Cyclodepsipeptides from a Kenyan marine cyanobacterium
- Authors: Dzeha, Thomas Mwambire
- Date: 2003
- Subjects: Cyanobacteria , Stereochemistry , Natural products -- Kenya
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4303 , http://hdl.handle.net/10962/d1004961 , Cyanobacteria , Stereochemistry , Natural products -- Kenya
- Description: An examination of an organic extract of the cyanobacterium Lyngbya majuscula collected from Wasini Island off the southern Kenyan coast led to the isolation of the known cyclodepsipeptide antanapeptin A (7), recently isolated from a Madagascan collection of L. majuscula, and a new bioactive cyclodepsipeptide, homodolastatin 16 (42). Although L. majuscula is a common, pantropical cyanobacterium this study represents the first investigation of the natural product chemistry of a Kenyan population of L. majuscula. The structures of the two cyclodepsipeptides were determined from 2D NMR and mass spectrometry data. The L- stereochemistry of the proline, valine, and N-methylphenylalanine amino acids in 7 and the L – proline configuration in 42, was confirmed by Marfey’s HPLC method. Chiral GC was used to determine the absolute stereochemistry of the hydroxyisovaleric acid moiety in 7 and 42, the lactate residue in 42 and tentatively propose an L-stereochemistry for the Nmethylisoleucine amino acid in 42. Homodolastatin 16, a higher homologue of the potential anti-cancer agent, dolastatin 16, exhibited moderate activity against two oesophageal cancer cell lines.
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- Date Issued: 2003
Effects of small-scale water movement on the settlement and growth rates of the brown mussel Perna perna, on the south-east coast of South Africa
- Authors: Mathagu, Tendamudzimu Titus
- Date: 2003
- Subjects: Mussels -- South Africa , Perna -- South Africa , Perna -- Growth
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5682 , http://hdl.handle.net/10962/d1005368 , Mussels -- South Africa , Perna -- South Africa , Perna -- Growth
- Description: The effects of small scale (cm) water movement on the settlement and growth rates of the brown mussel Perna perna were investigated on the south-east coast of South Africa (33°28′S, 27°10′E). L-shaped metal baffles attached to the substratum decreased the erosion rates of cement balls and it was concluded that the baffles decreased the water flow rate around cement balls. These L-shaped baffles were then used to decrease water flow rates around mussel patches and pot-scouring pads used as artificial substrata for the settlement of P.perna larvae. Anova indicated that settlement rate varied by date and site while decreased water flow rate significantly increased larval settlement (p<0.05), only on the site and day that had the overall highest number of settlers. Mussels in the low zone had significantly higher growth rates than those in the high zone. Decreased water flow rate significantly increased mussel growth rate in the lower zone (Anova, p<0.05), while it did not have a significant effect on the mussel in the high zone. Thus water flow manipulation increased growth rates in the zone, which already had high growth rate. It was concluded that small-scale (cm) water flow patterns have an effect on both Perna perna settlement and growth rates, but only under specific conditions. Larval settlement rate was significantly increased by water flow manipulation on the site and day that had the highest number of settlers. Growth rates were significantly increased by decreased water flow rate only in the low zone, where growth rates are the highest. Although water flow was manipulated in both zones its effect in the high zone was insignificant (Anova) compared to other factors affecting growth rates at this tidal level.
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- Date Issued: 2003
Finite element modelling of a magma chamber surrounded by country-rock, with particular reference to the groundwater flow in sections of different permeability
- Authors: Remsing, Carmen
- Date: 2003 , 2013-05-23
- Subjects: Magmas , Groundwater flow
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5508 , http://hdl.handle.net/10962/d1007537 , Magmas , Groundwater flow
- Description: This thesis presents results of two-dimensional finite element modelling of a magma chamber surrounded by country-rock containing a section of high permeability. The high permeability section in the country-rock simulates structure that is predominant in controlling the groundwater convection pattern and resulting mineral deposits. The models have analogies in nature: for instance the gold mines in the Massif Central of France, the Pogo mine in Alaska and the Pilgrim's Rest gold field in South Africa. This is a complicated coupled system involving fluid flow and heat transfer under extreme conditions. The magma in the chamber convects and as it cools the heat liberated causes convection in the groundwater contained in the surrounding country-rock. This convection in turn affects the rate of liberation of heat from the magma. The software used for the modelling, FLOTRAN, is the computational fluid dynamics component of the commercial ANSYS package. The results obtained describe in detail the flow pattern in the magma chamber, the country-rock and high permeability section thereof. During the cooling of the magma chamber the groundwater convects more vigorously in the high permeability section than elsewhere, and a convection cell is seen forming within this region. This provides a mechanism for hydrothermal formation of valuable mineral deposits in the structure near a magma chamber. It is found that the relationship between the velocity of the flow in the cell and the temperature of the magma chamber is well represented by a first order linear differential equation, providing a simple understanding of this process, , KMBT_363 , Adobe Acrobat 9.54 Paper Capture Plug-in
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- Date Issued: 2003
Food choices of tactile defensive children
- Authors: Smith, Anna Magdalena
- Date: 2003
- Subjects: Children -- Nutrition -- Psychological aspects
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:11034 , http://hdl.handle.net/10948/d1020371
- Description: This research explores a different view of the picky or fussy eater. Although occupational therapists and speech therapists are aware that children with sensory defensiveness and specific tactile defensiveness have different eating habits, this has not been described before.
