The effect of combined vitamin E succinate and ascorbic acid supplementation on growth and cyclooxygenase expression in murine melanoma (BL6) cells
- Authors: Van Rooyen, Megan Lynne
- Date: 1999
- Subjects: Vitamin E , Vitamin C , Melanoma
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3982 , http://hdl.handle.net/10962/d1004041 , Vitamin E , Vitamin C , Melanoma
- Description: This thesis examines the effect of combined vitamin E succinate and Asc supplementation on the in vitro growth of a non-malignant monkey kidney (LLCMK) and a malignant melanoma (BL6) cell line, with nutritional concentration ranges of 5-20µg/ml and 25-50µg/ml respectively. Vitamin E and C are thought to interact synergistically to inhibit tumour cell growth by virtue of their antioxidant properties, whereby they quench free radicals and terminate lipid peroxidation. Furthermore vitamin E and C are thought to modulate the biosynthetic pathways in arachidonic acid metabolism at a number of different points. This may also offer a means of regulating tumour cell growth. It is well documented that vitamin E and C are distributed in the lipid and aqueous phases in the cell respectively. However, the cells need to obtain the vitamins from the environment in which they are found in order to exert a growth inhibitory effect. Supplementation of combined vitamin E succinate and Asc on BL6 and LLCMK cells resulted in a significant increase in LLCMK cell growth, and a significant decrease in cell growth was observed in BL6 cells. Vitamin E succinate in its esterified form cannot function as an antioxidant and requires the cleavage of the succinate to become an active antioxidant. The metabolism of vitamin E succinate to form free vitamin E in LLCMK and BL6 cells resulted in the cleavage of the succinate group from the vitamin E molecule in BL6 cells only, thus suggesting that an esterase may be present in BL6 cells. This would allow for a synergistic interaction between the two vitamins. The arachidonic acid cascade generates a family of bioactive lipids that modulate diverse physiological and pathological responses including tumour growth and promotion. The enzyme prostaglandin endoperoxide synthase (PGHS) or cyclooxygenase (Cox) is the key enzyme in the biosynthetic pathway leading to the formation of prostaglandins. Two enzyme isoforms of Cox have been identified, Cox 1 and Cox 2. Supplementation with vitamin E succinate and Asc at a combination 20:25µg/ml respectively resulted in a trend of increasing Cox activity over 12 hours suggesting that vitamin E and Asc have a stimulatory effect on Cox activity in BL6 cells. The inhibitors of Cox 2, dexamethasone, showed a decreasing trend in Cox activity at the 20:25µg/ml combination, while cycloheximide showed an initial stimulatory effect and then a gradual decrease in Cox activity. The elimination of the Cox activity by dexamethasone suggests that transcriptional regulation may be occurring in BL6 cells. We examined by Northern blot analysis whether combined supplementation of vitamin E succinate and Asc caused an elevation of Cox 2 RNA expression in BL6 cells. An inducible effect of Cox 2 was observed after 2 hours of supplementation with a combination of vitamin E succinate and Asc in BL6 cells, however the results are inconclusive and further studies are required to substantiate this finding.
- Full Text:
- Authors: Van Rooyen, Megan Lynne
- Date: 1999
- Subjects: Vitamin E , Vitamin C , Melanoma
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3982 , http://hdl.handle.net/10962/d1004041 , Vitamin E , Vitamin C , Melanoma
- Description: This thesis examines the effect of combined vitamin E succinate and Asc supplementation on the in vitro growth of a non-malignant monkey kidney (LLCMK) and a malignant melanoma (BL6) cell line, with nutritional concentration ranges of 5-20µg/ml and 25-50µg/ml respectively. Vitamin E and C are thought to interact synergistically to inhibit tumour cell growth by virtue of their antioxidant properties, whereby they quench free radicals and terminate lipid peroxidation. Furthermore vitamin E and C are thought to modulate the biosynthetic pathways in arachidonic acid metabolism at a number of different points. This may also offer a means of regulating tumour cell growth. It is well documented that vitamin E and C are distributed in the lipid and aqueous phases in the cell respectively. However, the cells need to obtain the vitamins from the environment in which they are found in order to exert a growth inhibitory effect. Supplementation of combined vitamin E succinate and Asc on BL6 and LLCMK cells resulted in a significant increase in LLCMK cell growth, and a significant decrease in cell growth was observed in BL6 cells. Vitamin E succinate in its esterified form cannot function as an antioxidant and requires the cleavage of the succinate to become an active antioxidant. The metabolism of vitamin E succinate to form free vitamin E in LLCMK and BL6 cells resulted in the cleavage of the succinate group from the vitamin E molecule in BL6 cells only, thus suggesting that an esterase may be present in BL6 cells. This would allow for a synergistic interaction between the two vitamins. The arachidonic acid cascade generates a family of bioactive lipids that modulate diverse physiological and pathological responses including tumour growth and promotion. The enzyme prostaglandin endoperoxide synthase (PGHS) or cyclooxygenase (Cox) is the key enzyme in the biosynthetic pathway leading to the formation of prostaglandins. Two enzyme isoforms of Cox have been identified, Cox 1 and Cox 2. Supplementation with vitamin E succinate and Asc at a combination 20:25µg/ml respectively resulted in a trend of increasing Cox activity over 12 hours suggesting that vitamin E and Asc have a stimulatory effect on Cox activity in BL6 cells. The inhibitors of Cox 2, dexamethasone, showed a decreasing trend in Cox activity at the 20:25µg/ml combination, while cycloheximide showed an initial stimulatory effect and then a gradual decrease in Cox activity. The elimination of the Cox activity by dexamethasone suggests that transcriptional regulation may be occurring in BL6 cells. We examined by Northern blot analysis whether combined supplementation of vitamin E succinate and Asc caused an elevation of Cox 2 RNA expression in BL6 cells. An inducible effect of Cox 2 was observed after 2 hours of supplementation with a combination of vitamin E succinate and Asc in BL6 cells, however the results are inconclusive and further studies are required to substantiate this finding.
