Azadipyrromethenes for applications in photodynamic antimicrobial chemotherapy, photodynamic therapy and optical limiting
- Authors: Dubazana, Nadine
- Date: 2020
- Subjects: Dyes and dyeing -- Chemistry , Photochemotherapy , Cancer -- Photochemotherapy , Anti-infective agents , Staphylococcus aureus , Nonlinear optics , Azadipyrromethenes , BODIPY
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/166150 , vital:41333
- Description: Azadipyrromethenes, azaBODIPYs and zinc azadipyrromethene complexes were prepared and characterised to examine the effect on their photophysical properties of incorporating phenyl groups at the 1,3,5,7-positions with electron-donating and withdrawing groups at the para-positions. To enhance their ability to generate singlet oxygen, appropriate structural modifications were made through the addition of a Zn(II) ion or halogenation at the 2,6 positions. In vitro photodynamic therapy (PDT) studies targeting MCF-7 human breast cancer cells were carried out. To evaluate and understand the effectiveness of the dyes as photosensitisers, cellular uptake, phototoxicity and the half-maximal inhibitory concentration (IC50) values were analysed. Photodynamic antimicrobial chemotherapy (PACT) studies were also carried out to study the effectiveness of the dyes against Staphylococcus aureus (S. aureus). Dyes with donor-π-acceptor (D-π-A) properties were synthesised and tested against the second harmonic of the Nd:YAG laser in optical limiting (OL) studies. The second-order hyperpolarisability, third-order susceptibility and nonlinear absorption coefficient values were determined. The results suggest that 1,3,5,7-azaBODIPY dyes may be less suitable for use in this context than analogous D-π-A 3,5-distyrylBODIPY dyes. Molecular modelling was carried out to identify the structure-property relationships of the synthesised dyes by analysing trends in the energies of the frontier molecular orbitals (MOs) and spectroscopic properties.
- Full Text:
- Date Issued: 2020
- Authors: Dubazana, Nadine
- Date: 2020
- Subjects: Dyes and dyeing -- Chemistry , Photochemotherapy , Cancer -- Photochemotherapy , Anti-infective agents , Staphylococcus aureus , Nonlinear optics , Azadipyrromethenes , BODIPY
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/166150 , vital:41333
- Description: Azadipyrromethenes, azaBODIPYs and zinc azadipyrromethene complexes were prepared and characterised to examine the effect on their photophysical properties of incorporating phenyl groups at the 1,3,5,7-positions with electron-donating and withdrawing groups at the para-positions. To enhance their ability to generate singlet oxygen, appropriate structural modifications were made through the addition of a Zn(II) ion or halogenation at the 2,6 positions. In vitro photodynamic therapy (PDT) studies targeting MCF-7 human breast cancer cells were carried out. To evaluate and understand the effectiveness of the dyes as photosensitisers, cellular uptake, phototoxicity and the half-maximal inhibitory concentration (IC50) values were analysed. Photodynamic antimicrobial chemotherapy (PACT) studies were also carried out to study the effectiveness of the dyes against Staphylococcus aureus (S. aureus). Dyes with donor-π-acceptor (D-π-A) properties were synthesised and tested against the second harmonic of the Nd:YAG laser in optical limiting (OL) studies. The second-order hyperpolarisability, third-order susceptibility and nonlinear absorption coefficient values were determined. The results suggest that 1,3,5,7-azaBODIPY dyes may be less suitable for use in this context than analogous D-π-A 3,5-distyrylBODIPY dyes. Molecular modelling was carried out to identify the structure-property relationships of the synthesised dyes by analysing trends in the energies of the frontier molecular orbitals (MOs) and spectroscopic properties.
- Full Text:
- Date Issued: 2020
Bayesian accelerated life tests for the Weibull distribution under non-informative priors
- Authors: Mostert, Philip
- Date: 2020
- Subjects: Accelerated life testing -- Statistical methods , Accelerated life testing -- Mathematical models , Failure time data analysis , Bayesian statistical decision theory , Monte Carlo method , Weibull distribution
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/172181 , vital:42173
- Description: In a competitive world where products are designed to last for long periods of time, obtaining time-to-failure data is both difficult and costly. Hence for products with high reliability, accelerated life testing is required to obtain relevant life-data quickly. This is done by placing the products under higher-than-use stress levels, thereby causing the products to fail prematurely. Part of the analysis of accelerated life-data requires a life distribution that describes the lifetime of a product at a given stress level and a life-stress relationship – which is some function that describes the way in which the life distribution changes across different stress levels. In this thesis it is assumed that the underlying life distribution is the wellknown Weibull distribution, with shape parameter constant over all stress levels and scale parameter as a log-linear function of stress. The primary objective of this thesis is to obtain estimates from Bayesian analysis, and this thesis considers five types of non-informative prior distributions: Jeffreys’ prior, reference priors, maximal data information prior, uniform prior and probability matching priors. Since the associated posterior distribution under all the derived non-informative priors are of an unknown form, the propriety of the posterior distributions is assessed to ensure admissible results. For comparison purposes, estimates obtained via the method of maximum likelihood are also considered. Finding these estimates requires solving non-linear equations, hence the Newton-Raphson algorithm is used to obtain estimates. A simulation study based on the time-to-failure of accelerated data is conducted to compare results between maximum likelihood and Bayesian estimates. As a result of the Bayesian posterior distributions being analytically intractable, two methods to obtain Bayesian estimates are considered: Markov chain Monte Carlo methods and Lindley’s approximation technique. In the simulation study the posterior means and the root mean squared error values of the estimates under the symmetric squared error loss function and the two asymmetric loss functions: the LINEX loss function and general entropy loss function, are considered. Furthermore the coverage rates for the Bayesian Markov chain Monte Carlo and maximum likelihood estimates are found, and are compared by their average interval lengths. A case study using a dataset based on accelerated time-to-failure of an insulating fluid is considered. The fit of these data for the Weibull distribution is studied and is compared to that of other popular life distributions. A full simulation study is conducted to illustrate convergence of the proper posterior distributions. Both maximum likelihood and Bayesian estimates are found for these data. The deviance information criterion is used to compare Bayesian estimates between the prior distributions. The case study is concluded by finding reliability estimates of the data at use-stress levels.
