Multivariate morphometric analysis and behaviour of honeybees (Apis Mellifera L.) in the southern regions of Ethiopia
- Authors: Amssalu, Argaw Bezabeh
- Date: 2003
- Subjects: Honeybee -- Ethiopia -- Behavior Honeybee -- Ethiopia
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5616 , http://hdl.handle.net/10962/d1003130
- Description: Morphometric and behavioural characteristics of honeybees, Apis mellifera were analysed using multivariate and bivariate statistical methods to characterise honeybees of southern Ethiopian region. A total of 33800 morphometric character measurements were taken from 2600 individual worker honeybees of 130 honeybee colonies collected at 26 sampling localities with an average inter-locality distance of 89km to determine the occurrence of morphoclusters. 117 experienced farmer beekeepers and beekeeping experts were interviewed on pre-tested questionnaire to investigate the behavioural characteristics of these honeybees in their respective areas. Morphometric characters associated with pigmentation and body size exhibited a higher discriminant power while forewing venation angles (B4, N23 and 026) lack discriminatory power to segregate honeybees in the southern Ethiopian region. Principal components and discriminant analyses using the most discriminatory morphological characters delineated four statistically distinct morphoclusters in the southern Ethiopian region: the smallest and yellow honeybees, A. m. woyi-gambella which are different from all African honeybees, occur in the western and southern lowlands; the small and yellowiest honeybees, Apis mellifera jemenitica in the eastern escarpment; the largest and darkest honeybees, Apis mellifera bandasii in the central and eastern highlands; and dark honeybees, Apis mellifera scutellata in the wet tropical forests. High intracolonial and intercolonial variances within and between the boundaries of the honeybee groups were detected. The former suggests areas of ecological instability, while the latter areas of transitional or natural hybridisation zones. These areas are characterised by transitional ecological zones having intermediate climate and physiography. Even though honeybees of the southern Ethiopian region are generally dark and small, they show a high tendency to reproductive swarming, migration and aggressiveness, great variation in pigmentation, size and behaviour were observed both within and between the groups. A. m. jemenitica honeybees have a high propensity to migration and less a tendency for reproductive swarming. A. m. bandasii and A. m. scutellata have a high inclination to reproductive swarming and the former has a lesser and the later intermediate propensity to migration. A. m. woyi-gambella honeybees have intermediate swarming and migration tendencies. These results revealed that reproductive swarming and migration are higher in resource-rich and resource-poor areas respectively. Honey plants of the central highlands of Ethiopia are predominantly herbaceous in nature and mainly grow on open and cultivated lands. The bulk of pollen collected came comparatively from few genera. Strong correlation was observed between the intensities of flowering and rainfall. Reproductive swarming and migration occur during high and low intensity of flowering respectively.
- Full Text:
- Date Issued: 2003
- Authors: Amssalu, Argaw Bezabeh
- Date: 2003
- Subjects: Honeybee -- Ethiopia -- Behavior Honeybee -- Ethiopia
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5616 , http://hdl.handle.net/10962/d1003130
- Description: Morphometric and behavioural characteristics of honeybees, Apis mellifera were analysed using multivariate and bivariate statistical methods to characterise honeybees of southern Ethiopian region. A total of 33800 morphometric character measurements were taken from 2600 individual worker honeybees of 130 honeybee colonies collected at 26 sampling localities with an average inter-locality distance of 89km to determine the occurrence of morphoclusters. 117 experienced farmer beekeepers and beekeeping experts were interviewed on pre-tested questionnaire to investigate the behavioural characteristics of these honeybees in their respective areas. Morphometric characters associated with pigmentation and body size exhibited a higher discriminant power while forewing venation angles (B4, N23 and 026) lack discriminatory power to segregate honeybees in the southern Ethiopian region. Principal components and discriminant analyses using the most discriminatory morphological characters delineated four statistically distinct morphoclusters in the southern Ethiopian region: the smallest and yellow honeybees, A. m. woyi-gambella which are different from all African honeybees, occur in the western and southern lowlands; the small and yellowiest honeybees, Apis mellifera jemenitica in the eastern escarpment; the largest and darkest honeybees, Apis mellifera bandasii in the central and eastern highlands; and dark honeybees, Apis mellifera scutellata in the wet tropical forests. High intracolonial and intercolonial variances within and between the boundaries of the honeybee groups were detected. The former suggests areas of ecological instability, while the latter areas of transitional or natural hybridisation zones. These areas are characterised by transitional ecological zones having intermediate climate and physiography. Even though honeybees of the southern Ethiopian region are generally dark and small, they show a high tendency to reproductive swarming, migration and aggressiveness, great variation in pigmentation, size and behaviour were observed both within and between the groups. A. m. jemenitica honeybees have a high propensity to migration and less a tendency for reproductive swarming. A. m. bandasii and A. m. scutellata have a high inclination to reproductive swarming and the former has a lesser and the later intermediate propensity to migration. A. m. woyi-gambella honeybees have intermediate swarming and migration tendencies. These results revealed that reproductive swarming and migration are higher in resource-rich and resource-poor areas respectively. Honey plants of the central highlands of Ethiopia are predominantly herbaceous in nature and mainly grow on open and cultivated lands. The bulk of pollen collected came comparatively from few genera. Strong correlation was observed between the intensities of flowering and rainfall. Reproductive swarming and migration occur during high and low intensity of flowering respectively.
- Full Text:
- Date Issued: 2003
Pyrroloiminoquinone metabolites from South African Latrunculid sponges
- Authors: Antunes, Edith Martins
- Date: 2003
- Subjects: Sponges -- South Africa PQQ (Biochemistry) Marine metabolites -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4340 , http://hdl.handle.net/10962/d1005001
- Description: An in depth chemical investigation of the major and minor pyrroloiminoquinone metabolites produced by four species of endemic South African Latrunculid sponges, collected from Algoa Bay and the Tsitsikamma Marine Reserve off the south eastern coast of South Africa, yielded eleven new and twelve known pyrroloiminoquinone metabolites. The structures of the new metabolites were determined using standard spectroscopic techniques. Tsitsikamma pedunculata was shown to contain 7,8-dehydro-3-dihydro-discorhabdin C (2.1), 14-bromo-7,8-dehydro-3-dihydro-discorhabdin C (2.2), discorhabdin S (2.3), 14-bromo-1-hydroxy-discorhabdin S (2.4), 1-bromo-2-hydroxy-4-debromo-discorhabdin S (2.5), and 2,4-debromo-3-dihydro-discorhabdin C (2.6), together with the known compounds 14-bromo-discorhabdin C (1.51), 14-bromo-3-dihydro-discorhabdin C (1.52) and 3-dihydro-discorhabdin C. The metabolites from T. pedunculata were characterised by the presence of a reduced C-3 carbonyl and bromination at C-14. Compounds isolated from a second Latrunculid sponge, Latrunculia lorii, ranged from a substituted bicyclic pyrrolecarboxylic acid, makaluvic acid A (1.47), to the simple tricyclic known pyrroloiminoquinones makaluvamine C (1.33) and damirone B (1.20) and the more complex discorhabdin D type metabolites, discorhabdin M (3.2), 1-amino discorhabdin D (3.3), 1-methoxy discorhabdin D (3.4) and 1-alanyl discorhabdin D (3.5). Discorhabdin G* (3.1) was also isolated and characterised. This is the first reported occurrence of the known compounds 1.20, 1.33 and 1.47 in a Latrunculia sponge. Discorhabdin and bis-pyrroloiminoquinone type compounds predominated in Tsitsikamma favus. Three known, tsitsikammamines A (1.71) and B (1.72), 1.52, and five new pyrroloiminoquinones, tsitsikammamine N-oxime (4.1), tsitsikammamine B N-oxime (4.2), 2.1, 2.4 and 2.6, were isolated from this sponge. A fourth Latrunculid sponge (Strongylodesma sp.) yielded three known compounds, discorhabdins A (1.57), D (1.61) and 1.53, and one new pyrroloiminoquinone 3.3. The dual role of these metabolites as cytotoxic agents and pigments resulted in an attempt to relate the photochemical properties of these metabolites to their cytotoxicity. The pyrroloiminoquinone metabolites studied exhibited moderate singlet oxygen quantum yields, while three compounds (1.57, 4.1 and 4.2) were shown to be capable of producing radicals at a wavelength of 532 nm. The possibility of a correlation between the electrochemical properties and anti-cancer (HCT-116) activity of selected pyrroloiminoquinones was explored. A study of the oesophageal and ovarian cytotoxicities of two pyrroloiminoquinones (1.57 and 1.72), together with an investigation into the intercalation and topoisomerase I inhibitory activity of the bis-pyrroloiminoquinones (1.71, 1.72, 4.1 and 4.2), are presented.
- Full Text:
- Date Issued: 2003
- Authors: Antunes, Edith Martins
- Date: 2003
- Subjects: Sponges -- South Africa PQQ (Biochemistry) Marine metabolites -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4340 , http://hdl.handle.net/10962/d1005001
- Description: An in depth chemical investigation of the major and minor pyrroloiminoquinone metabolites produced by four species of endemic South African Latrunculid sponges, collected from Algoa Bay and the Tsitsikamma Marine Reserve off the south eastern coast of South Africa, yielded eleven new and twelve known pyrroloiminoquinone metabolites. The structures of the new metabolites were determined using standard spectroscopic techniques. Tsitsikamma pedunculata was shown to contain 7,8-dehydro-3-dihydro-discorhabdin C (2.1), 14-bromo-7,8-dehydro-3-dihydro-discorhabdin C (2.2), discorhabdin S (2.3), 14-bromo-1-hydroxy-discorhabdin S (2.4), 1-bromo-2-hydroxy-4-debromo-discorhabdin S (2.5), and 2,4-debromo-3-dihydro-discorhabdin C (2.6), together with the known compounds 14-bromo-discorhabdin C (1.51), 14-bromo-3-dihydro-discorhabdin C (1.52) and 3-dihydro-discorhabdin C. The metabolites from T. pedunculata were characterised by the presence of a reduced C-3 carbonyl and bromination at C-14. Compounds isolated from a second Latrunculid sponge, Latrunculia lorii, ranged from a substituted bicyclic pyrrolecarboxylic acid, makaluvic acid A (1.47), to the simple tricyclic known pyrroloiminoquinones makaluvamine C (1.33) and damirone B (1.20) and the more complex discorhabdin D type metabolites, discorhabdin M (3.2), 1-amino discorhabdin D (3.3), 1-methoxy discorhabdin D (3.4) and 1-alanyl discorhabdin D (3.5). Discorhabdin G* (3.1) was also isolated and characterised. This is the first reported occurrence of the known compounds 1.20, 1.33 and 1.47 in a Latrunculia sponge. Discorhabdin and bis-pyrroloiminoquinone type compounds predominated in Tsitsikamma favus. Three known, tsitsikammamines A (1.71) and B (1.72), 1.52, and five new pyrroloiminoquinones, tsitsikammamine N-oxime (4.1), tsitsikammamine B N-oxime (4.2), 2.1, 2.4 and 2.6, were isolated from this sponge. A fourth Latrunculid sponge (Strongylodesma sp.) yielded three known compounds, discorhabdins A (1.57), D (1.61) and 1.53, and one new pyrroloiminoquinone 3.3. The dual role of these metabolites as cytotoxic agents and pigments resulted in an attempt to relate the photochemical properties of these metabolites to their cytotoxicity. The pyrroloiminoquinone metabolites studied exhibited moderate singlet oxygen quantum yields, while three compounds (1.57, 4.1 and 4.2) were shown to be capable of producing radicals at a wavelength of 532 nm. The possibility of a correlation between the electrochemical properties and anti-cancer (HCT-116) activity of selected pyrroloiminoquinones was explored. A study of the oesophageal and ovarian cytotoxicities of two pyrroloiminoquinones (1.57 and 1.72), together with an investigation into the intercalation and topoisomerase I inhibitory activity of the bis-pyrroloiminoquinones (1.71, 1.72, 4.1 and 4.2), are presented.
- Full Text:
- Date Issued: 2003
Biological control initiatives against Lantana camara L. (Verbenaceae) in South Africa : an assessment of the present status of the programme, and an evaluation of Coelocephalapion camarae Kissinger (Coleoptera: Brentidae) and Falconia intermedia (Distant) (Heteroptera: Miridae), two new candidate natural enemies for release on the weed
- Authors: Baars, Jan-Robert
- Date: 2003
- Subjects: Lantana camara Lantana camera -- South Africa Biological pest control agents -- South Africa Beetles -- South Africa Hemiptera -- South Africa Weeds -- Biological control -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5647 , http://hdl.handle.net/10962/d1005329
- Description: Lantana camara (lantana), a thicket-forming shrub, a number of different varieties of which were introduced into South Africa as ornamental plants but which has become a serious invasive weed. Conventional control measures for lantana are expensive and ineffective and it has therefore been targeted for biological control since 1961. To date, eleven biological control agent species have become established on lantana in South Africa. However, most agents persist at low densities and only occasionally impact plant populations. Three species regularly cause significant damage, but only reach sufficiently high numbers by midsummer after populations crash during the winter. Overall, the impact of the biological control programme on the weed is negligible and this has been ascribed to the poor selection of agents for release, the accumulation of native parasitoids, differences in insect preference for different varieties of the weed and variable climatic conditions over the weed’s range. This study suggests that the importance of varietal preferences has been over-estimated. A predictive bioclimatic modelling technique showed that most of the agents established in South Africa have a wide climatic tolerance and that the redistribution and importation of new climatypes of these agents will not improve the level of control. Additional agents are required to improve the biocontrol in the temperate conditions, and also to increase damage in the sub-tropical areas where most of the agents are established and where the weed retains its leaves year round. New candidate agents that possess biological attributes that favour a high intrinsic rate of increase, a high impact per individual and that improve the synchrony between the weed and the agent in climatic conditions that promote the seasonal leaflessness of plants should receive prior consideration. A survey in Jamaica indicated that additional biological control agents are available in the region of origin but that care should be taken to prioritise the most effective agents. The various selection systems currently available in weed biocontrol produce contradictory results in the priority assigned to candidate agents and a new selection system is proposed. The biology and host range of two new candidate natural enemies, the leaf-galling weevil, Coelocephalapion camarae and the leaf-sucking mirid, Falconia intermedia were investigated for the biocontrol of lantana. The studies indicated that these have considerable biocontrol potential, in that the weevil has a wide climatic tolerance and has the potential to survive the host leaflessness typical of temperate conditions, while the mirid has a high intrinsic rate of increase, and the potential for several generations a year. Both agents caused a high level of damage to the leaves, with the weevil galling the vascular tissue in the leaf-petiole and the mirid causing chlorotic speckling of the leaves. During laboratory trials both agents accepted indigenous species in the genus Lippia. However, under multiple choice conditions these agents showed a significant and strong oviposition preference for lantana. A risk assessment and post release field trials indicated that F. intermedia is likely to attack some Lippia species in the presence of lantana, but the levels of damage are predicted to be relatively low. A possible low incidence of damage to indigenous species was considered a justifiable ‘trade-off’ for the potentially marked impact on L. camara. Preference and performance studies on the two candidate agents suggested that most of the South African lantana varieties are suitable host plants. The mirid preferred certain varieties in multiple choice experiments, but this is unlikely to affect its impact under field conditions. Permission for release was accordingly sought for both species. Finally, the challenges facing the biological control programme and the potential for improving the control of L. camara in South Africa are considered.
- Full Text:
- Date Issued: 2003
- Authors: Baars, Jan-Robert
- Date: 2003
- Subjects: Lantana camara Lantana camera -- South Africa Biological pest control agents -- South Africa Beetles -- South Africa Hemiptera -- South Africa Weeds -- Biological control -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5647 , http://hdl.handle.net/10962/d1005329
- Description: Lantana camara (lantana), a thicket-forming shrub, a number of different varieties of which were introduced into South Africa as ornamental plants but which has become a serious invasive weed. Conventional control measures for lantana are expensive and ineffective and it has therefore been targeted for biological control since 1961. To date, eleven biological control agent species have become established on lantana in South Africa. However, most agents persist at low densities and only occasionally impact plant populations. Three species regularly cause significant damage, but only reach sufficiently high numbers by midsummer after populations crash during the winter. Overall, the impact of the biological control programme on the weed is negligible and this has been ascribed to the poor selection of agents for release, the accumulation of native parasitoids, differences in insect preference for different varieties of the weed and variable climatic conditions over the weed’s range. This study suggests that the importance of varietal preferences has been over-estimated. A predictive bioclimatic modelling technique showed that most of the agents established in South Africa have a wide climatic tolerance and that the redistribution and importation of new climatypes of these agents will not improve the level of control. Additional agents are required to improve the biocontrol in the temperate conditions, and also to increase damage in the sub-tropical areas where most of the agents are established and where the weed retains its leaves year round. New candidate agents that possess biological attributes that favour a high intrinsic rate of increase, a high impact per individual and that improve the synchrony between the weed and the agent in climatic conditions that promote the seasonal leaflessness of plants should receive prior consideration. A survey in Jamaica indicated that additional biological control agents are available in the region of origin but that care should be taken to prioritise the most effective agents. The various selection systems currently available in weed biocontrol produce contradictory results in the priority assigned to candidate agents and a new selection system is proposed. The biology and host range of two new candidate natural enemies, the leaf-galling weevil, Coelocephalapion camarae and the leaf-sucking mirid, Falconia intermedia were investigated for the biocontrol of lantana. The studies indicated that these have considerable biocontrol potential, in that the weevil has a wide climatic tolerance and has the potential to survive the host leaflessness typical of temperate conditions, while the mirid has a high intrinsic rate of increase, and the potential for several generations a year. Both agents caused a high level of damage to the leaves, with the weevil galling the vascular tissue in the leaf-petiole and the mirid causing chlorotic speckling of the leaves. During laboratory trials both agents accepted indigenous species in the genus Lippia. However, under multiple choice conditions these agents showed a significant and strong oviposition preference for lantana. A risk assessment and post release field trials indicated that F. intermedia is likely to attack some Lippia species in the presence of lantana, but the levels of damage are predicted to be relatively low. A possible low incidence of damage to indigenous species was considered a justifiable ‘trade-off’ for the potentially marked impact on L. camara. Preference and performance studies on the two candidate agents suggested that most of the South African lantana varieties are suitable host plants. The mirid preferred certain varieties in multiple choice experiments, but this is unlikely to affect its impact under field conditions. Permission for release was accordingly sought for both species. Finally, the challenges facing the biological control programme and the potential for improving the control of L. camara in South Africa are considered.