- Full Text:
The effect of diet and age-at-weaning on growth and survival of clownfish Amphiprion percula (Pisces : Pomacentridae)
- Authors: Gordon, Andrew K
- Date: 1999
- Subjects: Anemonefishes , Pomacentridae , Fishes -- Growth
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5270 , http://hdl.handle.net/10962/d1005114 , Anemonefishes , Pomacentridae , Fishes -- Growth
- Description: The aim of this study was to improve the rearing of common clownfish Amphiprion percula, by weaning juveniles from a live feed to a formulated feed as early as possible, while still maintaining good growth and survival. The growth response of A. percula to a formulated dry feed was initially investigated. There was no difference in growth rate of juveniles fed a formulated feed, and the formulated feed supplemented with either Artemia or a Donax serra / Penaeus indicus combination. Amphiprion percula readily consumed the formulated feed, and the fishmeal/casein combination appeared an acceptable protein source. As the amount of protein included in a diet can have a profound effect on growth, the optimal dietary protein level for juvenile A. percula was investigated by feeding semipurified diets containing graded levels of protein, ranging from 40-65%. There was no difference in the growth rates of juveniles fed the various diets, however all diets promoted good growth with an average weight gain of 419%, and thus for the purposes of this study the diet formulation was deemed adequate. The histological study of the digestive system of larval A. percula revealed that the alimentary canal was advanced at hatching and that larvae start exogenous feeding immediately. Three days after hatch (DAH) the yolk sac is completely absorbed. In the hind-gut epithelium of 5-day-old larvae small supranuclear inclusion vacuoles appear, suggesting pinocytotic digestion, and by 7 DAH gastric glands are established in the epithelium of the stomach. Nine DAH supranuclear inclusion vacuoles appear in the epithelium of the mid-gut, indicating extracellular digestion and absorption across the lumen. As pinocytotic digestion of protein is less efficient than extracellular digestion, especially in the case of formulated feeds, it was hypothesised that the digestive system of A. percula could only effectively digest formulated feeds 9 DAH onwards. The two weaning experiments designed to test this hypothesis revealed that A. percula was able to utilise the formulated feed, without reduction in survival, from 7 DAH onwards. However, in terms of growth, the optimal time to wean juveniles from the live feed to the formulated dry feed was between 15 to 20 DAH. As A. percula accept a formulated feed and can benefit nutritionally from it, the dependence of larvae and juveniles on live feed can be reduced. This study has shown that the rearing of A. percula can been simplified and improved by weaning from 7 DAH with no reduction in survival, and from 15 to 20 DAH with no reduction in growth.
- Full Text:
- Authors: Gordon, Andrew K
- Date: 1999
- Subjects: Anemonefishes , Pomacentridae , Fishes -- Growth
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5270 , http://hdl.handle.net/10962/d1005114 , Anemonefishes , Pomacentridae , Fishes -- Growth
- Description: The aim of this study was to improve the rearing of common clownfish Amphiprion percula, by weaning juveniles from a live feed to a formulated feed as early as possible, while still maintaining good growth and survival. The growth response of A. percula to a formulated dry feed was initially investigated. There was no difference in growth rate of juveniles fed a formulated feed, and the formulated feed supplemented with either Artemia or a Donax serra / Penaeus indicus combination. Amphiprion percula readily consumed the formulated feed, and the fishmeal/casein combination appeared an acceptable protein source. As the amount of protein included in a diet can have a profound effect on growth, the optimal dietary protein level for juvenile A. percula was investigated by feeding semipurified diets containing graded levels of protein, ranging from 40-65%. There was no difference in the growth rates of juveniles fed the various diets, however all diets promoted good growth with an average weight gain of 419%, and thus for the purposes of this study the diet formulation was deemed adequate. The histological study of the digestive system of larval A. percula revealed that the alimentary canal was advanced at hatching and that larvae start exogenous feeding immediately. Three days after hatch (DAH) the yolk sac is completely absorbed. In the hind-gut epithelium of 5-day-old larvae small supranuclear inclusion vacuoles appear, suggesting pinocytotic digestion, and by 7 DAH gastric glands are established in the epithelium of the stomach. Nine DAH supranuclear inclusion vacuoles appear in the epithelium of the mid-gut, indicating extracellular digestion and absorption across the lumen. As pinocytotic digestion of protein is less efficient than extracellular digestion, especially in the case of formulated feeds, it was hypothesised that the digestive system of A. percula could only effectively digest formulated feeds 9 DAH onwards. The two weaning experiments designed to test this hypothesis revealed that A. percula was able to utilise the formulated feed, without reduction in survival, from 7 DAH onwards. However, in terms of growth, the optimal time to wean juveniles from the live feed to the formulated dry feed was between 15 to 20 DAH. As A. percula accept a formulated feed and can benefit nutritionally from it, the dependence of larvae and juveniles on live feed can be reduced. This study has shown that the rearing of A. percula can been simplified and improved by weaning from 7 DAH with no reduction in survival, and from 15 to 20 DAH with no reduction in growth.
