Cocrystals, salts and eutectics of anti-tuberculosis medicines
- Authors: Matlapeng, Tsebang Alice
- Date: 2024-10-11
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/464521 , vital:76518
- Description: Tuberculosis remains as a prominent cause of death worldwide. This infectious disease is treated with first and second line drugs. However, challenges of multi drug resistant tuberculosis and adverse side effects such as depletion of essential B group vitamins in the body by first line drugs, as well as poor physicochemical properties of second line drugs persist. Cocrystallisation of anti-tubercular drugs with various coformers has therefore been used as an alternative method to improve the physicochemical properties of active pharmaceutical ingredients (API) while maintaining their efficacy. The main objective of this study was to carry out cocrystal screening of anti-tubercular API and vitamin B coformers to make drug-drug or drug-vitamin multicomponent complexes. Preparation of the multicomponent complexes was carried out by mechanochemical grinding (neat grinding (NG), liquid assisted grinding (LAG) and slow evaporation. All complexes were characterised using Fourier-transform infrared spectroscopy (FTIR), powder X-ray diffraction (PXRD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) and crystal structures were elucidated using single-crystal X-ray diffraction (SCXRD). The cocrystal screening resulted in the formation of various solid forms, which included cocrystals, salts and eutectic products. Two cocrystals of 4-aminosalicylic acid (PAS) were isolated and characterised. The cocrystal of PAS with isoniazid (INH) demonstrated similar characteristics for both the bulk crystalline material and milled materials. The cocrystal of PAS and pyrazinamide (PYR) prepared using mechanochemical synthesis was met with challenges, while difficulties were also encountered in obtaining suitable crystals for SCXRD analysis. The ground and recrystallised samples of the PAS and pyrazinecarboxylic acid (PCBA) showed distinct differences in their thermal behaviour, with SCXRD revealing the decomposition product phenolammonium-pyrazinecarboxylate salt ‘sans’ the CO2 moiety. Salt formation involving pyridoxine (PN) yielded a salt hydrate with PAS (PN-PAS) and an anhydrous salt with PCBA (PN-PCBA). Both salts exhibited very complex packing arrangements with equally complex thermal behaviour depending on the solvent used during preparation, and the method of preparation. Three eutectic systems involving INH with PYR, PN and pyridoxine hydrochloride (PNꞏHCl) were identified, and their phase diagrams were constructed from DSC data. The eutectic compositions obtained were 1:1 for INH:PYR, 1:1 for INH:PN and 6:4 for INH:PNꞏHCl. Finally, a total of eight multicomponent complexes were prepared using selected API and vitamin B6 components. The results presented here provide motivation for further investigation and evaluation of the pharmacochemical properties of these API. , Thesis (MSc) -- Faculty of Science, Chemistry, 2024
- Full Text:
- Authors: Matlapeng, Tsebang Alice
- Date: 2024-10-11
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/464521 , vital:76518
- Description: Tuberculosis remains as a prominent cause of death worldwide. This infectious disease is treated with first and second line drugs. However, challenges of multi drug resistant tuberculosis and adverse side effects such as depletion of essential B group vitamins in the body by first line drugs, as well as poor physicochemical properties of second line drugs persist. Cocrystallisation of anti-tubercular drugs with various coformers has therefore been used as an alternative method to improve the physicochemical properties of active pharmaceutical ingredients (API) while maintaining their efficacy. The main objective of this study was to carry out cocrystal screening of anti-tubercular API and vitamin B coformers to make drug-drug or drug-vitamin multicomponent complexes. Preparation of the multicomponent complexes was carried out by mechanochemical grinding (neat grinding (NG), liquid assisted grinding (LAG) and slow evaporation. All complexes were characterised using Fourier-transform infrared spectroscopy (FTIR), powder X-ray diffraction (PXRD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) and crystal structures were elucidated using single-crystal X-ray diffraction (SCXRD). The cocrystal screening resulted in the formation of various solid forms, which included cocrystals, salts and eutectic products. Two cocrystals of 4-aminosalicylic acid (PAS) were isolated and characterised. The cocrystal of PAS with isoniazid (INH) demonstrated similar characteristics for both the bulk crystalline material and milled materials. The cocrystal of PAS and pyrazinamide (PYR) prepared using mechanochemical synthesis was met with challenges, while difficulties were also encountered in obtaining suitable crystals for SCXRD analysis. The ground and recrystallised samples of the PAS and pyrazinecarboxylic acid (PCBA) showed distinct differences in their thermal behaviour, with SCXRD revealing the decomposition product phenolammonium-pyrazinecarboxylate salt ‘sans’ the CO2 moiety. Salt formation involving pyridoxine (PN) yielded a salt hydrate with PAS (PN-PAS) and an anhydrous salt with PCBA (PN-PCBA). Both salts exhibited very complex packing arrangements with equally complex thermal behaviour depending on the solvent used during preparation, and the method of preparation. Three eutectic systems involving INH with PYR, PN and pyridoxine hydrochloride (PNꞏHCl) were identified, and their phase diagrams were constructed from DSC data. The eutectic compositions obtained were 1:1 for INH:PYR, 1:1 for INH:PN and 6:4 for INH:PNꞏHCl. Finally, a total of eight multicomponent complexes were prepared using selected API and vitamin B6 components. The results presented here provide motivation for further investigation and evaluation of the pharmacochemical properties of these API. , Thesis (MSc) -- Faculty of Science, Chemistry, 2024