- Full Text:
- Date Issued: 2020
- Authors: Mostert, Philip
- Date: 2020
- Subjects: Accelerated life testing -- Statistical methods , Accelerated life testing -- Mathematical models , Failure time data analysis , Bayesian statistical decision theory , Monte Carlo method , Weibull distribution
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/172181 , vital:42173
- Description: In a competitive world where products are designed to last for long periods of time, obtaining time-to-failure data is both difficult and costly. Hence for products with high reliability, accelerated life testing is required to obtain relevant life-data quickly. This is done by placing the products under higher-than-use stress levels, thereby causing the products to fail prematurely. Part of the analysis of accelerated life-data requires a life distribution that describes the lifetime of a product at a given stress level and a life-stress relationship – which is some function that describes the way in which the life distribution changes across different stress levels. In this thesis it is assumed that the underlying life distribution is the wellknown Weibull distribution, with shape parameter constant over all stress levels and scale parameter as a log-linear function of stress. The primary objective of this thesis is to obtain estimates from Bayesian analysis, and this thesis considers five types of non-informative prior distributions: Jeffreys’ prior, reference priors, maximal data information prior, uniform prior and probability matching priors. Since the associated posterior distribution under all the derived non-informative priors are of an unknown form, the propriety of the posterior distributions is assessed to ensure admissible results. For comparison purposes, estimates obtained via the method of maximum likelihood are also considered. Finding these estimates requires solving non-linear equations, hence the Newton-Raphson algorithm is used to obtain estimates. A simulation study based on the time-to-failure of accelerated data is conducted to compare results between maximum likelihood and Bayesian estimates. As a result of the Bayesian posterior distributions being analytically intractable, two methods to obtain Bayesian estimates are considered: Markov chain Monte Carlo methods and Lindley’s approximation technique. In the simulation study the posterior means and the root mean squared error values of the estimates under the symmetric squared error loss function and the two asymmetric loss functions: the LINEX loss function and general entropy loss function, are considered. Furthermore the coverage rates for the Bayesian Markov chain Monte Carlo and maximum likelihood estimates are found, and are compared by their average interval lengths. A case study using a dataset based on accelerated time-to-failure of an insulating fluid is considered. The fit of these data for the Weibull distribution is studied and is compared to that of other popular life distributions. A full simulation study is conducted to illustrate convergence of the proper posterior distributions. Both maximum likelihood and Bayesian estimates are found for these data. The deviance information criterion is used to compare Bayesian estimates between the prior distributions. The case study is concluded by finding reliability estimates of the data at use-stress levels.
- Full Text:
- Date Issued: 2020
BODIPY dyes for application in the photo-oxidation of pollutants, photodynamic antimicrobial chemotherapy, and nonlinear optics
- Authors: Kelechi, Lebechi Augustus
- Date: 2020
- Subjects: Dyes and dyeing -- Chemistry , Fluorescent probes , Fluorescence spectroscopy
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/140298 , vital:37859
- Description: The synthesis and structural characterization of a series of BODIPY dyes to analyze both the effects of halogenations at the 2,6-positions and the introduction of styryl groups at the 3,5-positions. The photophysical properties of these dyes were investigated to determine their suitability as singlet oxygen-generating photosensitiser dyes for application in photocatalytic degradation of azo dyes and in photodynamic antimicrobial chemotherapy (PACT). Upon halogenation, the dyes showed high to moderate singlet oxygen quantum yields. The potential utility of electrospun polystyrene (PS) nanofibres embedded with halogenated BODIPY dyes for the photocatalytic degradation of Orange G and Methyl Orange from textile industry effluents were investigated. A comparison of the singlet oxygen quantum yield of the BODIPY dyes in solution and when embedded in the PS nanofibres support demonstrates that its photosensitiser properties are maintained in the nanofibre mats. The photocatalytic degradation properties of the PS nanofibres for Orange G and Methyl Orange were determined by using a 530 nm and 660 nm light-emitting diodes. The rate of photodegradation increases with both the Orange G and Methyl Orange concentrations and follows pseudo-first-order kinetics. The PACT activities of brominated BODIPYs on Escherichia coli and Staphylococcus aureus were investigated. Log reduction values of over 9 were obtained during the photoinactivation of Staphylococcus aureus. To be able to red-shift the main spectral band of the BODIPY dyes into the therapeutic window, styryl groups were introduced at the 3,5-positions through a modified Knoevenagel condensation reaction. Because the red-shifted spectral band lies above 532 nm, the second harmonic of the Nd:YAG laser, there is very minute absorption at this wavelength. One of the novel brominated BODIPY dyes was investigated for its potential utility as optical limiting materials in nonlinear optics (NLO), and the dyes demonstrated typical nonlinear absorption behaviour characterised by reverse saturable absorption (RSA) in Z-scan measurements. Excellent optical limiting parameters were obtained for third-order susceptibility and hyperpolarisability.
