysis reaction, guanosine is converted into guanine and ribose-1-phosphate (R-1-P) in the presence of PNP enzyme. The reaction intermediate R-1-P is then coupled to thymine in the presence of PyNP enzyme during the synthesis reaction, producing 5-MU. The process was scaled up from lab-scale to bench-scale (10 - 20 L) and demonstrated to be robust and reproducible. This is evident from the average guanosine conversion (94.7 percent ± 2.03) and 5-MU yield (88.2 percent ± 6.21) and mole balance (104 percent ± 7.61) which were obtained at bench-scale (3 replicates, 10 L). The reaction was carried out at reactor productivities of between 7 – 11 g.L-1.h-1. The integration of the biocatalytic process and chemical processes was successfully carried out, showing that 5-MU produced using our novel biocatalytic process behaved similarly to commercially available 5- MU (ex. Dayang Chemicals, China). A PCT patent application (Ref. No. P44422PC01) on this chemo-enzymatic process has been filed and currently public private partnerships are being explored through Arvir Technologies to evaluate and validate this technology at one ton scale.
- Full Text:
- Date Issued: 2010
- Authors: Gordon, Gregory Ernest Robert
- Date: 2010
- Subjects: HIV infections -- Treatment -- South Africa , HIV infections -- South Africa -- Prevention , Antiretroviral agents
- Language: English
- Type: Thesis , Doctoral , DTech
- Identifier: vital:10423 , http://hdl.handle.net/10948/d1016217
- Description: The socio-economic impact of HIV/AIDS on South Africa has resulted in lower gross domestic product, loss of skills in key sectors such as education, and increased health-care costs in providing access to treatment. Currently active pharmaceutical ingredients (API’s) such as stavudine (d4T) and azidothymidine (AZT) are imported from India and China, while formulation is conducted locally. A strategy was initiated between CSIR Biosciences and LIFElab under the auspices of Arvir Technologies to investigate the feasibility of local antiretroviral manufacture (d4T and AZT) or the manufacture of a key intermediate such as β- thymidine (dT). Several advantages associated with successful implementation of this strategy include ensuring a local supply of API’s, thus reducing reliance on procurement from foreign sources and reducing the effect of foreign exchange rate fluctuations on providing cost effective access to treatment. A local supply source would also reduce the imports and thus aid the balance of payments deficit, and in addition to this, provide stimulus in the local pharmaceutical manufacturing industry (which has been in decline for several decades), resulting in increased skills and employment opportunities. This thesis describes the development of a superior chemo-enzymatic process for the production of β-thymidine (72 percent yield, prior to isolation), a key intermediate in the preparation of anti-retrovirals. Alternative processes based purely on chemical or bioprocess transformations to prepare either 5-methyluridine (5-MU) or dT suffer from several disadvantages: lengthy transformations due to protection/deprotection strategies, low selectivties and product yields (30 percent in the chemical process) and isolation of the product from dilute process streams requiring the use of large uneconomical reactors (bioprocesss). This contributes significantly to the cost of d4T and AZT manufacture. Our novel chemoenzymatic process comprises of a biocatalytic reaction for the production of 5-MU, with subsequent chemical transformation into dT (3 steps) negating and circumventing the limitations of the chemical or bioprocess routes. During the course of this project development, the β-thymidine selling price declined from 175 $/kg (2005) to 100 $/kg (2008). However, the process described in this work is still competitive based on the current β- thymidine selling price of 100 $/kg. The process economics show that with further optimization and increasing the isolated dT yield from 70 percent to 90 percent, the variable cost decreases from 136 $/kg to 110 $/kg. The increase in isolated yield is highly probable, based on solubility data of β-thymidine. The decrease in β-thymidine selling price and technological improvement in dT manufacture should translate into lower API manufacture costs and more cost effective access to treatment. Our novel biocatalytic process producing 5-MU uses a coupled enzyme system employing PNP, Purine Nucleoside Phosphorylase and PyNP, Pyrimidine Nucleoside Phosphorylase. The overall transglycosylation reaction may be decoupled into the phosphorolysis reaction (PNP) and synthesis reaction (PyNP). During the phosphorolysis reaction, guanosine is converted into guanine and ribose-1-phosphate (R-1-P) in the presence of PNP enzyme. The reaction intermediate R-1-P is then coupled to thymine in the presence of PyNP enzyme during the synthesis reaction, producing 5-MU. The process was scaled up from lab-scale to bench-scale (10 - 20 L) and demonstrated to be robust and reproducible. This is evident from the average guanosine conversion (94.7 percent ± 2.03) and 5-MU yield (88.2 percent ± 6.21) and mole balance (104 percent ± 7.61) which were obtained at bench-scale (3 replicates, 10 L). The reaction was carried out at reactor productivities of between 7 – 11 g.L-1.h-1. The integration of the biocatalytic process and chemical processes was successfully carried out, showing that 5-MU produced using our novel biocatalytic process behaved similarly to commercially available 5- MU (ex. Dayang Chemicals, China). A PCT patent application (Ref. No. P44422PC01) on this chemo-enzymatic process has been filed and currently public private partnerships are being explored through Arvir Technologies to evaluate and validate this technology at one ton scale.