- Full Text:
Towards a possible future solution against Multidrug Resistance: An in silico exploration of the Multidrug and Toxic compound Extrusion (MATE) transporter proteins as potential antimicrobial drug targets
- Authors: Damji, Amira Mahamood
- Date: 2024-04-04
- Subjects: Multidrug resistance , Multidrug and toxic compound extrusion family, eukaryotic , Docking , Molecular dynamics , Drug development , Transmembrane protein
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/435009 , vital:73123
- Description: The rise of multidrug resistance (MDR) has become a pressing global issue, hindering the treatment of cancers and infectious diseases, and imposing a burden on healthcare systems and the economy. The Multidrug and Toxic compound Extrusion (MATE) superfamily of membrane efflux transporters is one of the key players contributing to MDR due to their ability to export a wide range of cationic and hydrophilic xenobiotics, including treatment drugs, from cells, diminishing their efficacy. Targeting MATE transporters holds great promise in achieving some cellular control over MDR, but first, a deeper understanding of their structure-function-dynamics link is required. This study aimed to explore the MATE transporters as potential antimicrobial drug targets using a two-fold in silico approach. First, virtual screening of compounds from the South African Natural Compounds Database (SANCDB) was performed to identify prospective lead inhibitory compounds against the MATE transporters using molecular docking, and top hits were selected based on their binding energy and interaction with the active site on the N-lobe of the protein. Second, to investigate the molecular-level dynamics of their extrusion mechanism, the MATE transporter structures were embedded in a POPC membrane bilayer using the CHARMM-GUI online tool and then subjected to MD simulations for 100 ns with the CHARMM 36m force field using GROMACS. The resulting trajectories were evaluated using three standard metrics – RMSD, RMSF, and Rg; significant global structural changes were observed and key functional regions in both membrane- and non-membrane transmembrane (TM) segments were identified, containing more dynamic and flexible residues than other regions. Furthermore, the MATE transporters showed more of a loosely-packed structure, providing flexibility to allow for conformational switching during their substrate-transport cycle, which is typical for proteins whose secondary structures are composed of all α-helices. The scope of this study lied in the preliminary stages of the computer-aided drug design process, and provided insights that can be used to guide the development of strategies aimed at regulating or inhibiting the function of the MATE transporters, offering a possible future solution to the growing challenge of MDR. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2024
- Full Text:
- Authors: Damji, Amira Mahamood
- Date: 2024-04-04
- Subjects: Multidrug resistance , Multidrug and toxic compound extrusion family, eukaryotic , Docking , Molecular dynamics , Drug development , Transmembrane protein
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/435009 , vital:73123
- Description: The rise of multidrug resistance (MDR) has become a pressing global issue, hindering the treatment of cancers and infectious diseases, and imposing a burden on healthcare systems and the economy. The Multidrug and Toxic compound Extrusion (MATE) superfamily of membrane efflux transporters is one of the key players contributing to MDR due to their ability to export a wide range of cationic and hydrophilic xenobiotics, including treatment drugs, from cells, diminishing their efficacy. Targeting MATE transporters holds great promise in achieving some cellular control over MDR, but first, a deeper understanding of their structure-function-dynamics link is required. This study aimed to explore the MATE transporters as potential antimicrobial drug targets using a two-fold in silico approach. First, virtual screening of compounds from the South African Natural Compounds Database (SANCDB) was performed to identify prospective lead inhibitory compounds against the MATE transporters using molecular docking, and top hits were selected based on their binding energy and interaction with the active site on the N-lobe of the protein. Second, to investigate the molecular-level dynamics of their extrusion mechanism, the MATE transporter structures were embedded in a POPC membrane bilayer using the CHARMM-GUI online tool and then subjected to MD simulations for 100 ns with the CHARMM 36m force field using GROMACS. The resulting trajectories were evaluated using three standard metrics – RMSD, RMSF, and Rg; significant global structural changes were observed and key functional regions in both membrane- and non-membrane transmembrane (TM) segments were identified, containing more dynamic and flexible residues than other regions. Furthermore, the MATE transporters showed more of a loosely-packed structure, providing flexibility to allow for conformational switching during their substrate-transport cycle, which is typical for proteins whose secondary structures are composed of all α-helices. The scope of this study lied in the preliminary stages of the computer-aided drug design process, and provided insights that can be used to guide the development of strategies aimed at regulating or inhibiting the function of the MATE transporters, offering a possible future solution to the growing challenge of MDR. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2024
- Full Text:
Analysis of corporate failures: a case study of two South African banks
- Mqomboti, Xitshembiso Pronacia
- Authors: Mqomboti, Xitshembiso Pronacia
- Date: 2023-02
- Subjects: Business failures South Africa , Corporate governance South Africa , Risk management , Operational risk , Business ethics , Bank management South Africa , Banks and banking South Africa