- Full Text:
- Date Issued: 2003
Development of integrated algal ponding systems in the treatment of wine distillery wastewaters
- Authors: Dekker, Leendert Gideon
- Date: 2003
- Subjects: Sewage -- Purification -- Anaerobic treatment Wine and wine making -- Waste disposal -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4062 , http://hdl.handle.net/10962/d1004530
- Description: In South Africa, wastewater disposal in the wine and distilling industry is undergoing a profound transformation as a result of fundamental changes in regulations and license requirements. To deal with this problem conventional Waste Stabilisation Ponding systems have been used by the industry together with irrigation and evaporation disposal practises. Although effective in the evaporation and containment disposal functions, these pond systems are generally not properly designed and/or managed, resulting in overloading and, at times, the generation of seriously offensive odour problems. Preliminary studies on the feasibility of utilising the Advanced Integrated Wastewater Ponding System as a core treatment technology in winery wastewater treatment were conducted. Results indicated that specific problems had to be addressed before successful ponding treatment could be achieved. This research programme undertook an investigation of the performance of a demonstration ponding system treating household sewage, which formed the basis of the research due to limited experience reported on ponds treating wine industry wastewaters. Malfunctions identified were in correlation with the preliminary winery waste ponding survey, which included unstable fermentation pit functions and inadequate nutrient removal. Retrofitting the fermentation pit with a nylon net across the rising water column resulted in improved retention of active anaerobic sludge, especially during periods of system start-up and/or organic overloading. An investigation into nutrient removal utilising algal biomass provided a valuable contribution towards development of an independent nutrient removal system. Harvested algal biomass was passively manipulated to release polysaccharides under anoxic conditions, with subsequent use as a carbon source by denitrifying organisms. Following denitrification, the still viable algal cells were introduced into a High Rate Algal Pond raceway for photosynthetically produced alkalinity. This high pH environment resulted in induced calcium phosphate mineral formation and subsequent precipitation, as well as effective ammonia stripping from the water. Based on the novel positive research outcomes a decision was made to proceed to the construction of a pilot-scale integrated ponding system treating wastewater from a wine lees factory. The system linked the Anaerobic Baffle Reactor, for pre-treatment, with the improved Advanced Integrated Wastewater Ponding System. The potential of this system has shown that a Waste Stabilisation Ponding system can be engineered to treat wine industry wastewaters and thereby effectively reduce the organic and nutrient loads, by using low-cost retrofitted upgrading unit operations. Valuable algal biomass may also be recovered as a by-product of the treatment process.
- Full Text:
- Date Issued: 2003
- Authors: Dekker, Leendert Gideon
- Date: 2003
- Subjects: Sewage -- Purification -- Anaerobic treatment Wine and wine making -- Waste disposal -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4062 , http://hdl.handle.net/10962/d1004530
- Description: In South Africa, wastewater disposal in the wine and distilling industry is undergoing a profound transformation as a result of fundamental changes in regulations and license requirements. To deal with this problem conventional Waste Stabilisation Ponding systems have been used by the industry together with irrigation and evaporation disposal practises. Although effective in the evaporation and containment disposal functions, these pond systems are generally not properly designed and/or managed, resulting in overloading and, at times, the generation of seriously offensive odour problems. Preliminary studies on the feasibility of utilising the Advanced Integrated Wastewater Ponding System as a core treatment technology in winery wastewater treatment were conducted. Results indicated that specific problems had to be addressed before successful ponding treatment could be achieved. This research programme undertook an investigation of the performance of a demonstration ponding system treating household sewage, which formed the basis of the research due to limited experience reported on ponds treating wine industry wastewaters. Malfunctions identified were in correlation with the preliminary winery waste ponding survey, which included unstable fermentation pit functions and inadequate nutrient removal. Retrofitting the fermentation pit with a nylon net across the rising water column resulted in improved retention of active anaerobic sludge, especially during periods of system start-up and/or organic overloading. An investigation into nutrient removal utilising algal biomass provided a valuable contribution towards development of an independent nutrient removal system. Harvested algal biomass was passively manipulated to release polysaccharides under anoxic conditions, with subsequent use as a carbon source by denitrifying organisms. Following denitrification, the still viable algal cells were introduced into a High Rate Algal Pond raceway for photosynthetically produced alkalinity. This high pH environment resulted in induced calcium phosphate mineral formation and subsequent precipitation, as well as effective ammonia stripping from the water. Based on the novel positive research outcomes a decision was made to proceed to the construction of a pilot-scale integrated ponding system treating wastewater from a wine lees factory. The system linked the Anaerobic Baffle Reactor, for pre-treatment, with the improved Advanced Integrated Wastewater Ponding System. The potential of this system has shown that a Waste Stabilisation Ponding system can be engineered to treat wine industry wastewaters and thereby effectively reduce the organic and nutrient loads, by using low-cost retrofitted upgrading unit operations. Valuable algal biomass may also be recovered as a by-product of the treatment process.
- Full Text:
- Date Issued: 2003
An analysis of the impact of democratization on debt-led growth : the Nigerian experience, 1970-2000
- Authors: Dinneya, Godson Eze
- Date: 2013-05-22
- Subjects: Debts, External -- Nigeria Nigeria -- Politics and government -- 1960- Nigeria -- Economic policy Nigeria -- Economic conditions -- 1960-
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:1074 , http://hdl.handle.net/10962/d1007807
- Description: The debt-for democracy hypothesis is that undemocratic governments were largely responsible for not only the accumulation but also poor management of externally sourced capital resources. External borrowing had therefore failed to lead to growth of the economies of debtor countries under undemocratic political leadership. Despite this explanation of the debt problem conventional empirical analyses of the debt-growth relationship did not include political institutional variables. This study investigates the relationship between democratization and debt-led growth, using Nigeria, a typical debtor country whose politics was dominated by 'undemocratic ' governance, as a case study. Two broad research questions are investigated namely, whether available data support a negative or positive contribution of debt to the growth of the Nigeria economy during the period 1970-2000; and ifso was there any link between the levels of democratization in Nigeria and debt-led growth. Using a census of major political events in Nigeria around four dimensions of democratization, four primary indices of democratization and one composite index were constructed for the period. Using the Taylor (1983) marginal conditions to gauge the contribution of external debt to the growth of the Nigerian economy, the study found that external debt is capable of playing a double edged sword on the performance of the economy. Positive contributions coincided with the periods when Nigeria's oil dominated foreign exchange revenues were robust, and/ or when debt management strategies were better articulated and vice versa. The analyses of the link between democratization and debt-led growth using both correlation and regression techniques, yielded different results in two definitional contexts of debt-led growth. When defined purely in terms of the Taylor marginal conditions for a positive contribution of debt to the economy of a borrowing nation, the results support the pessimist view that democratization impeded growth. On the contrary, when debt-led growth was defined in a broader sense to incorporate variables such as domestic savings and investment, foreign direct investments, public and private consumption and debt burden, there was strong evidence that debt-led growth performed beller at higher levels of democratization than other wise. The result using the narrow definition was found to be a direct consequence of the overriding influence of export performance in the Taylor conditions. With Nigeria's exports almost entirely dominated by extractive industry the result derived using the narrow definition confirmed the theoretical links between natural resource endowment and regime type on the one hand, and external capital and the nature of the host country 's industry on the other. In the first resource dependence allowed the political leadership to be more detached and less accountable to the electorate since they did not need to levy taxes. Secondly foreign investors concerned with security of their sunk investments in the extractive oil induslly in particular favoured continuity of powerfol regimes with less democratic content. In both findings one thing was common: democratization was associated more with those factors whose decreases affect growth positively than with those whose increases improve growth. The conclusion from this is that the impact of democratization is stronger with negative than with positive growth factors. In other words, while democratization may be supportive of growth its greater impact appears to be in limiting the factors that themselves limit growth. To benefit from the favourable impact of democratisation on debt-led growth therefore the study suggests that improvements in the democratisation process in Nigeria is needed It identifies political education as central to this improvement. A model is developed to show how improvements in the political institutional framework may trickle down, through an enabling environment that is capable of engendering growth-enhancing domestic and international responses to lead in the direction of debt-led growth.
- Full Text:
An analysis of the impact of democratization on debt-led growth : the Nigerian experience, 1970-2000
- Authors: Dinneya, Godson Eze
- Date: 2013-05-22
- Subjects: Debts, External -- Nigeria Nigeria -- Politics and government -- 1960- Nigeria -- Economic policy Nigeria -- Economic conditions -- 1960-
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:1074 , http://hdl.handle.net/10962/d1007807
- Description: The debt-for democracy hypothesis is that undemocratic governments were largely responsible for not only the accumulation but also poor management of externally sourced capital resources. External borrowing had therefore failed to lead to growth of the economies of debtor countries under undemocratic political leadership. Despite this explanation of the debt problem conventional empirical analyses of the debt-growth relationship did not include political institutional variables. This study investigates the relationship between democratization and debt-led growth, using Nigeria, a typical debtor country whose politics was dominated by 'undemocratic ' governance, as a case study. Two broad research questions are investigated namely, whether available data support a negative or positive contribution of debt to the growth of the Nigeria economy during the period 1970-2000; and ifso was there any link between the levels of democratization in Nigeria and debt-led growth. Using a census of major political events in Nigeria around four dimensions of democratization, four primary indices of democratization and one composite index were constructed for the period. Using the Taylor (1983) marginal conditions to gauge the contribution of external debt to the growth of the Nigerian economy, the study found that external debt is capable of playing a double edged sword on the performance of the economy. Positive contributions coincided with the periods when Nigeria's oil dominated foreign exchange revenues were robust, and/ or when debt management strategies were better articulated and vice versa. The analyses of the link between democratization and debt-led growth using both correlation and regression techniques, yielded different results in two definitional contexts of debt-led growth. When defined purely in terms of the Taylor marginal conditions for a positive contribution of debt to the economy of a borrowing nation, the results support the pessimist view that democratization impeded growth. On the contrary, when debt-led growth was defined in a broader sense to incorporate variables such as domestic savings and investment, foreign direct investments, public and private consumption and debt burden, there was strong evidence that debt-led growth performed beller at higher levels of democratization than other wise. The result using the narrow definition was found to be a direct consequence of the overriding influence of export performance in the Taylor conditions. With Nigeria's exports almost entirely dominated by extractive industry the result derived using the narrow definition confirmed the theoretical links between natural resource endowment and regime type on the one hand, and external capital and the nature of the host country 's industry on the other. In the first resource dependence allowed the political leadership to be more detached and less accountable to the electorate since they did not need to levy taxes. Secondly foreign investors concerned with security of their sunk investments in the extractive oil induslly in particular favoured continuity of powerfol regimes with less democratic content. In both findings one thing was common: democratization was associated more with those factors whose decreases affect growth positively than with those whose increases improve growth. The conclusion from this is that the impact of democratization is stronger with negative than with positive growth factors. In other words, while democratization may be supportive of growth its greater impact appears to be in limiting the factors that themselves limit growth. To benefit from the favourable impact of democratisation on debt-led growth therefore the study suggests that improvements in the democratisation process in Nigeria is needed It identifies political education as central to this improvement. A model is developed to show how improvements in the political institutional framework may trickle down, through an enabling environment that is capable of engendering growth-enhancing domestic and international responses to lead in the direction of debt-led growth.
- Full Text:
Isolation of antigenic peptides of Cowdria ruminantium and their encoding genes using a genome-derived phage display library
- Authors: Fehrsen, Jeanni
- Date: 2003
- Subjects: Bacteriophages -- Genetics Ruminants -- Diseases Heartwater
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3920 , http://hdl.handle.net/10962/d1003979
- Description: The development of new and effective vaccines and immunodiagnostic reagents requires the characterisation of antigenically relevant proteins and their interactions with the products of the immune system. Phage display technology was investigated as a means of elucidating some of the antigenic properties of the rickettsial parasite, Cowdria ruminantium (Cowdria). Randomly fragmented gene-derived libraries have been useful in elucidating viral and other epitopes, but only limited work has been done with entire genomes. A phage display library expressing a repertoire of Cowdria peptides was constructed. It was sufficiently large to represent the organism's genome, but lacked phages displaying peptides coded for by genes containing a Pvu II restriction enzyme site, including the one coding for the major antigenic protein 1 (MAP1). This was considered advantageous since MAP1 is immunodominant and has already been well characterised. Affinity selection with antibodies against Cowdria proteins other than MAP1 allowed several antibody-reactive peptides to be isolated. These selected sequences were placed in the context of the genome by screening a lambda bacteriophage library and by comparison with Cowdria DNA sequences. Apart from showing that antigenic mimics were present in the phage display library, six open reading frames encoding putative Cowdria proteins were identified. All had similarities to, or motifs in common with, membrane proteins and are thus likely to be exposed to the host's humoral immune system. Some of the proteins identified were larger than the antigens used to elicit the antibodies used for selection, probably as a result of the presence of cross-reactive epitopes. Despite limitations experienced when extending a fragmented-gene approach for epitope location to genomes, it was possible to identify an antigenic region on MAP1 by comparison with selected mimics. In addition, binding peptide sequences were identified with two monoclonal antibodies that had been raised against non-Cowdria antigens. An epitope on the VP7 protein of bluetongue virus was identified and peptides were found that reacted with a monoclonal antibody directed against malignant catarrhal fever virus. Thus, apart from being able to identify several potentially important Cowdria epitopes and genes, the fragmented-genome library holds promise as a universal reagent for identifying useful mimics.
- Full Text:
- Date Issued: 2003
- Authors: Fehrsen, Jeanni
- Date: 2003
- Subjects: Bacteriophages -- Genetics Ruminants -- Diseases Heartwater
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3920 , http://hdl.handle.net/10962/d1003979
- Description: The development of new and effective vaccines and immunodiagnostic reagents requires the characterisation of antigenically relevant proteins and their interactions with the products of the immune system. Phage display technology was investigated as a means of elucidating some of the antigenic properties of the rickettsial parasite, Cowdria ruminantium (Cowdria). Randomly fragmented gene-derived libraries have been useful in elucidating viral and other epitopes, but only limited work has been done with entire genomes. A phage display library expressing a repertoire of Cowdria peptides was constructed. It was sufficiently large to represent the organism's genome, but lacked phages displaying peptides coded for by genes containing a Pvu II restriction enzyme site, including the one coding for the major antigenic protein 1 (MAP1). This was considered advantageous since MAP1 is immunodominant and has already been well characterised. Affinity selection with antibodies against Cowdria proteins other than MAP1 allowed several antibody-reactive peptides to be isolated. These selected sequences were placed in the context of the genome by screening a lambda bacteriophage library and by comparison with Cowdria DNA sequences. Apart from showing that antigenic mimics were present in the phage display library, six open reading frames encoding putative Cowdria proteins were identified. All had similarities to, or motifs in common with, membrane proteins and are thus likely to be exposed to the host's humoral immune system. Some of the proteins identified were larger than the antigens used to elicit the antibodies used for selection, probably as a result of the presence of cross-reactive epitopes. Despite limitations experienced when extending a fragmented-gene approach for epitope location to genomes, it was possible to identify an antigenic region on MAP1 by comparison with selected mimics. In addition, binding peptide sequences were identified with two monoclonal antibodies that had been raised against non-Cowdria antigens. An epitope on the VP7 protein of bluetongue virus was identified and peptides were found that reacted with a monoclonal antibody directed against malignant catarrhal fever virus. Thus, apart from being able to identify several potentially important Cowdria epitopes and genes, the fragmented-genome library holds promise as a universal reagent for identifying useful mimics.
- Full Text:
- Date Issued: 2003
Versions of confinement: Melville's bodies and the psychology of conquest
- Authors: Goddard, Kevin Graham
- Date: 2003
- Subjects: Melville, Herman, 1819-1891 -- Criticism and interpretation Human body in literature
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:2216 , http://hdl.handle.net/10962/d1002259
- Description: This thesis explores aspects of Melville’s presentation of both the whale and the human bodies in Moby-Dick and human bodies in other important novels. It argues that Melville uses his presentation of bodies to explore some of the versions of confinement those bodies experience, and by doing so, analyses the psychology which subtends that confinement. Throughout Melville’s works bodies are confined, both within literal spatial limits and by the psychology which creates and/or accepts these spatial limits. The thesis argues that perhaps the most important version of bodily confinement Melville addresses is the impulse to conquer bodies, both that of the other and one’s own. It adopts a largely psychoanalytic approach to interpreting bodies and their impulse to conquer, so that the body is seen to figure both in its actions and its external appearance the operations of the inner psyche. The figure of the body is equally prevalent in Melville’s exploration of nationalist conquest, where, as with Manifest Destiny and antebellum expansionism, the psychological and physical lack experienced by characters can be read as motivating factors in the ideology of conquest. A final important strand of the thesis is its argument in favour of a gradual shift in Melville’s interpretation of the value and possibility of genuine communion between human beings and between humans and the whale. One may read Typee as an attempt by Melville to explore the possibility of a this-worldly utopia in which human beings can return to a version of primitive interconnectedness. This exploration may be seen to be extended in Moby-Dick, particularly in Ishmael’s attempts to find communion with others and in some moments of encounter with the whales. The thesis uses phenomenology as a theory to interpret what Melville is trying to suggest in these moments of encounter. However, it argues, finally, that such encounter, or ‘intersubjectivity’ is eventually jettisoned, especially in the works after Moby-Dick. By the end of Melville’s life and work, any hope of an intersubjective utopia he may have harboured as a younger man have been removed in favour of a refusal actually to assert any final ‘truth’ about social, political or even religious experience. Billy Budd, his last body, is hanged, and his final word is silence.
