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Thesis (M.S.)--Duquesne University, 2009. / Title from document title page. Abstract included in electronic submission form. Includes bibliographical references (p. 82-90) and index.

Erythromycin, a macrolide antibiotic used mainly against gram-positive bacteria has been in clinical use since 1952 (1). Previous pharmacokinetic data published on this antibiotic have been derived predominantly from microbiological assay techniques. However, these techniques are relatively imprecise as well as being non-specific and extremely tedious to perform. A novel high performance liquid chromatographic analysis of erythromycin in human serum and urine using U.V. detection at 200 nm and/or electrochemical detection using both an amperometric and a coulometric electrochemical detector is presented. The method involves a solid phase extraction procedure followed by a simple phase separation step and chromatography on a reverse phase column. In order to select the optimum U.V. detector for this analysis, five "state of the art" detectors were compared in terms of their signal-to-noise ratios at U.V. wavelengths between 200 and 210 nm. A known metabolite des-N-methylerythromycin is readily detectable using U.V. detection, whilst another metabolite/degradation product anhydroerythromycin is not seen using U.V. detection but is readily observable using an electrochemical detector. The method has a limit of sensitivity of 0.25 μg/mL and 1.00 μg/mL in serum and urine respectively (U.V. detection) and is sufficiently sensitive to monitor serum and urine concentrations of erythromycin in man after administration of a single 500 mg erythromycin stearate tablet. / KMBT_363 / Adobe Acrobat 9.53 Paper Capture Plug-in

Magister Scientiae (Medical Bioscience) - MSc(MBS) / Even though Tulbaghia violacea has been used to treat and manage epilepsy in South Africa by traditional medicine practitioners, no evidence in any literature has shown any scientific scrutiny of the effectiveness of the plant species in therapy. This project was intended, therefore, to investigate the anticonvulsant effect of the leaf methanol extract of Tulbaghia violacea by studying its effect against tonic convulsion induced by either pentylenetetrazole (PTZ), bicuculline, picrotoxin, strychnine or N-methyl-DL-aspartic acid (NMDLA) in mice. Qualitative phytochemical analysis, acute toxicity and HPLC studies were also carried out on the plant species. The doses of 200 (mg/kg, i.p.) and 400 (mg/kg, i.p.) of the leaf methanol extract of T. Violacae significantly delayed the onset of PTZ (100 mg/kg, i.p.)-induced tonic convulsion in a dose dependent manner. Leaf methanol extract of the plant species (200 mg/kg, i.p.) did not affect the incidence of PTZ (100 mg/kg, i.p.)-induced tonic convulsion while 400 mg/kg (i.p.) protected only one mouse against the tonic convulsion. Leaf methanol extract of Tulbaghia violacea (100mg/kg, i.p.) did not significantly affect the onset or incidence of PTZ (100 mg/kg, i.p.)- induce tonic convulsion. Phenobarbitone (12 mg/kg, i.p.), diazepam (0.5 mg/kg, i.p.) and muscimol (2mg/kg, i.p.) significantly delayed the onset of PTZ (100 mg/kg, i.p.)-induced tonic convulsion and also significantly reduced the number of animals convulsing. Muscimol (0.2 mg/kg, i.p.) did not significantly affect the onset or incidence of PTZ (100 mg/kg, i.p.)