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/419151 , vital:71620
- Description: This study analysed the factors that contributed to the failure of Venda Building Society Mutual Bank (VBS) and African Bank Limited and the impact it had on their key stakeholders. The specific objectives of this study were to evaluate African Bank and VBS bank's operational risk management processes and controls, the role of ethical failures at VBS bank and African Bank; and assess how the failures affected their stakeholders. The population sample of the study included African Bank and VBS. The study adopted a qualitative research method. Existing reports from both African Bank and VBS were used to collect data. The study adopted a thematic data analysis method, which includes data coding and the development of themes. The data analysis framework was derived from a defined set of research propositions and seven (7) themes were derived from this analysis method. The failure in operational controls of both banks and ineffective risk management structures including unethical conduct by the executive management and board of VBS bank, irregular financial transactions and weakened external auditing function resulted in an unaccountable executive relationship and reckless lending decision-making. This research study will expand on the existing body of knowledge on the failures and near-failures of banks in the South African banking sector. The South African banking industry and its regulatory bodies will be better equipped to strengthen their corporate governance in risk controls to mitigate future collapses and near collapses of banks. , Thesis (MBA) -- Faculty of Commerce, Rhodes Business School, 2023
- Full Text:
- Authors: Mqomboti, Xitshembiso Pronacia
- Date: 2023-02
- Subjects: Business failures South Africa , Corporate governance South Africa , Risk management , Operational risk , Business ethics , Bank management South Africa , Banks and banking South Africa
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/419151 , vital:71620
- Description: This study analysed the factors that contributed to the failure of Venda Building Society Mutual Bank (VBS) and African Bank Limited and the impact it had on their key stakeholders. The specific objectives of this study were to evaluate African Bank and VBS bank's operational risk management processes and controls, the role of ethical failures at VBS bank and African Bank; and assess how the failures affected their stakeholders. The population sample of the study included African Bank and VBS. The study adopted a qualitative research method. Existing reports from both African Bank and VBS were used to collect data. The study adopted a thematic data analysis method, which includes data coding and the development of themes. The data analysis framework was derived from a defined set of research propositions and seven (7) themes were derived from this analysis method. The failure in operational controls of both banks and ineffective risk management structures including unethical conduct by the executive management and board of VBS bank, irregular financial transactions and weakened external auditing function resulted in an unaccountable executive relationship and reckless lending decision-making. This research study will expand on the existing body of knowledge on the failures and near-failures of banks in the South African banking sector. The South African banking industry and its regulatory bodies will be better equipped to strengthen their corporate governance in risk controls to mitigate future collapses and near collapses of banks. , Thesis (MBA) -- Faculty of Commerce, Rhodes Business School, 2023
- Full Text:
Variables associated with run out opportunities in cricket: coaches’ perceptions versus video analyses of the Indian Premier League 2018/2019 seasons
- Authors: Sholto-Douglas, Robert
- Date: 2022-10-14
- Subjects: Indian Premier League , Cricket Coaching , Video analysis , Fielding , Run out
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/365014 , vital:65670
- Description: The purpose of this investigation was to add a scientific element to the understanding of run out opportunities in T20 (twenty/twenty) formats, which would in turn, provide recommendations to players and coaches around the topic of run outs. Due to a lack of research in this field, the study was two-fold, as there is little research known on this topic. Firstly, surveying experienced coaches on what they would expect to take place during run out opportunities from different areas of the field in a T20 game. Secondly, the study looked at what happened during successful and unsuccessful run outs from different areas of the field in two Indian Premier League (IPL) seasons. The aim of the study was to determine what variables, within an area of fielding, led to a successful or non-successful run out from different zones in the field. Coaches were required to fill out a survey which asked them to give their expectations on run out opportunities during a T20 competition. The data from the questionnaire would be used to gain a greater understanding of the way coaches understand different characteristics of run outs and whether this matched the findings of the second part of this study. The second part of the study investigated run out opportunities in two consecutive IPL seasons, based on video footage, looking at how different variables impacted successful or marginally missed run out opportunities in different areas of the field and at different times of the game. The areas were put into four different zones. Zone 1 being close to the batter, zone 2 being in the inner ring, zone 3 being on the edge of the ring and zone 4 being boundary fielders. Most of the coaches believed that in zone 1 of the field, the bowler would have the most opportunities, where in fact, based on the video footage, these fell to the wicket-keeper. In