- Full Text:
The isolation and characterisation of thermostable hydantoinases from hydantoinase-producing bacteria
- Authors: Phehane, Vuyisile Ntosi
- Date: 1999
- Subjects: Hydantoin , Bacteria -- Physiology , Enzymes
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3998 , http://hdl.handle.net/10962/d1004058 , Hydantoin , Bacteria -- Physiology , Enzymes
- Description: In order to characterise thermostable hydantoin-hydrolysing enzymes from bacteria, locally-isolated thermophilic organisms were screened for the ability to convert hydantoin to N-carbamylglycine at 55°C using the hydantoinase enzyme. Cell disruption of a selected strain, RU-20-15, was conducted by French pressing to release enzyme from within the cell. In all of the experiments conducted, the amounts of product were low. In view of the low yields of products formed by the thermophiles, a previously-isolated Gram negative strain, RU-KM3L was selected from a number of mesophiles by screening for hydantoinase and carbamylase activity over a 40-55°C temperature range. Hydantoin conversion at 40°C using crude extract from pressed cells of this organism was similar to conversion at 50°C, and therefore subsequent assays were conducted at the higher temperature. The growth kinetics of RU-KM3L cells were studied and the enzyme activities of the extracts were compared in complete and chemically-defined media. The results suggested that the optimal time to harvest cells was at early stationary phase, when using complete medium for culture of cells; the specific activity of enzyme extracts produced by culture in complete medium was higher than that obtained in chemically-defined medium. 5-methylhydantoin was shown to be the preferred substrate for both the hydantoinase and carbamylase enzymes in the crude extract of RU-KM3L. The substrate specificity of the hydantoinase and carbamylase enzymes of the crude RU-KM3L extract was observed to be altered in the presence of increasing amounts of hydantoin, 5,5-dihydrouracil (DHU) and 5-thiouracil (TU) as inducers, showing selectivity for 5-methylhydantoin over hydantoin at inducer concentrations of 0.1 to 1%. A limiting effect on the hydrolysis of 5-methylhydantoin was observed when DHU and 5,5-dimethylhydantoin (DMH) were used as inducers, while the limiting effect on hydantoin specificity was observed when DHU and TU were used as inducers. The limiting effect was observed to be dependent upon the concentration of inducer, and was not observed when hydantoin was used as an inducer. The optimal time for assay of the hydantoinase enzyme in crude extract preparations at 50°C was observed to be 3h. Alkaline conditions were shown to be optimal for both the hydantoinase and carbamylase enzymes of RU-KM3L. Assay for enzyme activities of RU-KM3L extract in the presence of metal ions showed Mn²⁺ ions (and to a lesser extent, Co²⁺) to activate both the hydantoinase and carbamylase activities. Cu²⁺ ions were observed to inhibit the hydantoinase enzyme. In order to determine the location of the enzymes within the cell, cell debris from disrupted cells of RU-KM3L was removed by centrifugation. A decrease in enzyme activity in the supernatant was observed, and suggested association of the enzymes with the cell membrane. Ammonium sulfate fractionation experiments conducted on the crude extract provided further evidence for this result. Sonication of the crude enzyme extract was the only successful method for the releasing of membrane-associated enzyme. Of a number of strategies investigated, the use of sucrose at 50% (w/v) concentration was shown to preserve the hydantoinase and carbamylase enzyme activities during lyophilisation. Furthermore, assay for these enzyme activities showed the activities to be higher after lyophilisation in the presence of sucrose. However, sucrose did not increase the thermostability of lyophilised crude enzyme extracts. Water-miscible organic solvents at 1% concentration were shown to be inhibitory to the hydantoinase and carbamylase enzymes of RU-KM3L, and the inhibition was also observed to increase with increasing concentrations of these solvents. Hydantoinase activity in the presence of water-immiscible organic solvents was shown to increase with an increase in the hydrophobicity of these solvents, but the activity observed was not significantly higher than activity in the absence of solvent when hydantoin and 5-methylhydantoin were used as substrates. The possibility of reversing the hydantoinase enzyme reaction by water-immiscible organic solvents was investigated, and the results obtained suggested that the reaction could be reversed. It was thought that the partitioning of substrates or products into hydrophobic organic solvents could influence the reaction equilibrium, but the partitioning observed was not sufficient to affect reaction rates. Peptide synthesis was shown to have occurred in small amounts when the hydantoinase reaction was carried out in the presence of water-immiscible organic solvents. In conclusion, the hydantoin-hydrolyzing enzyme activity of a crude extract preparation from the bacterial strain RU-KM3L was characterised at elevated temperatures, and in the presence of watermiscible and -immiscible organic solvents.