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- Date Issued: 2003
Investigation of the comparative cost-effectiveness of different strategies for the management of multidrug-resistant tuberculosis
- Authors: Rockcliffe, Nicole
- Date: 2003
- Subjects: Tuberculosis , Multidrug resistance , Tuberculosis -- Treatment
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3788 , http://hdl.handle.net/10962/d1003266 , Tuberculosis , Multidrug resistance , Tuberculosis -- Treatment
- Description: The tuberculosis epidemic is escalating in South Africa as well as globally. This escalation is exacerbated by the increasing prevalence of multidrug-resistant tuberculosis (MDRTB), which is defined by the World Health Organisation (WHO) as resistance of Mycobacteria to at least isoniazid and rifampicin. Multi-drug resistant tuberculosis is estimated to occur in 1-2% of newly diagnosed tuberculosis (TB) patients and in 4-8% of previously treated patients. MDRTB is both difficult and expensive to treat, costing up to 126 times that of drug-sensitive TB. Resource constrained countries such as South Africa often lack both the money and the infrastructure to treat this disease. The aim of this project was to determine whether the performance of a systematic review with subsequent economic modelling could influence the decision making process for policy makers. Data was gathered and an economic evaluation of MDRTB treatment was performed from the perspective of the South African Department of Health. Three treatment alternatives were identified: a protocol regimen of second line anti-tuberculosis agents, as recommended in the South African guidelines for MDRTB, an appropriate regimen designed for each patient according to the results of culture and drug susceptibility tests, and non-drug management. A decision-analysis model using DATA 3.0 by Treeage® was developed to estimate the costs of each alternative. Outcomes were measured in terms of cost alone as well as the ‘number of cases cured’ and the number of ‘years of life saved’ for patients dying, being cured or failing treatment. Drug, hospital and laboratory costs incurred using each alternative were included in the analysis. A sensitivity analysis was performed on all variables in order to identify threshold values that would change the outcome of the evaluation. Results of the decision analysis indicate that the individualised regimen was both the cheaper and more cost-effective regimen of the two active treatment options, and was estimated to cost R50 661 per case cured and R2 070 per year of life saved. The protocol regimen was estimated to cost R73 609 per case cured and R2 741 per year of life saved. The outcome of the decision analysis was sensitive to changes in some of the variables used to model the disease, particularly the daily cost of drugs, the length of time spent in hospital and the length of treatment received by those patients dying or failing treatment. This modelling exercise highlighted significant deficiencies in the quality of evidence on MDRTB management available to policy makers. Pragmatic choices based on operational and other logistic concerns may need to be reviewed when further information becomes available. A case can be made for the establishment of a national database of costing and efficacy information to guide future policy revisions of the South African MDRTB treatment programme, which is resource intensive and of only moderate efficacy. However, due to the widely disparate range of studies on which this evaluation was based, the outcome of the study may not be credible. In this case, the use of a systematic review with subsequent economic modelling could not validly influence policy-makers to change the decision that they made on the basis of drug availability.
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- Date Issued: 2003
Investigations into the asymmetric reduction of ketones
- Authors: Bena, Luvuyo Clifford
- Date: 2003
- Subjects: Ketones , Asymmetric synthesis
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:11065 , http://hdl.handle.net/10948/323 , Ketones , Asymmetric synthesis
- Description: A six-step synthesis of salbutamol from methyl salicylate with an overall yield of 17% has been completed, although the yield was not optimised. In the process, Zn(BH4)2 was found to selectively reduce a ketone carbonyl group in the presence of an ester unit. In contrast, borane was found to reduce both the ketone and ester carbonyl groups. Reduction of phenacyl bromide with borane in the presence of chiral catalysts based on (R)-alaninol and (R,S)-ephidrine resulted a measure of enantioselectivity in the product. However, the configuration of the alcohol obtained in the case of (R)-alaninol was contrary to expectations based both on experimental trends observed elsewhere as well as our own theoretical predictions. The asymmetric reduction of methyl 5-bromoacetyl-2-benzyloxybenzoate was accomplished with both borane and Zn(BH4)2 in the presence of a range of chiral catalysts. Optically active products were obtained in all cases, although the optical rotations were significantly smaller in the case of Zn(BH4)2. Unfortunately, we were not successful in determining the enantiomeric excesses of these reactions. The use of a NMR lanthanide shift reagent resulted in a complex spectrum that was impossible to interpret unambiguously. This presumably arises from the presence of several Lewis base sites in the product at which complexation with the shift reagent can take place. It was also not possible to determine the optical rotation of salbutamol itself owing to the relatively small amount of material obtained. A conformational analysis of salbutamol, where NMR data was correlated with molecular modelling results, was successfully carried out and revealed a strong preference for that conformer family characterised by O–C–C–N and Ar–C–C–N torsion angles of ca. 60º and 180º, respectively. Interestingly, these conformers are found to be stabilised by OH…N rather than NH…O hydrogen bonding. This study has also confirmed the effectiveness of the MMFF94 force field for conformational analysis studies in compounds of this kind. Lastly, a relatively simple method for modelling the BH3/oxazaborolidine reduction of ketones at the PM3 semiempirical MO level of approximation was devised. This approach has provided insights into the mechanism of the reaction and has furthermore enabled us to predict the enantioselectivities likely to result from various catalysts and ketones. In comparing our theoretical and experimental findings, an anomalous result was observed in the case of (R)-alaninol; this will have to be investigated further, particularly at the experimental level. However, we believe that our approach provides a sound basis for aiding the design and screening of new, potentially better catalysts.