- Full Text:
- Date Issued: 2010
Social sustainability of biogas production in Sogwala village, lower Gweru district, Zimbabwe
- Authors: Dhliwayo, Nyaradzo
- Date: 2010
- Subjects: Biogas -- Zimbabwe , Biogas -- Developing countries , Sustainability -- Zimbabwe
- Language: English
- Type: Thesis , Masters , MA
- Identifier: vital:10648 , http://hdl.handle.net/10948/1459 , Biogas -- Zimbabwe , Biogas -- Developing countries , Sustainability -- Zimbabwe
- Description: The study has explored the social sustainability of biogas production in a local village of Sogwala which is located in the Lower Gweru District of Zimbabwe. The social sustainability assessment was based on the two concepts of social sustainability, namely human capital and social capital with particular emphasis on their respective elements and aspects. Qualitative and quantitative research methodology methods were used on a sample of 100 households using biogas and some key informants. Analysis of the variance was used to determine whether biogas production could account for any effects on the social and human capital elements of the Sogwala community. An overview and descriptive statistics analysis of the findings from the study have been provided where the biogas users’ experiences, perspectives are discussed and results were assessed. For instance, effects on the aspects of social capital elements which include increases in the social group membership of households after the introduction of biogas production the village; the performance of biogas energy as compared with other fuel sources in terms of accessibility, efficiency and the degree of labour needed for continual production of biogas. In view of the human capital element, assessment is done on the economic status per household before and after the production of biogas, where emphasis is placed on the aspects of employment, education and health indicators within the social sustainability context. The fundamental benefits from biogas production are considered and discussed with the technical issues surrounding biogas production also being presented. This is not just a technical unit for providing alternative energy supply, but it requires management, labour and knowledge, skills to operate on a daily basis for it to have an impact that can bring positive change to the daily social lives of both the young and old.
- Full Text:
- Date Issued: 2010
- Authors: Dhliwayo, Nyaradzo
- Date: 2010
- Subjects: Biogas -- Zimbabwe , Biogas -- Developing countries , Sustainability -- Zimbabwe
- Language: English
- Type: Thesis , Masters , MA
- Identifier: vital:10648 , http://hdl.handle.net/10948/1459 , Biogas -- Zimbabwe , Biogas -- Developing countries , Sustainability -- Zimbabwe
- Description: The study has explored the social sustainability of biogas production in a local village of Sogwala which is located in the Lower Gweru District of Zimbabwe. The social sustainability assessment was based on the two concepts of social sustainability, namely human capital and social capital with particular emphasis on their respective elements and aspects. Qualitative and quantitative research methodology methods were used on a sample of 100 households using biogas and some key informants. Analysis of the variance was used to determine whether biogas production could account for any effects on the social and human capital elements of the Sogwala community. An overview and descriptive statistics analysis of the findings from the study have been provided where the biogas users’ experiences, perspectives are discussed and results were assessed. For instance, effects on the aspects of social capital elements which include increases in the social group membership of households after the introduction of biogas production the village; the performance of biogas energy as compared with other fuel sources in terms of accessibility, efficiency and the degree of labour needed for continual production of biogas. In view of the human capital element, assessment is done on the economic status per household before and after the production of biogas, where emphasis is placed on the aspects of employment, education and health indicators within the social sustainability context. The fundamental benefits from biogas production are considered and discussed with the technical issues surrounding biogas production also being presented. This is not just a technical unit for providing alternative energy supply, but it requires management, labour and knowledge, skills to operate on a daily basis for it to have an impact that can bring positive change to the daily social lives of both the young and old.
- Full Text:
- Date Issued: 2010
Development of insulin resistance in a rat model and the effects of sutherlandia frutescens as treatment and prevention
- Authors: Mackenzie, Janine
- Date: 2010
- Subjects: Metabolic syndrome , Insulin resistance -- Animal models , Obesity -- Treatment
- Language: English
- Type: Thesis , Doctoral , DPhil
- Identifier: vital:10340 , http://hdl.handle.net/10948/d1016216
- Description: The global number of obese people has reached pandemic proportions. High caloric diets and reduced physical exercise are to blame for this growing epidemic. Obesity has a very complex association with several other metabolic disorders, such as insulin resistance (IR), diabetes mellitus type 2 (DMT2) and cardiovascular disease. This puts a huge burden on health care systems world wide and claims many lives. Sutherlandia frutescens is a traditionally used herb, which is known to have anti-diabetic properties. However, the direct mode of action of S. frutescens still remains to be elucidated. The aim of this study was to investigate the developmental stages of high fat diet (HFD)-induced IR, to illuminate the pathogenesis of IR with a focal point on modifications in the lipid metabolism. Furthermore, the effects of S. frutescens as a treatment or prevention drug for IR and associated metabolic changes were examined. Two sets of experiments were conducted on male Wistar rats. In the first experiment rats, one week post weaning received a low fat diet (LFD), high fat diet (HFD) or HFD supplemented with S. frutescens (50mg/kg BW/d). Rats were sacrificed at week 0, 1, 2, 4, 8 and 12 in the feeding regime. In a second experiment rats were fed with a LFD or a HFD for 12 weeks and treated thereafter with S. frutescens (50mg/kg BW/d), metformin (13mg/kg BW/d) or water (control) for 28 days. Rats in the second experiment were sacrificed at week 12 to confirm IR while concurrently run rats were sacrificed after 28 days of treatment. For all the experiments rats were anaesthetized, blood was removed and rats were dissected. Plasma samples were analyzed for insulin, glucose, blood lipid parameters and cytokines. Liver, muscle and adipose tissue were analyzed for glucose uptake, total lipid content, lipid profile and fatty acid profile. It was shown that the intake of HFD caused IR and hyperinsulinaemia. The developmental stages in experiment one confirmed that an increase in plasma free fatty acids preceeded the onset of IR. Plasma and tissue lipid parameters (free fatty acid-, triglyceride- and cholesterol concentrations) showed pathological modifications in the HFD group. An ectopic accumulation of fat was observed in muscle and liver, as well as a change in membrane fatty acid profile. The results for circulating cytokines were somewhat inconclusive. Rats supplemented with S. frutescens did not develop HFD-induced IR (study one) or IR was reversed (study two). S. frutescens treatment also resulted in positive changes in plasma and tissue lipid parameters. In summary, an animal model for HFD-induced IR was established and the detrimental effect of elevated plasma FFA on glucose and lipid metabolism was observed. A novel discovery suggests that the anti-diabetic mode of action of S. frutescens is through modulation of lipid metabolism. It was also established that S. frutescens has the potential to prevent IR in vivo.