- Full Text:
- Authors: Phehane, Vuyisile Ntosi
- Date: 1999
- Subjects: Hydantoin , Bacteria -- Physiology , Enzymes
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3998 , http://hdl.handle.net/10962/d1004058 , Hydantoin , Bacteria -- Physiology , Enzymes
- Description: In order to characterise thermostable hydantoin-hydrolysing enzymes from bacteria, locally-isolated thermophilic organisms were screened for the ability to convert hydantoin to N-carbamylglycine at 55°C using the hydantoinase enzyme. Cell disruption of a selected strain, RU-20-15, was conducted by French pressing to release enzyme from within the cell. In all of the experiments conducted, the amounts of product were low. In view of the low yields of products formed by the thermophiles, a previously-isolated Gram negative strain, RU-KM3L was selected from a number of mesophiles by screening for hydantoinase and carbamylase activity over a 40-55°C temperature range. Hydantoin conversion at 40°C using crude extract from pressed cells of this organism was similar to conversion at 50°C, and therefore subsequent assays were conducted at the higher temperature. The growth kinetics of RU-KM3L cells were studied and the enzyme activities of the extracts were compared in complete and chemically-defined media. The results suggested that the optimal time to harvest cells was at early stationary phase, when using complete medium for culture of cells; the specific activity of enzyme extracts produced by culture in complete medium was higher than that obtained in chemically-defined medium. 5-methylhydantoin was shown to be the preferred substrate for both the hydantoinase and carbamylase enzymes in the crude extract of RU-KM3L. The substrate specificity of the hydantoinase and carbamylase enzymes of the crude RU-KM3L extract was observed to be altered in the presence of increasing amounts of hydantoin, 5,5-dihydrouracil (DHU) and 5-thiouracil (TU) as inducers, showing selectivity for 5-methylhydantoin over hydantoin at inducer concentrations of 0.1 to 1%. A limiting effect on the hydrolysis of 5-methylhydantoin was observed when DHU and 5,5-dimethylhydantoin (DMH) were used as inducers, while the limiting effect on hydantoin specificity was observed when DHU and TU were used as inducers. The limiting effect was observed to be dependent upon the concentration of inducer, and was not observed when hydantoin was used as an inducer. The optimal time for assay of the hydantoinase enzyme in crude extract preparations at 50°C was observed to be 3h. Alkaline conditions were shown to be optimal for both the hydantoinase and carbamylase enzymes of RU-KM3L. Assay for enzyme activities of RU-KM3L extract in the presence of metal ions showed Mn²⁺ ions (and to a lesser extent, Co²⁺) to activate both the hydantoinase and carbamylase activities. Cu²⁺ ions were observed to inhibit the hydantoinase enzyme. In order to determine the location of the enzymes within the cell, cell debris from disrupted cells of RU-KM3L was removed by centrifugation. A decrease in enzyme activity in the supernatant was observed, and suggested association of the enzymes with the cell membrane. Ammonium sulfate fractionation experiments conducted on the crude extract provided further evidence for this result. Sonication of the crude enzyme extract was the only successful method for the releasing of membrane-associated enzyme. Of a number of strategies investigated, the use of sucrose at 50% (w/v) concentration was shown to preserve the hydantoinase and carbamylase enzyme activities during lyophilisation. Furthermore, assay for these enzyme activities showed the activities to be higher after lyophilisation in the presence of sucrose. However, sucrose did not increase the thermostability of lyophilised crude enzyme extracts. Water-miscible organic solvents at 1% concentration were shown to be inhibitory to the hydantoinase and carbamylase enzymes of RU-KM3L, and the inhibition was also observed to increase with increasing concentrations of these solvents. Hydantoinase activity in the presence of water-immiscible organic solvents was shown to increase with an increase in the hydrophobicity of these solvents, but the activity observed was not significantly higher than activity in the absence of solvent when hydantoin and 5-methylhydantoin were used as substrates. The possibility of reversing the hydantoinase enzyme reaction by water-immiscible organic solvents was investigated, and the results obtained suggested that the reaction could be reversed. It was thought that the partitioning of substrates or products into hydrophobic organic solvents could influence the reaction equilibrium, but the partitioning observed was not sufficient to affect reaction rates. Peptide synthesis was shown to have occurred in small amounts when the hydantoinase reaction was carried out in the presence of water-immiscible organic solvents. In conclusion, the hydantoin-hydrolyzing enzyme activity of a crude extract preparation from the bacterial strain RU-KM3L was characterised at elevated temperatures, and in the presence of watermiscible and -immiscible organic solvents.
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The relative influence of value priorities ethnicity and worries in the determination of political party affiliation amongst Ugandan university students
- Authors: Kibanja, Grace M
- Date: 1999
- Subjects: Political psychology , Uganda -- Politics and government -- 1979- , Students -- Uganda -- Political activity , College students -- Uganda -- Psychology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3003 , http://hdl.handle.net/10962/d1002512 , Political psychology , Uganda -- Politics and government -- 1979- , Students -- Uganda -- Political activity , College students -- Uganda -- Psychology
- Description: This study examines the relative influence of value priorities, ethnicity and 'worries' in the determination of political party affiliation amongst Ugandan university students. Schwartz's values questionnaire was administered to 309 male and 176 female first year students from the faculties of engineering, medicine, law, commerce, social sciences, sciences, education and mass communication at Makerere University Uganda. The sample included respondents from all of the six sub-ethnic groups in Uganda. Respondents also covered the major religious groups in the country and were also representative of the major political parties. Results from the statistical analysis on the data show that ethnicity and certain values playa role in the determination of these students' affiliation to a political party. Chi-square results show that achievement, benevolence, universalism, security, tradition and conformity values are given differing importance across political parties. And, Anova results show that the tradition value has a significant mean difference across parties. Other factors such as religion and course of study are also found to have significant influence on these student's affiliation to political parties. Although 'worries' are found not to have a significant influence on student's political party affiliation, findings show that students from different political parties differ in their ratings of the different types of ' worries'. Therefore results show that ethnicity and value priorities do infl4ence these students' affiliation to a political party but ' worries' do not. The discussion section explores these findings in the context of contemporary Ugandan politics.