- Full Text:
- Date Issued: 2020
- Authors: Kelechi, Lebechi Augustus
- Date: 2020
- Subjects: Dyes and dyeing -- Chemistry , Fluorescent probes , Fluorescence spectroscopy
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/140298 , vital:37859
- Description: The synthesis and structural characterization of a series of BODIPY dyes to analyze both the effects of halogenations at the 2,6-positions and the introduction of styryl groups at the 3,5-positions. The photophysical properties of these dyes were investigated to determine their suitability as singlet oxygen-generating photosensitiser dyes for application in photocatalytic degradation of azo dyes and in photodynamic antimicrobial chemotherapy (PACT). Upon halogenation, the dyes showed high to moderate singlet oxygen quantum yields. The potential utility of electrospun polystyrene (PS) nanofibres embedded with halogenated BODIPY dyes for the photocatalytic degradation of Orange G and Methyl Orange from textile industry effluents were investigated. A comparison of the singlet oxygen quantum yield of the BODIPY dyes in solution and when embedded in the PS nanofibres support demonstrates that its photosensitiser properties are maintained in the nanofibre mats. The photocatalytic degradation properties of the PS nanofibres for Orange G and Methyl Orange were determined by using a 530 nm and 660 nm light-emitting diodes. The rate of photodegradation increases with both the Orange G and Methyl Orange concentrations and follows pseudo-first-order kinetics. The PACT activities of brominated BODIPYs on Escherichia coli and Staphylococcus aureus were investigated. Log reduction values of over 9 were obtained during the photoinactivation of Staphylococcus aureus. To be able to red-shift the main spectral band of the BODIPY dyes into the therapeutic window, styryl groups were introduced at the 3,5-positions through a modified Knoevenagel condensation reaction. Because the red-shifted spectral band lies above 532 nm, the second harmonic of the Nd:YAG laser, there is very minute absorption at this wavelength. One of the novel brominated BODIPY dyes was investigated for its potential utility as optical limiting materials in nonlinear optics (NLO), and the dyes demonstrated typical nonlinear absorption behaviour characterised by reverse saturable absorption (RSA) in Z-scan measurements. Excellent optical limiting parameters were obtained for third-order susceptibility and hyperpolarisability.
- Full Text:
- Date Issued: 2020
Building a flexible and inexpensive multi-layer switch for software-defined networks
- Authors: Magwenzi, Tinashe
- Date: 2020
- Subjects: Software-defined networking (Computer network technology) , Telecommunication -- Switching systems , OpenFlow (Computer network protocol) , Local area networks (Computer networks)
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/142841 , vital:38122
- Description: Software-Defined Networking (SDN) is a paradigm which enables the realisation of programmable network through the separation of the control logic from the forwarding functions. This separation is a departure from the traditional architecture. Much of the work done in SDN enabled devices has concentrated on higher end, high speed networks (10s GBit/s 100s GBit/s), rather than the relatively low bandwidth links (10s MBit/s to a few GBit/s) which are seen, for example, in South Africa. As SDN is increasingly becoming more accepted, due to its advantages over the traditional networks, it has been adopted for industrial purposes such as networking in data centres and network providers. The demand for programmable networks is increasing but is limited by the ability of providers to upgrade their infrastructure. In addition, as access to the Internet has become less expensive, the use of Internet is increasing in academic institutions, NGOs, and small to medium enterprises. This thesis details a means of building and managing a small scale Software-Defined Network using commodity hardware and open source tools. Core to the SDN Network illustrated in this thesis is the prototype of a multi-layer SDN switch. The proposed device is targeted to serve lower bandwidth communication (in relation to commercially produced high speed SDN-enabled devices). The performance of the prototype multilayer switch had shown to achieve: data-rates of up to 99.998%, average latencies that are under 40µs during forwarding/switching and under 100µs during routing while using packet sizes between 64 bytes and 1518 bytes, and a jitter of less than 15µs during all tests. This research explores in detail the design, development, and management of a multi-layer switch and its placement and integration in small scale SDN network. This includes testing of Layer 2 forwarding and Layer 3 routing, OpenFlow compliance testing, the management of the switch using created SDN applications, and real life network functionality such as forwarding, routing and VLAN networking to demonstrate its real world applicability.