zone 2 there were a larger range of fielding positions (10) that run outs fell to when analysing video footage. The coaches supported this by often naming the same fielding positions. There was a divide in zone 3. Most run out opportunities from the video footage came to extra cover, where most coaches didn’t believe that would be the case and rather mentioned mid-off and mid-on. When analysing video footage in zone 4, it had two fielding positions (long on and deep mid-wicket) make up the majority of run out chances in this area and these two positions were quoted by most of the coaches as having the most run out opportunities fall to them. Throughout the findings, as concluded from the video footage, there were some results that matched the coaches’ expectation and some results where findings from the video footage differed from the coaches’ expectations. These findings are all useful. Firstly, it validates coaching practices. Secondly, it provides coaches with greater insight as to where they aren’t getting it correct in their fielding training. In conclusion, this study found that although coaches perceptions, for the most part, matched what took place on the field, there were instances where we saw the opposite action take place. This has important practical implications for coaching and practice. , Thesis (MSc) -- Faculty of Science, Human Kinetics and Ergonomics, 2022
- Full Text:
- Authors: Sholto-Douglas, Robert
- Date: 2022-10-14
- Subjects: Indian Premier League , Cricket Coaching , Video analysis , Fielding , Run out
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/365014 , vital:65670
- Description: The purpose of this investigation was to add a scientific element to the understanding of run out opportunities in T20 (twenty/twenty) formats, which would in turn, provide recommendations to players and coaches around the topic of run outs. Due to a lack of research in this field, the study was two-fold, as there is little research known on this topic. Firstly, surveying experienced coaches on what they would expect to take place during run out opportunities from different areas of the field in a T20 game. Secondly, the study looked at what happened during successful and unsuccessful run outs from different areas of the field in two Indian Premier League (IPL) seasons. The aim of the study was to determine what variables, within an area of fielding, led to a successful or non-successful run out from different zones in the field. Coaches were required to fill out a survey which asked them to give their expectations on run out opportunities during a T20 competition. The data from the questionnaire would be used to gain a greater understanding of the way coaches understand different characteristics of run outs and whether this matched the findings of the second part of this study. The second part of the study investigated run out opportunities in two consecutive IPL seasons, based on video footage, looking at how different variables impacted successful or marginally missed run out opportunities in different areas of the field and at different times of the game. The areas were put into four different zones. Zone 1 being close to the batter, zone 2 being in the inner ring, zone 3 being on the edge of the ring and zone 4 being boundary fielders. Most of the coaches believed that in zone 1 of the field, the bowler would have the most opportunities, where in fact, based on the video footage, these fell to the wicket-keeper. In zone 2 there were a larger range of fielding positions (10) that run outs fell to when analysing video footage. The coaches supported this by often naming the same fielding positions. There was a divide in zone 3. Most run out opportunities from the video footage came to extra cover, where most coaches didn’t believe that would be the case and rather mentioned mid-off and mid-on. When analysing video footage in zone 4, it had two fielding positions (long on and deep mid-wicket) make up the majority of run out chances in this area and these two positions were quoted by most of the coaches as having the most run out opportunities fall to them. Throughout the findings, as concluded from the video footage, there were some results that matched the coaches’ expectation and some results where findings from the video footage differed from the coaches’ expectations. These findings are all useful. Firstly, it validates coaching practices. Secondly, it provides coaches with greater insight as to where they aren’t getting it correct in their fielding training. In conclusion, this study found that although coaches perceptions, for the most part, matched what took place on the field, there were instances where we saw the opposite action take place. This has important practical implications for coaching and practice. , Thesis (MSc) -- Faculty of Science, Human Kinetics and Ergonomics, 2022
- Full Text:
Production, purification, and characterisation of proteases from an ericoid mycorrhizal fungus, Oidiodendron maius
- Authors: Manyumwa, Colleen Varaidzo
- Date: 2018
- Subjects: Ascomycetes , Mycorrhizal fungi , Ericaceae , Proteolytic enzymes , Silver Recycling
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/62833 , vital:28298