- Full Text:
- Date Issued: 2010
- Authors: Mackenzie, Janine
- Date: 2010
- Subjects: Metabolic syndrome , Insulin resistance -- Animal models , Obesity -- Treatment
- Language: English
- Type: Thesis , Doctoral , DPhil
- Identifier: vital:10340 , http://hdl.handle.net/10948/d1016216
- Description: The global number of obese people has reached pandemic proportions. High caloric diets and reduced physical exercise are to blame for this growing epidemic. Obesity has a very complex association with several other metabolic disorders, such as insulin resistance (IR), diabetes mellitus type 2 (DMT2) and cardiovascular disease. This puts a huge burden on health care systems world wide and claims many lives. Sutherlandia frutescens is a traditionally used herb, which is known to have anti-diabetic properties. However, the direct mode of action of S. frutescens still remains to be elucidated. The aim of this study was to investigate the developmental stages of high fat diet (HFD)-induced IR, to illuminate the pathogenesis of IR with a focal point on modifications in the lipid metabolism. Furthermore, the effects of S. frutescens as a treatment or prevention drug for IR and associated metabolic changes were examined. Two sets of experiments were conducted on male Wistar rats. In the first experiment rats, one week post weaning received a low fat diet (LFD), high fat diet (HFD) or HFD supplemented with S. frutescens (50mg/kg BW/d). Rats were sacrificed at week 0, 1, 2, 4, 8 and 12 in the feeding regime. In a second experiment rats were fed with a LFD or a HFD for 12 weeks and treated thereafter with S. frutescens (50mg/kg BW/d), metformin (13mg/kg BW/d) or water (control) for 28 days. Rats in the second experiment were sacrificed at week 12 to confirm IR while concurrently run rats were sacrificed after 28 days of treatment. For all the experiments rats were anaesthetized, blood was removed and rats were dissected. Plasma samples were analyzed for insulin, glucose, blood lipid parameters and cytokines. Liver, muscle and adipose tissue were analyzed for glucose uptake, total lipid content, lipid profile and fatty acid profile. It was shown that the intake of HFD caused IR and hyperinsulinaemia. The developmental stages in experiment one confirmed that an increase in plasma free fatty acids preceeded the onset of IR. Plasma and tissue lipid parameters (free fatty acid-, triglyceride- and cholesterol concentrations) showed pathological modifications in the HFD group. An ectopic accumulation of fat was observed in muscle and liver, as well as a change in membrane fatty acid profile. The results for circulating cytokines were somewhat inconclusive. Rats supplemented with S. frutescens did not develop HFD-induced IR (study one) or IR was reversed (study two). S. frutescens treatment also resulted in positive changes in plasma and tissue lipid parameters. In summary, an animal model for HFD-induced IR was established and the detrimental effect of elevated plasma FFA on glucose and lipid metabolism was observed. A novel discovery suggests that the anti-diabetic mode of action of S. frutescens is through modulation of lipid metabolism. It was also established that S. frutescens has the potential to prevent IR in vivo.
- 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
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
The management of extralimital giraffe (Giraffa Camelopardalis) in the mosaic thicket of Southern Cape, South Africa
- Authors: Cornelius, Andri Judith
- Date: 2010
- Subjects: Giraffe -- South Africa -- Eastern Cape , Giraffe -- Food -- South Africa -- Eastern Cape , Giraffe -- Ecology -- South Africa -- Eastern Cape , Animal behavior , Animal-plant relationships -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MTech
- Identifier: vital:10749 , http://hdl.handle.net/10948/1345 , Giraffe -- South Africa -- Eastern Cape , Giraffe -- Food -- South Africa -- Eastern Cape , Giraffe -- Ecology -- South Africa -- Eastern Cape , Animal behavior , Animal-plant relationships -- South Africa -- Eastern Cape
- Description: The giraffe at Nyaru were found to be browsers who made little use of graze during the study period. They utilized a diversity of 20 browse species. Two species, Acacia karroo and Acacia cyclops, formed the bulk of the giraffe diet throughout the year. A definite seasonal dietary shift was evident. A. karroo was favoured in summer and autumn and formed the main food species in spring, summer and autumn. A. cyclops was favoured throughout the study, but its contribution to the diet increased during winter when less A. karroo was consumed. This seasonal shift is related to the deciduous nature of Acacia karroo. Although a seasonal shift in species contribution to giraffe diet has been observed in many other giraffe feeding studies, no studies on giraffe feeding have been done in the Mosaic Thicket of the southern Cape. The ecological browsing capacity for giraffe in thicket was estimated to be between 0.020 BU/ha and 0.095 BU/ha. The browsing capacity for giraffe at Nyaru, based on the available phytomass 2–5 m above the ground, was estimated using those species that formed the bulk of the giraffe diet, and amounted to 0.063 BU/ha. A maximum of three giraffe could thus be stocked on the 157 ha of suitable giraffe habitat on Nyaru. This stocking rate recommendation lies within the range commonly recommended by local consultants for giraffe introductions into the southern Cape. Their recommendations are, however, not based on quantitative assessments such as performed in this study. The recommendation of this study should not be applied as a fixed ecological capacity for giraffe in thicket, but should be seen as a starting point in the adaptive management cycle. Ongoing monitoring of parameters, such as herbaceous composition and phytomass; as well as the condition of key browse species, is strongly advocated. A. karroo was browsed significantly more and carried significantly fewer pods per tree at a heavily used site compared to a lightly used site. Heavy browsing thus appears to affect the reproductive success of A. karroo significantly. Fewer pods are likely to lead to lower regeneration and thus reduced density of A. karroo. Whether A. karroo will maintain its dominance within the thicket vii community in the long run will be related to how individual plants survive and reproduce and if some can escape from herbivory. Fewer G. occidentalis were clumped with other species at the heavily used site compared to the lightly used site. This could possibly be attributed to the fact that intense browsing pressure at the heavily used site caused protective clumps to be eaten away, thus exposing G. occidentalis to higher ungulate browsing. Previous studies have found that nurse shrubs protect G. occidentalis against ungulate browsing. G. occidentalis was browsed significantly more at the heavily used site compared to the lightly used site. There was a general trend of fewer fruits at the heavily used site compared to the lightly used site, while fruits were absent on G. occidentalis growing alone at the heavily used site. This suggests a negative effect of heavy browsing on plant reproductive success and emphasizes the importance of nurse plants for the successful recruitment and hence long term prevalence of G. occidentalis in Mosaic Thicket.