- Full Text:
- Authors: Kibanja, Grace M
- Date: 1999
- Subjects: Political psychology , Uganda -- Politics and government -- 1979- , Students -- Uganda -- Political activity , College students -- Uganda -- Psychology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3003 , http://hdl.handle.net/10962/d1002512 , Political psychology , Uganda -- Politics and government -- 1979- , Students -- Uganda -- Political activity , College students -- Uganda -- Psychology
- Description: This study examines the relative influence of value priorities, ethnicity and 'worries' in the determination of political party affiliation amongst Ugandan university students. Schwartz's values questionnaire was administered to 309 male and 176 female first year students from the faculties of engineering, medicine, law, commerce, social sciences, sciences, education and mass communication at Makerere University Uganda. The sample included respondents from all of the six sub-ethnic groups in Uganda. Respondents also covered the major religious groups in the country and were also representative of the major political parties. Results from the statistical analysis on the data show that ethnicity and certain values playa role in the determination of these students' affiliation to a political party. Chi-square results show that achievement, benevolence, universalism, security, tradition and conformity values are given differing importance across political parties. And, Anova results show that the tradition value has a significant mean difference across parties. Other factors such as religion and course of study are also found to have significant influence on these student's affiliation to political parties. Although 'worries' are found not to have a significant influence on student's political party affiliation, findings show that students from different political parties differ in their ratings of the different types of ' worries'. Therefore results show that ethnicity and value priorities do infl4ence these students' affiliation to a political party but ' worries' do not. The discussion section explores these findings in the context of contemporary Ugandan politics.
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The removal of toxic heavy metals from aqueous solutions by algal extracellular polysaccharides
- Authors: Selepe, Mamaropeng Marcus
- Date: 1999
- Subjects: Heavy metals -- Absorption and adsorption , Copper , Lead , Algae -- Biotechnology , Polysaccharides -- Biotechnology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3994 , http://hdl.handle.net/10962/d1004054 , Heavy metals -- Absorption and adsorption , Copper , Lead , Algae -- Biotechnology , Polysaccharides -- Biotechnology
- Description: This study investigated the possible use of algal extracellular polysaccharide as a biosorbent for removal of heavy metals (copper and lead) from aqueous solutions as a means of bioremediation for metal containing effluents. This biopolymer has good biosorbent properties and a potential to provide a cost effective, selective and efficient purification system. A variety of environmental conditions induce the production of extracellular polysaccharides in algae. The production of exopolysaccharides by Dunaliella cultures was induced by nitrogen deficient conditions. A high ratio of carbon to nitrogen source considerably enhanced the polysaccharide release. Purified extracellular polysaccharide samples exhibited a monosaccharide composition consisting of the following sugars: xylose, arabinose, 2-0-methyl mannose, mannose, glucose and galactose. The relative abundance (%) of these sugars were calculated relative to xylose. The major sugar constituent was 2-0-methyl mannose, which was present at approximately 160% relative to xylose. The percentage relative abundance of other sugars was as follows: 18.8; 86.8; 85.3 and 22.3% for arabinose; mannose; glucose and galactose respectively. The identity of the various constituents were confirmed by mass spectrometry. The ability of Dunaliella exopolysaccharides to accumulate metals was investigated. The following parameters were studied because they affect metal uptake: solution pH, biomass concentration, temperature, time and metal concentration. The uptake of both copper and lead were pH dependent. However, metal uptake was not significantly affected by temperature. Kinetic studies showed that Dunaliella extracellular polysaccharides exhibit good bioremediation properties. Metal uptake was rapid. In addition, the exopolysaccharide has good metal binding capacity with an uptake capacity for lead of 80 mg/g from a solution containing initial lead concentration of approximately 40 mg/l. Competition studies revealed that the presence of a second metal in solution inhibits uptake of the other metal compared to uptake in single metal solution of that particular metal. The presence of lead inhibited the uptake of copper from approximately 65% in single metal solution to 10% in binary metal solution. The presence of copper also inhibited lead uptake, though not to the same extent. Higher concentrations of lead could not completely prevent removal of copper from solution and visa versa. The same was true for lead which could not be displaced by a four-fold concentration of copper. Instead, a certain percentage of copper was always removed showing that lead did not compete with copper for these binding sites. In conclusion it appears that, copper and lead bind to different sites on Dunaliella exopolysaccharides and that they exhibit selective or preferential removal of lead.