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- Date Issued: 2003
Isolation, expression and purification of the hydantoin hydrolysing enzymes of agrobacterium tumefaciens
- Authors: Clark, Sally-Ann
- Date: 2003
- Subjects: Agrobacterium tumefaciens , Amino acids Hydantoin Enzymes
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4140 , http://hdl.handle.net/10962/d1016233
- Description: The production of enantiomerically pure amino acids is of industrial importance as they are used in the synthesis of a number of pharmaceuticals, insecticides and herbicides and biologically active peptides and hormones. A number of microorganisms have been identified which possess hydantoin hydrolysing enzymes that stereoselectively convert racemic hydantoins into anantiomerically pure amino acids. Consequently these microorganisms and their enzymes are sought after as biocatalysts for the production of amino acids. The isolation of novel hydantoin hydrolising enzymes with unique or improved biocatalytic characteristics is of importance for the development of potential biocatalysts to be used in the production of enantiomerically pure amino acids. The genes encoding an N-carbamoyl-amino acid amidohydrolase, an enzyme involved in the hydrolysis of hydantoin, was isolated by screening a genomic DNA library of Agrobacterium tumefacience RU-AE01. Nucleotide sequence analysis of the region upstream of this gene revealed a fragment of a gene encoding the hydantoinase enzyme. I this study, a DNA probe consisting of the gene encoding the N-carbamoyl amino acid amidohydrolase, on a large enough fragment of the genomic DNA library which would allow for the simultaneous isolation the hydantoinase gene located upstream. Recombinant expression of the genes encoding hydantoin hydrolysing enzymes has been used to facilitate the production and purification of these enzymes for their use as biocatalysts. Two genes (ncaR1 and ncaR2) encoding different N-carbamoyl-amino acid amidohydrolases with distinct nucleotide and deduced amino acid sequences were isolated from the genome of A, tumefaciens RU-OR. In this study, the heterologous expression of ncaR1 and ncaR2 was explored. Investigation into the optimisation of the heterologous expression of ncaR1 showed that reducing the growth temperature of the recombinant E. coli producing NcaR1 resulted in a two-fold increase in N-carbamoyl-amino acid amidohydrolase activity and solubility. Furthermore, NcaR1 was produced with a C-terminal 6xHis tag, but NcaR1-6xHis did not possess N-carbamoyl amino acid amidohydrolase activity. Furthermore, purification of NcaR-6xHis under native conditions using affinity chromatography performed, and used for the production of antibodies.
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- Date Issued: 2003
Mesozooplankton community structure and grazing impact in the polar frontal zone of the Southern Ocean
- Authors: Bernard, Kim Sarah
- Date: 2003
- Subjects: Zooplankton -- Antarctic Ocean , Copepoda -- Feeding and feeds
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5731 , http://hdl.handle.net/10962/d1005417 , Zooplankton -- Antarctic Ocean , Copepoda -- Feeding and feeds
- Description: Mesozooplankton community structure and grazing impact in the Polar Frontal Zone (PFZ) of the Southern Ocean were investigated during two cruises of the South African National Antarctic Programme (SANAP), the Marion Offshore Ecosystem Variability Study I & II (MOEVS). During the first cruise (MOEVS I), a meso-scale oceanographic grid survey was conducted in the upstream region of the Prince Edward Islands (PEI) in austral autumn (April) 2001. Mesozooplankton samples, collected using a Bongo net (fitted with 200 and 300µm mesh nets) at depths between 200 and 300 m, were separated into three size fractions: 200-500 µm; 500-1000 µm; 1000-2000 µm by reverse filtration. Total surface (depth <5 m) chlorophyll-a (chl-a) concentration (measured fluorometrically) during the study ranged between 0.11 and 0.34 µg 1^(-1) and was always dominated by picophytoplankton (<2.0 µm). Total mesozooplankton abundance and biomass during the survey ranged between 49 and 1512 ind. m^(-3) and between 0.7 and 25 mg Dwt. m^(-3), respectively. Throughout the survey, the 200-500 µm class numerically dominated the mesozooplankton community, comprising an average of ~ 69% (SD = ± 12.3%). The dominant species in the 200-500 µm size fraction were the copepods Oithona similis, Calanus simillimus and Metridia lucens and the pteropod, Limacina retroversa. However, in terms of biomass, the 1000-2000 µm group was predominant, with dry weight values constituting an average of ~ 66% (SD = ± 10.2%). Biomass was dominated by carnivorous zooplankton, particularly the euphausiids, Euphausia vallentini and Thysanoessa vicina and the chaetognaths, Sagitta gazellae and Eukrohnia hamata. Three distinct groupings of stations were identified by multivariate analysis. The different station groupings identified reflect changes in the relative contributions of the rather than different species assemblages. During the second cruise (MOEVS II), conducted in April 2002 (austral autumn), mesozooplankton community structure and grazing impact were investigated at 13 stations in the west Indian sector of the PFZ. Total integrated chl-a biomass ranged between 11.17 and 28.34 mg chl-a m^(-2) and was always dominated by nano- and picophytoplankton (<20 µm). Throughout the study, small copepods, mainly Oithona similis and Ctenocalanus vanus, numerically dominated the mesozooplankton community comprising up to 85% (range 30 to 85%) of the total abundance. Grazing activity of the four most abundant copepods (O. similis, C. vanus, Calanus simillimus and Clausocalanus spp.), which comprised up to 93% of total mesozooplankton abundance, was investigated using the gut fluorescent technique. Results of gut fluorescence analyses indicated that C. simillimus, Clausocalanus spp. and Ctenocalanus vanus exhibited diel variability in gut pigments, with maximum values at various stages of the night. In contrast, O. similis did not demonstrate diel variation in gut pigment contents. Ingestion rates of the four copepods ranged from 23.23 to 1462.02 ng (pigm.) ind^(-1) day^(-1), depending on the species. The combined grazing impact of the four copepods, ranged between 1 and 36% of the phytoplankton standing stock per day, with the highest daily impact (~ 35.86%) occurring at stations in the vicinity of the Antarctic Polar Front. Among the copepods, O. similis and C. vanus were generally the most important consumers of phytoplankton biomass; together they were responsible for up to 89% (range 15 to 89%) of the total daily grazing impact. Carbon specific ingestion rates of the copepods varied between 42 and 320% body carbon per day, depending on the species. The study highlights the importance of small copepods in terms of both their significant contribution to total mesozooplankton numbers and their grazing impact on the phytoplankton standing stocks in the PFZ during austral autumn.