- Full Text:
- Date Issued: 2003
- Authors: Goddard, Kevin Graham
- Date: 2003
- Subjects: Melville, Herman, 1819-1891 -- Criticism and interpretation Human body in literature
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:2216 , http://hdl.handle.net/10962/d1002259
- Description: This thesis explores aspects of Melville’s presentation of both the whale and the human bodies in Moby-Dick and human bodies in other important novels. It argues that Melville uses his presentation of bodies to explore some of the versions of confinement those bodies experience, and by doing so, analyses the psychology which subtends that confinement. Throughout Melville’s works bodies are confined, both within literal spatial limits and by the psychology which creates and/or accepts these spatial limits. The thesis argues that perhaps the most important version of bodily confinement Melville addresses is the impulse to conquer bodies, both that of the other and one’s own. It adopts a largely psychoanalytic approach to interpreting bodies and their impulse to conquer, so that the body is seen to figure both in its actions and its external appearance the operations of the inner psyche. The figure of the body is equally prevalent in Melville’s exploration of nationalist conquest, where, as with Manifest Destiny and antebellum expansionism, the psychological and physical lack experienced by characters can be read as motivating factors in the ideology of conquest. A final important strand of the thesis is its argument in favour of a gradual shift in Melville’s interpretation of the value and possibility of genuine communion between human beings and between humans and the whale. One may read Typee as an attempt by Melville to explore the possibility of a this-worldly utopia in which human beings can return to a version of primitive interconnectedness. This exploration may be seen to be extended in Moby-Dick, particularly in Ishmael’s attempts to find communion with others and in some moments of encounter with the whales. The thesis uses phenomenology as a theory to interpret what Melville is trying to suggest in these moments of encounter. However, it argues, finally, that such encounter, or ‘intersubjectivity’ is eventually jettisoned, especially in the works after Moby-Dick. By the end of Melville’s life and work, any hope of an intersubjective utopia he may have harboured as a younger man have been removed in favour of a refusal actually to assert any final ‘truth’ about social, political or even religious experience. Billy Budd, his last body, is hanged, and his final word is silence.
- Full Text:
- Date Issued: 2003
Structural and synthetic investigations of diterpenoid natural products from southern African marine invertebrates
- Authors: Gray, Christopher Anthony
- Date: 2003
- Subjects: Ethyl acetoacetate Diterpenes Limpets -- South Africa Natural products -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4349 , http://hdl.handle.net/10962/d1005014
- Description: This thesis is divided into two parts. The first part (Chapter Two) documents a bioassay guided investigation of the ethyl acetate extracts of four marine invertebrates from Mozambique (an Irciniid sponge, a Haliclona sp. sponge, an ascidian tentatively identified as Diplosoma sp., and the soft coral Cladiella kashmani). Eight known compounds [ilimaquinone (2.1), renierone (2.7), N-formyl-1,2-dihydrorenierone (2.8), 1,6-dimethyl-7-methoxy-5,8-dihydroisoquinoline-5,8-dione (2.9), mimosamycin (2.10) 7Z-allylidene-5-hydroxy-7,7a-dihydro-2H-cyclopenta[b]pyran-6-one (2.11), flaccidoxide (2.18) and 11S,12S-epoxycembra-1Z,3E,7E-trien-14S-ol (2.19)] and a new diterpene [13S,14R-diacetoxy-11S,12R-epoxycembra-1Z,3E,7E-triene (2.20)] were isolated and identified using standard spectroscopic techniques. Anomalies in the published spectral data of 2.1 and 2.8 were exposed and corrected, and the absolute stereochemistry of the cembrane diterpenes 2.18 and 2.20 established using the modified Mosher’s method. The comparative activities of the nine natural products against four cancer cell lines (A549, LOX, OVCAR3, SNB19) are reported. The second part of the thesis (Chapter Three – Chapter Six) is concerned with an ecological, structural and synthetic study of diterpenes from the endemic South African pulmonate limpet Trimusculus costatus. Two new labdane diterpenes [6b,7a-diacetoxylabda-8,13E-dien-15-ol (3.10) and 2a,6b,7a-triacetoxylabda-8,13E-dien-15-ol (3.11)] were isolated from T. costatus and evaluated for anti-feeding activity against the common predatory fish Pomadasys commersonnii. A strategy for the semi-synthesis of 3.10 from rhinocerotinoic acid (4.14), a diterpene reportedly present in the ubiquitous South African shrub Elytropappus rhinocerotis, was devised in order to allow further bioactivity tests to be performed and unequivocally assign the unknown absolute stereochemistry of the T. costatus metabolites. Attempts to isolate rhinocerotinoic acid from local specimens of Elytropappus rhinocerotis were unsuccessful, and as the repetition of a published synthesis of 4.14 from (-)-sclareol (4.15) gave rhinocerotinoic acid in unacceptably low yields with poor stereoselectivity, an improved synthesis of 4.14 is presented. Comprehensive studies using hispanone (5.1) as a model compound showed that 6,7-dioxygenated labda-8-enes could be prepared from compounds possessing a 7-oxo-labda-8-ene skeleton with some degree of stereocontrol. In the process, fourteen new hispanone analogues were prepared and most of these were tested for activity in a suite of ten agro-chemical assays. The novel compound 7b-hydroxy-9a-carbonitrile-15,16-epoxylabda-13(16),14-dien-6-one (5.34) exhibited significant activity against the crop fungus Phytophthora infestans and is currently being subjected to further agro-chemical tests. Unfortunately, the results from the oxygenation study performed on the model compound 5.1 could not be directly extrapolated to rhinocerotinoic acid. Attempts to prepare the naturally occurring 3.10 from 4.14 via an alternative route were unsuccessful but yielded an analogue of 3.10 in which the substituents at C-6 and C-7 are in a diequatorial rather than a diaxial configuration.
- Full Text:
- Date Issued: 2003
- Authors: Gray, Christopher Anthony
- Date: 2003
- Subjects: Ethyl acetoacetate Diterpenes Limpets -- South Africa Natural products -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4349 , http://hdl.handle.net/10962/d1005014
- Description: This thesis is divided into two parts. The first part (Chapter Two) documents a bioassay guided investigation of the ethyl acetate extracts of four marine invertebrates from Mozambique (an Irciniid sponge, a Haliclona sp. sponge, an ascidian tentatively identified as Diplosoma sp., and the soft coral Cladiella kashmani). Eight known compounds [ilimaquinone (2.1), renierone (2.7), N-formyl-1,2-dihydrorenierone (2.8), 1,6-dimethyl-7-methoxy-5,8-dihydroisoquinoline-5,8-dione (2.9), mimosamycin (2.10) 7Z-allylidene-5-hydroxy-7,7a-dihydro-2H-cyclopenta[b]pyran-6-one (2.11), flaccidoxide (2.18) and 11S,12S-epoxycembra-1Z,3E,7E-trien-14S-ol (2.19)] and a new diterpene [13S,14R-diacetoxy-11S,12R-epoxycembra-1Z,3E,7E-triene (2.20)] were isolated and identified using standard spectroscopic techniques. Anomalies in the published spectral data of 2.1 and 2.8 were exposed and corrected, and the absolute stereochemistry of the cembrane diterpenes 2.18 and 2.20 established using the modified Mosher’s method. The comparative activities of the nine natural products against four cancer cell lines (A549, LOX, OVCAR3, SNB19) are reported. The second part of the thesis (Chapter Three – Chapter Six) is concerned with an ecological, structural and synthetic study of diterpenes from the endemic South African pulmonate limpet Trimusculus costatus. Two new labdane diterpenes [6b,7a-diacetoxylabda-8,13E-dien-15-ol (3.10) and 2a,6b,7a-triacetoxylabda-8,13E-dien-15-ol (3.11)] were isolated from T. costatus and evaluated for anti-feeding activity against the common predatory fish Pomadasys commersonnii. A strategy for the semi-synthesis of 3.10 from rhinocerotinoic acid (4.14), a diterpene reportedly present in the ubiquitous South African shrub Elytropappus rhinocerotis, was devised in order to allow further bioactivity tests to be performed and unequivocally assign the unknown absolute stereochemistry of the T. costatus metabolites. Attempts to isolate rhinocerotinoic acid from local specimens of Elytropappus rhinocerotis were unsuccessful, and as the repetition of a published synthesis of 4.14 from (-)-sclareol (4.15) gave rhinocerotinoic acid in unacceptably low yields with poor stereoselectivity, an improved synthesis of 4.14 is presented. Comprehensive studies using hispanone (5.1) as a model compound showed that 6,7-dioxygenated labda-8-enes could be prepared from compounds possessing a 7-oxo-labda-8-ene skeleton with some degree of stereocontrol. In the process, fourteen new hispanone analogues were prepared and most of these were tested for activity in a suite of ten agro-chemical assays. The novel compound 7b-hydroxy-9a-carbonitrile-15,16-epoxylabda-13(16),14-dien-6-one (5.34) exhibited significant activity against the crop fungus Phytophthora infestans and is currently being subjected to further agro-chemical tests. Unfortunately, the results from the oxygenation study performed on the model compound 5.1 could not be directly extrapolated to rhinocerotinoic acid. Attempts to prepare the naturally occurring 3.10 from 4.14 via an alternative route were unsuccessful but yielded an analogue of 3.10 in which the substituents at C-6 and C-7 are in a diequatorial rather than a diaxial configuration.
- Full Text:
- Date Issued: 2003
An analysis of selected World Trade Organisation agreements to determine whether they discriminate unfairly against developing economices
- Authors: Grimett, Leticia Anthea
- Date: 2013-07-29
- Subjects: World Trade Organization Foreign trade regulation General Agreement on Tariffs and Trade (Organization) Free trade -- Developing countries Foreign trade regulation -- Developing countries Competition, Unfair -- Developing countries
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3718 , http://hdl.handle.net/10962/d1008368
- Description: The focus of this thesis is the question whether or not the WTO discriminates unfairly against developing economies. In the absence of a test of guidelines for detennining unfairness or fairness of WTO provisions or Agreements has been drawn up using welfare economic and constitutional law principles as a foundation. Unfairness is therefore determined by asking whether the provisions of each Agreement are rational, proportional, efficient and whether they prevent the abuse of power amongst states. In addition, the economic effects of the provisions of the selected Agreements have been analysed to determine whether the relevant provisions are welfare enhancing and conclusive to promoting growth and development within developing economies. The Agreements chosed for analysis are the Agreements on Trade-related Investment Measures (TRIMS), Trade-related Intellectual Property (TRIPS), Agriculture and Services (GATS). The dispute settlement and negotiating process, labour standards and the impact of decreasing most-favoured nation rates on developing economy competitiveness is also discussed. Application of the test has shown that the WTO provisions do not reflect the interests of all members. Even though most member states are developing economies, the3 Agreements constantly cater foe developed country concerns and interests. Where provision is made for developing country interests, it is the LDC's who are favoured, with nonnal developing economies being bound by the same provisions as the developed economies. A fonnal, as opposed to a substantive, defmition has been adopted by the WTO, with a result that the process of equality is placed above the outcomes. While concessions have been made to development, members have not gone for enough. A main reason for the imbalance can be attributed to the negotiating process, which is based upon concessionary bargaining and trade-off. Those states with greater economic power are therefore at an advantage as they have the leverage needed to influence the outcomes of negotiations and hence the provisions of the various Agreements. Even with the LDC's, the WTO has been found to discriminate unfairly against developing economies because it does not adequately address developing country concerns. , KMBT_363 , Adobe Acrobat 9.54 Paper Capture Plug-in
- Full Text:
- Authors: Grimett, Leticia Anthea
- Date: 2013-07-29
- Subjects: World Trade Organization Foreign trade regulation General Agreement on Tariffs and Trade (Organization) Free trade -- Developing countries Foreign trade regulation -- Developing countries Competition, Unfair -- Developing countries
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3718 , http://hdl.handle.net/10962/d1008368
- Description: The focus of this thesis is the question whether or not the WTO discriminates unfairly against developing economies. In the absence of a test of guidelines for detennining unfairness or fairness of WTO provisions or Agreements has been drawn up using welfare economic and constitutional law principles as a foundation. Unfairness is therefore determined by asking whether the provisions of each Agreement are rational, proportional, efficient and whether they prevent the abuse of power amongst states. In addition, the economic effects of the provisions of the selected Agreements have been analysed to determine whether the relevant provisions are welfare enhancing and conclusive to promoting growth and development within developing economies. The Agreements chosed for analysis are the Agreements on Trade-related Investment Measures (TRIMS), Trade-related Intellectual Property (TRIPS), Agriculture and Services (GATS). The dispute settlement and negotiating process, labour standards and the impact of decreasing most-favoured nation rates on developing economy competitiveness is also discussed. Application of the test has shown that the WTO provisions do not reflect the interests of all members. Even though most member states are developing economies, the3 Agreements constantly cater foe developed country concerns and interests. Where provision is made for developing country interests, it is the LDC's who are favoured, with nonnal developing economies being bound by the same provisions as the developed economies. A fonnal, as opposed to a substantive, defmition has been adopted by the WTO, with a result that the process of equality is placed above the outcomes. While concessions have been made to development, members have not gone for enough. A main reason for the imbalance can be attributed to the negotiating process, which is based upon concessionary bargaining and trade-off. Those states with greater economic power are therefore at an advantage as they have the leverage needed to influence the outcomes of negotiations and hence the provisions of the various Agreements. Even with the LDC's, the WTO has been found to discriminate unfairly against developing economies because it does not adequately address developing country concerns. , KMBT_363 , Adobe Acrobat 9.54 Paper Capture Plug-in
- Full Text:
The geochemistry of ore fluids and control of gold mineralization in banded iron-formation at the Kalahari Goldridge deposit, Kraaipan greenstone belt, South Africa
- Authors: Hammond, Napoleon Quaye
- Date: 2003
- Subjects: Gold ores -- Geology -- South Africa -- North-West Greenstone belts -- South Africa -- North-West Ore deposits -- South Africa -- North-West Geochemistry -- South Africa -- North-West
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5048 , http://hdl.handle.net/10962/d1008370
- Description: The Kalahari Goldridge mine is located within the Archaean Kraaipan Greenstone Belt about 60 km SW of Mafikeng in the Northwestern Province, South Africa. Several gold deposits are located within approximately north - south-striking banded iron-formation (BIF). Current opencast mining operations are focused on the largest of these (D Zone). The orebody is stratabound and hosted primarily in the BIF, which consists of alternating chert and magnetite-chloritestilpnomelane-sulphide-carbonate bands ranging from mm to cm scale. The ore body varies in thickness from 15 to 45 m along a strike length of about 1.5 km. The BlF is sandwiched between a sericite-carbonate-chlorite schist at the immediate footwall and carbonaceous meta-pelites in the hanging-wall. Further west in the footwall, the schists are underlain by mafic meta-volcanic amphibolite. Overlying the hanging-wall carbonaceous metapeiites are schist units and meta-greywackes that become increasingly conglomeratic up the stratigraphy. Stilpnomelane-, chlorite- and minnesotaite-bearing assemblages in the BlFs indicate metamorphic temperatures of 300 - 450°C and pressures of less than 5 kbars. The BIF generally strikes approximately 3400 and dips from 60 to 75°E. Brittle-ductile deformation is evidenced by small-scale isoclinal folds, brecciation, extension fractures and boudinaging of cherty BIF units. Fold axial planes are sub-parallel to the foliation orientation with sub-vertical plunges parallel to prominent rodding and mineral lineation in the footwall. Gold mineralization at the Kalahari Goldridge deposit is associated with two generations of subhorizontal quartz-carbonate veins dips approximately 20 to 40°W. The first generation consists of ladder vein sets (Group lIA) preferentially developed in Fe-rich meso bands, whilst the second generation consists of large quartz-carbonate veins (Group lIB), which crosscut the entire ore body extending into the footwall and hanging-wall in places. Major structures that control the ore body are related to meso-scale isoclinal folds with fold axes subparallel to mineral elongation lineations, which plunge approximately 067°E. These linear structures form orthogonal orientation with the plane of the mineralized shallowdipping veins indicating stretching and development of fluid - focusing conduits. A second-order controlling feature corresponds to the intersection of the mineralized veins and foliation planes of host rock, plunging approximately 008°N and trending 341°. G0ld is closely associated with sulphides, mainly pyrite and pyrrhotite and to a lesser extent with bismuth tellurides, and carbonate gangue. The ore fluid responsible for the gold deposition is in the C-O-H system with increased CH₄ contents attributed to localized hydrolysis reaction between interbedded carbonaceous sediment and ore fluid. The fluid is characterized by significant C0₂ contents and low salinities below 7.0 wt % NaCl equivalent (averages of 3.5 and 3.0 wt % NaCl equivalent for the first and second episodes of the mineralization respectively) . Calculated values of f0₂. ranging from 10⁻²⁹·⁹⁸ to 10⁻³²·⁹⁶ bars, bracket the C0₂-CH₄ and pyrite-pyrrhotite-magnetite buffer boundaries and reveal the reducing nature of the ore fluid at deposition. Calculated total sulphur content in the ore fluid (mΣs), ranges from 0.011 to 0.018M and is consistent with the range (10⁻³·⁵ to 10⁻¹M) reported for subamphibolite facies ore fluids. The close association of sulphides with the Au and nature of the fluid also give credence that the Au was carried in solution by the Au(HS)₂ - complex. Extensive epigenetic replacement of magnetite and chlorite in BIF and other meta-pelitic sediments in the deposit by sulphides and carbonates, both on meso scopic and microscopic scales gives evidence of an interaction by a CO₂- and H₂S-bearing fluid with the Fe-rich host rocks in the deposit. This facilitated Au precipitation due to changes in the physico-chemical conditions of the ore fluid such as a decrease in the mΣs and pH leading to the destabilization of the reduced sulphur complexes. Local gradients in f0₂ may account for gold precipitation in places within carbonaceous sediments. The fineness of the gold grams (1000*Au/(Au + Ag) ranges from 823 to 921. This compares favourably with the fineness reported for some Archaean BIFhosced deposits (851 - 970). Mass balance transfer calculations indicate that major chemical changes associated with the hydrothermal alteration of BIF include enrichment of Au, Ag, Bi, Te, volatiles (S and CO₂), MgO, Ba, K and Rb but significant depletion of SiO₂ and minor losses of Fe₂O₃. In addition, anomalous enrichment of Sc (average, 1247%) suggests its possible use as an exploration tool in the ferruginous sediments in the Kraaipan greenstone terrane. Evidence from light stable isotopes and fluid inclusions suggests that the mineralized veins crystallized from a single homogeneous fluid source during the two episodes of mineralization under the similar physicochemical conditions. Deposition occurred at temperatures rangmg from 350 to 400°C and fluid pressures ranging from 0.7 to 2.0kbars. Stable isotope constraints indicate the following range for the hydrothermal fluid; θ¹⁸H₂O = 6.65 to 10.48%0, 8¹³CΣc = -6.0 to -8.0 %0 and 8³⁴SΣs = + 1.69 to + 4.0%0 . These data do not offer conclusive evidence for the source of fluid associated with the mineralization at the Kalahari Goldridge deposit as they overlap the range prescribed for fluid derived from devolatization of deep-seated volcano-sedimentary piles near the brittle-ductile transition in greenstone belts during prograde metamorphism, and magmatic hydrothermal fluids. , KMBT_363 , Adobe Acrobat 9.54 Paper Capture Plug-in