- Full Text:
- Authors: Selepe, Mamaropeng Marcus
- Date: 1999
- Subjects: Heavy metals -- Absorption and adsorption , Copper , Lead , Algae -- Biotechnology , Polysaccharides -- Biotechnology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3994 , http://hdl.handle.net/10962/d1004054 , Heavy metals -- Absorption and adsorption , Copper , Lead , Algae -- Biotechnology , Polysaccharides -- Biotechnology
- Description: This study investigated the possible use of algal extracellular polysaccharide as a biosorbent for removal of heavy metals (copper and lead) from aqueous solutions as a means of bioremediation for metal containing effluents. This biopolymer has good biosorbent properties and a potential to provide a cost effective, selective and efficient purification system. A variety of environmental conditions induce the production of extracellular polysaccharides in algae. The production of exopolysaccharides by Dunaliella cultures was induced by nitrogen deficient conditions. A high ratio of carbon to nitrogen source considerably enhanced the polysaccharide release. Purified extracellular polysaccharide samples exhibited a monosaccharide composition consisting of the following sugars: xylose, arabinose, 2-0-methyl mannose, mannose, glucose and galactose. The relative abundance (%) of these sugars were calculated relative to xylose. The major sugar constituent was 2-0-methyl mannose, which was present at approximately 160% relative to xylose. The percentage relative abundance of other sugars was as follows: 18.8; 86.8; 85.3 and 22.3% for arabinose; mannose; glucose and galactose respectively. The identity of the various constituents were confirmed by mass spectrometry. The ability of Dunaliella exopolysaccharides to accumulate metals was investigated. The following parameters were studied because they affect metal uptake: solution pH, biomass concentration, temperature, time and metal concentration. The uptake of both copper and lead were pH dependent. However, metal uptake was not significantly affected by temperature. Kinetic studies showed that Dunaliella extracellular polysaccharides exhibit good bioremediation properties. Metal uptake was rapid. In addition, the exopolysaccharide has good metal binding capacity with an uptake capacity for lead of 80 mg/g from a solution containing initial lead concentration of approximately 40 mg/l. Competition studies revealed that the presence of a second metal in solution inhibits uptake of the other metal compared to uptake in single metal solution of that particular metal. The presence of lead inhibited the uptake of copper from approximately 65% in single metal solution to 10% in binary metal solution. The presence of copper also inhibited lead uptake, though not to the same extent. Higher concentrations of lead could not completely prevent removal of copper from solution and visa versa. The same was true for lead which could not be displaced by a four-fold concentration of copper. Instead, a certain percentage of copper was always removed showing that lead did not compete with copper for these binding sites. In conclusion it appears that, copper and lead bind to different sites on Dunaliella exopolysaccharides and that they exhibit selective or preferential removal of lead.
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The role of vitamin E succinate in regulation of growth and cyclooxygenase expression in B16 murine melanoma cells
- Van der Merwe, Adele Shanette
- Authors: Van der Merwe, Adele Shanette
- Date: 1999
- Subjects: Melanoma , Vitamin E
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3988 , http://hdl.handle.net/10962/d1004047 , Melanoma , Vitamin E
- Description: This study was undertaken to determine the effects and possible mechanism of action of vitamin E succinate supplementation on B16 murine melanoma cell growth in vitro. Studies revealed that supplementation of 5, 7 and 10µg/ml of this vitamin significantly inhibited growth of B16 cells. Non-malignant LLCMK cells supplemented with the same concentrations of vitamin E succinate resulted in similar inhibition of cell growth. The actual mechanism by which vitamin E succinate inhibits B16 cell growth is unclear, though there has been much speculation about its possible role as an antioxidant. Vitamin E succinate is not a physiological antioxidant and for this ester to behave as an antioxidant, cleavage of the ester bond must occur, releasing the antioxidant vitamin E part of the molecule. To determine whether the observed inhibitory effects on B16 cell growth were due to the intact vitamin E succinate or the vitamin E cleavage product, cleavage studies were undertaken. Results from these studies revealed that in B16 cells vitamin E succinate cleavage did not occur suggesting that the observed inhibitory effects of vitamin E succinate on B16 cells were due to the intact compound. In contrast vitamin E succinate cleavage was shown to occur in LLCMK cells, suggesting that these cells may contain an esterase capable of liberating succinic acid and vitamin E. Further studies focussed on the possible role of vitamin E succinate in regulation of cyclooxygenase activity in B16 cells as vitamin E succinate was found to effect the activity of various enzymes involved in the arachidonic acid cascade, notably cyclooxygenase, the rate-limiting enzyme in prostaglandin synthesis. Time course studies were used to determine when the cyclooxygenase protein was being produced, thus allowing an estimation of when the gene was being 'switched on'. These studies revealed that vitamin E succinate does not significantly effect cyclooxygenase activity in B16 cells over a period of 2 to 12 hours as compared to the OE control cultures. Further studies using RNA techniques investigated whether vitamin E succinate was having an effect on cyclooxygenase activity at a molecular level. These investigations were unsuccessful for the 6 day supplementation for a number of possible reasons, the main reason being RNA stability. Subsequent studies revealed an increase in COX mRNA after 2 hours, suggesting that the gene was 'switched on' soon after supplementation with vitamin E succinate, and further increases in COX mRNA were observed after 8 to 12 hours. The molecular studies were, however, inconclusive. Previous studies suggested that vitamin E succinate was indirectly causing growth inhibition of B16 cells via regulation of cyclooxygenase activity, however, this study does not support these findings and it would seem unlikely that regulation of cyclooxygenase expression in B16 cells by vitamin E succinate has a role to play in the mechanism by which vitamin E succinate inhibits growth in B16 cells.