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- Date Issued: 2003
Morphometrics and reproduction of Terebrasabella heterouncinata (Polychaeta:Sabellidae), infesting abalone (Haliotis midae) from different culture environments
- Authors: Gray, Michael
- Date: 2003
- Subjects: Polychaeta , Sabellidae , Abalones -- Physiology , Abalones -- Nutrition , Abalone culture
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5384 , http://hdl.handle.net/10962/d1018231
- Description: In the late 1980's abalone culturalists noticed reduced growth rate and shell deformities in some abalone stocks. These problems were the result of infestations by a shell boring polychaete, Terebrasabella heterouncinata. Under intensive abalone culture conditions the level of infestation can reach epidemic proportions and there are often severe consequences for the host abalone. Heavy sabellid infestation levels have placed the economic viability of several South African farms under threat. This study formed part of an ongoing project that is aimed at investigating the basic biology of Terebrasabella heterouncinata. The majority of abalone farmers in South Africa feed their abalone either naturally occurring kelp (Ecklonia maxima) or the formulated abalone feed, Abfeed. Farmers have suggested that the use of Abfeed is associated with higher sabellid infestation levels and changing the abalone diet from Abfeed to kelp helps reduce sabellid infestation. Speculation has arisen indicating that older, slower growing abalone are more susceptible to sabellid infestation. The effect of host abalone diet history and their growth on sabellid settlement success, morphometries and reproduction was quantified. To better understand the plasticity of the expression of life history traits the variability of morphometric and reproductive characteristics was compared between different farm environments. And more...
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- Date Issued: 2003
Novel approaches to the monitoring of computer networks
- Authors: Halse, G A
- Date: 2003
- Subjects: Computer networks , Computer networks -- Management , Computer networks -- South Africa -- Grahamstown , Rhodes University -- Information Technology Division
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4645 , http://hdl.handle.net/10962/d1006601
- Description: Traditional network monitoring techniques suffer from a number of limitations. They are usually designed to solve the most general case, and as a result often fall short of expectation. This project sets out to provide the network administrator with a set of alternative tools to solve specific, but common, problems. It uses the network at Rhodes University as a case study and addresses a number of issues that arise on this network. Four problematic areas are identified within this network: the automatic determination of network topology and layout, the tracking of network growth, the determination of the physical and logical locations of hosts on the network, and the need for intelligent fault reporting systems. These areas are chosen because other network monitoring techniques have failed to adequately address these problems, and because they present problems that are common across a large number of networks. Each area is examined separately and a solution is sought for each of the problems identified. As a result, a set of tools is developed to solve these problems using a number of novel network monitoring techniques. These tools are designed to be as portable as possible so as not to limit their use to the case study network. Their use within Rhodes, as well as their applicability to other situations is discussed. In all cases, any limitations and shortfalls in the approaches that were employed are examined.
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- Date Issued: 2003
Observed metabolic changes in male Wistar rats after treatment with an antidepressant implied in undesirable weight gain, or Sutherlandia frutescens for Type II diabetes
- Authors: Chadwick, Wayne
- Date: 2003
- Subjects: Rats -- Metabolism , Non-insulin-dependent diabetes -- Research , Rats as laboratory animals
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:11068 , http://hdl.handle.net/10948/313 , Rats -- Metabolism , Non-insulin-dependent diabetes -- Research , Rats as laboratory animals
- Description: Type II diabetes is fast becoming a growing problem in developed countries worldwide. Traditionally the median age for diagnosis was around sixty, but recent surveys have shown that the entire age distribution curve has shifted to the left. Western countries boast the worst statistics in which type II diabetes is being reported in children under the age of ten. At such a young age the disease often goes undiagnosed for long periods of time allowing considerable damage to occur. The incidence of type II diabetes is thought to be parallel with the growing rate of obesity associated with a characteristically unhealthy western diet. Type II diabetes is an extremely expensive disease to manage, and with the rapid growth of this pandemic our country will soon feel the economic burden of this disease. It is for this reason that cheaper medication needs to be investigated in the form of traditional plants, such as Sutherlandia frutescens. Prescription medication, such as tricyclic antidepressants, may also increase body weight or appetite thereby playing a role in obesity. The cause of weight gain in such cases may go unrecognized or lead to cessation of the medication with or without the practitioner’s knowledge or approval. It is therefore necessary to investigate the causative agents responsible for the excessive weight gain. Drinking water containing extracts of the S. frutescens, metformin (a well known type II diabetes medication) and amitriptyline (a common tricyclic antidepressant) was administered to three groups of ten male Wistar rats. The control group received water without any medication. The rat’s weight and food consumption was monitored throughout the trial and their oxygen consumption was also determined. Rats were sacrificed after four months of medicinal compliance and glucose uptake, in the presence and absence of insulin, was tested in epididymal fat, liver and muscle. Fasting plasma glucose levels, lipoprotein, cholesterol and triglyceride concentrations were also determined.