- Full Text:
- Date Issued: 2003
- Authors: Hammond, Napoleon Quaye
- Date: 2003
- Subjects: Gold ores -- Geology -- South Africa -- North-West Greenstone belts -- South Africa -- North-West Ore deposits -- South Africa -- North-West Geochemistry -- South Africa -- North-West
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5048 , http://hdl.handle.net/10962/d1008370
- Description: The Kalahari Goldridge mine is located within the Archaean Kraaipan Greenstone Belt about 60 km SW of Mafikeng in the Northwestern Province, South Africa. Several gold deposits are located within approximately north - south-striking banded iron-formation (BIF). Current opencast mining operations are focused on the largest of these (D Zone). The orebody is stratabound and hosted primarily in the BIF, which consists of alternating chert and magnetite-chloritestilpnomelane-sulphide-carbonate bands ranging from mm to cm scale. The ore body varies in thickness from 15 to 45 m along a strike length of about 1.5 km. The BlF is sandwiched between a sericite-carbonate-chlorite schist at the immediate footwall and carbonaceous meta-pelites in the hanging-wall. Further west in the footwall, the schists are underlain by mafic meta-volcanic amphibolite. Overlying the hanging-wall carbonaceous metapeiites are schist units and meta-greywackes that become increasingly conglomeratic up the stratigraphy. Stilpnomelane-, chlorite- and minnesotaite-bearing assemblages in the BlFs indicate metamorphic temperatures of 300 - 450°C and pressures of less than 5 kbars. The BIF generally strikes approximately 3400 and dips from 60 to 75°E. Brittle-ductile deformation is evidenced by small-scale isoclinal folds, brecciation, extension fractures and boudinaging of cherty BIF units. Fold axial planes are sub-parallel to the foliation orientation with sub-vertical plunges parallel to prominent rodding and mineral lineation in the footwall. Gold mineralization at the Kalahari Goldridge deposit is associated with two generations of subhorizontal quartz-carbonate veins dips approximately 20 to 40°W. The first generation consists of ladder vein sets (Group lIA) preferentially developed in Fe-rich meso bands, whilst the second generation consists of large quartz-carbonate veins (Group lIB), which crosscut the entire ore body extending into the footwall and hanging-wall in places. Major structures that control the ore body are related to meso-scale isoclinal folds with fold axes subparallel to mineral elongation lineations, which plunge approximately 067°E. These linear structures form orthogonal orientation with the plane of the mineralized shallowdipping veins indicating stretching and development of fluid - focusing conduits. A second-order controlling feature corresponds to the intersection of the mineralized veins and foliation planes of host rock, plunging approximately 008°N and trending 341°. G0ld is closely associated with sulphides, mainly pyrite and pyrrhotite and to a lesser extent with bismuth tellurides, and carbonate gangue. The ore fluid responsible for the gold deposition is in the C-O-H system with increased CH₄ contents attributed to localized hydrolysis reaction between interbedded carbonaceous sediment and ore fluid. The fluid is characterized by significant C0₂ contents and low salinities below 7.0 wt % NaCl equivalent (averages of 3.5 and 3.0 wt % NaCl equivalent for the first and second episodes of the mineralization respectively) . Calculated values of f0₂. ranging from 10⁻²⁹·⁹⁸ to 10⁻³²·⁹⁶ bars, bracket the C0₂-CH₄ and pyrite-pyrrhotite-magnetite buffer boundaries and reveal the reducing nature of the ore fluid at deposition. Calculated total sulphur content in the ore fluid (mΣs), ranges from 0.011 to 0.018M and is consistent with the range (10⁻³·⁵ to 10⁻¹M) reported for subamphibolite facies ore fluids. The close association of sulphides with the Au and nature of the fluid also give credence that the Au was carried in solution by the Au(HS)₂ - complex. Extensive epigenetic replacement of magnetite and chlorite in BIF and other meta-pelitic sediments in the deposit by sulphides and carbonates, both on meso scopic and microscopic scales gives evidence of an interaction by a CO₂- and H₂S-bearing fluid with the Fe-rich host rocks in the deposit. This facilitated Au precipitation due to changes in the physico-chemical conditions of the ore fluid such as a decrease in the mΣs and pH leading to the destabilization of the reduced sulphur complexes. Local gradients in f0₂ may account for gold precipitation in places within carbonaceous sediments. The fineness of the gold grams (1000*Au/(Au + Ag) ranges from 823 to 921. This compares favourably with the fineness reported for some Archaean BIFhosced deposits (851 - 970). Mass balance transfer calculations indicate that major chemical changes associated with the hydrothermal alteration of BIF include enrichment of Au, Ag, Bi, Te, volatiles (S and CO₂), MgO, Ba, K and Rb but significant depletion of SiO₂ and minor losses of Fe₂O₃. In addition, anomalous enrichment of Sc (average, 1247%) suggests its possible use as an exploration tool in the ferruginous sediments in the Kraaipan greenstone terrane. Evidence from light stable isotopes and fluid inclusions suggests that the mineralized veins crystallized from a single homogeneous fluid source during the two episodes of mineralization under the similar physicochemical conditions. Deposition occurred at temperatures rangmg from 350 to 400°C and fluid pressures ranging from 0.7 to 2.0kbars. Stable isotope constraints indicate the following range for the hydrothermal fluid; θ¹⁸H₂O = 6.65 to 10.48%0, 8¹³CΣc = -6.0 to -8.0 %0 and 8³⁴SΣs = + 1.69 to + 4.0%0 . These data do not offer conclusive evidence for the source of fluid associated with the mineralization at the Kalahari Goldridge deposit as they overlap the range prescribed for fluid derived from devolatization of deep-seated volcano-sedimentary piles near the brittle-ductile transition in greenstone belts during prograde metamorphism, and magmatic hydrothermal fluids. , KMBT_363 , Adobe Acrobat 9.54 Paper Capture Plug-in
- Full Text:
- Date Issued: 2003
Biogeography and community structure of fishes in South African estuaries
- Authors: Harrison, Trevor D
- Date: 2003
- Subjects: Estuarine ecology -- South Africa Estuarine fishes -- South Africa Fish communities -- South Africa Biogeography -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5235 , http://hdl.handle.net/10962/d1005078
- Description: The biogeography and community structure of the fishes of South African estuaries was investigated. In all, 109 systems were examined representing two broad types: temporarily closed and permanently open estuaries. Multivariate analyses of the fish communities identified three biogeographic regions. A cool-temperate region extended along the west and southwest coasts; a warmtemperate zone stretched along the south, southeast and east coasts and a subtropical region occurred along the east coast. The boundaries of these biogeographic regions were also delineated. The general physico-chemical characteristics of the estuaries within the three biogeographic regions also reflected regional differences in climate, rainfall and ocean conditions. Estuarine temperatures followed the trend for marine coastal waters, decreasing from subtropical estuaries toward cool-temperate systems. The low rainfall and runoff in the warm-temperate region together with high evaporation rates and strong seawater input resulted in higher salinities in these estuaries. These factors also accounted for the predominantly clearer waters in warm-temperate estuaries. The estuaries in the three biogeographic regions were also shown to contain somewhat distinctive fish assemblages. Temperature and salinity appeared to be the two main factors affecting the distribution and abundance of fishes in South African estuaries. Subtropical systems were characterised by fishes mostly of tropical origin as well as certain south coast endemic species. Warm-temperate estuaries were dominated by endemic taxa with some tropical species also present. The fish fauna of cooltemperate estuaries mostly comprised south coast endemic species with cosmopolitan and temperate taxa also present. Certain functional components of the ichthyofauna also exhibited slight differences between regions. Freshwater fishes were a major component of closed subtropical estuaries while estuarine resident species were more abundant in warm-temperate estuaries. Overall, estuarine-dependent marine species dominated the fish fauna of the estuaries in all biogeographic regions, signifying that South African estuaries perform a vital nursery function for this group of fishes. Slight differences were also apparent in the trophic structure of the fishes; these were related to environmental differences between regions. Zooplanktivores and fishes that feed on aquatic macrophytes/invertebrates assumed a relatively higher importance in warm-temperate systems. Overall, detritivores dominated the estuarine fish fauna in all regions, indicating that detritus forms the main energy source in South African estuaries.
- Full Text:
- Date Issued: 2003
- Authors: Harrison, Trevor D
- Date: 2003
- Subjects: Estuarine ecology -- South Africa Estuarine fishes -- South Africa Fish communities -- South Africa Biogeography -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5235 , http://hdl.handle.net/10962/d1005078
- Description: The biogeography and community structure of the fishes of South African estuaries was investigated. In all, 109 systems were examined representing two broad types: temporarily closed and permanently open estuaries. Multivariate analyses of the fish communities identified three biogeographic regions. A cool-temperate region extended along the west and southwest coasts; a warmtemperate zone stretched along the south, southeast and east coasts and a subtropical region occurred along the east coast. The boundaries of these biogeographic regions were also delineated. The general physico-chemical characteristics of the estuaries within the three biogeographic regions also reflected regional differences in climate, rainfall and ocean conditions. Estuarine temperatures followed the trend for marine coastal waters, decreasing from subtropical estuaries toward cool-temperate systems. The low rainfall and runoff in the warm-temperate region together with high evaporation rates and strong seawater input resulted in higher salinities in these estuaries. These factors also accounted for the predominantly clearer waters in warm-temperate estuaries. The estuaries in the three biogeographic regions were also shown to contain somewhat distinctive fish assemblages. Temperature and salinity appeared to be the two main factors affecting the distribution and abundance of fishes in South African estuaries. Subtropical systems were characterised by fishes mostly of tropical origin as well as certain south coast endemic species. Warm-temperate estuaries were dominated by endemic taxa with some tropical species also present. The fish fauna of cooltemperate estuaries mostly comprised south coast endemic species with cosmopolitan and temperate taxa also present. Certain functional components of the ichthyofauna also exhibited slight differences between regions. Freshwater fishes were a major component of closed subtropical estuaries while estuarine resident species were more abundant in warm-temperate estuaries. Overall, estuarine-dependent marine species dominated the fish fauna of the estuaries in all biogeographic regions, signifying that South African estuaries perform a vital nursery function for this group of fishes. Slight differences were also apparent in the trophic structure of the fishes; these were related to environmental differences between regions. Zooplanktivores and fishes that feed on aquatic macrophytes/invertebrates assumed a relatively higher importance in warm-temperate systems. Overall, detritivores dominated the estuarine fish fauna in all regions, indicating that detritus forms the main energy source in South African estuaries.
- Full Text:
- Date Issued: 2003
Binding and transcriptional activation by Uga3p, a zinc binuclear cluster protein of Saccharomyces cerevisiae redefining the UAS [subscript GABA] and the Uga3p binding site
- Authors: Idicula, Anu Mary
- Date: 2003
- Subjects: Saccharomyces cerevisiae GABA
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3933 , http://hdl.handle.net/10962/d1003992
- Description: Uga3p, a member of the zinc binuclear cluster transcription factor family, is required for [gamma]-aminobutyric acid-dependent transcription of the UGA genes in Saccharomyces cerevisiae. Crystallographic data of some of the protein-DNA complexes of this family reveal that members of this family bind to CGG triplets. A conserved 19-nucleotide activation element in certain UGA gene promoter regions contains a CCG-N4-CGG everted repeat, proposed to be the binding site of Uga3p, UAS[subscript GABA]. The spacer region (N4) between the CGG triplets has been suggested to be the specificity determinant for binding to UAS[subscript GABA]. The data available from the Saccharomyces genome database indicates that there are multiple repeats of -CCG-N4-CGG- regions within the genome. These transcription factors are involved in the activation of specific pathways and the question arises as to how their specificity of binding is determined. The aim of this study was to understand the binding characteristics of Uga3p to UAS[subscript GABA] and to determine the affinity and specificity of this interaction. In this study, full-length (tagged and untagged) and truncated (1-124 a.a.) Uga3p was produced in a heterologous expression system (E. coli). The interaction of Uga3p with UAS[subscript GABA] in Saccharomyces cerevisiae was characterized in terms of binding in vitro and the transcriptional activation of lacZ reporter genes in vivo. The Uga3p was capable of binding to these sites in vitro independent of exogenous GABA. Electrophoretic mobility shift assays (EMSA) of the full-length Uga3p with the wild type UAS[subscript GABA] sequences produced two distinct mobility complexes. The complexes formed in the EMSA of the full-length Uga3p were those specific to the interaction of the Uga3p to UAS[subscript GABA]. The truncated Uga3p(1-124 a.a.), which has the DNA-binding zinc cluster domain, the linker region and the putative coiled-coil domain was not functionally equivalent to the full-length protein with respect to binding in vitro because the EMSAs of the UAS[subscript GABA] with the truncated Uga3p produced indistinct complexes. EMSAs using mutant UAS[subscript GABA] sequences and heterologously-produced full-length Uga3p, demonstrated that UAS[subscript GABA] consists of two, independent Uga3p-binding sites. This work presents evidence that the two Uga3p molecules bound to UAS[subscript GABA] most likely interact with each other. Unlike other zinc cluster binding sites the Uga3p-binding site is an asymmetric site of 5’-SGCGGNWWT-3’ (S= G or C, W = A or T and N = no nucleotide or G or C). UAS[subscript GABA] is a palindrome containing the two asymmetric Uga3p-binding sites. The two-site consensus sequence required for the binding of Uga3p to the UAS[subscript GABA] is present upstream of UGA1 (region -387 to -370) and UGA4 (region -403 to -387). Furthermore, a single Uga3p-binding site was identified in the 5’ untranslated regions of UGA2 (region -219 to -211). GABA-dependent transcriptional activation by UAS[subscript GABA] in vivo could be directly correlated to a high affinity, specific interaction of two Uga3p molecules to this UAS. Binding with high affinity required the conserved sequences flanking the everted repeat. This study provided evidence that the binding pattern of Uga3p is novel compared to other zinc cluster motifs investigated, as the sequences flanking the everted repeat are important regions for recognition by Uga3p. The studies with the truncated Uga3p (1 –124 a.a.), also suggested that the regions C-terminal to the DNA-binding motif and putative coiled-coil area of this protein are important for Uga3p-specific interactions with UAS[subscript GABA]. Investigation of regions C-terminal to the zinc cluster, linker and putative coiledcoil revealed an eight-motif regulatory region similar to that in other zinc cluster proteins. This indicated that the regions C-terminal to these domains are important for the regulation and activity of these proteins. A putative seven repeat WD40-like motif was identified within this region. This putative domain has been speculated to be important for protein-protein interactions. Phosphorylation and dephosphorylation in other proteins of this class have been indicated to be important for the regulation of the activity of these proteins. The bioinformatic analysis of Uga3p revealed two possible cAMP/cGMP-dependent protein kinase phosphorylation sites, four putative protein kinase C phosphorylation motifs and four putative casein kinase II phosphorylation motifs. This study has contributed to the understanding of the nature of interactions between Uga3p and its specific UAS [subscript GABA] and how the regions flanking the everted repeat determine its specificity. The comparison of the nature of the binding of truncated and full-length Uga3p in vitro provided evidence for the role played by the full-length protein in determining this specific interaction. This evidence suggested that the in vitro binding evidence for other proteins of this family, using truncated peptides that carry the DNA-binding domain, might not reflect the true nature of interactions between the proteins of this class and their specific UASs in vivo.