- Full Text:
- Date Issued: 2010
- Authors: Cornelius, Andri Judith
- Date: 2010
- Subjects: Giraffe -- South Africa -- Eastern Cape , Giraffe -- Food -- South Africa -- Eastern Cape , Giraffe -- Ecology -- South Africa -- Eastern Cape , Animal behavior , Animal-plant relationships -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MTech
- Identifier: vital:10749 , http://hdl.handle.net/10948/1345 , Giraffe -- South Africa -- Eastern Cape , Giraffe -- Food -- South Africa -- Eastern Cape , Giraffe -- Ecology -- South Africa -- Eastern Cape , Animal behavior , Animal-plant relationships -- South Africa -- Eastern Cape
- Description: The giraffe at Nyaru were found to be browsers who made little use of graze during the study period. They utilized a diversity of 20 browse species. Two species, Acacia karroo and Acacia cyclops, formed the bulk of the giraffe diet throughout the year. A definite seasonal dietary shift was evident. A. karroo was favoured in summer and autumn and formed the main food species in spring, summer and autumn. A. cyclops was favoured throughout the study, but its contribution to the diet increased during winter when less A. karroo was consumed. This seasonal shift is related to the deciduous nature of Acacia karroo. Although a seasonal shift in species contribution to giraffe diet has been observed in many other giraffe feeding studies, no studies on giraffe feeding have been done in the Mosaic Thicket of the southern Cape. The ecological browsing capacity for giraffe in thicket was estimated to be between 0.020 BU/ha and 0.095 BU/ha. The browsing capacity for giraffe at Nyaru, based on the available phytomass 2–5 m above the ground, was estimated using those species that formed the bulk of the giraffe diet, and amounted to 0.063 BU/ha. A maximum of three giraffe could thus be stocked on the 157 ha of suitable giraffe habitat on Nyaru. This stocking rate recommendation lies within the range commonly recommended by local consultants for giraffe introductions into the southern Cape. Their recommendations are, however, not based on quantitative assessments such as performed in this study. The recommendation of this study should not be applied as a fixed ecological capacity for giraffe in thicket, but should be seen as a starting point in the adaptive management cycle. Ongoing monitoring of parameters, such as herbaceous composition and phytomass; as well as the condition of key browse species, is strongly advocated. A. karroo was browsed significantly more and carried significantly fewer pods per tree at a heavily used site compared to a lightly used site. Heavy browsing thus appears to affect the reproductive success of A. karroo significantly. Fewer pods are likely to lead to lower regeneration and thus reduced density of A. karroo. Whether A. karroo will maintain its dominance within the thicket vii community in the long run will be related to how individual plants survive and reproduce and if some can escape from herbivory. Fewer G. occidentalis were clumped with other species at the heavily used site compared to the lightly used site. This could possibly be attributed to the fact that intense browsing pressure at the heavily used site caused protective clumps to be eaten away, thus exposing G. occidentalis to higher ungulate browsing. Previous studies have found that nurse shrubs protect G. occidentalis against ungulate browsing. G. occidentalis was browsed significantly more at the heavily used site compared to the lightly used site. There was a general trend of fewer fruits at the heavily used site compared to the lightly used site, while fruits were absent on G. occidentalis growing alone at the heavily used site. This suggests a negative effect of heavy browsing on plant reproductive success and emphasizes the importance of nurse plants for the successful recruitment and hence long term prevalence of G. occidentalis in Mosaic Thicket.