- Full Text:
- Date Issued: 2020
- Authors: Magwenzi, Tinashe
- Date: 2020
- Subjects: Software-defined networking (Computer network technology) , Telecommunication -- Switching systems , OpenFlow (Computer network protocol) , Local area networks (Computer networks)
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/142841 , vital:38122
- Description: Software-Defined Networking (SDN) is a paradigm which enables the realisation of programmable network through the separation of the control logic from the forwarding functions. This separation is a departure from the traditional architecture. Much of the work done in SDN enabled devices has concentrated on higher end, high speed networks (10s GBit/s 100s GBit/s), rather than the relatively low bandwidth links (10s MBit/s to a few GBit/s) which are seen, for example, in South Africa. As SDN is increasingly becoming more accepted, due to its advantages over the traditional networks, it has been adopted for industrial purposes such as networking in data centres and network providers. The demand for programmable networks is increasing but is limited by the ability of providers to upgrade their infrastructure. In addition, as access to the Internet has become less expensive, the use of Internet is increasing in academic institutions, NGOs, and small to medium enterprises. This thesis details a means of building and managing a small scale Software-Defined Network using commodity hardware and open source tools. Core to the SDN Network illustrated in this thesis is the prototype of a multi-layer SDN switch. The proposed device is targeted to serve lower bandwidth communication (in relation to commercially produced high speed SDN-enabled devices). The performance of the prototype multilayer switch had shown to achieve: data-rates of up to 99.998%, average latencies that are under 40µs during forwarding/switching and under 100µs during routing while using packet sizes between 64 bytes and 1518 bytes, and a jitter of less than 15µs during all tests. This research explores in detail the design, development, and management of a multi-layer switch and its placement and integration in small scale SDN network. This includes testing of Layer 2 forwarding and Layer 3 routing, OpenFlow compliance testing, the management of the switch using created SDN applications, and real life network functionality such as forwarding, routing and VLAN networking to demonstrate its real world applicability.
- Full Text:
- Date Issued: 2020
Combination ecosystem green engineering and early life history processes to enhance the intertidal biodiversity in the Port of East London
- Authors: Mafanya, Sandisiwer
- Date: 2020
- Subjects: Biotic communities -- South Africa -- East London , Perna -- Ecology --South Africa -- South Africa-- East London , Mexilhao mussel -- Ecology -- South Africa -- East London , Habitat (Ecology) -- Modification -- South Africa -- East London , Benthic animals -- Effect of habitat modification on -- South Africa -- East London , Harbors -- Environmental aspects -- South Africa -- East London
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/166376 , vital:41357
- Description: Marine coastal ecosystems are highly impacted by anthropogenic activities, including the development of, and practices within, harbours and/or ports. Artificial structures associated with harbours have a different chemical composition, texture and orientation than that of the natural benthic habitats they have replaced, and are therefore not generally favourable for the settlement and occurrence of indigenous species. Attempts are being made to identify what structures and materials can be used to aid in the rehabilitation of native species within harbours. The aim of this study was to investigate the effects of utilising environmentally-friendly artificial structures (tiles) with varying complexity (crevices and ridges with different depths) with ecological engineers (the indigenous bivalve Perna perna), on the associated benthic biodiversity within an international port in South Africa (Port of East London). The objectives of the study were to test the effects of artificial habitat complexity and the presence of bivalves on 1) the growth, mortality and biomass of a selected model species of ecosystem engineer (P. perna), and 2) the associated intertidal biodiversity. Tiles seeded with bivalves (P. perna) were deployed at two sites in the Port of East London and monitored monthly over a period of 12 months (November 2016-October 2017) to assess survival, growth and associated biodiversity. Environmental variables were also measured every month. The results of the three-way repeated measures ANOVAs indicated that tile treatment (especially high complexity of 2.5 and 5 cm) had an effect on the diversity of mobile species, length, height and biomass of the bivalves used as model ecosystem engineer, indicating the potentials for protection from wave dislodgement and refuge provision. Treatment (especially flat tiles) also had an effect on sessile/sedentary species diversity, indicating the greater proportion of space provided by these non-complex tiles. In addition, month had an effect on the mortality, height and biomass of the bivalves. Mortality was highest in October 2017, while the growth in height and biomass of P. perna were largest in January and October 2017 respectively. The use of artificial tiles also had a positive effect on the survival, growth, and biomass of bivalves (especially the length, weight and dry weight in 2.5 cm and 5 cm complexity). Additionally, this research investigated patterns of settlement and recruitment of the local bivalve population (P. perna) in the Port of East London and adjacent natural coastline to understand the early life history temporal and spatial dynamics of this model ecosystem engineer species. Settlement and recruitment were assessed every month by deploying and replacing artificial collectors for a total period of 19 months (November 2016-May 2018). The results of the two-way ANOVAs indicated that month and site (port vs natural) had an effect on settlement and recruitment of bivalves. Settlement and recruitment of bivalves were highest in July 2017 (port) and March 2018 (natural rocky shore). This study has highlighted that the use of artificial concrete tiles with increased complexity, as well as the investigation of the early stages of mussel populations could be important to consider in a framework of rehabilitation of urban coastal environments such as the Port of East London. Ecological engineering (in terms of increased complexity and heterogeneity) has indeed the potentials to be incorporated in South African programmes aiming at improving natural biodiversity in coastal urban environments. Nonetheless, the spatio-temporal variability of early driver of mussel populations (settlement and recruitment) is also an important feature to be closely monitored if biodiversity in South African coastal armouring is to be enhanced effectively and in the long term.