- Full Text:
- Date Issued: 2003
- Authors: Idicula, Anu Mary
- Date: 2003
- Subjects: Saccharomyces cerevisiae GABA
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3933 , http://hdl.handle.net/10962/d1003992
- Description: Uga3p, a member of the zinc binuclear cluster transcription factor family, is required for [gamma]-aminobutyric acid-dependent transcription of the UGA genes in Saccharomyces cerevisiae. Crystallographic data of some of the protein-DNA complexes of this family reveal that members of this family bind to CGG triplets. A conserved 19-nucleotide activation element in certain UGA gene promoter regions contains a CCG-N4-CGG everted repeat, proposed to be the binding site of Uga3p, UAS[subscript GABA]. The spacer region (N4) between the CGG triplets has been suggested to be the specificity determinant for binding to UAS[subscript GABA]. The data available from the Saccharomyces genome database indicates that there are multiple repeats of -CCG-N4-CGG- regions within the genome. These transcription factors are involved in the activation of specific pathways and the question arises as to how their specificity of binding is determined. The aim of this study was to understand the binding characteristics of Uga3p to UAS[subscript GABA] and to determine the affinity and specificity of this interaction. In this study, full-length (tagged and untagged) and truncated (1-124 a.a.) Uga3p was produced in a heterologous expression system (E. coli). The interaction of Uga3p with UAS[subscript GABA] in Saccharomyces cerevisiae was characterized in terms of binding in vitro and the transcriptional activation of lacZ reporter genes in vivo. The Uga3p was capable of binding to these sites in vitro independent of exogenous GABA. Electrophoretic mobility shift assays (EMSA) of the full-length Uga3p with the wild type UAS[subscript GABA] sequences produced two distinct mobility complexes. The complexes formed in the EMSA of the full-length Uga3p were those specific to the interaction of the Uga3p to UAS[subscript GABA]. The truncated Uga3p(1-124 a.a.), which has the DNA-binding zinc cluster domain, the linker region and the putative coiled-coil domain was not functionally equivalent to the full-length protein with respect to binding in vitro because the EMSAs of the UAS[subscript GABA] with the truncated Uga3p produced indistinct complexes. EMSAs using mutant UAS[subscript GABA] sequences and heterologously-produced full-length Uga3p, demonstrated that UAS[subscript GABA] consists of two, independent Uga3p-binding sites. This work presents evidence that the two Uga3p molecules bound to UAS[subscript GABA] most likely interact with each other. Unlike other zinc cluster binding sites the Uga3p-binding site is an asymmetric site of 5’-SGCGGNWWT-3’ (S= G or C, W = A or T and N = no nucleotide or G or C). UAS[subscript GABA] is a palindrome containing the two asymmetric Uga3p-binding sites. The two-site consensus sequence required for the binding of Uga3p to the UAS[subscript GABA] is present upstream of UGA1 (region -387 to -370) and UGA4 (region -403 to -387). Furthermore, a single Uga3p-binding site was identified in the 5’ untranslated regions of UGA2 (region -219 to -211). GABA-dependent transcriptional activation by UAS[subscript GABA] in vivo could be directly correlated to a high affinity, specific interaction of two Uga3p molecules to this UAS. Binding with high affinity required the conserved sequences flanking the everted repeat. This study provided evidence that the binding pattern of Uga3p is novel compared to other zinc cluster motifs investigated, as the sequences flanking the everted repeat are important regions for recognition by Uga3p. The studies with the truncated Uga3p (1 –124 a.a.), also suggested that the regions C-terminal to the DNA-binding motif and putative coiled-coil area of this protein are important for Uga3p-specific interactions with UAS[subscript GABA]. Investigation of regions C-terminal to the zinc cluster, linker and putative coiledcoil revealed an eight-motif regulatory region similar to that in other zinc cluster proteins. This indicated that the regions C-terminal to these domains are important for the regulation and activity of these proteins. A putative seven repeat WD40-like motif was identified within this region. This putative domain has been speculated to be important for protein-protein interactions. Phosphorylation and dephosphorylation in other proteins of this class have been indicated to be important for the regulation of the activity of these proteins. The bioinformatic analysis of Uga3p revealed two possible cAMP/cGMP-dependent protein kinase phosphorylation sites, four putative protein kinase C phosphorylation motifs and four putative casein kinase II phosphorylation motifs. This study has contributed to the understanding of the nature of interactions between Uga3p and its specific UAS [subscript GABA] and how the regions flanking the everted repeat determine its specificity. The comparison of the nature of the binding of truncated and full-length Uga3p in vitro provided evidence for the role played by the full-length protein in determining this specific interaction. This evidence suggested that the in vitro binding evidence for other proteins of this family, using truncated peptides that carry the DNA-binding domain, might not reflect the true nature of interactions between the proteins of this class and their specific UASs in vivo.
- Full Text:
- Date Issued: 2003
Studies of the environmental and endocrine control of reproduction in the four striped field mouse, Rhabdomys pumilio
- Authors: Jackson, Claire
- Date: 2003
- Subjects: Microtus Microtus -- South Africa -- Reproduction Mice -- Reproduction
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5777 , http://hdl.handle.net/10962/d1005465
- Description: Previous studies of the control of reproduction in Rhabdomys pumilio have shown that day length alone does not inhibit spermatogenesis, that a reduction in food availability and ambient temperature results in an inhibition of gametogenesis, that females are more susceptible to inhibition than are males, and that mice that are able to maintain a body fat store in the face of an energetic challenge, are less likely to show reproductive inhibition than those that lose their fat store. In the present study, field and laboratory experiments were conducted to examine the effects of winter food supplementation on reproduction and population dynamics, and the effects of exogenous GnRH, leptin and mercaptoacetate (MA) on reproductive activity of Rhabdomys pumilio exposed to an energetic challenge. In the field food supplementation experiments in Thomas Baines Nature Reserve (2000, 2001), there was no winter inhibition of reproduction and provision of supplementary food had little effect. While at Mountain Zebra National Park (2002) winter was harsher, females became reproductively inactive, spermatogenesis continued and the provision of extra food resulted in higher rates of individual growth and larger reproductive organs. Treatment of mice that had been exposed to a prolonged energetic challenge, with exogenous GnRH (1µg/mouse/treatment) resulted in an increase in the masses of the testes and epididymides, and in the activity of the reproductive organs. Treatment with exogenous leptin (40µg/mouse/treatment), concurrently with an energetic challenge, countered the negative effects of the energetic challenge, and treated males had larger reproductive organs. MA (600µmol/kg body mass), given concurrently with an energetic challenge, did not inhibit fat metabolism, although the high-fat diet countered the effects of the energetic challenge. Results suggest that the first response of male Rhabdomys pumilio to an energetic challenge is a reduction in the size of the reproductive organs, without an inhibition of spermatogenesis. It is likely that this effect is mediated via white fat and leptin, and leptin’s influence on the hypothalamic-pituitary-gonad axis. Results of the study support the suggestion that females are more sensitive to reproductive inhibition than males and that reproduction in Rhabdomys pumilio is truly opportunistic.
- Full Text:
- Date Issued: 2003
- Authors: Jackson, Claire
- Date: 2003
- Subjects: Microtus Microtus -- South Africa -- Reproduction Mice -- Reproduction
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5777 , http://hdl.handle.net/10962/d1005465
- Description: Previous studies of the control of reproduction in Rhabdomys pumilio have shown that day length alone does not inhibit spermatogenesis, that a reduction in food availability and ambient temperature results in an inhibition of gametogenesis, that females are more susceptible to inhibition than are males, and that mice that are able to maintain a body fat store in the face of an energetic challenge, are less likely to show reproductive inhibition than those that lose their fat store. In the present study, field and laboratory experiments were conducted to examine the effects of winter food supplementation on reproduction and population dynamics, and the effects of exogenous GnRH, leptin and mercaptoacetate (MA) on reproductive activity of Rhabdomys pumilio exposed to an energetic challenge. In the field food supplementation experiments in Thomas Baines Nature Reserve (2000, 2001), there was no winter inhibition of reproduction and provision of supplementary food had little effect. While at Mountain Zebra National Park (2002) winter was harsher, females became reproductively inactive, spermatogenesis continued and the provision of extra food resulted in higher rates of individual growth and larger reproductive organs. Treatment of mice that had been exposed to a prolonged energetic challenge, with exogenous GnRH (1µg/mouse/treatment) resulted in an increase in the masses of the testes and epididymides, and in the activity of the reproductive organs. Treatment with exogenous leptin (40µg/mouse/treatment), concurrently with an energetic challenge, countered the negative effects of the energetic challenge, and treated males had larger reproductive organs. MA (600µmol/kg body mass), given concurrently with an energetic challenge, did not inhibit fat metabolism, although the high-fat diet countered the effects of the energetic challenge. Results suggest that the first response of male Rhabdomys pumilio to an energetic challenge is a reduction in the size of the reproductive organs, without an inhibition of spermatogenesis. It is likely that this effect is mediated via white fat and leptin, and leptin’s influence on the hypothalamic-pituitary-gonad axis. Results of the study support the suggestion that females are more sensitive to reproductive inhibition than males and that reproduction in Rhabdomys pumilio is truly opportunistic.
- Full Text:
- Date Issued: 2003
Intercohort cannibalism and parturition-associated behaviour of captive-bred swordtail, Xiphophorus helleri (Pisces: Poeciliidae)
- Jones, Clifford Louis Wilshire
- Authors: Jones, Clifford Louis Wilshire
- Date: 2003
- Subjects: Poeciliidae -- Behavior Xiphophorus helleri Fish culture
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5351 , http://hdl.handle.net/10962/d1007812
- Description: Adult fish that belong to the family Poeciliidae cannibalise juveniles, both in the wild and under captive conditions, but this behaviour has only been partly investigated in the Poeciliidae in some of the commercially valuable species. The objective of the research is to develop an understanding of intercohort cannibalism and parturition-associated behaviour in captive-bred swordtail (Xiphophorus helleri), with applications to industry and future research of other poeciliids. Experiments investigating the effect of adult stocking density and sex ratio on the production of juveniles were used to determine if cannibalism occurs under culture conditions. The average rate of intercohort cannibalism ranged from (5.5 to 53.9%), and was positively density dependent and independent of sex ratio, indicating that males and females were probably equally cannibalistic. The highest number of juveniles (1725.7±141.4) produced per tank over 70 days was obtained from two males and eight females. To develop a better understanding of adult and juvenile behaviour during parturition, fish were observed under controlled laboratory conditions using video and behaviours such as attack (burst of speed by an adult in the direction of a juvenile), escape (avoidance of cannibalism after attack) and cannibalism (predation of a live juvenile by an adult), for example, were identified. Under laboratory conditions most young escaped in downward direction after attack (49%) and most utilised the refuge made that was made available. Furthermore, most attacks (62-65%) and cannibalism (57-84%) occurred at the bottom. Since the presence of refuge significantly increased the rate of juvenile survival under culture conditions, it was hypothesised that the rate of cannibalism could be reduced under farming conditions if juveniles were protected when they escaped downwards. This hypothesis was accepted as it was found that refuge at the bottom of the water column or the inclusion of a false-bottom reduced the rate of cannibalism by 49% and 72%, respectively. Similarly, the hypothesise that the rate of cannibalism could be reduced if juveniles where given protection when escaping sideways (32% of juveniles escaped sideways in the laboratory) was also accepted when tested under farm-scale conditions because a false-side reduced the rate of cannibalism by an average of 45%. Since males and females were equally responsible for cannibalising juveniles in the laboratory, it was hypothesised that the rate of cannibalism would decrease proportionately with the removal of males (Le. 20% of the cannibals) from the population; the removal of males under farming conditions resulted in a 19.5% reduction in the rate of cannibalism. Since older juveniles were better able to escape cannibalism than neonates and since adults habituate to stimuli that previously resulted in attack behaviour, it was hypothesised that the rate of cannibalism would remain unaffected by the length of time that juveniles were exposed to adults in the breeding tanks. This hypothesis was also accepted when tested under farm conditions. However, some hypotheses based on laboratory observations were not accepted. For example, a constant low light intensity did not appear to decrease the rate of cannibalism under farm conditions; also, the occurrence of dead and deformed juveniles went unnoticed in the laboratory, and under farm conditions, where adults did not have access to the bottom of the tank, 10% of the harvest consisted of dead and deformed juveniles. It is concluded that technologies, such as bottom-refuge or a false-side, that increase the size of the liveharvest and allow for the removal of potentially less viable offspring are recommended for the commercial production of poeciliids. The overall similarity of X. helleri behaviour between the laboratory experiments and the farm-scale trials suggests that the post-partum behaviour of X. helleri remains consistent under these different conditions; thus, behaviour under one set of conditions may be used to predict behaviour under other conditions. The application and significance of extrapolations to industry and future research of X. helleri and possibly other poeciliids were discussed and the most applicable laboratory observations with the highest extrapolation capacity were proposed. Furthermore, techniques were developed to aid industry and future researchers in making predictions relating to behaviour of X. helleri under different conditions based on laboratory observations. The results were used to develop a model indicating that selection pressures against cannibalism are not likely to exist at the rate of cannibalism observed here since the potential genetic gain through kin survival and inclusive fitness was shown to be greater than any potential genetiC loss experienced by a victim of cannibalism. The model was successfully tested under a range of social conditions. Other possible explanations for cannibalism in poeciliids, such as parental manipulation, nutritional advantages, opportunistic predation and the recovery of energy are discussed. It is suggested that the most likely proximate cause of cannibalism under captive conditions is opportunistic predation. The theory that cannibalism ensures that only viable genes of the victim are expressed, through inclusive fitness, is a possible ultimate cause of cannibalism, which may have been inherited from feral ancestors of captive-bred X. helleri.
- Full Text:
- Date Issued: 2003
- Authors: Jones, Clifford Louis Wilshire
- Date: 2003
- Subjects: Poeciliidae -- Behavior Xiphophorus helleri Fish culture
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5351 , http://hdl.handle.net/10962/d1007812
- Description: Adult fish that belong to the family Poeciliidae cannibalise juveniles, both in the wild and under captive conditions, but this behaviour has only been partly investigated in the Poeciliidae in some of the commercially valuable species. The objective of the research is to develop an understanding of intercohort cannibalism and parturition-associated behaviour in captive-bred swordtail (Xiphophorus helleri), with applications to industry and future research of other poeciliids. Experiments investigating the effect of adult stocking density and sex ratio on the production of juveniles were used to determine if cannibalism occurs under culture conditions. The average rate of intercohort cannibalism ranged from (5.5 to 53.9%), and was positively density dependent and independent of sex ratio, indicating that males and females were probably equally cannibalistic. The highest number of juveniles (1725.7±141.4) produced per tank over 70 days was obtained from two males and eight females. To develop a better understanding of adult and juvenile behaviour during parturition, fish were observed under controlled laboratory conditions using video and behaviours such as attack (burst of speed by an adult in the direction of a juvenile), escape (avoidance of cannibalism after attack) and cannibalism (predation of a live juvenile by an adult), for example, were identified. Under laboratory conditions most young escaped in downward direction after attack (49%) and most utilised the refuge made that was made available. Furthermore, most attacks (62-65%) and cannibalism (57-84%) occurred at the bottom. Since the presence of refuge significantly increased the rate of juvenile survival under culture conditions, it was hypothesised that the rate of cannibalism could be reduced under farming conditions if juveniles were protected when they escaped downwards. This hypothesis was accepted as it was found that refuge at the bottom of the water column or the inclusion of a false-bottom reduced the rate of cannibalism by 49% and 72%, respectively. Similarly, the hypothesise that the rate of cannibalism could be reduced if juveniles where given protection when escaping sideways (32% of juveniles escaped sideways in the laboratory) was also accepted when tested under farm-scale conditions because a false-side reduced the rate of cannibalism by an average of 45%. Since males and females were equally responsible for cannibalising juveniles in the laboratory, it was hypothesised that the rate of cannibalism would decrease proportionately with the removal of males (Le. 20% of the cannibals) from the population; the removal of males under farming conditions resulted in a 19.5% reduction in the rate of cannibalism. Since older juveniles were better able to escape cannibalism than neonates and since adults habituate to stimuli that previously resulted in attack behaviour, it was hypothesised that the rate of cannibalism would remain unaffected by the length of time that juveniles were exposed to adults in the breeding tanks. This hypothesis was also accepted when tested under farm conditions. However, some hypotheses based on laboratory observations were not accepted. For example, a constant low light intensity did not appear to decrease the rate of cannibalism under farm conditions; also, the occurrence of dead and deformed juveniles went unnoticed in the laboratory, and under farm conditions, where adults did not have access to the bottom of the tank, 10% of the harvest consisted of dead and deformed juveniles. It is concluded that technologies, such as bottom-refuge or a false-side, that increase the size of the liveharvest and allow for the removal of potentially less viable offspring are recommended for the commercial production of poeciliids. The overall similarity of X. helleri behaviour between the laboratory experiments and the farm-scale trials suggests that the post-partum behaviour of X. helleri remains consistent under these different conditions; thus, behaviour under one set of conditions may be used to predict behaviour under other conditions. The application and significance of extrapolations to industry and future research of X. helleri and possibly other poeciliids were discussed and the most applicable laboratory observations with the highest extrapolation capacity were proposed. Furthermore, techniques were developed to aid industry and future researchers in making predictions relating to behaviour of X. helleri under different conditions based on laboratory observations. The results were used to develop a model indicating that selection pressures against cannibalism are not likely to exist at the rate of cannibalism observed here since the potential genetic gain through kin survival and inclusive fitness was shown to be greater than any potential genetiC loss experienced by a victim of cannibalism. The model was successfully tested under a range of social conditions. Other possible explanations for cannibalism in poeciliids, such as parental manipulation, nutritional advantages, opportunistic predation and the recovery of energy are discussed. It is suggested that the most likely proximate cause of cannibalism under captive conditions is opportunistic predation. The theory that cannibalism ensures that only viable genes of the victim are expressed, through inclusive fitness, is a possible ultimate cause of cannibalism, which may have been inherited from feral ancestors of captive-bred X. helleri.
- Full Text:
- Date Issued: 2003
Language and value : the place of evaluation in linguistic theory
- Authors: Kilpert, Diana Mary
- Date: 2003
- Subjects: Linguistics -- Philosophy Systemic grammar Discourse analysis English language -- Standardization Linguistic analysis (Linguistics) Functionalism (Linguistics) Halliday, M. A. K. (Michael Alexander Kirkwood), 1925- Chomsky, Noam Labov, William Pinker, Steven, 1954- Harris, Roy, 1931-
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:2353 , http://hdl.handle.net/10962/d1002635
- Description: It is a central claim of modern linguistic theory that linguists do not prescribe, but describe language as it is, without pronouncing on correctness or judging one variety better than another. This attempt to exclude evaluation is motivated by a desire to be ' politically correct', which hinders objective analysis of language, and by an ill-advised imitation of the natural sciences, which obstructs the discipline's progress towards becoming a science in its own right. It involves linguists, as users of a valued variety, in self-deception and disingenuousness, distances them from the concerns of the ordinary language user, and betrays a failure to understand the involvement of social values in language, the nature of language itself, and the limits of linguistic science. On a wider scale, linguistics reflects society's devaluing and mechanisation of language. Despite growing concern expressed in the literature, and the incoherence that becomes apparent when linguists attempt to address social problems using a theory that regards language as an autonomous object, newcomers to the discipline continue to be taught that anti-prescriptivism is the natural corollary of a scientific approach to language. This thesis suggests that the way out of these difficulties is to rethink the meaning of ' theory' in linguistics. If we take the reflexivity of language seriously, building on M.A.K. Halliday's notion of 'linguistics as metaphor', we are reminded that a linguistic theory is made of language. Metalanguage must use the experiential and interpersonal meaning-making resources of everyday language. It follows that a linguistic theory cannot escape being evaluative, because evaluation is an inherent part of interpersonal meaning. If we fail to notice our own metalinguistic evaluation, this is because language disguises its evaluative meanings, or perhaps we are just not used to thinking of them as part of the grammar. To achieve clarity about the involvement of value in language, we need to turn our metalanguage back on itself - 'using the grammar to think with about the grammar' . Some ways of doing this are demonstrated here, turning the resources of systemic functional linguistics on linguists' own language. The circularity of this process should be seen not as a drawback but as a salutary reminder that linguistics is an interpretive rather than a discovery process. This knowledge should help us revalue language and make a place for evaluation in linguistic theory, paving the way for a socially responsible and productive linguistics.