- 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
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 geology of the Witteberg group, Cape supergroup, with specific focus on the Perdepoort member as a potential silica source
- Authors: Olivier, Wernich Corné
- Date: 2010
- Subjects: Geology -- South Africa -- Witteberg Group , Stratiography -- South Africa -- Witteberg Group , Silicate minerals -- South Africa -- Witteberg Group
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10660 , http://hdl.handle.net/10948/1386 , http://hdl.handle.net/10948/d1012889 , Geology -- South Africa -- Witteberg Group , Stratiography -- South Africa -- Witteberg Group , Silicate minerals -- South Africa -- Witteberg Group
- Description: Selected outcrops of the Upper Devonian to Lower Carboniferous, Witteberg Group, Cape Supergroup were mineralogically and structurally analyzed. The study area is located approximately 30km northwest of Kirkwood and 10km south of Darlington Dam, Eastern Cape, South Africa. Strata predominantly consist of arenaceous Witpoort Formation, which includes the Perdepoort, and Rooirand Members. The Perdepoort Member is a thinly bedded quartzite also known as the "white streak". The Rooirand Member quartzite is a highly iron stained red-brown quartzite. The dark-grey, pyritic rich shales of the Kweekvlei Formation overlie the Witpoort Formation in the southern half of the study site. These shales are highly deformed and display closely spaced thrust faults and close folds. The study area encapsulates a range of folding from tight to open folds. Faulting consists of low angle north verging thrust fault, south verging back thrusts, south and north dipping normal faults, and strike-slip faults. Closely spaced, fore-land verging thrusts faults predominate over hinterland verging back thrusts. Normal faulting post-dates thrust faulting and utilized weaknesses in axial planar cleavage and in certain instances existing thrust fault planes. Strike-slip faulting post-dates thrusting and has in places reactivated pre-existing thrust fault planes. Macro scale folding includes overturned synclines and large anticlines which have been eroded, exposing older strata. Fold axes plunge at low to moderate angles west-southwest. This correlates with tension gashes which indicate north westward directed forces. Eastward directed forces are confirmed by the presence of tension gashes and strike-slip movement. The local geology displays north westward directed compression followed by strike-slip movement. Normal faulting post-dates all other structures and is associated with the Mesozoic break-up of Gondwana. The Perdepoort Member was sampled along strike, at different outcrop latitudes. Seven samples were selected for scanning electron microscope analysis. Samples are composed almost entirely of quartz; accessories include, biotite, muscovite, sericite, baryte, and apatite. Epigenetic hematite is present along cracks within certain samples Epigenetic hematite occur along cracks with oxides and phosphates in the form of rutile, apatite and monazite present in a number of samples. When compared to other silica extraction operations the Perdepoort Member appears viable for explotation. However, for the solar cell industry the purity of this horizon is clearly far below that required for industy.
- Full Text:
- Date Issued: 2010
- Authors: Olivier, Wernich Corné
- Date: 2010
- Subjects: Geology -- South Africa -- Witteberg Group , Stratiography -- South Africa -- Witteberg Group , Silicate minerals -- South Africa -- Witteberg Group
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10660 , http://hdl.handle.net/10948/1386 , http://hdl.handle.net/10948/d1012889 , Geology -- South Africa -- Witteberg Group , Stratiography -- South Africa -- Witteberg Group , Silicate minerals -- South Africa -- Witteberg Group
- Description: Selected outcrops of the Upper Devonian to Lower Carboniferous, Witteberg Group, Cape Supergroup were mineralogically and structurally analyzed. The study area is located approximately 30km northwest of Kirkwood and 10km south of Darlington Dam, Eastern Cape, South Africa. Strata predominantly consist of arenaceous Witpoort Formation, which includes the Perdepoort, and Rooirand Members. The Perdepoort Member is a thinly bedded quartzite also known as the "white streak". The Rooirand Member quartzite is a highly iron stained red-brown quartzite. The dark-grey, pyritic rich shales of the Kweekvlei Formation overlie the Witpoort Formation in the southern half of the study site. These shales are highly deformed and display closely spaced thrust faults and close folds. The study area encapsulates a range of folding from tight to open folds. Faulting consists of low angle north verging thrust fault, south verging back thrusts, south and north dipping normal faults, and strike-slip faults. Closely spaced, fore-land verging thrusts faults predominate over hinterland verging back thrusts. Normal faulting post-dates thrust faulting and utilized weaknesses in axial planar cleavage and in certain instances existing thrust fault planes. Strike-slip faulting post-dates thrusting and has in places reactivated pre-existing thrust fault planes. Macro scale folding includes overturned synclines and large anticlines which have been eroded, exposing older strata. Fold axes plunge at low to moderate angles west-southwest. This correlates with tension gashes which indicate north westward directed forces. Eastward directed forces are confirmed by the presence of tension gashes and strike-slip movement. The local geology displays north westward directed compression followed by strike-slip movement. Normal faulting post-dates all other structures and is associated with the Mesozoic break-up of Gondwana. The Perdepoort Member was sampled along strike, at different outcrop latitudes. Seven samples were selected for scanning electron microscope analysis. Samples are composed almost entirely of quartz; accessories include, biotite, muscovite, sericite, baryte, and apatite. Epigenetic hematite is present along cracks within certain samples Epigenetic hematite occur along cracks with oxides and phosphates in the form of rutile, apatite and monazite present in a number of samples. When compared to other silica extraction operations the Perdepoort Member appears viable for explotation. However, for the solar cell industry the purity of this horizon is clearly far below that required for industy.