- Full Text:
- Date Issued: 2020
- Authors: Mafanya, Sandisiwer
- Date: 2020
- Subjects: Biotic communities -- South Africa -- East London , Perna -- Ecology --South Africa -- South Africa-- East London , Mexilhao mussel -- Ecology -- South Africa -- East London , Habitat (Ecology) -- Modification -- South Africa -- East London , Benthic animals -- Effect of habitat modification on -- South Africa -- East London , Harbors -- Environmental aspects -- South Africa -- East London
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/166376 , vital:41357
- Description: Marine coastal ecosystems are highly impacted by anthropogenic activities, including the development of, and practices within, harbours and/or ports. Artificial structures associated with harbours have a different chemical composition, texture and orientation than that of the natural benthic habitats they have replaced, and are therefore not generally favourable for the settlement and occurrence of indigenous species. Attempts are being made to identify what structures and materials can be used to aid in the rehabilitation of native species within harbours. The aim of this study was to investigate the effects of utilising environmentally-friendly artificial structures (tiles) with varying complexity (crevices and ridges with different depths) with ecological engineers (the indigenous bivalve Perna perna), on the associated benthic biodiversity within an international port in South Africa (Port of East London). The objectives of the study were to test the effects of artificial habitat complexity and the presence of bivalves on 1) the growth, mortality and biomass of a selected model species of ecosystem engineer (P. perna), and 2) the associated intertidal biodiversity. Tiles seeded with bivalves (P. perna) were deployed at two sites in the Port of East London and monitored monthly over a period of 12 months (November 2016-October 2017) to assess survival, growth and associated biodiversity. Environmental variables were also measured every month. The results of the three-way repeated measures ANOVAs indicated that tile treatment (especially high complexity of 2.5 and 5 cm) had an effect on the diversity of mobile species, length, height and biomass of the bivalves used as model ecosystem engineer, indicating the potentials for protection from wave dislodgement and refuge provision. Treatment (especially flat tiles) also had an effect on sessile/sedentary species diversity, indicating the greater proportion of space provided by these non-complex tiles. In addition, month had an effect on the mortality, height and biomass of the bivalves. Mortality was highest in October 2017, while the growth in height and biomass of P. perna were largest in January and October 2017 respectively. The use of artificial tiles also had a positive effect on the survival, growth, and biomass of bivalves (especially the length, weight and dry weight in 2.5 cm and 5 cm complexity). Additionally, this research investigated patterns of settlement and recruitment of the local bivalve population (P. perna) in the Port of East London and adjacent natural coastline to understand the early life history temporal and spatial dynamics of this model ecosystem engineer species. Settlement and recruitment were assessed every month by deploying and replacing artificial collectors for a total period of 19 months (November 2016-May 2018). The results of the two-way ANOVAs indicated that month and site (port vs natural) had an effect on settlement and recruitment of bivalves. Settlement and recruitment of bivalves were highest in July 2017 (port) and March 2018 (natural rocky shore). This study has highlighted that the use of artificial concrete tiles with increased complexity, as well as the investigation of the early stages of mussel populations could be important to consider in a framework of rehabilitation of urban coastal environments such as the Port of East London. Ecological engineering (in terms of increased complexity and heterogeneity) has indeed the potentials to be incorporated in South African programmes aiming at improving natural biodiversity in coastal urban environments. Nonetheless, the spatio-temporal variability of early driver of mussel populations (settlement and recruitment) is also an important feature to be closely monitored if biodiversity in South African coastal armouring is to be enhanced effectively and in the long term.