- Full Text:
- Date Issued: 2003
- Authors: Kilpert, Diana Mary
- Date: 2003
- Subjects: Linguistics -- Philosophy Systemic grammar Discourse analysis English language -- Standardization Linguistic analysis (Linguistics) Functionalism (Linguistics) Halliday, M. A. K. (Michael Alexander Kirkwood), 1925- Chomsky, Noam Labov, William Pinker, Steven, 1954- Harris, Roy, 1931-
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:2353 , http://hdl.handle.net/10962/d1002635
- Description: It is a central claim of modern linguistic theory that linguists do not prescribe, but describe language as it is, without pronouncing on correctness or judging one variety better than another. This attempt to exclude evaluation is motivated by a desire to be ' politically correct', which hinders objective analysis of language, and by an ill-advised imitation of the natural sciences, which obstructs the discipline's progress towards becoming a science in its own right. It involves linguists, as users of a valued variety, in self-deception and disingenuousness, distances them from the concerns of the ordinary language user, and betrays a failure to understand the involvement of social values in language, the nature of language itself, and the limits of linguistic science. On a wider scale, linguistics reflects society's devaluing and mechanisation of language. Despite growing concern expressed in the literature, and the incoherence that becomes apparent when linguists attempt to address social problems using a theory that regards language as an autonomous object, newcomers to the discipline continue to be taught that anti-prescriptivism is the natural corollary of a scientific approach to language. This thesis suggests that the way out of these difficulties is to rethink the meaning of ' theory' in linguistics. If we take the reflexivity of language seriously, building on M.A.K. Halliday's notion of 'linguistics as metaphor', we are reminded that a linguistic theory is made of language. Metalanguage must use the experiential and interpersonal meaning-making resources of everyday language. It follows that a linguistic theory cannot escape being evaluative, because evaluation is an inherent part of interpersonal meaning. If we fail to notice our own metalinguistic evaluation, this is because language disguises its evaluative meanings, or perhaps we are just not used to thinking of them as part of the grammar. To achieve clarity about the involvement of value in language, we need to turn our metalanguage back on itself - 'using the grammar to think with about the grammar' . Some ways of doing this are demonstrated here, turning the resources of systemic functional linguistics on linguists' own language. The circularity of this process should be seen not as a drawback but as a salutary reminder that linguistics is an interpretive rather than a discovery process. This knowledge should help us revalue language and make a place for evaluation in linguistic theory, paving the way for a socially responsible and productive linguistics.
- Full Text:
- Date Issued: 2003
An investigation of the isolation, characterisation and application of hydantoinases for the industrial production of amino acids
- Authors: Kirchmann, Shaun
- Date: 2003
- Subjects: Hydantoin Amino acids Hydrolysis
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3969 , http://hdl.handle.net/10962/d1004028
- Description: This thesis describes a series of investigations into the hydantoin-hydrolysing activity of bacterial strains RU-KM1 and RU-OR, which were previously isolated for their ability to hydrolyse hydantoins to amino acids. The main aim of the study was to develop biotransformations with potential application in the production of enantiomerically pure amino acids using a bioreactor based system utilising the hydantoin hydrolysing enzymes of the two isolated microorganisms. Different substituted hydantoins may be used as substrates by these enzymes for the production of a variety of amino acids. These are not only important for amino acid production, but they may be used for production of other industrially important compounds, such as semisynthetic penicillin/ampicillin, L-aspartame (sweetener), Fluvalinate (insecticide), Enalapril (ACE inhibitor). Thus, the ability of the above-mentioned strains to hydrolyse these substrates was investigated, with the view to utilizing the maximum potential of these biocatalysts. Hydantoin conversion involves a two-step hydrolysis reaction which yields, initially, an N-carbamylamino acid intermediate, and subsequently, an amino acid. The hydantoin-hydrolysing enzymes of a Pseudomonas sp. RU-KM1, and an Agrobacterium sp. RU-OR were characterised as whole cells and in a crude extract preparation, and reaction conditions for its biocatalytic application were optimised. The optimum conditions for conversion of hydantoin to glycine were found to be 1 hour at 40 °C, with conversion yields greater than 30 % achieved. The enzymes of RU-KM1 demonstrated considerable stability, retaining 80 % of their activity after storage for 2 weeks at 4 °C. The activities of the enzymes were increased by the addition of a detergent to the extraction medium, suggesting that the enzymes might be membrane-bound. The results of the determination of the metal-dependence of the hydantoinase and N-carbamoylase of RU-KM1 suggested that these enzymes required metal ions for activity, with metal ions such as Cu[superscript (2+)], Fe[superscript (2+)], and Co[superscript (2+)] resulting in no significant change in enzyme activity, however there was an activation of the enzymes when Mn[superscript (2+)] was added to the enzymes. The stereoselectivity of the enzymes was investigated, and the results suggested that the hydantoinase was D-selective, whereas the N-carbamoylase was shown to be L-selective by other researchers. The hydantoin substrate selectivity of RU-KM1 and RU-OR was investigated, and the organisms were shown to be able to hydrolyse all of the seven substrates tested. However, there was a difference in activity levels between crude extract preparations and whole cells, with crude extracts generally showing slightly lower activity than whole cells in RU-KM1, and the whole cells or RU-OR showing the lower activity than its crude extract. Some difference was also observed in the order of preference of substrates between whole cells and crude extracts. The preferred substrate for RU-KM1 whole cells was isopropylhydantoin, whereas the crude extract preparation preferentially hydrolysed p-hydroxyphenylhydantoin, achieving 57 % and 52 % conversions respectively. RU-OR whole cells preferred methylhydantoin where as the crude extract preferred isopropylhydantoin, and showed 49 % and 51 % conversions respectively. The enzymes were characterised in terms of their temperature and pH optima, inducer requirements, and product inhibition studies. The hydantoinase of RU-KM1 was shown to be inducible with low levels of hydantoin, and thermostable upto 75 °C with its optima between 60 and 70 °C. The N-carbamoylase was shown to have its optima at 50 °C. The addition of ATP (0.5 mM), DTT (1 mM) and a protease inhibitor (2 mg.mL[superscript (-1)]) all increased the hydantoinase activity of RU-KM1 crude extract, however they had very little effect on the N-carbamoylase activity. The hydantoinase enzyme from extracts of RU-KM1 was partially purified by development of cell disruption methods using mechanical and lysing enzymes, followed by precipitation and chromatographic resolution. The results obtained showed a hydantoinase enzyme of between 48 and 66 kDa. RU-KM1 was grown under fermentation conditions using different minimal media. The activity and yields under these conditions were low. Previous attempt to grow the organism in a rich medium had resulted in an increase in biomass but no hydantoinase activity. A rich medium was developed by carbon and nitrogen optimisation and yielded biomass up to 30 g.L[superscript (-1)] dry cell weight. The hydantoinase activity was restored by nitrogen starvation in stationary phase. This resulted in high biomass with increased activity. This data is currently in press. Crude extract and whole cells were immobilised on flat sheet membranes, hollow fibre membranes and in alginate beads. Low hydantoinase activity was measured in bioreactors using membranes in different configurations. A significant increase in hydantoinase activity was measured when the crude extract was immobilised in sodium alginate, as a result of stabilisation of the N-carbamoylase. Temperature and pH optima were unaffected by the immobilisation procedure, however the durability of the enzymes increased 2-fold. Different configurations of the bioreactor were investigated, as well as a hydroxyphenylhydantoin as an alternative substrate in this study. The bioreactors showed a near 95 % conversion of the hydantoin to glycine, and a 99 % conversion using HPG. In conclusion, the hydantoin-hydrolysing enzymes of RU-KM1 have been shown to be possibly membrane associated, which is a novel finding. This study has shown that the hydantoinase of RU-KM1 is D-stereoselective, with high temperature stability. A growth medium was developed for the rapid production of active biomass. A bioreactor was developed using a single and a dual biocatalyst configuration, which was capable of hydrolysing hydantoin and monosubstituted hydantoins to produce amino acids. To our knowledge this system is the first such dual biocatalyst system reported for the production of amino acids.
- Full Text:
- Date Issued: 2003
- Authors: Kirchmann, Shaun
- Date: 2003
- Subjects: Hydantoin Amino acids Hydrolysis
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3969 , http://hdl.handle.net/10962/d1004028
- Description: This thesis describes a series of investigations into the hydantoin-hydrolysing activity of bacterial strains RU-KM1 and RU-OR, which were previously isolated for their ability to hydrolyse hydantoins to amino acids. The main aim of the study was to develop biotransformations with potential application in the production of enantiomerically pure amino acids using a bioreactor based system utilising the hydantoin hydrolysing enzymes of the two isolated microorganisms. Different substituted hydantoins may be used as substrates by these enzymes for the production of a variety of amino acids. These are not only important for amino acid production, but they may be used for production of other industrially important compounds, such as semisynthetic penicillin/ampicillin, L-aspartame (sweetener), Fluvalinate (insecticide), Enalapril (ACE inhibitor). Thus, the ability of the above-mentioned strains to hydrolyse these substrates was investigated, with the view to utilizing the maximum potential of these biocatalysts. Hydantoin conversion involves a two-step hydrolysis reaction which yields, initially, an N-carbamylamino acid intermediate, and subsequently, an amino acid. The hydantoin-hydrolysing enzymes of a Pseudomonas sp. RU-KM1, and an Agrobacterium sp. RU-OR were characterised as whole cells and in a crude extract preparation, and reaction conditions for its biocatalytic application were optimised. The optimum conditions for conversion of hydantoin to glycine were found to be 1 hour at 40 °C, with conversion yields greater than 30 % achieved. The enzymes of RU-KM1 demonstrated considerable stability, retaining 80 % of their activity after storage for 2 weeks at 4 °C. The activities of the enzymes were increased by the addition of a detergent to the extraction medium, suggesting that the enzymes might be membrane-bound. The results of the determination of the metal-dependence of the hydantoinase and N-carbamoylase of RU-KM1 suggested that these enzymes required metal ions for activity, with metal ions such as Cu[superscript (2+)], Fe[superscript (2+)], and Co[superscript (2+)] resulting in no significant change in enzyme activity, however there was an activation of the enzymes when Mn[superscript (2+)] was added to the enzymes. The stereoselectivity of the enzymes was investigated, and the results suggested that the hydantoinase was D-selective, whereas the N-carbamoylase was shown to be L-selective by other researchers. The hydantoin substrate selectivity of RU-KM1 and RU-OR was investigated, and the organisms were shown to be able to hydrolyse all of the seven substrates tested. However, there was a difference in activity levels between crude extract preparations and whole cells, with crude extracts generally showing slightly lower activity than whole cells in RU-KM1, and the whole cells or RU-OR showing the lower activity than its crude extract. Some difference was also observed in the order of preference of substrates between whole cells and crude extracts. The preferred substrate for RU-KM1 whole cells was isopropylhydantoin, whereas the crude extract preparation preferentially hydrolysed p-hydroxyphenylhydantoin, achieving 57 % and 52 % conversions respectively. RU-OR whole cells preferred methylhydantoin where as the crude extract preferred isopropylhydantoin, and showed 49 % and 51 % conversions respectively. The enzymes were characterised in terms of their temperature and pH optima, inducer requirements, and product inhibition studies. The hydantoinase of RU-KM1 was shown to be inducible with low levels of hydantoin, and thermostable upto 75 °C with its optima between 60 and 70 °C. The N-carbamoylase was shown to have its optima at 50 °C. The addition of ATP (0.5 mM), DTT (1 mM) and a protease inhibitor (2 mg.mL[superscript (-1)]) all increased the hydantoinase activity of RU-KM1 crude extract, however they had very little effect on the N-carbamoylase activity. The hydantoinase enzyme from extracts of RU-KM1 was partially purified by development of cell disruption methods using mechanical and lysing enzymes, followed by precipitation and chromatographic resolution. The results obtained showed a hydantoinase enzyme of between 48 and 66 kDa. RU-KM1 was grown under fermentation conditions using different minimal media. The activity and yields under these conditions were low. Previous attempt to grow the organism in a rich medium had resulted in an increase in biomass but no hydantoinase activity. A rich medium was developed by carbon and nitrogen optimisation and yielded biomass up to 30 g.L[superscript (-1)] dry cell weight. The hydantoinase activity was restored by nitrogen starvation in stationary phase. This resulted in high biomass with increased activity. This data is currently in press. Crude extract and whole cells were immobilised on flat sheet membranes, hollow fibre membranes and in alginate beads. Low hydantoinase activity was measured in bioreactors using membranes in different configurations. A significant increase in hydantoinase activity was measured when the crude extract was immobilised in sodium alginate, as a result of stabilisation of the N-carbamoylase. Temperature and pH optima were unaffected by the immobilisation procedure, however the durability of the enzymes increased 2-fold. Different configurations of the bioreactor were investigated, as well as a hydroxyphenylhydantoin as an alternative substrate in this study. The bioreactors showed a near 95 % conversion of the hydantoin to glycine, and a 99 % conversion using HPG. In conclusion, the hydantoin-hydrolysing enzymes of RU-KM1 have been shown to be possibly membrane associated, which is a novel finding. This study has shown that the hydantoinase of RU-KM1 is D-stereoselective, with high temperature stability. A growth medium was developed for the rapid production of active biomass. A bioreactor was developed using a single and a dual biocatalyst configuration, which was capable of hydrolysing hydantoin and monosubstituted hydantoins to produce amino acids. To our knowledge this system is the first such dual biocatalyst system reported for the production of amino acids.
- Full Text:
- Date Issued: 2003
Metal interactions with neural substrates and their role in neurodegeneration
- Authors: Lack, Barbara Anne
- Date: 2003
- Subjects: Nervous system -- Degeneration Neurotoxicology Chemical reactions Metals -- Physiological effect Melatonin -- Physiological effect
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4391 , http://hdl.handle.net/10962/d1005709
- Description: "Life" may be characterized as a controlled stationary flow equilibrium, maintained by energy consuming chemical reactions. The physiological functioning of these life systems include at least 28 of the elements isolated on the periodic table thus far, most of which are metals. However, as with Paracelsus Principle: "The dose makes the poison", there exists a definite link between metal levels, essential and toxic, and the onset of neurodegenerative diseases. The economic costs of brain dysfunction are enormous, but this pales in comparison to the staggering emotional toll on the victims themselves and their families. In an attempt to improve the understanding of the causes of neurodegeneration, this study focuses on one potential aspect: the possible link between metals and neurotransmitter homeostasis utilising a variety of electronanalytical techniques. Adsorptive cathodic stripping voltammetry was employed to investigate the binding affinities and complex formation of melatonin and its precursor serotonin with calcium, potassium, sodium, lithium and aluminium. The results showed that all the metals studied formed complexes with both pineal indoleamines. However, the stability and affmity of the ligands toward the various metals varied greatly. The study suggests a further role for melatonin, that of metalloregulator and possible metal detoxifier in the brain, the in vivo studies which followed will further substantiate this notion. This research additionally focused on the cholinergic system, in particular acetylcholine complex formation studies with mercury, lead, cadmium, copper and zinc using the adsorptive cathodic stripping voltammetry method. The formation and characterisation of a solid mercury-acetylcholine complex lent further strength to the in situ electrochemical complex formation observed. The results showed the preference of acetylcholine for environmentally toxic heavy metals (such as Cd²⁺) over those divalent cations that occur naturally in the body. The possible metalloregulatory role melatonin played in the three brain regIOns: cerebellum, cortex and corpus striatum of male Wistar rats was studied as an in vivo extension of the earlier in vitro studies. Anodic stripping voltammetry was employed to detect metal levels present. The results showed that daily injections of melatonin was responsible for significantly decreasing copper(I), cadmium(II) and lead(II) levels in various regions of the rat brain of those animals that had undergone a pinealectomy in comparison to the saline injected group having undergone the same treatment. Histological and electrochemical stripping techniques were applied to investigate the implications of high A1³⁺ levels in the brain regions, particularly the hippocampus. Melatonin showed signs of promise in indirect symptom alleviation and by significantly decreasing A1³⁺ levels in rats that had been dosed with melatonin prior to A1³⁺ treatments in comparison with the control groups. Finally a preliminary study outlining a method for the production of a calcium selective microelectrode was undertaken. Further work is still needed to optimise the microelectrode production as well as its possible applications. However, whilst the overall conclusions of this entire multidisciplinary study may indeed only be in effect one piece of a very large puzzle on neurodegenerative diseases, this piece will no doubt serve as a building block for further ideas and work in this field.