- Full Text:
- Date Issued: 2010
White rhinoceros populations in the Eastern Cape: distribution, performance and diet
- Authors: Kraai, Manqhai
- Date: 2010
- Subjects: Rhinoceroses -- South Africa -- Eastern Cape , Rhinoceroses -- Habitat -- South Africa -- Eastern Cape , Rhinoceroses -- Behavior -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10692 , http://hdl.handle.net/10948/1297 , Rhinoceroses -- South Africa -- Eastern Cape , Rhinoceroses -- Habitat -- South Africa -- Eastern Cape , Rhinoceroses -- Behavior -- South Africa -- Eastern Cape
- Description: White rhino populations did not occur historically in the Eastern Cape, South Africa (Skead 2007). They have, however, been introduced into a number of reserves within the Eastern Cape for reasons such as conservation, ecotourism and hunting. Based on the literature on the species, it was hypothesized that white rhinoceros would have been introduced in grass-rich habitats, as they are bulk grazers. White rhinoceros populations were further assessed in terms of population establishment by outlining their distribution and performance since introductions and this was done by extracting intercalving intervals from the literature of native populations and comparing such to the Eastern Cape intercalving intervals, extracted from data obtained from landowners. Finally, the diet of white rhinoceros was quantified, in order to identify plant species potentially at risk from white rhino herbivory using Shamwari Private Game Reserve as a study site. The distribution of white rhino in the Eastern Cape varies in terms of habitat and vegetation types. The majority (62%) of white rhino are located in grass-poor habitats and are primarily there for ecotourism purposes with only a small percentage kept for conservation purposes in the state reserves. Limited data for intercalving intervals of the white rhino in Eastern Cape are comparable to those of populations in the natural range. The ex situ conservation of white rhino in the Eastern Cape proved to be a success as population numbers have increased markedly since the first survey was conducted (Buijs 1999). The diet section of this study showed that white rhino are highly selective grazers even in a grass-limiting habitat which is dominated by browse. Twenty-five principal dietary items were identified in their diet, with only six being preferred in autumn. These are thus the plant species that could be the most vulnerable to white rhino herbivory. Supplementary food provision plays a big part of the feeding scheme of white rhino in Shamwari Private Game Reserve during the winter season, revealing the little confidence that the management has on natural resource availability to the animals at this time. These findings support the value of the Eastern Cape for ex situ conservation of white rhino, and highlight plant species potentially at risk. There is a need for further research on the population performance of these animals, and the plants identified here need to be monitored to assess white rhino impact on the vegetation
- Full Text:
- Date Issued: 2010
- Authors: Kraai, Manqhai
- Date: 2010
- Subjects: Rhinoceroses -- South Africa -- Eastern Cape , Rhinoceroses -- Habitat -- South Africa -- Eastern Cape , Rhinoceroses -- Behavior -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10692 , http://hdl.handle.net/10948/1297 , Rhinoceroses -- South Africa -- Eastern Cape , Rhinoceroses -- Habitat -- South Africa -- Eastern Cape , Rhinoceroses -- Behavior -- South Africa -- Eastern Cape
- Description: White rhino populations did not occur historically in the Eastern Cape, South Africa (Skead 2007). They have, however, been introduced into a number of reserves within the Eastern Cape for reasons such as conservation, ecotourism and hunting. Based on the literature on the species, it was hypothesized that white rhinoceros would have been introduced in grass-rich habitats, as they are bulk grazers. White rhinoceros populations were further assessed in terms of population establishment by outlining their distribution and performance since introductions and this was done by extracting intercalving intervals from the literature of native populations and comparing such to the Eastern Cape intercalving intervals, extracted from data obtained from landowners. Finally, the diet of white rhinoceros was quantified, in order to identify plant species potentially at risk from white rhino herbivory using Shamwari Private Game Reserve as a study site. The distribution of white rhino in the Eastern Cape varies in terms of habitat and vegetation types. The majority (62%) of white rhino are located in grass-poor habitats and are primarily there for ecotourism purposes with only a small percentage kept for conservation purposes in the state reserves. Limited data for intercalving intervals of the white rhino in Eastern Cape are comparable to those of populations in the natural range. The ex situ conservation of white rhino in the Eastern Cape proved to be a success as population numbers have increased markedly since the first survey was conducted (Buijs 1999). The diet section of this study showed that white rhino are highly selective grazers even in a grass-limiting habitat which is dominated by browse. Twenty-five principal dietary items were identified in their diet, with only six being preferred in autumn. These are thus the plant species that could be the most vulnerable to white rhino herbivory. Supplementary food provision plays a big part of the feeding scheme of white rhino in Shamwari Private Game Reserve during the winter season, revealing the little confidence that the management has on natural resource availability to the animals at this time. These findings support the value of the Eastern Cape for ex situ conservation of white rhino, and highlight plant species potentially at risk. There is a need for further research on the population performance of these animals, and the plants identified here need to be monitored to assess white rhino impact on the vegetation
- Full Text:
- Date Issued: 2010
Customer profiling using a service-orientated architecture
- Authors: Ntawanga, Felix
- Date: 2010
- Subjects: Consumer profiling -- South Africa , Consumer behavior -- South Africa , Service-oriented architecture (Computer science) , Electronic commerce -- South Africa -- Computer programs
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10464 , http://hdl.handle.net/10948/1146 , Consumer profiling -- South Africa , Consumer behavior -- South Africa , Service-oriented architecture (Computer science) , Electronic commerce -- South Africa -- Computer programs
- Description: Customer profiling has recently gained much recognition in the e-commerce domain because of the benefits it is capable of bringing to online business. Customer profiling has been implemented in various systems development approaches such as in a client-server environment. Recently there has been an increase in the number of organisations adopting and implementing e-commerce systems using service-oriented architecture (SOA) principles. This research set out to determine how a customer profile can be implemented using open source SOA implementation tools, and how SOA-based customer profiles can be utilised to provide appropriate personalisation in an SOA environment. The research further endeavoured to complete a comparative study on customer profile implementation in two different architectures, namely SOA and client-server. An extensive literature review was conducted on SOA, customer profiling and e-commerce systems development. SOA enabling technologies, such as, web services, enterprise service bus (ESB) and open source Sun Java SOA implementation tools, for example, Open ESB, GlassFish application server and Netbeans IDE were analysed. A Java web services-based customer profiling system was prototyped following SOA design principles. An end-user evaluation survey was conducted using eye tracking with a sample of 30 participants. The evaluation was done on two e-commerce systems with the same interface but running on two different customer profile back-ends, SOA and client-server. The results show that participants did not experience significant difference between the two systems, however, eye tracking results showed a significant difference between the two systems. The research concluded that customer profiling using SOA offers more benefits than implementations using other architectures such as client-server. SOA component-based development proved to be easier to manage, develop, integrate and improves interoperability between different technologies. The research brought together necessary techniques and technologies that organisations can use to implement SOA. Using SOA, organisations can integrate and utilise different technologies seamlessly to achieve business goals.