- Description: The aim of this study was to produce, purify and characterise proteases from the ericoid mycorrhizal fungus, Oidiodendron maius (CafRU082b/KP119480), as well as to explore their potential application in the recovery of silver from X-ray film. Firstly, the growth of the ericoid mycorrhizal fungus, Oidiodendron maius (CafRU082b), was studied, and its ability to produce proteolytic enzymes was investigated. O. maius proved to grow well in the dark, submerged in Modified Melin Norkran’s liquid medium at a pH of 5 and at 25°C. Pure cultures of the fungus were maintained on Potato Dextrose Agar (PDA). The fungus grew on PDA plates containing different substrates including haemoglobin, casein, gelatin as well as azocasein. Zones of clearance, however, were only observed on plates containing gelatin after treatment with mercuric chloride, HgCl2. Proteases were successfully produced after 14 days when gelatin was incorporated into the growth medium. After production of the proteases, purification and characterisation of the enzymes was performed. Purification of the enzymes was performed by acetone precipitation followed by ultrafiltration with 50 kDa and 30 kDa cut off membrane filters. A final purification fold of approximately 37.6 was achieved. Unusual yields of above 100% were observed after each purification step with the final yield achieved being 196% with a final specific activity of 2707 U/mg. SDS-PAGE revealed a protease band of 35 kDa which was also visible on the zymogram at approximately 36 kDa. The zymogram showed clear hydrolysis bands against a blue background after staining with Coomassie Brilliant Blue. Physico-chemical characterisation of the protease revealed its pH optimum to be pH 3.0 and its temperature optimum 68°C. Another peak was observed on the pH profile at pH 7.0. The protease exhibited high thermostability at temperatures 37°C, 80°C as well as 100°C with the enzyme retaining close to 50% of its initial activity after 4 h of exposure to all three temperatures. All ions tested for their effects on the proteases, except Ca2+, enhanced protease activity. Ca2+ did not exhibit any significant effect on the enzyme’s activity while Zn2+ had the highest effect, enhancing enzyme activity by 305%. The proteases, however, were not significantly inhibited by EDTA, a metal chelating agent and a known metalloprotease inhibitor. The enzyme was classified as an aspartic protease due to complete inhibition by 25 μM of pepstatin A, coupled to its low pH optimum of 3.0. Addition of trans-Epoxysuccinyl-L-leucylamido-(4-guanidino)butane (E-64), a cysteine protease inhibitor, and 2-mercaptoethanol increased protease activity. The proteases exhibited a narrow substrate specificity towards gelatin and no other substrate. Substrate kinetics values were plotted on a Michaelis-Menten Graph and showed that the enzyme had a Vmax of 55.25 U/ml and a Km of 2.7 mg/ml gelatin. A low Km indicated that the protease had a high affinity for gelatin. Silver recovery studies from X-ray film revealed the proteases’ capability to remove silver from X-ray film, leaving the film intact. The recovery of silver was perceived visually, by film observation, as well as by scan electron microscopy (SEM) images, where clearance of the film was observed after incubation with the enzyme. Energy dispersive X-ray spectroscopy (EDS) profiles also confirmed removal of silver from the film, with a Ag peak showing on the profile of the film before treatment with the proteases and no peak after treatment. The crude protease sample was, however, catalytically more efficient compared to the partially purified sample. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2018
- Full Text:
- Authors: Manyumwa, Colleen Varaidzo
- Date: 2018
- Subjects: Ascomycetes , Mycorrhizal fungi , Ericaceae , Proteolytic enzymes , Silver Recycling
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/62833 , vital:28298
- Description: The aim of this study was to produce, purify and characterise proteases from the ericoid mycorrhizal fungus, Oidiodendron maius (CafRU082b/KP119480), as well as to explore their potential application in the recovery of silver from X-ray film. Firstly, the growth of the ericoid mycorrhizal fungus, Oidiodendron maius (CafRU082b), was studied, and its ability to produce proteolytic enzymes was investigated. O. maius proved to grow well in the dark, submerged in Modified Melin Norkran’s liquid medium at a pH of 5 and at 25°C. Pure cultures of the fungus were maintained on Potato Dextrose Agar (PDA). The fungus grew on PDA plates containing different substrates including haemoglobin, casein, gelatin as well as azocasein. Zones of clearance, however, were only observed on plates containing gelatin after treatment with mercuric chloride, HgCl2. Proteases were successfully produced after 14 days when gelatin was incorporated into the growth medium. After production of the proteases, purification and characterisation of the enzymes was performed. Purification of the enzymes was performed by acetone precipitation followed by ultrafiltration with 50 kDa and 30 kDa cut off membrane filters. A final purification fold of approximately 37.6 was achieved. Unusual yields of above 100% were observed after each purification step with the final yield achieved being 196% with a final specific activity of 2707 U/mg. SDS-PAGE revealed a protease band of 35 kDa which was also visible on the zymogram at approximately 36 kDa. The zymogram showed clear hydrolysis bands against a blue background after staining with Coomassie Brilliant Blue. Physico-chemical characterisation of the protease revealed its pH optimum to be pH 3.0 and its temperature optimum 68°C. Another peak was observed on the pH profile at pH 7.0. The protease exhibited high thermostability at temperatures 37°C, 80°C as well as 100°C with the enzyme retaining close to 50% of its initial activity after 4 h of exposure to all three temperatures. All ions tested for their effects on the proteases, except Ca2+, enhanced protease activity. Ca2+ did not exhibit any significant effect on the enzyme’s activity while Zn2+ had the highest effect, enhancing enzyme activity by 305%. The proteases, however, were not significantly inhibited by EDTA, a metal chelating agent and a known metalloprotease inhibitor. The enzyme was classified as an aspartic protease due to complete inhibition by 25 μM of pepstatin A, coupled to its low pH optimum of 3.0. Addition of trans-Epoxysuccinyl-L-leucylamido-(4-guanidino)butane (E-64), a cysteine protease inhibitor, and 2-mercaptoethanol increased protease activity. The proteases exhibited a narrow substrate specificity towards gelatin and no other substrate. Substrate kinetics values were plotted on a Michaelis-Menten Graph and showed that the enzyme had a Vmax of 55.25 U/ml and a Km of 2.7 mg/ml gelatin. A low Km indicated that the protease had a high affinity for gelatin. Silver recovery studies from X-ray film revealed the proteases’ capability to remove silver from X-ray film, leaving the film intact. The recovery of silver was perceived visually, by film observation, as well as by scan electron microscopy (SEM) images, where clearance of the film was observed after incubation with the enzyme. Energy dispersive X-ray spectroscopy (EDS) profiles also confirmed removal of silver from the film, with a Ag peak showing on the profile of the film before treatment with the proteases and no peak after treatment. The crude protease sample was, however, catalytically more efficient compared to the partially purified sample. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2018
- Full Text:
Assessing the potential role of microorganisms in the production of seedlings for the restoration of Albany Thicket
- Authors: Mpama, Nelisa
- Date: 2017
- Subjects: Vesicular-arbuscular mycorrhizas , Rhizobacteria , Restoration ecology South Africa Albany , Microorganisms
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/64555 , vital:28558
- Description: The role of microorganisms in restoration of the Albany Thicket has not been well documented, although the benefits to plants of these various interactions has been well documented. Microorganisms are chief ecological engineers and assist in resolving environmental problems and act to restore degraded ecosystem function by forming mutual relationships with the roots of the plants. The aim of this study was to assess the potential of microorganisms for the improved biomass production of selected woody and succulent seedlings used in mesic thicket restoration. Three tree species were selected for propagation in this study namely; Mystroxylon aethiopicum Scutia myrtina and Aloe ferox. Soil samples were collected from a degraded and intact thicket site from Bathurst, South Africa. Soils were evaluated for number of arbuscular mycorrhizal (AM) spores, mycorrhizal infectivity potential and nutrient availability both before and after seedling propagation. Pasteurized soil from the degraded site was used in a pot trial. Ten replicates seedling for plant species were planted and subjected to four treatments which included inoculation with AM fungi and the rhizobacterium, Enterobacter sp., alone and in combination; the fourth treatment was an un-inouclated control. Plant growth parameters were recorded at regular intervals where appropriate and seedlings were harvested after 24 weeks for biomass measurements and AM colonisation assessments. Although generally low (< 1 spore per gram) the density of AM fungal spores was significantly higher in soils from the intact site when compared with soils from the degraded site. The mycorrhizal potential of the soils was however not significantly different. Mystroxylon aethiopicum seedling shoot height, canopy diameter and shoot biomass showed a significant increase when inoculated with AM fungi while S. myrtina seedlings showed increased shoot height when inoculated with both AM fungi and Enterobacter sp. Aloe ferox seedlings did not respond to microbial inoculation. The concentration of soil P and Na increased in treatments with Enterobacter sp. alone and in combination with AM fungi. Mystroxylon aethiopicum and S. myrtina seedlings showed a dependency on microbial inoculants indicating the importance of inoculation in the nursery before planting out into the field. Overall AM fungal inoculants applied to seedlings can be used to compensate for nutrient deficiency in soils. Although the Enterobacter isolate used was known to have various plant growth promoting capabilities. It is recommended that other rhizobacterial isolates be investigated. , Thesis (MSc) -- Faculty of Science, Environmental Science, 2017
- Full Text:
- Authors: Mpama, Nelisa
- Date: 2017
- Subjects: Vesicular-arbuscular mycorrhizas , Rhizobacteria , Restoration ecology South Africa Albany , Microorganisms
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/64555 , vital:28558
- Description: The role of microorganisms in restoration of the Albany Thicket has not been well documented, although the benefits to plants of these various interactions has been well documented. Microorganisms are chief ecological engineers and assist in resolving environmental problems and act to restore degraded ecosystem function by forming mutual relationships with the roots of the plants. The aim of this study was to assess the potential of microorganisms for the improved biomass production of selected woody and succulent seedlings used in mesic thicket restoration. Three tree species were selected for propagation in this study namely; Mystroxylon aethiopicum Scutia myrtina and Aloe ferox. Soil samples were collected from a degraded and intact thicket site from Bathurst, South Africa. Soils were evaluated for number of arbuscular mycorrhizal (AM) spores, mycorrhizal infectivity potential and nutrient availability both before and after seedling propagation. Pasteurized soil from the degraded site was used in a pot trial. Ten replicates seedling for plant species were planted and subjected to four treatments which included inoculation with AM fungi and the rhizobacterium, Enterobacter sp., alone and in combination; the fourth treatment was an un-inouclated control. Plant growth parameters were recorded at regular intervals where appropriate and seedlings were harvested after 24 weeks for biomass measurements and AM colonisation assessments. Although generally low (< 1 spore per gram) the density of AM fungal spores was significantly higher in soils from the intact site when compared with soils from the degraded site. The mycorrhizal potential of the soils was however not significantly different. Mystroxylon aethiopicum seedling shoot height, canopy