- Full Text:
- Date Issued: 2020
Combining DNA barcoding and morphology to identify larval fishes from the nearshore environment off the south-east coast of South Africa
- Authors: Somana, Zinzi Sinazo
- Date: 2020
- Subjects: Fishes -- Larvae -- South Africa -- Identification , Fishes -- Genetics -- Research -- Technique , Fishes -- South Africa -- Classification , Genetic markers
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/144605 , vital:38362
- Description: The early life history stages of most marine fish species are undescribed. The problem is, most of these fishes have pelagic larvae which are minute, delicate forms. Linking the larval stage to an adult counterpart is extremely challenging as larvae are morphologically different from the adults. Historically, larval fish identification relied solely on distinguishing morphological characteristics and meristic measurements, which has resulted in taxonomic confusion and misidentification. The introduction of the deoxyribonucleic acid (DNA) barcoding technique as an alternative approach has been successful in positively identifying larval fishes. The correct identification of larval specimens is the key to a better understanding of larval ecology, which underpins the success of any adult fish population. This study aimed to positively identify larval fishes of the south-east coast of South Africa using morphological characteristics and DNA barcoding. Larval and eggs specimens for this study were collected from the shallow nearshore waters of the south-east coast of South Africa. A total of 177 larval specimens were used for morphological analysis. Body shape, gut shape, pigmentation and morphometric measurements (such as body depth, preanal length and total body length) were used to identify each specimen to the family level. In addition, a fragment of mitochondrial cytochrome c oxidase subunit 1 gene (COI) was adopted for sequencing to identify larval fish specimens and fish eggs. Sequences generated from this study were compared to those in the Barcode of Life Database (BOLD). When there were no close matches to a sequence, the GenBank nucleic acid sequence database, maintained by the National Center for Biotechnology Information (NCBI), was used as an alternative. A total of 18 different families were identified through morphology. Seventy-seven of the 177 larval specimens were not subjected to morphological identification due to physical damage. The majority of larvae identified using morphological characteristics belonged to either the Sparidae, Tripterygiidae or Gobiesocidae fish families. Through DNA barcoding, 12 fish families, 16 genera and 18 different species were identified. Ten DNA barcodes (categorised as ‘no match’) from 10 different larval specimens were not identified through any of the online databases. Therefore, the 2% threshold value was used to identify members of the same species. The K2P genetic distance relationships were calculated among the no match sequences and downloaded probability matches from NCBI. This resulted in two unknown specimens assigned to the Blenniidae and Gobiidae. All other taxa were identified to species level, except specimens representing the Gobiidae and Tripterygiidae families. Based on the K2P genetic distances Gobiidae representatives were categorised as members of the Caffrogobius genus. Twenty-eight barcodes represented specimens from the Tripterygiidae. DNA barcode data from COI was analysed using the standard phylogenetic procedures in MEGA6 to examine relationships and differentiation among sequences. These could not be identified to the lowest taxonomic rank due to limited sequence data to compare them with. The sequence data from these specimens gave different results in the two online databases. BOLD results were to family level (Tripterygiidae) and NCBI to the species level (Clinidae: Pavoclinus profundus). Results in this study confirmed the efficiency of the DNA barcoding technique in species level identification of fish larvae. The evidence from genetic barcodes of the Tripterygiidae specimens, supported by morphological characteristics, suggests the need for thorough research to identify the individuals to the species level. The fact that this study identified taxonomically problematic Gobiidae and Tripterygiidae specimens suggests that studies similar to this may highlight additional diversity and help to resolve the taxonomy of other species in these families. However, the lack of reference sequence data from the adult specimens, and especially those with cryptic diversity, were both shortcomings for the positive identification of larvae. With that being said, it shows the necessity for more research to be conducted on barcoding of larvae in general as to accommodate all kinds of species from biodiversity to economic perspectives.
- Full Text:
- Date Issued: 2020
- Authors: Somana, Zinzi Sinazo
- Date: 2020
- Subjects: Fishes -- Larvae -- South Africa -- Identification , Fishes -- Genetics -- Research -- Technique , Fishes -- South Africa -- Classification , Genetic markers
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/144605 , vital:38362
- Description: The early life history stages of most marine fish species are undescribed. The problem is, most of these fishes have pelagic larvae which are minute, delicate forms. Linking the larval stage to an adult counterpart is extremely challenging as larvae are morphologically different from the adults. Historically, larval fish identification relied solely on distinguishing morphological characteristics and meristic measurements, which has resulted in taxonomic confusion and misidentification. The introduction of the deoxyribonucleic acid (DNA) barcoding technique as an alternative approach has been successful in positively identifying larval fishes. The correct identification of larval specimens is the key to a better understanding of larval ecology, which underpins the success of any adult fish population. This study aimed to positively identify larval fishes of the south-east coast of South Africa using morphological characteristics and DNA barcoding. Larval and eggs specimens for this study were collected from the shallow nearshore waters of the south-east coast of South Africa. A total of 177 larval specimens were used for morphological analysis. Body shape, gut shape, pigmentation and morphometric measurements (such as body depth, preanal length and total body length) were used to identify each specimen to the family level. In addition, a fragment of mitochondrial cytochrome c oxidase subunit 1 gene (COI) was adopted for sequencing to identify larval fish specimens and fish eggs. Sequences generated from this study were compared to those in the Barcode of Life Database (BOLD). When there were no close matches to a sequence, the GenBank nucleic acid sequence database, maintained by the National Center for Biotechnology Information (NCBI), was used as an alternative. A total of 18 different families were identified through morphology. Seventy-seven of the 177 larval specimens were not subjected to morphological identification due to physical damage. The majority of larvae identified using morphological characteristics belonged to either the Sparidae, Tripterygiidae or Gobiesocidae fish families. Through DNA barcoding, 12 fish families, 16 genera and 18 different species were identified. Ten DNA barcodes (categorised as ‘no match’) from 10 different larval specimens were not identified through any of the online databases. Therefore, the 2% threshold value was used to identify members of the same species. The K2P genetic distance relationships were calculated among the no match sequences and downloaded probability matches from NCBI. This resulted in two unknown specimens assigned to the Blenniidae and Gobiidae. All other taxa were identified to species level, except specimens representing the Gobiidae and Tripterygiidae families. Based on the K2P genetic distances Gobiidae representatives were categorised as members of the Caffrogobius genus. Twenty-eight barcodes represented specimens from the Tripterygiidae. DNA barcode data from COI was analysed using the standard phylogenetic procedures in MEGA6 to examine relationships and differentiation among sequences. These could not be identified to the lowest taxonomic rank due to limited sequence data to compare them with. The sequence data from these specimens gave different results in the two online databases. BOLD results were to family level (Tripterygiidae) and NCBI to the species level (Clinidae: Pavoclinus profundus). Results in this study confirmed the efficiency of the DNA barcoding technique in species level identification of fish larvae. The evidence from genetic barcodes of the Tripterygiidae specimens, supported by morphological characteristics, suggests the need for thorough research to identify the individuals to the species level. The fact that this study identified taxonomically problematic Gobiidae and Tripterygiidae specimens suggests that studies similar to this may highlight additional diversity and help to resolve the taxonomy of other species in these families. However, the lack of reference sequence data from the adult specimens, and especially those with cryptic diversity, were both shortcomings for the positive identification of larvae. With that being said, it shows the necessity for more research to be conducted on barcoding of larvae in general as to accommodate all kinds of species from biodiversity to economic perspectives.