- Full Text:
- Date Issued: 2010
- Authors: Ntawanga, Felix
- Date: 2010
- Subjects: Consumer profiling -- South Africa , Consumer behavior -- South Africa , Service-oriented architecture (Computer science) , Electronic commerce -- South Africa -- Computer programs
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10464 , http://hdl.handle.net/10948/1146 , Consumer profiling -- South Africa , Consumer behavior -- South Africa , Service-oriented architecture (Computer science) , Electronic commerce -- South Africa -- Computer programs
- Description: Customer profiling has recently gained much recognition in the e-commerce domain because of the benefits it is capable of bringing to online business. Customer profiling has been implemented in various systems development approaches such as in a client-server environment. Recently there has been an increase in the number of organisations adopting and implementing e-commerce systems using service-oriented architecture (SOA) principles. This research set out to determine how a customer profile can be implemented using open source SOA implementation tools, and how SOA-based customer profiles can be utilised to provide appropriate personalisation in an SOA environment. The research further endeavoured to complete a comparative study on customer profile implementation in two different architectures, namely SOA and client-server. An extensive literature review was conducted on SOA, customer profiling and e-commerce systems development. SOA enabling technologies, such as, web services, enterprise service bus (ESB) and open source Sun Java SOA implementation tools, for example, Open ESB, GlassFish application server and Netbeans IDE were analysed. A Java web services-based customer profiling system was prototyped following SOA design principles. An end-user evaluation survey was conducted using eye tracking with a sample of 30 participants. The evaluation was done on two e-commerce systems with the same interface but running on two different customer profile back-ends, SOA and client-server. The results show that participants did not experience significant difference between the two systems, however, eye tracking results showed a significant difference between the two systems. The research concluded that customer profiling using SOA offers more benefits than implementations using other architectures such as client-server. SOA component-based development proved to be easier to manage, develop, integrate and improves interoperability between different technologies. The research brought together necessary techniques and technologies that organisations can use to implement SOA. Using SOA, organisations can integrate and utilise different technologies seamlessly to achieve business goals.
- Full Text:
- Date Issued: 2010
The dynamics of Larval fish and Zooplankton assemblages in the Sundays Estuary, South Africa
- Authors: Sutherland, Kate
- Date: 2010
- Subjects: Marine zooplankton -- South Africa , Zooplankton -- South Africa , Fishes -- Larvae -- South Africa , Estuarine ecology -- South Africa , Estuaries -- South Africa -- Management , Environmental monitoring -- South Africa , Coastal zone management -- South Africa , Estuarine area conservation -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10697 , http://hdl.handle.net/10948/1119 , Marine zooplankton -- South Africa , Zooplankton -- South Africa , Fishes -- Larvae -- South Africa , Estuarine ecology -- South Africa , Estuaries -- South Africa -- Management , Environmental monitoring -- South Africa , Coastal zone management -- South Africa , Estuarine area conservation -- South Africa
- Description: The larval fish and zooplankton assemblages were studied in the permanently open Sundays Estuary on the south-east coast of South Africa, using standard boat-based plankton towing methods. A total of 8174 larval and early juvenile fishes were caught, representing 12 families and 23 taxa. The Clupeidae, Gobiidae and Blenniidae were the dominant fish families. Common species included Gilchristella aestuaria, Caffrogobius gilchristi, Omobranchus woodi, Liza dumerilii, Glossogobius callidus and Myxus capensis. Estuarine resident species (Category I) predominantly in the preflexion developmental stage, dominated the system. A total of 19 zooplankton taxa were recorded. Copepoda dominated the zooplankton community. Dominant species included Pseudodiaptomus hessei, Acartia longipatella, Halicyclops sp., Mesopodopsis wooldridgei, and the larvae of Paratylodiplax edwardsii and Hymenosoma orbiculare. Mean larval fish density showed similar trends seasonally, spatially and across salinity zones, with mean zooplankton density in the Sundays Estuary. Gut content analysis of five larval fish species: Gilchristella aestuaria, Pomadasys commersonnii, Monodactylus falciformis, Myxus capensis and Rhabdosargus holubi, revealed species specific diet and prey selection. Although larval fish diet contained a variety of prey items, guts were dominated by P. hessei, chironomid larvae, Corophium triaenonyx, copepod eggs and insect larvae. Physico-chemical drivers and the interactions between these two plankton communities provide information that enables a more holistic view of the dynamics occurring in the Sundays Estuary planktonic ecosystem.
- Full Text:
- Date Issued: 2010
- Authors: Sutherland, Kate
- Date: 2010
- Subjects: Marine zooplankton -- South Africa , Zooplankton -- South Africa , Fishes -- Larvae -- South Africa , Estuarine ecology -- South Africa , Estuaries -- South Africa -- Management , Environmental monitoring -- South Africa , Coastal zone management -- South Africa , Estuarine area conservation -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10697 , http://hdl.handle.net/10948/1119 , Marine zooplankton -- South Africa , Zooplankton -- South Africa , Fishes -- Larvae -- South Africa , Estuarine ecology -- South Africa , Estuaries -- South Africa -- Management , Environmental monitoring -- South Africa , Coastal zone management -- South Africa , Estuarine area conservation -- South Africa
- Description: The larval fish and zooplankton assemblages were studied in the permanently open Sundays Estuary on the south-east coast of South Africa, using standard boat-based plankton towing methods. A total of 8174 larval and early juvenile fishes were caught, representing 12 families and 23 taxa. The Clupeidae, Gobiidae and Blenniidae were the dominant fish families. Common species included Gilchristella aestuaria, Caffrogobius gilchristi, Omobranchus woodi, Liza dumerilii, Glossogobius callidus and Myxus capensis. Estuarine resident species (Category I) predominantly in the preflexion developmental stage, dominated the system. A total of 19 zooplankton taxa were recorded. Copepoda dominated the zooplankton community. Dominant species included Pseudodiaptomus hessei, Acartia longipatella, Halicyclops sp., Mesopodopsis wooldridgei, and the larvae of Paratylodiplax edwardsii and Hymenosoma orbiculare. Mean larval fish density showed similar trends seasonally, spatially and across salinity zones, with mean zooplankton density in the Sundays Estuary. Gut content analysis of five larval fish species: Gilchristella aestuaria, Pomadasys commersonnii, Monodactylus falciformis, Myxus capensis and Rhabdosargus holubi, revealed species specific diet and prey selection. Although larval fish diet contained a variety of prey items, guts were dominated by P. hessei, chironomid larvae, Corophium triaenonyx, copepod eggs and insect larvae. Physico-chemical drivers and the interactions between these two plankton communities provide information that enables a more holistic view of the dynamics occurring in the Sundays Estuary planktonic ecosystem.