diameter and shoot biomass showed a significant increase when inoculated with AM fungi while S. myrtina seedlings showed increased shoot height when inoculated with both AM fungi and Enterobacter sp. Aloe ferox seedlings did not respond to microbial inoculation. The concentration of soil P and Na increased in treatments with Enterobacter sp. alone and in combination with AM fungi. Mystroxylon aethiopicum and S. myrtina seedlings showed a dependency on microbial inoculants indicating the importance of inoculation in the nursery before planting out into the field. Overall AM fungal inoculants applied to seedlings can be used to compensate for nutrient deficiency in soils. Although the Enterobacter isolate used was known to have various plant growth promoting capabilities. It is recommended that other rhizobacterial isolates be investigated. , Thesis (MSc) -- Faculty of Science, Environmental Science, 2017
- Full Text:
Synthesis, characterisation and evaluation of benzoxaborole-based hybrids as antiplasmodial agents
- Authors: Gumbo, Maureen
- Date: 2017
- Subjects: Malaria Chemotherapy , Antimalarials , Boron compounds , Drug resistance , Plasmodium falciparum , Drug development
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/59193 , vital:27456
- Description: Malaria is a mosquito-borne disease, which continues to pose a threat to the entire humanity. About 40% of the world population is estimated to be at risk of infections by malaria. Despite efforts undertaken by scientific community, government entities and international organizations, malaria is still rampant. The major problem is drug resistance, where the Plasmodium spp have over the past decades developed drug resistance against available drugs. In order to counter this problem, novel antimalarial drugs that are efficacious and with novel mode of action are of great necessity. Benzoxaborole derivatives have been shown to exhibit promising antimalarial activity against Plasmodium falciparum strains. Previous studies reported on the compounds such as 6-(2- (alkoxycarbonyl)pyrazinyl-5-oxy)-1,3-dihydro-1-hydroxy-2,1-benzoxaboroles, which showed good antimalarial activity against both W7 and 3D7 strains without significant toxicity. On the other hand, chloroquine (CQ) and cinnamic acids have a wide variety of biological activity including antimalarial activity. Herein, a hybridisation strategy was employed to synthesise new CQ-benzoxaborole and cinnamoyl-benzoxaborole hybrids. CQ-Benzoxaborole 2.12a-c and cinnamoylbenzoxaborole 2.11a-g hydrid molecules were synthesised in low to good yields. Their structural identities were confirmed using conventional spectroscopic techniques (1H and 13C NMR, and mass spectrometry). CQ-benzoxaborole compounds, however, showed instability, and only 2.12b was used for in vitro biological assay and showed activity comparable to CQ. Furthermore, in vitro biological assay revealed that compounds 2.11a-g poorly inhibited the growth of P. falciparum parasites. Interestingly, these compounds, however, exhibited satisfactory activity against Trypanosoma brucei with IC50 = 0.052 μM for compound 2.11g. The cell cytotoxicity assay of all final compounds confirmed that all CQ-benzoxaborole 2.12b and cinnamoyl-benzoxaborole 2.11a-g hybrids were non-toxic against HeLa cell lines. However, efforts to further expand the structure-activity relationship (SAR) of CQbenzoxaborole by increasing the length of the linker with one extra carbon (Scheme 2.10) were not possible as an important precursor 6-formylbenzoxaborole 2.29 could not be synthesized in sufficient yields. , Thesis (MSc) -- Faculty of Faculty of Science, Chemistry, 2017
- Full Text:
- Authors: Gumbo, Maureen
- Date: 2017
- Subjects: Malaria Chemotherapy , Antimalarials , Boron compounds , Drug resistance , Plasmodium falciparum , Drug development
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/59193 , vital:27456
- Description: Malaria is a mosquito-borne disease, which continues to pose a threat to the entire humanity. About 40% of the world population is estimated to be at risk of infections by malaria. Despite efforts undertaken by scientific community, government entities and international organizations, malaria is still rampant. The major problem is drug resistance, where the Plasmodium spp have over the past decades developed drug resistance against available drugs. In order to counter this problem, novel antimalarial drugs that are efficacious and with novel mode of action are of great necessity. Benzoxaborole derivatives have been shown to exhibit promising antimalarial activity against Plasmodium falciparum strains. Previous studies reported on the compounds such as 6-(2- (alkoxycarbonyl)pyrazinyl-5-oxy)-1,3-dihydro-1-hydroxy-2,1-benzoxaboroles, which showed good antimalarial activity against both W7 and 3D7 strains without significant toxicity. On the other hand, chloroquine (CQ) and cinnamic acids have a wide variety of biological activity including antimalarial activity. Herein, a hybridisation strategy was employed to synthesise new CQ-benzoxaborole and cinnamoyl-benzoxaborole hybrids. CQ-Benzoxaborole 2.12a-c and cinnamoylbenzoxaborole 2.11a-g hydrid molecules were synthesised in low to good yields. Their structural identities were confirmed using conventional spectroscopic techniques (1H and 13C NMR, and mass spectrometry). CQ-benzoxaborole compounds, however, showed instability, and only 2.12b was used for in vitro biological assay and showed activity comparable to CQ. Furthermore, in vitro biological assay revealed that compounds 2.11a-g poorly inhibited the growth of P. falciparum parasites. Interestingly, these compounds, however, exhibited satisfactory activity against Trypanosoma brucei with IC50 = 0.052 μM for compound 2.11g. The cell cytotoxicity assay of all final compounds confirmed that all CQ-benzoxaborole 2.12b and cinnamoyl-benzoxaborole 2.11a-g hybrids were non-toxic against HeLa cell lines. However, efforts to further expand the structure-activity relationship (SAR) of CQbenzoxaborole by increasing the length of the linker with one extra carbon (Scheme 2.10) were not possible as an important precursor 6-formylbenzoxaborole 2.29 could not be synthesized in sufficient yields. , Thesis (MSc) -- Faculty of Faculty of Science, Chemistry, 2017