-induced tonic convulsion. However, combined therapy of sub effective doses of the leaf methanol extract of T. Violacea (100 mg/kg, i.p.) and muscimol (0.2 mg/kg, i.p.) significantly delayed the onset of PTZ (100mg/kg, i.p.)-induced tonic convulsion and but did not significantly reduce the number of animals convulsing. The combined therapy of sub effective doses of the leaf methanol extract of T. violacea (100 mg/kg, i.p.) and muscimol (0.2 mg/kg, i.p.) protected two of the mice against the tonic convulsion. Leaf methanol extract of Tulbaghia violacea (100-400 mg/kg, i.p.) significantly and dose dependently delayed the onset of tonic convulsion produced by bicuculline (30 mg/kg, i.p.), picrotoxin (20 mg/kg, i.p.) or NMDLA (400 mg/kg, i.p.)-induced tonic convulsion but did not affect the incidence of any of the convulsions. Phenobarbitone (12 mg/kg, i.p.), diazepam (0.5 mg/kg, i.p.) or muscimol (2 mg/kg, i.p.) significantly delayed the onset of bicuculline (30 mg/kg, i.p.) or picrotoxin (20 mg/kg, i.p.)-induced tonic convulsion and also significantly reduced the number of animals convulsing. Phenobarbitone (12 mg/kg, i.p.) or diazepam (0.5 mg/kg, i.p.) did affect significantly affect the onset or incidence of NMDLA (400 mg/kg, i.p.)-induced tonic convulsion. LY233053 (5 mg/kg, i.p.) significantly delayed the onset of tonic convulsion produced by NMDLA (400 mg/kg, i.p.) and also significantly reduced the number of animals convulsing. Leaf methanol extract of Tulbaghia violacea (200 and 400 mg/kg, i.p.) significantly delayed the onset of strychnine (2 mg/kg, i.p.)-induced tonic convulsion but did not significantly affect the number of mice convulsing. The dose of 100 mg/kg (i.p.) of leaf methanol extract of T. violacea did not significantly affect the onset or incidence of strychnine (2 mg/kg, i.p.)-induced tonic convulsion. Phenobarbitone (12 mg/kg, i.p.) significantly delayed the onset of strychnine (2 mg/kg, i.p.)-induced tonic convulsion and also significantly reduced the number of animals convulsing. Diazepam (0.5 mg/kg, i.p.) did not significantly delay the onset of strychnine (2 mg/kg, i.p.)-induced tonic convulsion and also did not significantly affect the number of mice convulsing. Phenytoin (30 mg/kg, i.p.) or DMSO (0.25 ml, i.p.) did not significantly affect the onset or incidence of bicuculline (30 mg/kg, i.p.), picrotoxin, strychnine or NMDLA-induced tonic convulsion. The qualitative phytochemical analysis of the plant species showed the presence of alkaloids, saponins, cardiac glycosides, flavonoids, triterpene steroids, quinones and tannins. The LD50 value obtained following oral administration of the leaf methanol extract of Tulbaghia violacea may be greater than 4000 mg/kg. The HPLC fingerprint of the leaf methanol extract of Tulbaghia violacea showed distinct peaks at the following retention times, 2.911, 3.269, 4.010, 7.597, and 15.122 min. The results obtained in this study indicate that the leaf methanol extract of Tulbaghia violacea has anticonvulsant activity. The results obtained also indicate that GABA, glutamic acid and glycine mechanisms may probably be involved in the anticonvulsant activity of the plant extract. The relatively high LD50 obtained for the plant species, given orally, indicate that it is safe in mice.