- Full Text:
- Authors: Van der Merwe, Adele Shanette
- Date: 1999
- Subjects: Melanoma , Vitamin E
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3988 , http://hdl.handle.net/10962/d1004047 , Melanoma , Vitamin E
- Description: This study was undertaken to determine the effects and possible mechanism of action of vitamin E succinate supplementation on B16 murine melanoma cell growth in vitro. Studies revealed that supplementation of 5, 7 and 10µg/ml of this vitamin significantly inhibited growth of B16 cells. Non-malignant LLCMK cells supplemented with the same concentrations of vitamin E succinate resulted in similar inhibition of cell growth. The actual mechanism by which vitamin E succinate inhibits B16 cell growth is unclear, though there has been much speculation about its possible role as an antioxidant. Vitamin E succinate is not a physiological antioxidant and for this ester to behave as an antioxidant, cleavage of the ester bond must occur, releasing the antioxidant vitamin E part of the molecule. To determine whether the observed inhibitory effects on B16 cell growth were due to the intact vitamin E succinate or the vitamin E cleavage product, cleavage studies were undertaken. Results from these studies revealed that in B16 cells vitamin E succinate cleavage did not occur suggesting that the observed inhibitory effects of vitamin E succinate on B16 cells were due to the intact compound. In contrast vitamin E succinate cleavage was shown to occur in LLCMK cells, suggesting that these cells may contain an esterase capable of liberating succinic acid and vitamin E. Further studies focussed on the possible role of vitamin E succinate in regulation of cyclooxygenase activity in B16 cells as vitamin E succinate was found to effect the activity of various enzymes involved in the arachidonic acid cascade, notably cyclooxygenase, the rate-limiting enzyme in prostaglandin synthesis. Time course studies were used to determine when the cyclooxygenase protein was being produced, thus allowing an estimation of when the gene was being 'switched on'. These studies revealed that vitamin E succinate does not significantly effect cyclooxygenase activity in B16 cells over a period of 2 to 12 hours as compared to the OE control cultures. Further studies using RNA techniques investigated whether vitamin E succinate was having an effect on cyclooxygenase activity at a molecular level. These investigations were unsuccessful for the 6 day supplementation for a number of possible reasons, the main reason being RNA stability. Subsequent studies revealed an increase in COX mRNA after 2 hours, suggesting that the gene was 'switched on' soon after supplementation with vitamin E succinate, and further increases in COX mRNA were observed after 8 to 12 hours. The molecular studies were, however, inconclusive. Previous studies suggested that vitamin E succinate was indirectly causing growth inhibition of B16 cells via regulation of cyclooxygenase activity, however, this study does not support these findings and it would seem unlikely that regulation of cyclooxygenase expression in B16 cells by vitamin E succinate has a role to play in the mechanism by which vitamin E succinate inhibits growth in B16 cells.
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The social construction of 'capacity building': a grounded theory study of organisation development consultants' accounts
- Authors: Eagar, Ryan
- Date: 1999
- Subjects: Organizational change -- South Africa , Business consultants -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:2969 , http://hdl.handle.net/10962/d1002478 , Organizational change -- South Africa , Business consultants -- South Africa
- Description: The primary aim of this thesis is to explore, through organisation development (OD) consultants' accounts, the meaning of 'capacity building' in the South African development context. The need for theory development in this area is apparent from two interrelated vantage points. Firstly, while 'capacity building' is an increasingly espoused development approach, it is seen to be a confusing and ill-defined concept, for which there exists no adequate theory. Secondly, there is an growing call within the OD field to perfonn 'recOIlllaissance' (Weick,1990) on OD as it is being practised in different socio-historical and organisational contexts, so as to discern future trends for this discipline. Due to the lack of documented debate which grounds OD issues in the South African development sector, OD practitioners' 'capacity building' interventions were seen to provide suitably unchartered terrain for this study. Given that theory generation was intended, the general epistemological principles provided by Glaser and Strauss's (1967) 'grounded theory' methodology were adopted. As a way of avoiding some of the criticisms and limitations of this approach, this thesis followed later conceptualisations of this method, in particular its reframing within the social constructionist idiom. In accordance with this perspective, this study directed attention to the ways in which OD consultants, in a non-governmental organisational (NGO) sector known as 'intermediary' NGOs, accounted for their 'capacity building' role in this development context. The results, based on in-depth interviews with ten OD consultants, indicate that 'capacity building' is an elusive and inchoate concept with more than one meaning for the participants. Their narrative account variously constructs 'capacity building' as value-driven OD process facilitation; funder and market regulated service provision; and people-driven product delivery. As a result of this multi-vocal construction, the participants' accounts reveal that OD consultancy in this sector is primarily concerned with 'managing the tensions' of the consultants' ambiguous and contradictory roles. By examining how the tensions articulated by the consultants inhere in their relationship to the environment in which they operate, this thesis firstly explores how the contradiction and anlbiguity attached to this concept can be traced to different stakeholder expectations of 'capacity building'. Secondly, it exanlines how these different stakeholder discourses conflict with each other and with an OD perspective. Thirdly, through an explication of the core category of 'managing tensions', it explores the image of OD consulting as a 'shifting and inconstant balancing act'. Fourthly, it shows how there exist wider contextual forces operating in the development sector which serve to throw these consultants 'off balance' and into delimited and 'received' service provision roles which run counter to their raison d'etre. Finally, the research examines new ways of approaching the 'capacity building' question and of understanding the nature of OD consultancy. It concludes with an attempt to respond to a conceptual aporia in OD literature by examining possible alternative images and metaphors for the role of the OD consultant.