- Full Text:
- Date Issued: 2003
- Authors: Lack, Barbara Anne
- Date: 2003
- Subjects: Nervous system -- Degeneration Neurotoxicology Chemical reactions Metals -- Physiological effect Melatonin -- Physiological effect
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4391 , http://hdl.handle.net/10962/d1005709
- Description: "Life" may be characterized as a controlled stationary flow equilibrium, maintained by energy consuming chemical reactions. The physiological functioning of these life systems include at least 28 of the elements isolated on the periodic table thus far, most of which are metals. However, as with Paracelsus Principle: "The dose makes the poison", there exists a definite link between metal levels, essential and toxic, and the onset of neurodegenerative diseases. The economic costs of brain dysfunction are enormous, but this pales in comparison to the staggering emotional toll on the victims themselves and their families. In an attempt to improve the understanding of the causes of neurodegeneration, this study focuses on one potential aspect: the possible link between metals and neurotransmitter homeostasis utilising a variety of electronanalytical techniques. Adsorptive cathodic stripping voltammetry was employed to investigate the binding affinities and complex formation of melatonin and its precursor serotonin with calcium, potassium, sodium, lithium and aluminium. The results showed that all the metals studied formed complexes with both pineal indoleamines. However, the stability and affmity of the ligands toward the various metals varied greatly. The study suggests a further role for melatonin, that of metalloregulator and possible metal detoxifier in the brain, the in vivo studies which followed will further substantiate this notion. This research additionally focused on the cholinergic system, in particular acetylcholine complex formation studies with mercury, lead, cadmium, copper and zinc using the adsorptive cathodic stripping voltammetry method. The formation and characterisation of a solid mercury-acetylcholine complex lent further strength to the in situ electrochemical complex formation observed. The results showed the preference of acetylcholine for environmentally toxic heavy metals (such as Cd²⁺) over those divalent cations that occur naturally in the body. The possible metalloregulatory role melatonin played in the three brain regIOns: cerebellum, cortex and corpus striatum of male Wistar rats was studied as an in vivo extension of the earlier in vitro studies. Anodic stripping voltammetry was employed to detect metal levels present. The results showed that daily injections of melatonin was responsible for significantly decreasing copper(I), cadmium(II) and lead(II) levels in various regions of the rat brain of those animals that had undergone a pinealectomy in comparison to the saline injected group having undergone the same treatment. Histological and electrochemical stripping techniques were applied to investigate the implications of high A1³⁺ levels in the brain regions, particularly the hippocampus. Melatonin showed signs of promise in indirect symptom alleviation and by significantly decreasing A1³⁺ levels in rats that had been dosed with melatonin prior to A1³⁺ treatments in comparison with the control groups. Finally a preliminary study outlining a method for the production of a calcium selective microelectrode was undertaken. Further work is still needed to optimise the microelectrode production as well as its possible applications. However, whilst the overall conclusions of this entire multidisciplinary study may indeed only be in effect one piece of a very large puzzle on neurodegenerative diseases, this piece will no doubt serve as a building block for further ideas and work in this field.
- Full Text:
- Date Issued: 2003
The effects of goat browsing on ecosystem patterns and processes in succulent thicket, South Africa
- Lechmere-Oertel, Richard Geoffrey
- Authors: Lechmere-Oertel, Richard Geoffrey
- Date: 2003
- Subjects: Succulent plants -- South Africa -- Eastern Cape , Landscape ecology -- South Africa -- Eastern Cape , Goats -- Feeding and feeds
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:11074 , http://hdl.handle.net/10948/309 , Succulent plants -- South Africa -- Eastern Cape , Landscape ecology -- South Africa -- Eastern Cape , Goats -- Feeding and feeds
- Description: Transformation in the arid succulent thicket of the Eastern Cape of South Africa in response to unsustainable livestock production has been widespread, with less than 10 percent remaining intact. Transformation in succulent thicket has resulted in large areas of dense thicket (comprising a two-phase mosaic of perennial-vegetated patches separated by animal paths and bare patches) being replaced with a ‘pseudo-savanna’ of remnant canopy trees with a structurally simple field layer of ephemeral and short- lived perennial grasses and forbs. There is an extensive literature describing the transformation of succulent thicket, with many speculative statements about the underlying mechanisms of transformation. The central focus of this study was to improve our mechanistic understanding of transformation in succulent thicket using field experiments. Hopefully these results will set another foundation upon which future management of succulent thicket can be improved and large-scale restoration initiated. This study comprises four themes that are linked to the concept of landscape function. The central premise of landscape function is that functional landscapes have mechanisms that capture and retain scarce resources. Conversely, as landscapes become increasingly dysfunctional, so these mechanisms become disrupted. In succulent thicket, dysfunctio n appears to be linked to the reduced ability to harvest water, cycle carbon and a loss of organic carbon. In this thesis I examined some of the key processes that influence water and organic carbon fluxes: perennial vegetation cover, soil fertility, litter fall and decomposition, and runoff and soil erosion. The experimental design that was used for all this work was a factorial ANOVA based on replicated fenceline contrasts that reflect differences in long-term management history. The main objectives of this thesis were to: quantify the patterns of transformation in an arid form of succulent thicket, including changes in the biomass, cover and structure of the dominant vegetation guilds; test the stability of the transformed succulent thicket ecosystem to show whether it is a new stable state or an intermediate stage in a trajectory towards a highly desertified state where only the ephemeral grasses and forbs persist; describe and compare soil fertility across transformation contrasts, concentrating on changes in the spatial patterns of soil resources and the ability of the soil to harvest precipitation; to compare litter fall and decomposition of leaf material from the dominant plants in intact and transformed succulent thicket; to quantify and compare run-off and erosion from run-off plots in intact and transformed succulent thicket. Transformation and stability I quantified the changes in plant diversity, physiognomy and biomass that occur across transformation contrasts. Thicket transformation results in a significant loss of plant diversity and functional types. There is also a significant reduction in the biomass (c. 80 t.ha-1) and structural complexity of the vegetation, both vertically and horizontally. These results were interpreted in terms of their implications for ecosystem functioning and stability. To test the stability of the transformed succulent thicket I used aerial photographs and ground-truthing to track the survivorship of canopy trees over 60 years in pseudo-savanna landscapes. I also measured seedling establishment in different habitats. I show that the pseudo-savanna is not a stable state owing to ongoing adult mortality and no recruitment of canopy trees. Soil fertility and water status I hypothesised that the above-ground changes in ve getation would be accompanied by similar trends in the pattern and levels of soil nutrient resources and the ability of the landscape to harvest precipitation. I compared soil fertility (organic carbon, available nitrogen and phosphorus), texture, matric potential, and surface micro-topography in the two main micro- habitats on either side of the replicated fenceline contrasts. The results show that intact spekboom thicket has a distinct spatial pattern of soil fertility where nutrients and organic carbon are concentrated under the patches of perennial shrubs, compared to under canopy trees and open spaces. Transformation results in a significant homogenisation out of this pattern and an overall reduction in the fertility of the landscape. The proportion of the landscape surface that would promote infiltration of water decreases from 60 – 0.6 percent. Soil moisture retention (matric potential) also decreases with transformation. I interpreted these patterns in terms of the ability of the landscape to harvest and release water after rainfall events. Litter fall and decomposition Surface litter and soil organic matter are critical components to wooded ecosystems; contributing to several ecosystem functions. The rates of litter fall and decomposition are ratelimiting steps in nutrient cycling and incorporation of organic matter into the soil. The ecological mechanisms behind the collapse of succulent thicket in the face of domestic herbivory are not fully understood, but are believed to include the breakdown of several ecosystem processes, including litter fall and decomposition. I quantified the changes in litter fall and litter decomposition of four of the dominant perennial woody plants (Euclea undulata, Pappea capensis, Portulacaria afra and Rhus longispina) across the replicated fenceline. Litter fall was measured over 14 months using mesh traps. Decomposition was measured over 15 months using a combination of litterbags and unprotected leaf packs. I also quantified soil microclimate during the experimental period; hypothesising that transformation would lead to soil conditions less amenable for biotic activity. Litter fall in succulent thicket was very high for a semi-arid system, comparing more to temperate forests. The leaf-succulent P. afra contributed the largest single component of the total litter production at a landscape scale. The effect of transformation on litter fall was species specific. Deep-rooted or drought-adapted species showed no change in litter yield with transformation; shallow-rooted species showed a significant decrease. There were few significant differences in decomposition rates across the transformation gradient and between litter types. Portulacaria afra litter had the steepest rate of mass loss, and was most affected by transformation. The more recalcitrant (high C:N ratio) leaves of P. capensis remained largely unaffected by transformation. These results indicate the critical role of the perennial vegetation in incorporating organic carbon into the soil. Transformation of succulent thicket leads to a disruption of the flow of carbon into the soil, reinforcing the cycle of transformation through reduced fertility. Rehabilitation of this ecosystem will require the active establishment of species, such as P. afra, that will restart the flow of carbon into the soil. Run-off & Erosion The landscape function model predicts that functional semi-arid shrublands efficiently conserve limiting resources such as water and water-bourn sediments (soil and organic matter). As these rangelands become transformed through unsustainable livestock production, so their ability to conserve resources decreases. The primary determinant of landscape function and conservation of resources appears to be the proportional cover of perennial vegetation. I hypothesised that the switch from a two-phase mosaic dominated by perennial succulent and woody shrubs to a single phase system dominated by an ephemeral field layer would be accompanied by disruption of the mechanisms that conserve resources. Specifically, I tested the hypothesis that transformation of succulent thicket increases runoff volume, sediment concentration of runoff, soil erosion and loss of organic matter at a patch scale (c. 100 m2). Runoff and water-borne sediment were measured from runoff plots established across replicated fenceline contrasts. Data were collected from eight extreme weather events over two years. There were no significant differences between runoff and erosion across the transformation contrast, as each extreme weather event was unique in terms of its runoff response. The transformed runoff plots alone also gave inconsistent results, largely due to differences in the cover of ephemeral forbs and weakly perennial grasses. Runoff and erosion were not predictable from the data across the transformation gradient due to complex interactions between the nature of the above-ground vegetation, soil micro-topography and land use history. The results highlighted the need for longer-term catchment experiments to generate a predictive understanding of the effect of transformation on runoff and erosion in succulent thicket.
- Full Text:
- Date Issued: 2003
- Authors: Lechmere-Oertel, Richard Geoffrey
- Date: 2003
- Subjects: Succulent plants -- South Africa -- Eastern Cape , Landscape ecology -- South Africa -- Eastern Cape , Goats -- Feeding and feeds
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:11074 , http://hdl.handle.net/10948/309 , Succulent plants -- South Africa -- Eastern Cape , Landscape ecology -- South Africa -- Eastern Cape , Goats -- Feeding and feeds
- Description: Transformation in the arid succulent thicket of the Eastern Cape of South Africa in response to unsustainable livestock production has been widespread, with less than 10 percent remaining intact. Transformation in succulent thicket has resulted in large areas of dense thicket (comprising a two-phase mosaic of perennial-vegetated patches separated by animal paths and bare patches) being replaced with a ‘pseudo-savanna’ of remnant canopy trees with a structurally simple field layer of ephemeral and short- lived perennial grasses and forbs. There is an extensive literature describing the transformation of succulent thicket, with many speculative statements about the underlying mechanisms of transformation. The central focus of this study was to improve our mechanistic understanding of transformation in succulent thicket using field experiments. Hopefully these results will set another foundation upon which future management of succulent thicket can be improved and large-scale restoration initiated. This study comprises four themes that are linked to the concept of landscape function. The central premise of landscape function is that functional landscapes have mechanisms that capture and retain scarce resources. Conversely, as landscapes become increasingly dysfunctional, so these mechanisms become disrupted. In succulent thicket, dysfunctio n appears to be linked to the reduced ability to harvest water, cycle carbon and a loss of organic carbon. In this thesis I examined some of the key processes that influence water and organic carbon fluxes: perennial vegetation cover, soil fertility, litter fall and decomposition, and runoff and soil erosion. The experimental design that was used for all this work was a factorial ANOVA based on replicated fenceline contrasts that reflect differences in long-term management history. The main objectives of this thesis were to: quantify the patterns of transformation in an arid form of succulent thicket, including changes in the biomass, cover and structure of the dominant vegetation guilds; test the stability of the transformed succulent thicket ecosystem to show whether it is a new stable state or an intermediate stage in a trajectory towards a highly desertified state where only the ephemeral grasses and forbs persist; describe and compare soil fertility across transformation contrasts, concentrating on changes in the spatial patterns of soil resources and the ability of the soil to harvest precipitation; to compare litter fall and decomposition of leaf material from the dominant plants in intact and transformed succulent thicket; to quantify and compare run-off and erosion from run-off plots in intact and transformed succulent thicket. Transformation and stability I quantified the changes in plant diversity, physiognomy and biomass that occur across transformation contrasts. Thicket transformation results in a significant loss of plant diversity and functional types. There is also a significant reduction in the biomass (c. 80 t.ha-1) and structural complexity of the vegetation, both vertically and horizontally. These results were interpreted in terms of their implications for ecosystem functioning and stability. To test the stability of the transformed succulent thicket I used aerial photographs and ground-truthing to track the survivorship of canopy trees over 60 years in pseudo-savanna landscapes. I also measured seedling establishment in different habitats. I show that the pseudo-savanna is not a stable state owing to ongoing adult mortality and no recruitment of canopy trees. Soil fertility and water status I hypothesised that the above-ground changes in ve getation would be accompanied by similar trends in the pattern and levels of soil nutrient resources and the ability of the landscape to harvest precipitation. I compared soil fertility (organic carbon, available nitrogen and phosphorus), texture, matric potential, and surface micro-topography in the two main micro- habitats on either side of the replicated fenceline contrasts. The results show that intact spekboom thicket has a distinct spatial pattern of soil fertility where nutrients and organic carbon are concentrated under the patches of perennial shrubs, compared to under canopy trees and open spaces. Transformation results in a significant homogenisation out of this pattern and an overall reduction in the fertility of the landscape. The proportion of the landscape surface that would promote infiltration of water decreases from 60 – 0.6 percent. Soil moisture retention (matric potential) also decreases with transformation. I interpreted these patterns in terms of the ability of the landscape to harvest and release water after rainfall events. Litter fall and decomposition Surface litter and soil organic matter are critical components to wooded ecosystems; contributing to several ecosystem functions. The rates of litter fall and decomposition are ratelimiting steps in nutrient cycling and incorporation of organic matter into the soil. The ecological mechanisms behind the collapse of succulent thicket in the face of domestic herbivory are not fully understood, but are believed to include the breakdown of several ecosystem processes, including litter fall and decomposition. I quantified the changes in litter fall and litter decomposition of four of the dominant perennial woody plants (Euclea undulata, Pappea capensis, Portulacaria afra and Rhus longispina) across the replicated fenceline. Litter fall was measured over 14 months using mesh traps. Decomposition was measured over 15 months using a combination of litterbags and unprotected leaf packs. I also quantified soil microclimate during the experimental period; hypothesising that transformation would lead to soil conditions less amenable for biotic activity. Litter fall in succulent thicket was very high for a semi-arid system, comparing more to temperate forests. The leaf-succulent P. afra contributed the largest single component of the total litter production at a landscape scale. The effect of transformation on litter fall was species specific. Deep-rooted or drought-adapted species showed no change in litter yield with transformation; shallow-rooted species showed a significant decrease. There were few significant differences in decomposition rates across the transformation gradient and between litter types. Portulacaria afra litter had the steepest rate of mass loss, and was most affected by transformation. The more recalcitrant (high C:N ratio) leaves of P. capensis remained largely unaffected by transformation. These results indicate the critical role of the perennial vegetation in incorporating organic carbon into the soil. Transformation of succulent thicket leads to a disruption of the flow of carbon into the soil, reinforcing the cycle of transformation through reduced fertility. Rehabilitation of this ecosystem will require the active establishment of species, such as P. afra, that will restart the flow of carbon into the soil. Run-off & Erosion The landscape function model predicts that functional semi-arid shrublands efficiently conserve limiting resources such as water and water-bourn sediments (soil and organic matter). As these rangelands become transformed through unsustainable livestock production, so their ability to conserve resources decreases. The primary determinant of landscape function and conservation of resources appears to be the proportional cover of perennial vegetation. I hypothesised that the switch from a two-phase mosaic dominated by perennial succulent and woody shrubs to a single phase system dominated by an ephemeral field layer would be accompanied by disruption of the mechanisms that conserve resources. Specifically, I tested the hypothesis that transformation of succulent thicket increases runoff volume, sediment concentration of runoff, soil erosion and loss of organic matter at a patch scale (c. 100 m2). Runoff and water-borne sediment were measured from runoff plots established across replicated fenceline contrasts. Data were collected from eight extreme weather events over two years. There were no significant differences between runoff and erosion across the transformation contrast, as each extreme weather event was unique in terms of its runoff response. The transformed runoff plots alone also gave inconsistent results, largely due to differences in the cover of ephemeral forbs and weakly perennial grasses. Runoff and erosion were not predictable from the data across the transformation gradient due to complex interactions between the nature of the above-ground vegetation, soil micro-topography and land use history. The results highlighted the need for longer-term catchment experiments to generate a predictive understanding of the effect of transformation on runoff and erosion in succulent thicket.