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- Date Issued: 2003
Over-expression, purification and biochemical characterisation of trypanosomal heat shock protein
- Authors: Edkins, Adrienne Lesley
- Date: 2003
- Subjects: Heat shock proteins , Heat shock proteins -- Structure activity relationships
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4106 , http://hdl.handle.net/10962/d1011736 , Heat shock proteins , Heat shock proteins -- Structure activity relationships
- Description: The molecular chaperone process of assisted protein folding, characteristic of members of the Heat Shock Protein 70 kDa (Hsp70) and Heat Shock Protein 40kDa (Hsp40) families, is essential for cytoprotection in stressful cellular conditions. Examples of such conditions are heat shock or invasion by pathogens. The Hsp70/Hsp40 process of assisted protein folding is dependent on ATP (governed by the intrinsic ATPase activity of Hsp70) and the ability of molecular chaperones to recognise and bind non-native protein conformations. Here, we analyse and attempt to characterise the molecular chaperone activity of an inducible, cytoplasmic Hsp70 (TcHsp70) from Trypanosoma cruzi and its interactions with its potential partner Hsp40s, Tcj 1, Tcj2, Tcj3 and Tcj4. A bioinformatic analyses of the primary sequences of the trypanosomal proteins revealed that they all contained the canonical domains that define other members of the Hsp70 and Hsp40 family. Tcj2 and Tcj4 showed deviations from the consensus sequence in their substrate binding regions, which may have implications for their substrate binding specificities. TcHsp70, Tcj 1, Tcj2, Tcj3 and Tcj4 were over-expressed recombinantly as 6xHis-tag fusion proteins in Escherichia coli. His-TcHsp70, Tcjl-His and His-Tcj2 were successfully purified by Nickel-affinity chromatography for functional analyses to assess the molecular chaperone activity of His-TcHsp70 in terms of its ATPase activity and substrate binding ability. The basal ATPase activity of His-TcHsp70 was determined as 40 nmol Pi/min/mg, significantly higher than that reported for other Hsp70s. This basal ATPase activity was stimulated to a maximal level of 60 nmol Pi/min/mg in the presence of His-Tcj2 and a model non-native substrate, reduced carboxymethylated αx-lactalbumin (RCMLA). Using native polyacrylamide gel electrophoresis and Western analysis, His-TcHsp70 was shown to form discrete complexes when in the presence of Tcj 1- His, His-Tcj2 and/or RCMLA. These complexes potentially represent His-TcHsp70 - RCMLA or His-TcHsp70 - Tcj interactions, that may be indicative of chaperone activity. In vivo complementation assays showed that Tcj2, but not Tcj3, was able to overcome the temperature sensitivity of the ydjJ mutant Saccharomyces cerevisiae strain JJ160, suggesting that Tcj2 may be functionally equivalent to the yeast Hsp40 Ydj1.
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- Date Issued: 2003
Pharmaceutical analysis and aspects of the quality control of St. John's Wort products
- Authors: Wild, Tracy Joy
- Date: 2003
- Subjects: Hypericum perforatum , Hypericum perforatum -- Analysis , Hypericum perforatum -- Therapeutic use , Hypericum perforatum -- Physiological effect , Flavonoids -- Analysis
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3804 , http://hdl.handle.net/10962/d1003282 , Hypericum perforatum , Hypericum perforatum -- Analysis , Hypericum perforatum -- Therapeutic use , Hypericum perforatum -- Physiological effect , Flavonoids -- Analysis
- Description: Most complementary medicines contain a multitude of chemical components, some of which are claimed to contribute to the biological activity of such products. Use of complementary medicines for preventative and therapeutic purposes is increasing rapidly worldwide. Unfortunately, although control of these products is essential to ensure quality, safety, and efficacy, the quality control of most herbal preparations is currently poor to non-existent, with little or no safety and efficacy data required to support the marketing and use of these products. The objective of this study was therefore to develop suitable analytical methods to qualitatively and quantitatively analyse the relevant components (rutin, isoquercitrin, hyperoside, quercitrin, quercetin, kaempferol, hypericin, pseudohypericin and hyperforin) in St John's Wort dosage forms for quality control purposes. A gradient HPLC method using a Luna 5·mC₁₈(2) 150 x 2.00mm internal diameter (i.d.) column and UV detection, was developed for the separation of six of the relevant flavonoid compounds in St John's Wort, namely rutin, isoquercitrin, hyperoside, quercitrin, quercetin and kaempferol. The development process involved a systematic investigation of gradient conditions, flow rate, and temperature. This method was subsequently applied to assay selected commercially available St John's Wort products. This system provided the necessary accuracy, precision and reproducibility and was associated with several advantages when compared to using standard bore (4.60 mm i.d.) HPLC columns. The method developed is currently the only known method that separates all six relevant flavonoids in a reasonable run time (less than 20 minutes). It is also one of the few methods that has sufficient separation between rutin, isoquercitrin and hyperoside. A qualitative method for the fingerprinting of flavonoid components was also developed, using capillary electrophoresis (CE). CE is a rapidly growing powerful analytical technique for the separation of charged compounds. Micellar electrokinetic chromatography (MEKC) is a very powerful electrophoretic technique that is capable of selectively resolving both neutral and ionic solutes in a single run. A MEKC method suitable for the separation and determination of various flavonoid constituents used as marker compounds in Hypericum perforatum was developed. Investigations into the effect of pH, ionic strength, applied voltage and capillary dimensions on separation were performed. The optimised method was then applied to qualitatively analyse various St John's Wort products on the market. This method was found to be advantageous in that it was simple, cost-effective, required minimal sample preparation and utilised very small quantities of sample. Due to the vast differences in chemical properties between the various marker and active components in St John's Wort, it was necessary to develop separate analytical methods for the flavonoids and for the other three relevant compounds (hypericin, pseudohypericin and hyperforin). An isocratic HPLC method using a Luna 5·mC₁₈(2) 150 x 2.00mm (i.d.) column and UV detection