Envelope proteins of the human immunodefiency virus (HIV) use the cell surface CD-4 molecule of target cells to initiate infection which eventually lead to the acquired immunodeficiency syndrome (AIDS). HIV-1 strains form three groups, namely the M, N and O, with the former group further divided into at least ten equidistant subtypes or clades (i.e. A through J) classified on the basis of sequence homologies in the envelope gene. Recombinant envelope proteins expressed in transfected Chinese hamster ovary (CHO) cells were isolated and purified here (~ 0.01 mg yield). An economical but efficient purification procedure using affinity chromatography and freeze-drying was developed. The results obtained through SDS-PAGE, western blotting, specific ELISA (using Galanthus nivalis a lectin with affinity for ENV glycoproteins) and partial sequencing confirmed the purity (~ 85 - 90 %) and identity of the proteins. Since these proteins were derived in a clade A (Uganda) and B (USA) environment we anticipated limited crossreactivity with immune responses induced in a subtype C (RSA) environment. This was assessed using ELISA (titers of 1000) and western blot analysis. The ability to induce apoptosis was used to demonstrate functionality of the purified protein (Results showed that in-vitro induction of apoptosis (65 %) using the continuous cell line PM1 was achieved). / Dr. Debra Meyer

M.Sc. / The continued use and popularity of plant based traditional medicine necessitates scientific validation of the therapeutic potential of medicinal plants through phytochemical and pharmacological screening as well as the isolation and identification of bioactive compounds. Since the pharmacological effectiveness of medicinal plants is affected by several intrinsic and extrinsic factors, studies on the variations on chemical composition and biological activity are important as well. These provide a scientific rationale for using plants from different regions for the same medicinal purposes and allow traditional healers and consumers to make informed decisions with regard to the collection and use. Rapanea melanophloeos is a popular medicinal plant from the Myrsinaceae family widely distributed in southern Africa. It‟s bark, fruits and rarely the leaves are used traditionally for ailments ranging from stomach disorders, respiratory problems to disorders of the nervous system. Available chemical information reports on the accumulation of benzoquinones as major compounds, as well as some triterpenoid saponins and tannins. In view of the plant‟s wide distribution and medicinal use of different plant parts, this study comparatively evaluated the chemical composition of various crude extracts of the leaves, fruits and bark of plants collected from six localities. This was coupled with antibacterial tests to evaluate the therapeutic potential of different solvent extracts of the leaves, fruits and bark as well as the isolation of bioactive compounds from the fruits. Plants were collected from six different localities between Swaziland and South Africa and sequentially extracted with petroleum ether, ethyl acetate, methanol and water as a series of increasingly polar solvents. Thin layer chromatography (TLC), Gas-Chromatography/Mass spectrometry (GC-MS) and High Performance Liquid Chromatography (HPLC) were used to obtain a semi-quantitative chemical composition profile of different extracts. The TLC fingerprints of petroleum ether, ethyl acetate and methanol extracts showed the accumulation of non-polar terpenes, benzoquinones, saponins, tannins and flavonoids in the three plant parts. Slight variations in the chemical composition of the leaves, bark, and fruits were noted. Some components occurred in specific plant parts and others occurred in all three plants. The chemical profile of the leaves and bark were quite comparable in most instances with the fruits showing a generally different profile. This confirms previous literature reports on the comparability of leaves and the bark accounting for interchangeable use in traditional medicine.

M.Sc. (Chemistry) / The storage as solid Ag12BI(s) of reactor produced 12BI (t~ = 1.6 x 107 a) in underground respositories can present an environmental problem due to the potential release of radioactive 12BI into the environment over extended storage periods. One process that causes concern is the solubilization of AgI through complexation when brought into contact with underground water, particularly those with high salt content. In this work the charaqterization of the complexation equilibria in the AgI(s)/I-(aq),CI-(aq) system was studied. Solu- I tions containing high CI- and low 1- concentrations were of special interest with regard to the possible formation of ternary complexes. The presence of AgICI22- was indicated experimentally but no evidence of other ternary species like AgI2CI2- or AgIsCIS- could be established. Solubilities were determined radiochemically by using specially prepared 110mAgI. Solubility curves for AgI in different I-/CI- mixtures were adequately explained in terms of models consisting of the species AgCls2-, AgC14S-, AgIs2-, AgI4s- and AgICI22-. Distribution curves were calculated for different I-/CI- mixtures. As an alternative detection mode for silver-determination a flow injection system incorporating a micro-column of activated alumina was used in conjuntion with flame atomic absorption spectrometry for the pre-concentration and determination of silver in water. The procedure was successfully applied to a range of water samples, but no determination of silver in halide or perchlorate media was possible. The limit of detection based on a sample volume of 15 cms was 4.5 ~gdm-3 and the relative standard deviations at 50 and 5 ~gdm-3 were 5.4 and 19%, respectively. In part B of this work a method for the determination of chloride in organic compounds (oils provided by the Atomic Energy Corporation) by using an Parr oxygen bomb and ion chromatography was studied. The combustion products were absorbed in H20. The solution was injected into an ion chromatograph equipped with an anion analytical column, membrane suppressor and conductivity detector. The instrument response was calibrated using different standard solutions. These were prepared from several organic solutions, an organic salt and from sodiumchloride. Calibration curves were linear over a wide range and presented good reproducibility. It differed however according to the type of standard solution used. The limit of detection was 255 ~gdm-3 and the relative standard deviations at 10, 1 and 0.5 ugcm-3 were 0.52, 1.64 and 3.5%, respectively. As an alternative decomposition method wet decomposition within a sealed system (autoclave) was used. Due to the high blank values in this method no analysis of substances could be made in the lower ugcm-3 range.

Ph.D. / The main objective of this study was to investigate the presence, variations and the fate of lipophilic extractives during all the stages in the dissolving pulp production process. Lipophilic wood extractives which include fatty acids, sterols, hydrocarbons, steroid hydrocarbons and ketones are organic compounds present in the wood and dissolving pulp which are soluble in organic solvents. Their presence in wood and pulp has negative effects on both the pulping process and the quality of the produced dissolving pulp. Comparison of two Eucalyptus species (E. grandis and E. dunnii) at three different sites (Richmond, Umvoti and Ixopo in KwaZulu-Natal Province, South Africa) revealed that lipophilic extractives were higher in E. dunnii than in E. grandis in all the sampled sites and that the woods at the Ixopo site were found to have the highest amount of lipophilic extractives. The quantitative variation of lipophilic extractives in wood was found to be significantly affected by respective sites and tree species. The principal component analysis (PCA) model, which was used to correlate soil parameters and amount of lipophilic extractives in woods per each site, revealed that high amounts of lipophilic extractives were found in trees grown at sites with a high composition of clay soil and organic matter. Thus if the industry is to avoid the high levels of lipophilic extractives in wood, sandy sites should be the choice to grow the trees. Partial least squares (PLS) modelling revealed that sterols and saturated fatty acids significantly influence the quality parameters (Kappa number, viscosity, copper number, cellulose yield and retained carbohydrates) of dissolving pulp, whereas unsaturated fatty acids and unsaturated sterols in particular were found to positively correlate with the Kappa number, thus affecting its measurement.The main objective of this study was to investigate the presence, variations and the fate of lipophilic extractives during all the stages in the dissolving pulp production process. Lipophilic wood extractives which include fatty acids, sterols, hydrocarbons, steroid hydrocarbons and ketones are organic compounds present in the wood and dissolving pulp which are soluble in organic solvents. Their presence in wood and pulp has negative effects on both the pulping process and the quality of the produced dissolving pulp. Comparison of two Eucalyptus species (E. grandis and E. dunnii) at three different sites (Richmond, Umvoti and Ixopo in KwaZulu-Natal Province, South Africa) revealed that lipophilic extractives were higher in E. dunnii than in E. grandis in all the sampled sites and that the woods at the Ixopo site were found to have the highest amount of lipophilic extractives. The quantitative variation of lipophilic extractives in wood was found to be significantly affected by respective sites and tree species. The principal component analysis (PCA) model, which was used to correlate soil parameters and amount of lipophilic extractives in woods per each site, revealed that high amounts of lipophilic extractives were found in trees grown at sites with a high composition of clay soil and organic matter. Thus if the industry is to avoid the high levels of lipophilic extractives in wood, sandy sites should be the choice to grow the trees. Partial least squares (PLS) modelling revealed that sterols and saturated fatty acids significantly influence the quality parameters (Kappa number, viscosity, copper number, cellulose yield and retained carbohydrates) of dissolving pulp, whereas unsaturated fatty acids and unsaturated sterols in particular were found to positively correlate with the Kappa number, thus affecting its measurement. Viscosity of pulp which is an important quality parameter was also found to be positively influenced by sterol residues, whereas the reducing properties of pulp were found to be affected by sterols and saturated fatty acids with a carbon chain length of less than C20.

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A thesis submitted to the Faculty of Science, University of the Witwatersrand, Johannesburg, in fulfilment of the requirements for the degree of Doctor of Philosophy / In this dissertation the physical and chemical alterations induced by mastication and manipulation of wax by the worker bee in honeycomb construction, and the subsequent contribution afforded the structural integrity of the nest, are elucidated. In comb building, the freshly secreted wax scales are mandibulated together with a frothy salivary emulsion, and added piece-meal to form honeycomb. Textural modifications were revealed using X-ray crystallography. While virgin scale wax is highly structured, with the crystallites aligned approximately perpendicular to the planar surface, comb wax has a random crystallographic arrangement. This reflects a disruption of the crystallite structure following the mechanical insult of mastication. Chemical analyses included investigation of both lipid and proteinaceous elements. Lipid composition was evaluated by enzyme-catalyzed as well as thin-layer and gas-liquid chromatographic methods. The results indicate a reduction in scale diacylglycerols with a corresponding increase in comb saturated monoaeylglycerols. Such modifications are highly suggestive of lipase activity within the salivary addition. The proteins of comb and scale wax were analyzed electrophoretically, under reduced conditions. Each wax possesses unique polypeptide fractions, in addition to sharing common protein species, It is speculated that those in common represent integral proteins, such as transport molecules, while the disparities noted may be due to salivary enzymatic degradation, or even glycosylation. The effects of these textural and chemical alterations on the mechanical behaviour of the waxes was assessed. Tensile tests were performed on a variety of scale and comb wax preparations over the range of temperatures likely to impinge on the honeybee nest. These investigations reveal the specific structural contributions made by each of the physico-chemical alterations described. Further, they demonstrate that while scales are ideal moulding materials due to their high distensibility and low stiffness, the greater resistance to deformation and lower potential for extension makes comb wax a superior structural material. The mechanical advantage for including propolis and cocoon silk within the comb structure was also investigated. Tensile testing indicates that the resultant composite material is structurally superior, largely due to the presence of silk reinforcement. / Andrew Chakane 2018