- Full Text:
- Date Issued: 2003
The phosphorylation and nuclear localization of the co-chaperone murine stress-inducible protein 1
- Authors: Longshaw, Victoria Mary
- Date: 2003
- Subjects: Phosphorylation Proteins Heat shock proteins
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3979 , http://hdl.handle.net/10962/d1004038
- Description: The co-chaperone murine stress-inducible protein 1 (mSTI1), a heat shock protein 70 (Hsp70)/ heat shock protein 90 (Hsp90) organizing protein (Hop) homologue, mediates the assembly of the Hsp70/Hsp90 chaperone heterocomplex. mSTI1 is phosphorylated in vitro by cell cycle kinases, proximal to a putative nuclear localization signal (NLS), substantiating a predicted CKII-cdc2-NLS (CcN) motif at position 189-239. Stable transfectants of NIH 3T3 fibroblasts that expressed mSTI1-EGFP, NLSmSTI1-EGFP and EGFP, were prepared. Fluorescence microscopy revealed mSTI1 was cytoplasmically localized, and that this localization was not affected by the fusion of mSTI1 with the EGFP moiety. NLSmSTI1-EGFP was targeted to the nucleus compared to EGFP, suggesting that the NLSmSTI1 was a functional NLS. The localization of mSTI1 was determined under normal and heat shock conditions, inhibition of nuclear export (leptomycin B), inhibition of CKII 5,6-dichlorobenzimidazole riboside, DRB), inhibition of cdc2 kinase (olomoucine), and G1/S phase arrest (hydroxyurea). mSTI1-EGFP and mSTI1 were excluded from the nucleus in the majority of resting cells, but accumulated in the nucleus following leptomycin B treatment, implying that mSTI1 possibly undergoes a functional import process, and export via the chromosomal region maintenance 1 (CRM-1)-mediated export pathway. Hydroxyurea and olomoucine (but not DRB or heat shock) treatment increased the proportion of cells in which mSTI1-EGFP exhibited cytoplasmic and nuclear localization. 2D gel electrophoresis detected three endogenous mSTI1 isoforms, which changed following hydroxyurea treatment. Furthermore, point inactivation and mimicking of phosphorylatable residues in mSTI1 altered the translocation of the protein and the isoform composition. Modification of mSTI1 at S189 and T198 decreased the number of isoforms of mSTI1-EGFP, suggesting that the protein is modified at these sites in vivo. The removal of the in vitro cdc2 kinase site at T198 promoted a nuclear localization during G1/S phase arrest. Therefore active cdc2 kinase, but not CKII, may be required for cytoplasmic localization of mSTI1. The CKII site appears to have no regulatory role under heat shock conditions or during the cell cycle. In vitro phosphorylation studies on untagged mSTI1 further supported the prediction that S189 is the only site recognised by CKII. The cdc2 kinase site at T198, however, although the major site, was not the only site phosphorylated in vitro. However, mSTI1 and cdc2 kinase did not interact in a detectable stable complex. Bioinformatic analysis of mSTI1 revealed NLS residues were conserved in STI1 proteins, and the NLS and TPR2A motifs were in close proximity. This may have mechanistic implications for the formation of the Hsp90-mSTI1 heterocomplex. The cytoplasmic or nuclear localization of mSTI1 is predicted to be the result of a dynamic equilibrium between nuclear import and nuclear export, the fulcrum of which may be shifted under different cell cycle conditions. These data provide the first evidence of regulated nuclear import/export of a major Hsp70/Hsp90 co-chaperone, and the regulation of this nuclear import by cell cycle status and cell cycle kinases.
- Full Text:
- Date Issued: 2003
- Authors: Longshaw, Victoria Mary
- Date: 2003
- Subjects: Phosphorylation Proteins Heat shock proteins
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3979 , http://hdl.handle.net/10962/d1004038
- Description: The co-chaperone murine stress-inducible protein 1 (mSTI1), a heat shock protein 70 (Hsp70)/ heat shock protein 90 (Hsp90) organizing protein (Hop) homologue, mediates the assembly of the Hsp70/Hsp90 chaperone heterocomplex. mSTI1 is phosphorylated in vitro by cell cycle kinases, proximal to a putative nuclear localization signal (NLS), substantiating a predicted CKII-cdc2-NLS (CcN) motif at position 189-239. Stable transfectants of NIH 3T3 fibroblasts that expressed mSTI1-EGFP, NLSmSTI1-EGFP and EGFP, were prepared. Fluorescence microscopy revealed mSTI1 was cytoplasmically localized, and that this localization was not affected by the fusion of mSTI1 with the EGFP moiety. NLSmSTI1-EGFP was targeted to the nucleus compared to EGFP, suggesting that the NLSmSTI1 was a functional NLS. The localization of mSTI1 was determined under normal and heat shock conditions, inhibition of nuclear export (leptomycin B), inhibition of CKII 5,6-dichlorobenzimidazole riboside, DRB), inhibition of cdc2 kinase (olomoucine), and G1/S phase arrest (hydroxyurea). mSTI1-EGFP and mSTI1 were excluded from the nucleus in the majority of resting cells, but accumulated in the nucleus following leptomycin B treatment, implying that mSTI1 possibly undergoes a functional import process, and export via the chromosomal region maintenance 1 (CRM-1)-mediated export pathway. Hydroxyurea and olomoucine (but not DRB or heat shock) treatment increased the proportion of cells in which mSTI1-EGFP exhibited cytoplasmic and nuclear localization. 2D gel electrophoresis detected three endogenous mSTI1 isoforms, which changed following hydroxyurea treatment. Furthermore, point inactivation and mimicking of phosphorylatable residues in mSTI1 altered the translocation of the protein and the isoform composition. Modification of mSTI1 at S189 and T198 decreased the number of isoforms of mSTI1-EGFP, suggesting that the protein is modified at these sites in vivo. The removal of the in vitro cdc2 kinase site at T198 promoted a nuclear localization during G1/S phase arrest. Therefore active cdc2 kinase, but not CKII, may be required for cytoplasmic localization of mSTI1. The CKII site appears to have no regulatory role under heat shock conditions or during the cell cycle. In vitro phosphorylation studies on untagged mSTI1 further supported the prediction that S189 is the only site recognised by CKII. The cdc2 kinase site at T198, however, although the major site, was not the only site phosphorylated in vitro. However, mSTI1 and cdc2 kinase did not interact in a detectable stable complex. Bioinformatic analysis of mSTI1 revealed NLS residues were conserved in STI1 proteins, and the NLS and TPR2A motifs were in close proximity. This may have mechanistic implications for the formation of the Hsp90-mSTI1 heterocomplex. The cytoplasmic or nuclear localization of mSTI1 is predicted to be the result of a dynamic equilibrium between nuclear import and nuclear export, the fulcrum of which may be shifted under different cell cycle conditions. These data provide the first evidence of regulated nuclear import/export of a major Hsp70/Hsp90 co-chaperone, and the regulation of this nuclear import by cell cycle status and cell cycle kinases.
- Full Text:
- Date Issued: 2003
An investigation into the physico-chemical and neuroprotective properties of melatonin and 6-hydroxymelatonin
- Authors: Maharaj, Deepa Sukhdev
- Date: 2003
- Subjects: Melatonin Nervous system -- Degeneration -- Treatment
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3768 , http://hdl.handle.net/10962/d1003246
- Description: Until the beginning of this decade the antioxidant, melatonin, had been considered as little more than a tranquilizing hormone, responsible for regulating certain circadian and circannual rhythms. However, it is the discovery of melatonin as a free radical scavenger that has generated the most interest in recent years. The reduction of melatonin with age has been associated with neurodegenerative diseases such as Alzheimer’s disease (AD)and therefore, melatonin has been implicated to have an important clinical role in neuroprotection. Thus, for several years melatonin has attracted increasing attention from the general press with many advertisements touting this indoleamine to act as an aphrodisiac, rejuvenator, protector against diseases and a general wonder drug. However, melatonin formulations appear with no labelling for the correct storage conditions, dosage and side effects, as well as no control for purity and self-medicating with an unregulated product. In addition, there is much controversy surrounding the antioxidative properties of the indolemaine, 6-hydroxymelatonin (6-OHM). Therefore, the first part of this study aims to elucidate the physico-chemical and various stability characteristics of the pineal antioxidant, melatonin, while the second part is devoted to investigating the neuroprotective properties of the primary hepatic metabolite of melatonin, 6-OHM. The physical properties of melatonin were determined using various chemical techniques. This information served to both characterize and confirm the identity of melatonin raw material used in this study. In addition, this information serves to be essential as the physical properties of melatonin have not been reported in detail in literature, to date. Thereafter, using a validated high performance liquid chromatography (HPLC) method, the various physico-chemical and stability characteristics of melatonin were determined. Melatonin was shown to be extremely lipophilic, while the hygroscopic study indicates that melatonin raw material is extremely hygroscopic at temperatures above 40°C, whereas melatonin tablets are hygroscopic when left out of the original container. This study highlights the need for consumers to be aware of the proper storage of melatonin tablets to improve the stability and ensure long term integrity of the compound. Since, melatonin is most often administered orally, thus exposing it to a large variations in pH, within the gastrointestinal tract, it was decided to investigate the stability of melatonin over a range of pH’s and temperatures. The findings imply that melatonin is relatively stable at body temperature when ingested orally and that orally administered slow release preparations of melatonin should be relatively stable and therefore exhibit favourable bioavailability. However melatonin was shown to be unstable in solution. This provides important information and a challenge to the formulators of this drug substance in a liquid dosage form. An assessment of the photostability of melatonin dosage forms using International Committee on Harmonization (ICH) conditions revealed melatonin to be light sensitive and thus indicates a need for careful consideration of the packaging of these drug products. In addition a detailed assessment of the photochemistry and photoproducts formed during the UV photodegradation of melatonin is reported. Melatonin is shown to rapidly degrade in the presence of UV light, with the presence of oxygen accelerating the photodegradation. N1-acetyl-N2-formyl-5-methoxykynurenamine(AFMK) and 6-OHM were identified as the major photoproducts formed and these agents have been shown previously to retain antioxidant activity. One of the concerns of using melatonin in sunscreens is its photostability. However, it is reported in this study that the degraded solution of melatonin still possesses equipotent free radical scavenging ability as melatonin, despite the absence of melatonin in solution. In addition, melatonin is shown to reduce UV-induced oxidative stress in rat skin homogenate. Thus, these results make melatonin a likely candidate for inclusion in sunscreen preparations. Neuronal damage due to oxidative stress has been implicated in several neurodegenerative disorders. 6-OHM is not only formed as the major hepatic metabolite of melatonin, but also when melatonin reacts with toxic radicals as well as UV light. Thus the second part of the study aims to elucidate and further characterize the mechanism behind 6-OHM’s neuroprotection. The results show 6-OHM to be a more potent singlet oxygen and superoxide anion scavenger than melatonin. In addition, the results show 6-OHM to offer protection against, oxidative stress and lipid peroxidation induced by several neurotoxins in the rat brain and hippocampus. The hippocampus is an important region of the brain responsible for the formation of memory and any agent that induces stress in this area has detrimental effects and could lead to various types of dementia. Such agents include quinolinic acid (QA) and iron (II). Histological studies undertaken reveal that 6-OHM is able to protect hippocampal neurons against QA and iron (II) induced necrotic cell death. Immunohistochemical investigations showed that QA moderately induces apoptotic cell death in the hippocampus which is inhibited by both melatonin and 6-OHM. The study sought to elucidate possible mechanisms by which 6-OHM exerts its neuroprotective capabilities and the results show 6-OHM to inhibit the action of cyanide on the mitochondrial electron transport chain (ETC), one of the most common sources of free radicals. In addition, 6-OHM treatment alone, increased ETC activity above basal control levels and the results show 6-OHM to increase complex I activity in the mitochondrial ETC. Electrochemical, ultraviolet/visible spectroscopy (UV/Vis) and HPLC assessment show that an interaction exists between 6-OHM and iron (III) and 6-OHM is able to reduce iron (III) to a more biologically usable form viz. iron (II) which can be incorporated into important biomolecules such as heme. One dire consequence of this interaction is the ready provision of iron (II) to drive the Fenton reaction. However the biological and histological assessments show 6-OHM to prevent iron (II)-induced lipid peroxidation and necrotic cell death and thus, provide evidence of its antioxidant properties. The results also show 6-OHM to promote Hsp70 induction in the hippocampus. Heat shock proteins, especially Hsp 70 plays a role in cytoprotection by capturing denatured proteins and facilitating the refolding of these proteins once the stress has been relieved. 6-OHM treatment alone and together with QA was shown to increase the level of expression of Hsp70, both inducible and cognate forms of the protein. This suggests that 6-OHM helps to protect against cellular protein damage induced by any form of stress the cell may encounter. Melatonin treatment alone and in combination with QA was shown to prevent increases in the level of Hsp70 in the hippocampus, indicating that melatonin was able to reduce oxidative stress induced by QA such that Hsp70 expression was not required. The discovery of neuroprotective agents, such as melatonin and 6-OHM, is becoming important considering the rapid rise in the elderly population and the proportionate increase in neurological disorders. The findings of this study indicate the need for important information regarding the correct storage conditions and stability characteristics of melatonin dosage forms. In addition, the results indicate that 6-OHM has a definite role to play as an antioxidant. Thus further research may favour the use of these agents in the treatment of several neurodegenerative disorders.
- Full Text:
- Date Issued: 2003
- Authors: Maharaj, Deepa Sukhdev
- Date: 2003
- Subjects: Melatonin Nervous system -- Degeneration -- Treatment
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3768 , http://hdl.handle.net/10962/d1003246
- Description: Until the beginning of this decade the antioxidant, melatonin, had been considered as little more than a tranquilizing hormone, responsible for regulating certain circadian and circannual rhythms. However, it is the discovery of melatonin as a free radical scavenger that has generated the most interest in recent years. The reduction of melatonin with age has been associated with neurodegenerative diseases such as Alzheimer’s disease (AD)and therefore, melatonin has been implicated to have an important clinical role in neuroprotection. Thus, for several years melatonin has attracted increasing attention from the general press with many advertisements touting this indoleamine to act as an aphrodisiac, rejuvenator, protector against diseases and a general wonder drug. However, melatonin formulations appear with no labelling for the correct storage conditions, dosage and side effects, as well as no control for purity and self-medicating with an unregulated product. In addition, there is much controversy surrounding the antioxidative properties of the indolemaine, 6-hydroxymelatonin (6-OHM). Therefore, the first part of this study aims to elucidate the physico-chemical and various stability characteristics of the pineal antioxidant, melatonin, while the second part is devoted to investigating the neuroprotective properties of the primary hepatic metabolite of melatonin, 6-OHM. The physical properties of melatonin were determined using various chemical techniques. This information served to both characterize and confirm the identity of melatonin raw material used in this study. In addition, this information serves to be essential as the physical properties of melatonin have not been reported in detail in literature, to date. Thereafter, using a validated high performance liquid chromatography (HPLC) method, the various physico-chemical and stability characteristics of melatonin were determined. Melatonin was shown to be extremely lipophilic, while the hygroscopic study indicates that melatonin raw material is extremely hygroscopic at temperatures above 40°C, whereas melatonin tablets are hygroscopic when left out of the original container. This study highlights the need for consumers to be aware of the proper storage of melatonin tablets to improve the stability and ensure long term integrity of the compound. Since, melatonin is most often administered orally, thus exposing it to a large variations in pH, within the gastrointestinal tract, it was decided to investigate the stability of melatonin over a range of pH’s and temperatures. The findings imply that melatonin is relatively stable at body temperature when ingested orally and that orally administered slow release preparations of melatonin should be relatively stable and therefore exhibit favourable bioavailability. However melatonin was shown to be unstable in solution. This provides important information and a challenge to the formulators of this drug substance in a liquid dosage form. An assessment of the photostability of melatonin dosage forms using International Committee on Harmonization (ICH) conditions revealed melatonin to be light sensitive and thus indicates a need for careful consideration of the packaging of these drug products. In addition a detailed assessment of the photochemistry and photoproducts formed during the UV photodegradation of melatonin is reported. Melatonin is shown to rapidly degrade in the presence of UV light, with the presence of oxygen accelerating the photodegradation. N1-acetyl-N2-formyl-5-methoxykynurenamine(AFMK) and 6-OHM were identified as the major photoproducts formed and these agents have been shown previously to retain antioxidant activity. One of the concerns of using melatonin in sunscreens is its photostability. However, it is reported in this study that the degraded solution of melatonin still possesses equipotent free radical scavenging ability as melatonin, despite the absence of melatonin in solution. In addition, melatonin is shown to reduce UV-induced oxidative stress in rat skin homogenate. Thus, these results make melatonin a likely candidate for inclusion in sunscreen preparations. Neuronal damage due to oxidative stress has been implicated in several neurodegenerative disorders. 6-OHM is not only formed as the major hepatic metabolite of melatonin, but also when melatonin reacts with toxic radicals as well as UV light. Thus the second part of the study aims to elucidate and further characterize the mechanism behind 6-OHM’s neuroprotection. The results show 6-OHM to be a more potent singlet oxygen and superoxide anion scavenger than melatonin. In addition, the results show 6-OHM to offer protection against, oxidative stress and lipid peroxidation induced by several neurotoxins in the rat brain and hippocampus. The hippocampus is an important region of the brain responsible for the formation of memory and any agent that induces stress in this area has detrimental effects and could lead to various types of dementia. Such agents include quinolinic acid (QA) and iron (II). Histological studies undertaken reveal that 6-OHM is able to protect hippocampal neurons against QA and iron (II) induced necrotic cell death. Immunohistochemical investigations showed that QA moderately induces apoptotic cell death in the hippocampus which is inhibited by both melatonin and 6-OHM. The study sought to elucidate possible mechanisms by which 6-OHM exerts its neuroprotective capabilities and the results show 6-OHM to inhibit the action of cyanide on the mitochondrial electron transport chain (ETC), one of the most common sources of free radicals. In addition, 6-OHM treatment alone, increased ETC activity above basal control levels and the results show 6-OHM to increase complex I activity in the mitochondrial ETC. Electrochemical, ultraviolet/visible spectroscopy (UV/Vis) and HPLC assessment show that an interaction exists between 6-OHM and iron (III) and 6-OHM is able to reduce iron (III) to a more biologically usable form viz. iron (II) which can be incorporated into important biomolecules such as heme. One dire consequence of this interaction is the ready provision of iron (II) to drive the Fenton reaction. However the biological and histological assessments show 6-OHM to prevent iron (II)-induced lipid peroxidation and necrotic cell death and thus, provide evidence of its antioxidant properties. The results also show 6-OHM to promote Hsp70 induction in the hippocampus. Heat shock proteins, especially Hsp 70 plays a role in cytoprotection by capturing denatured proteins and facilitating the refolding of these proteins once the stress has been relieved. 6-OHM treatment alone and together with QA was shown to increase the level of expression of Hsp70, both inducible and cognate forms of the protein. This suggests that 6-OHM helps to protect against cellular protein damage induced by any form of stress the cell may encounter. Melatonin treatment alone and in combination with QA was shown to prevent increases in the level of Hsp70 in the hippocampus, indicating that melatonin was able to reduce oxidative stress induced by QA such that Hsp70 expression was not required. The discovery of neuroprotective agents, such as melatonin and 6-OHM, is becoming important considering the rapid rise in the elderly population and the proportionate increase in neurological disorders. The findings of this study indicate the need for important information regarding the correct storage conditions and stability characteristics of melatonin dosage forms. In addition, the results indicate that 6-OHM has a definite role to play as an antioxidant. Thus further research may favour the use of these agents in the treatment of several neurodegenerative disorders.
- Full Text:
- Date Issued: 2003