- Full Text:
An investigation into the biological treatment of platinum refinery effluent using the plant Azolla Filiculoides
- Authors: Marran, Vernon Edward
- Date: 2003
- Subjects: Uncatalogued
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/193453 , vital:45333
- Description: In order to understand the effects of metals contained in effluent and to define effluent quality suitable for safe discharge to natural water streams, it is essential to understand the effects of the interaction of metal ions with plants. The availability of metal ions and their ability to bind to plants are dependent on the chemical speciation of metals and on the biological factors governing the availability of metals within the plant cells. This thesis will address both aspects and thereby propose a combination of an appropriate chemical and biological approach to the investigation of bioaccumulation of the plant Azolla Filiculoides. Laboratory studies have shown that varying concentrations of free metal ions in solution determine efficiency of metal uptake and that metal toxicity can also be detrimental to plant life and efficiency of metal recovery from solution. Many questions however, remain unanswered with regard to the application of a biological treatment for effluent discharge. This thesis includes the determination of metal speciation combined with the study of bioaccumulation of metals in plants and their effects from test- work utilising effluent generated from a Precious Metals Refinery (PMR). Plant species are known to differ widely in their tolerance to metals, however despite an abundant knowledge on molecular, biochemical and physiological effects of metals to plants, only a few general principles have been proposed to guide the prediction of tolerance differences. The properties of protective cellular responses as well as of the molecular target sites are important components in determining the intrinsic tolerance of a particular species to a metal. The role of the whole assembly of cellular ligands in buffering metal ions within the cells will be evaluated. Standard preparation methods combined with use of Inductively Coupled Plasma Emission Spectrophotometer (1CP) used for analytical analysis will be included to reflect analytical data in providing evidence to support a conclusion. The outcome of the test work utilising the aquatic plant Azoila has proven that it can be used as a process step to re-mediate effluent generated from Precious Metal Refining operations. This process offers an alternative to the classical chemical methods widely used in the Precious Metals Refining industry proving economically viable and ensuring environmental sustainability in comparison to the current known methods of effluent treatment. , Thesis (MSc) -- Faculty of Science, Biochemistry, Microbiology and Biotechnology, 2003
- Full Text:
- Authors: Marran, Vernon Edward
- Date: 2003
- Subjects: Uncatalogued
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/193453 , vital:45333
- Description: In order to understand the effects of metals contained in effluent and to define effluent quality suitable for safe discharge to natural water streams, it is essential to understand the effects of the interaction of metal ions with plants. The availability of metal ions and their ability to bind to plants are dependent on the chemical speciation of metals and on the biological factors governing the availability of metals within the plant cells. This thesis will address both aspects and thereby propose a combination of an appropriate chemical and biological approach to the investigation of bioaccumulation of the plant Azolla Filiculoides. Laboratory studies have shown that varying concentrations of free metal ions in solution determine efficiency of metal uptake and that metal toxicity can also be detrimental to plant life and efficiency of metal recovery from solution. Many questions however, remain unanswered with regard to the application of a biological treatment for effluent discharge. This thesis includes the determination of metal speciation combined with the study of bioaccumulation of metals in plants and their effects from test- work utilising effluent generated from a Precious Metals Refinery (PMR). Plant species are known to differ widely in their tolerance to metals, however despite an abundant knowledge on molecular, biochemical and physiological effects of metals to plants, only a few general principles have been proposed to guide the prediction of tolerance differences. The properties of protective cellular responses as well as of the molecular target sites are important components in determining the intrinsic tolerance of a particular species to a metal. The role of the whole assembly of cellular ligands in buffering metal ions within the cells will be evaluated. Standard preparation methods combined with use of Inductively Coupled Plasma Emission Spectrophotometer (1CP) used for analytical analysis will be included to reflect analytical data in providing evidence to support a conclusion. The outcome of the test work utilising the aquatic plant Azoila has proven that it can be used as a process step to re-mediate effluent generated from Precious Metal Refining operations. This process offers an alternative to the classical chemical methods widely used in the Precious Metals Refining industry proving economically viable and ensuring environmental sustainability in comparison to the current known methods of effluent treatment. , Thesis (MSc) -- Faculty of Science, Biochemistry, Microbiology and Biotechnology, 2003
- Full Text:
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