- Full Text:
- Date Issued: 2020
Cyclooxygenase-1 as an anti-stroke target: potential inhibitor identification and non-synonymous single nucleotide polymorphism analysis
- Authors: Muronzi, Tendai
- Date: 2020
- Subjects: Cerebrovascular disease , Cerebrovascular disease -- Treatment , Cerebrovascular disease -- Chemotherapy , Cyclooxygenases , High throughput screening (Drug development) , Drug development , Molecular dynamics , South African Natural Compounds Database , ZINC database
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/143404 , vital:38243
- Description: Stroke is the third leading cause of death worldwide, with 87% of cases being ischemic stroke. The two primary therapeutic strategies to reduce post-ischemic brain damage are cellular and vascular approaches. The vascular strategy aims to rapidly re-open obstructed blood vessels, while the cellular approach aims to interfere with the signalling pathways that facilitate neuron damage and death. Unfortunately, popular vascular treatments have adverse side effects, necessitating the need for alternative chemotherapeutics. In this study, cyclooxygenase-1 (COX-1), which plays a significant role in the post- ischemic neuroinflammation and neuronal death, was targeted for identification of novel drug compounds and to assess the effect of nsSNPs on its structure and function. In a drug discovery part, ligands from the South African Natural Compounds Database (SANCDB-https://sancdb.rubi.ru.ac.za/) and ZINC database (http://zinc15.docking.org/) were used for high-throughput virtual screening (HVTS) against COX-1. Additionally, five nsSNPs were being investigated to assess their impact on protein structure and function. Three of these SNPs were in the COX-1 dimer interface. Molecular docking and molecular dynamics simulations revealed asymmetric nature of the protein. Several ligands, peculiar to each monomer, exhibited favourable binding energies in the respective active sites. SNP analysis indicated effects on inter-monomer interactions and protein stability.
- Full Text:
- Date Issued: 2020
- Authors: Muronzi, Tendai
- Date: 2020
- Subjects: Cerebrovascular disease , Cerebrovascular disease -- Treatment , Cerebrovascular disease -- Chemotherapy , Cyclooxygenases , High throughput screening (Drug development) , Drug development , Molecular dynamics , South African Natural Compounds Database , ZINC database
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/143404 , vital:38243
- Description: Stroke is the third leading cause of death worldwide, with 87% of cases being ischemic stroke. The two primary therapeutic strategies to reduce post-ischemic brain damage are cellular and vascular approaches. The vascular strategy aims to rapidly re-open obstructed blood vessels, while the cellular approach aims to interfere with the signalling pathways that facilitate neuron damage and death. Unfortunately, popular vascular treatments have adverse side effects, necessitating the need for alternative chemotherapeutics. In this study, cyclooxygenase-1 (COX-1), which plays a significant role in the post- ischemic neuroinflammation and neuronal death, was targeted for identification of novel drug compounds and to assess the effect of nsSNPs on its structure and function. In a drug discovery part, ligands from the South African Natural Compounds Database (SANCDB-https://sancdb.rubi.ru.ac.za/) and ZINC database (http://zinc15.docking.org/) were used for high-throughput virtual screening (HVTS) against COX-1. Additionally, five nsSNPs were being investigated to assess their impact on protein structure and function. Three of these SNPs were in the COX-1 dimer interface. Molecular docking and molecular dynamics simulations revealed asymmetric nature of the protein. Several ligands, peculiar to each monomer, exhibited favourable binding energies in the respective active sites. SNP analysis indicated effects on inter-monomer interactions and protein stability.