was developed for the separation of hypericin, pseudohypericin and hyperforin. The development process involved a systematic investigation of buffer molarity, mobile phase composition, pH, flow rate, and temperature. This method was subsequently applied to assay selected commercially available St John's Wort products on the South African market. This system also provided the necessary accuracy, precision and reproducibility, as well as the advantages associated with the use of a narrow bore column as opposed to the use of the more commonly used wider bore columns. This method was validated and used to quantitate these three compounds in various commercial St John's Wort products. By applying this method to liquid chromatography – tandem mass spectrometry (LC-MS-MS), qualitative analyses of the same products was performed to obtain confirmation of the quantitative HPLC results. Mass spectrometry is a powerful detection tool that is more selective and specific than many detection systems used with HPLC. Natural medicines usually constitute a multitude of constituents with much potential interference. In this regard LC-MS-MS is a powerful tool, with its ability to unequivocally identify target analytes regardless of the presence of interferences or complex matrices. ESI-MS-MS was used for the qualitative analysis of the content of the naphthodianthrones and hyperforin in the respective tablet products assayed with HPLC. LC-MS-MS analyses were performed in order to identify the constituents and to verify the specificity of the HPLC method. High inter-product and inter-batch variability was observed for all nine compounds assayed. These quantitative results were confirmed with the respective qualitative analyses. This study confirms the need for strict quality control of herbal medicinal products commercially available to consumers.
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- Date Issued: 2003
Physiological, perceptual and performance responses during cricket activity
- Authors: King, Gregory Allen
- Date: 2003
- Subjects: Cricket , Sports -- Psychological aspects , Sports -- Physiological aspects
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5131 , http://hdl.handle.net/10962/d1005210 , Cricket , Sports -- Psychological aspects , Sports -- Physiological aspects
- Description: The present study sought to determine the influence of environmental conditions and protective clothing on physiological, perceptual and performance responses during batting activity. The investigation examined mean skin temperature, average heart rate, estimated sweat rate, rating of perceived exertion, thermal sensation rating, average sprint time and pre-post choice reaction time. Twenty-five cricketers (18-22 yr, 73.1 ± 9.6 kg, 1768 ± 75 mm, 12.6 ± 3.1% body fat, 1.89 ± 0.16 m2) performed a work-bout consisting of a seven-Over batting period, during which time they faced deliveries from a bowling machine and performed two shuttle runs every third ball to total four sprints per Over. Trials were carried out under High-stress (23.8 ± 2.2 °C) and Low-stress (13.3 ± 1.9 °C) environmental conditions (WBGT). Within each environmental condition subjects performed the test wearing full protective batting gear and no protective gear. Thus, four specific conditions were examined; high full-gear (HFG), high no-gear (HNG), low full-gear (LFG) and low no-gear (LNG). Two-way ANOVAs were calculated to determine whether there were differences between environmental conditions and whether differences existed between the clothing conditions. One-way ANOVAs were utilised to compute differences between the four specific conditions combining clothing and environment. High environmental stress and wearing protective clothing caused batsmen to experience significant physiological strain. The environment was the greatest stressor, with the protective gear exacerbating these effects. However, when padding covered skin areas directly, this was the primary skin temperature stressor, particularly later in the activity. For skin temperature and heart rate, the strain was the most pronounced at the end of the trials. Perceptual responses indicated that the protective gear had no influence on effort sense, thermal sensation or thermal comfort. However, environmental conditions had an effect, and High-stress conditions resulted in significantly higher perceptions of effort, elevated sensations of heat and greater thermal discomfort. Effort was perceived to be greatest towards the end of the trial. There were mixed findings for performance factors. In general sprint performance was not hindered by environmental stress, but protective clothing caused a reduction in several sprint times. Choice reaction times were for the most part unaffected by either environment or clothing and few differences were observed between pre and post times. It is contended that intense short duration batting activity, likely encountered during one-day participation, imposes a stress on batsmen. The stress is greater when conditions are warmer and protective padding is worn, although it is not sufficient to impede choice reaction time. However, protective gear did have a deleterious effect on sprint performance.
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- Date Issued: 2003
Progress towards the development and implementation of an unambiguous copper wire fingerprinting system
- Authors: Poole, Martin
- Date: 2003
- Subjects: Electroplating , Copper , Telecommunication , Theft -- Prevention
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5484 , http://hdl.handle.net/10962/d1005270 , Electroplating , Copper , Telecommunication , Theft -- Prevention
- Description: The Telecommunications industry in Southern Africa is faced with the problem of theft of the signal carrying copper wire, both from the ground and from telephone poles. In many cases, if the offenders are caught, the prosecuting party has no way of proving that the wire is the property of any one Telecommunication company, as any inked markings on the insulating sheaths have been burned off along with the insulation and protective coatings themselves. Through this work we * describe the problem, * specify the necessary and preferred technical properties of a viable solution, * report the preliminary investigations into the devising of an unambiguous "fingerprinting" of the 0.5 mm wires, including some of those solutions that, upon investigation, appear non-viable, * describe the development and implementation of an electrochemical marker with detection mechanism which has shown in proof-of-principle to work, * outline the road-map of necessary future work.
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- Date Issued: 2003
Spatial and temporal occurrence of forensically important South African blowflies (Diptera: Calliphorida)
- Authors: Williams, Kirstin Alexa
- Date: 2003
- Subjects: Blowflies -- South Africa , Forensic entomology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5617 , http://hdl.handle.net/10962/d1003217 , Blowflies -- South Africa , Forensic entomology
- Description: Forensic entomology is an emergjng field in South Africa. Little is known about South African blowflies and factors that affect their use in a forensic context. This work provides a review and synthesis of previous work in South Africa and supplements some of the background and basic knowledge required for forensic entomology in South Africa. The seasonal occurrence of eight forensicaIIy important blowfly species was quantified by fortnightly trapping in Grahamstown, South Africa. The spatial distribution of each species was related to seasonal occurrence and habitat preference. Seasonal distributions of blowflies in carcasses in South Africa were obtained from the literature and compared to the seasonal trapping. By mapping South African locality records of forensicaIIy important blowflies and analyzing these records in a modified Principal Components Analysis of climatic data, the potential geographic distributions of each fly species was modeIIed. Most species were widespread, but Calliphora croceipalpis, Jaennicke, 1867, was found in cold places. This information is important for determining where certain species are likely to occur in forensic investigations. Nocturnal oviposition was examined in both field and laboratory experiments. Lucilia species could oviposit nocturnaIIy in the field, while Lucilia species, Chrysomya chloropyga, (Weidemann, 1818) and C. putoria (Weidemann, 1830) could oviposit nocturnaIIy in the laboratory. These findings are important factors in affecting the precision of estimates of a post mortem interval (PM!) by up to 12 hours. The thermophysiological ranges of four species of adult blowflies were determined by measuring onset temperatures of four significant behaviours: onset of neural activity; onset of coordinated movement; shade-seeking and death. There was a sexual size dimorphism in Lucilia species, Chrysomya chloropyga and Calliphora croceipalpis with females being larger than males. Chrysomya megacephala (Fabricius, 1794) had an unexpectedly high death threshold, while Calliphora croceipalpis had the lowest death threshold of the flies tested. These points were related to the seasonal and geographic occurrence of each species, to nocturnal activity and placed in a forensic context.
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- Date Issued: 2003
Symplasmic pathway in phloem loading and unloading in source and sink leaves of Zea mays L. as evidenced under normal and elevated CO₂ conditions
- Authors: Nogemane, Noluyolo
- Date: 2003
- Subjects: Phloem , Plant translocation , Plant cells and tissues , Corn -- Metabolsim
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4254 , http://hdl.handle.net/10962/d1007813
- Description: Zea mays plants kept at ambient (ca 375ppm) and elevated CO₂ (ca 650 to 700ppm) were used to examine the possibility of a symplasmic loading, unloading and transport pathway in dark-adapted and illuminated (200μmolm⁻²sec⁻¹ ) sink and source leaves. 5,6-carboxyfluorescein diacetate was introduced into the mesophyll cells and symplasmic transfer observed 3h after application. In sink and source leaves exposed to ambient CO₂ and illuminated at 200 molm-2sec-1, the fluorescence front was observed approximately 3cm from the point of application, while in dark-adapted plants, the fluorescence front was observed approximately 1cm from the point of application. Under elevated CO₂ conditions the fluorescence front in illuminated plants appeared to transport faster moving approximately 5cm from the point of application, and in dark-adapted plants, only 3cm from the point of application. Based on the increase in 5,6-CF accumulation under elevated CO₂ conditions, the present study suggests that there was an increase in capacity for assimilate loading and transport under elevated CO₂ conditions. In source leaves, 5,6-CFDA was taken up into the mesophyll cells, loaded symplasmically and transported basipetally. In sink leaves 5,6- CFDA was taken up from basal mesophyll and after symplasmic loading, was transported acropetally where it was offloaded into the younger immature sink region. Transport in the sieve tubes was confirmed by using aniline blue, which was applied 3h after 5,6-CF transport. Aniline blue coupled with 5,6-CF transport studies showed that the sieve tubes of both cross and longitudinal veins are involved in symplasmic unloading, loading and transport processes in sink and source leaves. Apoplasmic uptake of 5,6-CFDA by cut leaves showed that after apoplasmic transport via the transpiration stream, 5,6-CFDA was offioaded to the xylem parenchyma where it was metabolically cleaved , releasing fluorescent 5,6-CF into the xylem parenchyma. Transverse sections cut after 3h of uptake were observed after 120 and 180 min suggesting that a retrieval of solutes occurs from the xylem to the xylem parenchyma, bundle sheath, phloem parenchyma and to the th in-walled sieve tubes. It was not possible to determine if the thick-walled sieve tubes were involved or if they took up 5,6-CF. Given the available data on loading and offioading of assimilates in sink and source leaves respectively, this study demonstrated that a slow symplasmic pathway exists from the mesophyll to the phloem, and that offloading from the phloem in sink leaves can occur via a symplasmic route.
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- Date Issued: 2003
Synthesis of zinc phthalocyanine derivatives for possible use in photodynamic therapy
- Authors: Matlaba, Pulane Maseleka
- Date: 2003
- Subjects: Photochemotherapy , Electrochemistry , Phthalocyanines , Zinc
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4374 , http://hdl.handle.net/10962/d1005039 , Photochemotherapy , Electrochemistry , Phthalocyanines , Zinc
- Description: The synthesis of symmetrically and unsymmetrically substituted zinc phthalocyanines (ZnPc) derivatives is done according to reported procedures. The unsymmetrical ZnPc derivatives are synthesized by ring expansion of sub-phthalocyanine complexes. Ring substitution is effected with tert-butyl phenol, naphthol, and hydroxybenzoic acid. Comparison of the redox potentials for the complexes substituted with varying numbers of tert-butyl phenol: 1, 2, 3, 6 and 8 show that the complex with the highest number of substituents are more difficult to oxidize and easier to reduce. Water soluble sulphonated ZnPc (ZnPcSn) was prepared. The possibility of using axial ligation to increase the solubility and the photochemical activity of sulphotnated ZnPc in aqueous solutions was investigated. Pyridine, aminopyridyl and bipyridyl were used as axial ligands. When bipyridyl was used as the axial ligand, solubility of the ZnPcSn increased, shown by the increase in the Q-band of the monomer species in solution and the singlet oxygen quantum yields was relatively higher than that of the unligated ZnPcSn. The singlet oxygen quantum yields by the various complexes in DMF using diphenylisobenzofuran as a chemical quencher for organic solvent were determined. Singlet oxygen quantum yields for the unsymmetrically ring substituted complexes range from 0.22 to 0.68. Photobleaching quantum yields are in the order of 10-5, which means that the complexes are relatively photostable.
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- Date Issued: 2003
The bioaccumulation of platinum (IV) from aqueous solution using sulphate reducing bacteria: role of a hydrogenase enzyme
- Authors: Rashamuse, Konanani Justice
- Date: 2003
- Subjects: Sulfur bacteria , Bioremediation , Enzymes -- Metabolism , Platinum , Platinum compounds , Reduction (Chemistry) , Hydrogenation
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4003 , http://hdl.handle.net/10962/d1004063 , Sulfur bacteria , Bioremediation , Enzymes -- Metabolism , Platinum , Platinum compounds , Reduction (Chemistry) , Hydrogenation
- Description: The enzymatic reduction of a high-valence form of metals to a low-valence reduced form has been proposed as a strategy to treat water contaminated with a range of metals and radionuclides. Metal reduction by sulphate reducing bacteria (SRB) is carried out either chemically (involving reduction by hydrogen sulphide) or enzymatically (involving redox enzymes such as the hydrogenases). While reduction of metal ions by hydrogen sulphide is well known, the enzymatic mechanism for metal reduction is poorly understood. The aims of this study were to investigate the role of SRB in facilitating platinum removal, and to investigate the role of a hydrogenase in platinum reduction in vitro. In order to avoid precipitation of platinum as platinum sulphide, a resting (non-growing) mixed SRB culture was used. The maximum initial concentration of platinum (IV), which SRB can effectively remove from solution was shown to be 50 mg.l⁻¹. Electron donor studies showed high platinum (IV) uptake in the presence of hydrogen, suggesting that platinum (IV) uptake from solution by SRB requires careful optimization with respect to the correct electron donor. Transmission electron microscopy (TEM) and energy dispersive X-ray (EDX) analysis indicated that platinum was being precipitated in the periplasm, a major area of hydrogenase activity in SRB. Purification of the hydrogenase by ammonium sulphate precipitation (65%), Toyopearl-Super Q 650S ion exchange and Sephacry 1 S-100 size exclusion chromatography revealed that the hydrogenase was monomeric with a molecular weight of 58 KDa, when analyzed by 12% SDS-PAGE. The purified hydrogenase showed optimal temperature and pH at 35°C and 7.5 respectively, and a poor thermal stability. In vitro investigation of platinum reduction by purified hydrogenase from mixed SRB culture showed that hydrogenase reduces platinum only in the presence of hydrogen. Major platinum (IV) reduction was observed when hydrogenase was incubated with cytochrome C₃ (physiological electron carrier in vivo) under hydrogen. The same observations were also noted with industrial effluent. Collectively these findings suggest that in vitro platinum reduction is mediated by hydrogenase with a concerted action of cytochrome C₃ required to shuttle the electron from hydrogenase.
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- Date Issued: 2003
The development of a baculovirus expression system for the production of Helicoverpa armigera stunt virus capsids for use in the encapsidation of foreign molecules
- Authors: Mosisili, Kekeletso Mpho Thakane
- Date: 2003
- Subjects: Helicoverpa armigera , Fall armyworm , Baculoviruses , Insects -- Viruses , RNA -- Analysis
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4088 , http://hdl.handle.net/10962/d1007700 , Helicoverpa armigera , Fall armyworm , Baculoviruses , Insects -- Viruses , RNA -- Analysis
- Description: The capsid protein of Helicoverpa armigera stunt virus (HaSV) a T=4 insect virus was expressed in Spodoptera frugiperda 9 cells using a baculovirus vector. When the insect cells were infected at a high MOl the expressed coat protein assembled into virus-like particles (VLPs) that spontaneously underwent maturation and were morphologically indistinguishable from wild-type HaSV. The VLPs were electron dense when viewed under EM and encapsidated their coat protein mRNA. When Sf9 cells were infected at a low multiplicity of infection (MOl) the expressed capsid protein assembled into procapsids that did not spontaneously undergo maturation. These procapsids underwent autoproteolytic maturation cleavage when they were treated with an acidic buffer. The procapsids were used in the encapsidation of a FITC labelled peptide. The peptide encapsidating VLPs showed an increase in their buoyant density that was not collaborated by an increase in the concentration of the FITC labelled peptide detected when these samples were compared to control samples with similar buoyant densities.
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- Date Issued: 2003