- Full Text:
- Authors: Eagar, Ryan
- Date: 1999
- Subjects: Organizational change -- South Africa , Business consultants -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:2969 , http://hdl.handle.net/10962/d1002478 , Organizational change -- South Africa , Business consultants -- South Africa
- Description: The primary aim of this thesis is to explore, through organisation development (OD) consultants' accounts, the meaning of 'capacity building' in the South African development context. The need for theory development in this area is apparent from two interrelated vantage points. Firstly, while 'capacity building' is an increasingly espoused development approach, it is seen to be a confusing and ill-defined concept, for which there exists no adequate theory. Secondly, there is an growing call within the OD field to perfonn 'recOIlllaissance' (Weick,1990) on OD as it is being practised in different socio-historical and organisational contexts, so as to discern future trends for this discipline. Due to the lack of documented debate which grounds OD issues in the South African development sector, OD practitioners' 'capacity building' interventions were seen to provide suitably unchartered terrain for this study. Given that theory generation was intended, the general epistemological principles provided by Glaser and Strauss's (1967) 'grounded theory' methodology were adopted. As a way of avoiding some of the criticisms and limitations of this approach, this thesis followed later conceptualisations of this method, in particular its reframing within the social constructionist idiom. In accordance with this perspective, this study directed attention to the ways in which OD consultants, in a non-governmental organisational (NGO) sector known as 'intermediary' NGOs, accounted for their 'capacity building' role in this development context. The results, based on in-depth interviews with ten OD consultants, indicate that 'capacity building' is an elusive and inchoate concept with more than one meaning for the participants. Their narrative account variously constructs 'capacity building' as value-driven OD process facilitation; funder and market regulated service provision; and people-driven product delivery. As a result of this multi-vocal construction, the participants' accounts reveal that OD consultancy in this sector is primarily concerned with 'managing the tensions' of the consultants' ambiguous and contradictory roles. By examining how the tensions articulated by the consultants inhere in their relationship to the environment in which they operate, this thesis firstly explores how the contradiction and anlbiguity attached to this concept can be traced to different stakeholder expectations of 'capacity building'. Secondly, it exanlines how these different stakeholder discourses conflict with each other and with an OD perspective. Thirdly, through an explication of the core category of 'managing tensions', it explores the image of OD consulting as a 'shifting and inconstant balancing act'. Fourthly, it shows how there exist wider contextual forces operating in the development sector which serve to throw these consultants 'off balance' and into delimited and 'received' service provision roles which run counter to their raison d'etre. Finally, the research examines new ways of approaching the 'capacity building' question and of understanding the nature of OD consultancy. It concludes with an attempt to respond to a conceptual aporia in OD literature by examining possible alternative images and metaphors for the role of the OD consultant.
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Towards modelling the formation of ore bodies initial results dealing with the fluid mechanical aspects of magma chamber convection
- Authors: Botha, André Erasmus
- Date: 1999
- Subjects: Ore deposits , Fluid mechanics , Magmatism
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5492 , http://hdl.handle.net/10962/d1005278 , Ore deposits , Fluid mechanics , Magmatism
- Description: This thesis forms part of a larger effort which aims to establish the means of assessing the fluid mechanical behaviour of magma 1 as it cools inside a magma chamber surrounded by porous country rock. The reason for doing so is to advance the understanding of some types of mineral deposits; for example,the Platinum Group Elements (PGEs). The magma is modelled with the governing equations for a single-phase incompressible Newtonian fluid with variable viscosity and density. In this thesis, thermal conductivity and specific heat are approximated as constants and the country rock is treated as a conducting solid so as to save on computational time in the initial phases of the project. A basic review of the relevant literature is presented as background material and three basic models of magma chambers are discussed: crystal settling, compositional convection and double diffusive convection.The results presented in this thesis are from finite element calculations by a commercial computer code: ANSYS 5.4. This code has been employed in industry for over 26 years and has a long and successful benchmark history. In this context, finite element methods that are applicable to the code are discussed in chapter 5. In chapter 6, results that were obtained in the course of this research are presented. The thesis concludes with an indication of the possible geological significance of the results and various refinements that should be made to future models.
- Full Text:
- Authors: Botha, André Erasmus
- Date: 1999
- Subjects: Ore deposits , Fluid mechanics , Magmatism
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5492 , http://hdl.handle.net/10962/d1005278 , Ore deposits , Fluid mechanics , Magmatism
- Description: This thesis forms part of a larger effort which aims to establish the means of assessing the fluid mechanical behaviour of magma 1 as it cools inside a magma chamber surrounded by porous country rock. The reason for doing so is to advance the understanding of some types of mineral deposits; for example,the Platinum Group Elements (PGEs). The magma is modelled with the governing equations for a single-phase incompressible Newtonian fluid with variable viscosity and density. In this thesis, thermal conductivity and specific heat are approximated as constants and the country rock is treated as a conducting solid so as to save on computational time in the initial phases of the project. A basic review of the relevant literature is presented as background material and three basic models of magma chambers are discussed: crystal settling, compositional convection and double diffusive convection.The results presented in this thesis are from finite element calculations by a commercial computer code: ANSYS 5.4. This code has been employed in industry for over 26 years and has a long and successful benchmark history. In this context, finite element methods that are applicable to the code are discussed in chapter 5. In chapter 6, results that were obtained in the course of this research are presented. The thesis concludes with an indication of the possible geological significance of the results and various refinements that should be made to future models.
- Full Text: