Thesis (M.S.)--University of North Carolina at Wilmington, 2006. / Includes bibliographical references (leaf: 26)

This work is an account of the phytochemical analysis of two genera, Vernonia and Vepris which are used as remedies for illness by the Kalenjin community of Kenya. Species of Vernonia are known to yield sesquiterpene lactones, which typify the genus whereas Vepris is rich in alkaloids and limonoids which have a wide range of biological activities. The species studied in this work were Vernonia auriculifera, Vernonia urticifolia, Vepris glomerata and Vepris uguenensis. Phytochemical studies revealed a range of compounds being present in the four species. From Vernonia, triterpenoids, a sesquiterpene amine, a carotenoid and a polyene were isolated. This was the first account of a sesquiterpene amine from a plant species and the first account of the novel polyene. The triterpenoids showed moderate antibacterial activity, with b-amyrin acetate and oleanolic acid being effective at decreasing adhesion of selected gram-negative and gram-positive bacteria. Lutein and urticifolene showed good antibacterial activity against Enterococcus feacium and Pseudomonas aeruginosa. In Vepris, a range of compounds were isolated, belonging to the furoquinoline alkaloids, coumarins, flavonoids, cinnamic acid derivatives, lignins, cinnamaldehydes, triterpenoids and limonoids. Five new compounds; a cinnamaldehyde derivative (glomeral), two flavonoids (veprisinol, uguenenprenol) and two A, D-seco-limonoids (uguenensene and uguenensone) were amongst the compounds isolated. Antibacterial studies showed that glomeral inhibited the growth of Staphylococcus aureus and Shigella dysentrieae at low concentrations (MIC of 2 μg mLˉ¹ and 0.4 μg mLˉ¹ respectively). Antioxidant assays of several compounds revealed that, veprisinol, isohaplopine-3,3’-dimethylallyl ether, uguenenprenol and 7-O-methylaromadenrin are good antioxidant agents. The limonoids isolated from Vepris uguenensis also make up an interesting biogenetic relationship. Structural elucidation was carried out by 1D and 2D NMR spectroscopy in conjuction with mass spectrometry, infrared, ultraviolet and circular dichroism analysis where applicable. Biological assays were carried out using standard methods at laboratories in the University of KwaZulu-Natal and Kenya Medical Research Institute (KEMRI-Nairobi). / Thesis (Ph.D.)-University of KwaZulu-Natal, Westville, 2012.

Thesis (M.S.)--University of North Carolina at Wilmington, 2006. / Includes bibliographical references (leaves: 71-75)

Thesis (doctoral)--Swedish University of Agricultural Sciences, 2003. / Appendix consists of reprints and manuscripts of five papers co-authored with others. Includes bibliographical references. Also partially available online in PDF format; online version lacks appendix.

M.Sc. / Plants have the ability to continuously respond to various stimuli which alter their physiology, morphology and development. These stimuli may be abiotic or biotic and range from essential to toxic in their effects. One of these stimuli is a steroid from fungal membranes, ergosterol (C28H44O), which does not occur in plants. Ergosterol acts as a pathogen-associated molecular pattern molecule and triggers defence mechanisms in plants, characterised by highly regulated and interrelated events that include the elicitation of the oxidative burst and expression of a number of defencerelated genes. However, the ergosterol-induced global cellular reprogramming of the host has not been fully investigated in all aspects. No metabolomic study has previously been conducted to elucidate, for instance, the effect of ergosterol on plant metabolism. A clear and broader understanding of the molecular mechanisms involved in plant : ergosterol interactions is of paramount importance, for it would open up possibilities of developing novel, more effective and sustainable strategies to control or eradicate fungal diseases in plants. In plants, the metabolome is a compilation of all primary and secondary metabolites. The latter are the final recipients of genetic information, and their levels can influence gene expression and protein stability. Metabolite patterns reveal the actual cellular dynamic environment. Hence, qualitative and quantitative measurements of extra- and intracellular metabolites yield insights into the cellular processes that control the biochemical phenotype of the cell, tissue or whole organism. Metabolomics, the most recent of the ‘omics’ approaches, is the holistic analysis of metabolites present within a biological system under specific physiological conditions. In the present study a metabolomic approach was used to elucidate and analyse changes in the metabolism of tobacco (Nicotiana tabacum) cells following ergosterol treatment. Special attention is given to sesquiterpenoids since the antimicrobial compounds (phytoalexins) isolated from plants within the Solanaceae are mostly bicyclic sesquiterpenoids. Suspension of tobacco cells were treated with different concentrations (0 - 1000 nM) of ergosterol and incubated for different time periods (0 - 24 h). A viability assay, based on the ability of viable cells to reduce 2,3,5- triphenyltetrazolium chloride (TTC), was used to determine whether cell death occurred due to ergosterol treatment. No loss of cell viability was observed over the concentration range and time periods used in this study, indicating that the observed responses were due to the treatment alone and possible secondary responses due to cell death could be excluded. Intracellular metabolites were extracted with two methods: a selective dispersive liquid-liquid micro extraction and a general methanol extraction. Chromatographic techniques (TLC/HPTLC, GC-FID, GC-MS, GC×GC-TOF-MS, UPLC-MS) and 1H NMR spectroscopy were used for quantitative and qualitative analyses. Multivariate data analyses (PCA and OPLS-DA models) were used to extract interpretable information from the multidimensional data generated from the aforementioned techniques.

No description available.

published_or_final_version / Pharmacology and Pharmacy / Master / Master of Medical Sciences

No description available.

No description available.

Investigations are described which were carried out to analyse the way in which certain mineral element deficiencies restrict the growth and development of plants. The plant system used in this work was excised pea roots grown in sterile culture media, and the deficiencies studied were those of iron, magnesium and molybdenum. Growth was measured at the cell level and related to other characteristics of the system; two different experimental designs being employed to assess the effects of deficiencies. In the first, roots were grown in full nutrient and in deficient media and growth was measured on samples taken after growing periods of 0, 3, 5, 7, 9 and 11 days; while in the second, roots were grown for 7 days in full nutrient and in deficient media and serial one centimetre sections taken from these roots were compared. The first approach assessed the effects of the deficiency on overall growth, and the second gave an indication of the effects of the deficiency on the longitudinal differentiation of pea roots. Both experimental approaches were employed when examining iron and magnesium deficient roots, but only the second when examining molybdenum deficient roots. Roots deficient in iron and magnesium were obtained by culturing tips cut from germinated seeds in deficient media, but two successive tip passages were necessary to obtain roots deficient in molybdenum. Growth was assessed basically in terms of length, cell volume, protein nitrogen, and rate of oxygen uptake. However with iron deficient roots measurements of invertase activity, sensitivity of the oxygen uptake to cyanide, and the frequencies of cells in the different stages of division were also made. The techniques involved in the culture of deficient and full nutrient roots, and the analytical techniques are described. It has been shown that iron deficiency markedly affects the growth and development of excised pea roots. Growth in terms of length and cell number per root is stopped after 7 days and no further increases occur between days 7 and 11. Although iron deficiency stops cell division, measurements made at day 7 indicate that this deficiency does not restrict the process of cell expansion. In fact 7 day old iron deficient roots carry larger cells in the terminal centimetre than full nutrient roots. By 11 days the iron deficient roots have a pronounced swelling at the terminal end, and it is suggested that this is brought about by an abnormal expansion of the cells in the lateral direction. Some cells containing mitotic figures are present in the tips of 7 and 11 day old iron deficient roots. However there are fewer cells in the division stages of prophase and metaphase and practically no cells in the stages of telophase and anaphase in the deficient roots when comparisons were made with full nutrient roots. The protein nitrogen content of iron deficient roots is lower than that of full nutrient roots at day 7, but there is a considerable increase in both deficient and full nutrient roots between days 7 and 11. The trend of the derived quantity, average protein nitrogen content per cell, is the same in both groups of roots up to day 7, but from day 7 to day 11 it increases sharply in the deficient roots but does not change in the full nutrient roots. This result indicates that cell division was not stopped in the deficient roots by a shortage of protein nitrogen as such. At the day 7 stage the distribution of protein nitrogen along the length of deficient roots is different to that in full nutrient roots. The front sections of the deficient roots contain an increased content and the back sections a decreased content when compared with full nutrient roots. On a per cell basis the situation is the same, as the cells in the front sections of deficient roots have a higher average protein content and those in the back sections a lower content when compared with the cells of full nutrient roots. The accumulation of protein nitrogen in the front sections of iron deficient roots is most probably associated with the cessation of active cell division in the meristem. Evidence is available which suggests that under normal conditions the formation and development of cells in the apex of the root is dependent on substrates synthesised in the mature regions of the root and translocated forward. It is considered that in iron deficient roots precursors of protein are no longer removed by the demands of the meristem and they condense to form protein in the regions adjacent to the apex. Invertase activity per unit protein nitrogen is the same in both full nutrient and iron deficient roots at all stages. Further, there is no difference in invertase activity when the corresponding sections of full nutrient and deficient roots are compared at day 7. It is clear that in this one respect the protein of iron deficient roots is similar to that of full nutrient roots. The rate of oxygen intake per root of iron deficient roots is lower than that of full nutrient roots at the early day 3 stage, but there are large increases in the rates in both deficient and full nutrient roots between days 3 and 11. It is of some significance that iron deficiency clearly reduces the rate of oxygen uptake at a stage before the process of cell division is stopped. On a per unit protein nitrogen basis the rate of oxygen uptake of deficient roots is lower than that of full nutrient roots after day 3. It is suggested that the effects of days 3 and 5 are a direct effect of iron deficiency but the effects at days 9 and 11 are influenced by the fact that cell division stops at day 7. The results from 7 day roots show that the effect of iron deficiency in reducing the rate of oxygen intake per unit protein nitrogen is confined to the front three sections of the root as iron deficiency does not alter the rates in the back three sections. Iron recovery experiments show that iron deficient roots 7, 9 and 11 days old can resume cell division and grow when they are transferred to a full nutrient medium. It is of interest that in these experiments the recovery in terms of an increased rate of oxygen uptake is greater than the recovery in terms of length and protein nitrogen. Experiments in which the rate of oxygen uptake of deficient and full nutrient roots were measured in the presence and absence of cyanide show that in both groups of roots there is a large fraction of the respiration insensitive to cyanide. The activity of this cyanide insensitive system increases considerably from day 3 to day 11 in both the full nutrient and iron deficient roots. Increases, after day 3 in the activity of this cyanide insensitive system, which would not contain iron, account for the large increase in the total rate of oxygen uptake of iron deficient roots between days 3 and 11. The activity of the cyanide sensitive system involved in respiration decreases in both groups of roots between days 0 and 5. It increases from day 5 to 11 in full nutrient roots, but does not increase in deficient roots over this period. That synthesis of a cyanide sensitive system involved in respiration stops at or about the same stage as cell division in iron deficient roots is considered to be highly important. This cyanide sensitive system most probably corresponds to the iron containing cytochrome/cytochrome oxidase system, and there is other circumstantial evidence that this system is important in the process of cell division. It is important to note that the activity of the cyanide sensitive system was the same in the tips of deficient and full nutrient roots at the day 7 stage. It may be that a certain minimum level of activity per cell is necessary to maintain division; a slight reduction stopping cell division completely, but not being capable of detection by the method of measurement.