- 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
Bioaccumulation and ecotoxicology of b-methylamino-l-alanine (BMAA) in model crop plants
- Niyonzima, Francois Niyongabo
- Authors: Niyonzima, Francois Niyongabo
- Date: 2010
- Subjects: Cyanobacteria , Environmental toxicology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10312 , http://hdl.handle.net/10948/1475 , Cyanobacteria , Environmental toxicology
- Description: Cyanobacteria are known to produce a variety of toxic compounds. β-N-methylamino-L-alanine (BMAA) is one of the neurotoxins produced by most cyanobacteria. BMAA has been implicated in amyotrophic lateral sclerosis / Parkinsonism dementia complex (ALS / PDC) and was suggested to contribute to this pathology after biomagnification and slow release of BMAA from a protein associated form. The uptake and accumulation of BMAA by the aquatic macrophyte Ceratophyllum demersum has recently been shown, but the consumption of aquatic macrophytes by humans is not typical. The uptake by, and accumulation in, crop plants (Nasturtium officinale and Daucus carota) was therefore investigated so as to establish the existence of any risk to humans from the consumption of plants irrigated with water from dams with high cyanobacterial biomass and therefore high BMAA levels. After the exposure to the BMAA through the growth medium, BMAA had no effect on growth and development of N. officinale and D. carota. The uptake and bioaccumulation of BMAA was observed in N. officinale and D. carota, and was found to be concentration-dependent. Both free and bound cellular BMAA was detected following BMAA exposure through the growth medium. The photosynthetic apparatus of N. officinale was not significantly damaged. The uptake and accumulation of BMAA in edible terrestrial plants may constitute another route of human exposure to BMAA; it may now be prudent to avoid spray irrigation of edible plants with waters from dams with high cyanobacterial biomass and therefore high BMAA levels. After uptake by plants, the cyanotoxins may induce oxidative stress. A recent study showed that BMAA has a significant inhibitory effect on the oxidative stress enzymes in C. demersum. Therefore, the toxicological effects on selected plants were investigated by a range of biochemical enzyme assays in order to establish the plant stress response to exogenous BMAA. The inhibition of antioxidant enzymes upon exposure of N. officinale to BMAA through the growth medium was observed. The inhibition of antioxidant defence enzymes by BMAA correlated with the BMAA bioaccumulation in N. officinale. Further investigations are needed to analyze the uptake, accumulation, and ecotoxicology of BMAA in other crop plants, and to examine the fate of BMAA in these plants particularly its distribution and metabolism.
- Full Text:
- Date Issued: 2010
- Authors: Niyonzima, Francois Niyongabo
- Date: 2010
- Subjects: Cyanobacteria , Environmental toxicology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10312 , http://hdl.handle.net/10948/1475 , Cyanobacteria , Environmental toxicology
- Description: Cyanobacteria are known to produce a variety of toxic compounds. β-N-methylamino-L-alanine (BMAA) is one of the neurotoxins produced by most cyanobacteria. BMAA has been implicated in amyotrophic lateral sclerosis / Parkinsonism dementia complex (ALS / PDC) and was suggested to contribute to this pathology after biomagnification and slow release of BMAA from a protein associated form. The uptake and accumulation of BMAA by the aquatic macrophyte Ceratophyllum demersum has recently been shown, but the consumption of aquatic macrophytes by humans is not typical. The uptake by, and accumulation in, crop plants (Nasturtium officinale and Daucus carota) was therefore investigated so as to establish the existence of any risk to humans from the consumption of plants irrigated with water from dams with high cyanobacterial biomass and therefore high BMAA levels. After the exposure to the BMAA through the growth medium, BMAA had no effect on growth and development of N. officinale and D. carota. The uptake and bioaccumulation of BMAA was observed in N. officinale and D. carota, and was found to be concentration-dependent. Both free and bound cellular BMAA was detected following BMAA exposure through the growth medium. The photosynthetic apparatus of N. officinale was not significantly damaged. The uptake and accumulation of BMAA in edible terrestrial plants may constitute another route of human exposure to BMAA; it may now be prudent to avoid spray irrigation of edible plants with waters from dams with high cyanobacterial biomass and therefore high BMAA levels. After uptake by plants, the cyanotoxins may induce oxidative stress. A recent study showed that BMAA has a significant inhibitory effect on the oxidative stress enzymes in C. demersum. Therefore, the toxicological effects on selected plants were investigated by a range of biochemical enzyme assays in order to establish the plant stress response to exogenous BMAA. The inhibition of antioxidant enzymes upon exposure of N. officinale to BMAA through the growth medium was observed. The inhibition of antioxidant defence enzymes by BMAA correlated with the BMAA bioaccumulation in N. officinale. Further investigations are needed to analyze the uptake, accumulation, and ecotoxicology of BMAA in other crop plants, and to examine the fate of BMAA in these plants particularly its distribution and metabolism.
- Full Text:
- Date Issued: 2010