- Full Text:
- Date Issued: 2020
Default in payment, an application of statistical learning techniques
- Authors: Gcakasi, Lulama
- Date: 2020
- Subjects: Credit -- South Africa -- Risk assessment , Risk management -- Statistical methods -- South Africa , Credit -- Management -- Statistical methods , Commercial statistics
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/141547 , vital:37984
- Description: The ability of financial institutions to detect whether a customer will default on their credit card payment is essential for its profitability. To that effect, financial institutions have credit scoring systems in place to be able to estimate the credit risk associated with a customer. Various classification models are used to develop credit scoring systems such as k-nearest neighbours, logistic regression and classification trees. This study aims to assess the performance of different classification models on the prediction of credit card payment default. Credit data is usually of high dimension and as a result dimension reduction techniques, namely principal component analysis and linear discriminant analysis, are used in this study as a means to improve model performance. Two classification models are used, namely neural networks and support vector machines. Model performance is evaluated using accuracy and area under the curve (AUC). The neuarl network classifier performed better than the support vector machine classifier as it produced higher accuracy rates and AUC values. Dimension reduction techniques were not effective in improving model performance but did result in less computationally expensive models.
- Full Text:
- Date Issued: 2020
- Authors: Gcakasi, Lulama
- Date: 2020
- Subjects: Credit -- South Africa -- Risk assessment , Risk management -- Statistical methods -- South Africa , Credit -- Management -- Statistical methods , Commercial statistics
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/141547 , vital:37984
- Description: The ability of financial institutions to detect whether a customer will default on their credit card payment is essential for its profitability. To that effect, financial institutions have credit scoring systems in place to be able to estimate the credit risk associated with a customer. Various classification models are used to develop credit scoring systems such as k-nearest neighbours, logistic regression and classification trees. This study aims to assess the performance of different classification models on the prediction of credit card payment default. Credit data is usually of high dimension and as a result dimension reduction techniques, namely principal component analysis and linear discriminant analysis, are used in this study as a means to improve model performance. Two classification models are used, namely neural networks and support vector machines. Model performance is evaluated using accuracy and area under the curve (AUC). The neuarl network classifier performed better than the support vector machine classifier as it produced higher accuracy rates and AUC values. Dimension reduction techniques were not effective in improving model performance but did result in less computationally expensive models.
- Full Text:
- Date Issued: 2020
Design, formulation and evalauation of liposomes co-loaded with human serum Albumin and Rifampicin
- Authors: Bapolisi, Alain Murhimalika
- Date: 2020
- Subjects: Liposomes , Serum albumin , Rifampin , Mycobacterium tuberculosis
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/161780 , vital:40670
- Description: Tuberculosis (TB) is a devastating infectious disease caused by Mycobacterium tuberculosis and is the leading cause of death from a single infectious agent. The high morbidity and mortality rates of TB are partly due to factors such as the lengthy regimen (of 6–24 months), the development of drug resistance, and the pathogen location within the macrophages. These, with poor physiochemical properties of existing drugs hamper the effectiveness of the treatment despite the existence of potent antibiotics such as Rifampicin (Rif). Hydrophobicity plagues many drugs, including Rif, which are then particularly affected due to inherently poor intracellular availability. Novel drug delivery approaches are therefore needed in order to optimize the cytotoxic potential of said antitubercular drugs. To improve the bioavailability of hydrophobic drugs, numerous delivery strategies have been developed. Amongst these, the coordination of cytotoxic drugs to therapeutic proteins have shown some success for improved efficacy in the management of illnesses including infectious diseases. Of therapeutic proteins, Human Serum Albumin (HSA) is an attractive drug carrier with interestingbenefits such as low immunogenicity, antioxidant properties and improving cellular uptake ofdrugs through HSA-specific binding sites which are expressed on most cells including macrophages, where M. tuberculosis often resides. Hence, combination of Rif to HSA (Rif-HSA)seems a promising approach for improved intracellular delivery of Rif. However, the in vivo stability of colloidal protein-based therapeutics is mostly challenging and an effective vehicle is needed to control the biological fate of such conjugates.Liposomes seem to be appropriate carriers for the Rif-HSA complex due to their reputable applicability for encapsulating diverse materials (i.e., hydrophobic and hydrophilic compounds or small and complex molecules) and preventing chemical and biological degradation of the cargo. Therefore, the main objective of this study was to simultaneously encapsulate Rif and HSA in liposomes, which, to the best of our knowledge, has not been done before. The dual liposomes (Rif-HSA-lip) were made by a modified “Reverse Phase Evaporation” method (REV), following a Design of Experiments (DOE) approach to determine which factors impact the formulation. In addition, liposomes were made from crude soybean lecithin (CSL), rather than expensive and highly purified lipids. The liposomes were fully characterised, and the encapsulation efficiency (î) was monitored using high-performance liquid chromatography (HPLC). The results were correlated with factors such as organic and aqueous phase composition, as well as the in vitro release profile of Rif. Transmission electron microscopy (TEM) results confirmed the formation of spherical dual liposomes nanoparticles of roughly 200 nm. Dynamic light scattering (DLS) and Zeta potential measurements showed a negative charge (<–45 mV) and with satisfactory polydispersity (PDI<0.5). HSA dramatically improved the aqueous solubility of Rif (from1.9 mg/ml in water to around 4.3 mg/ml in HSA 10% solution) mainly due to Rif-HSA hydrophobic interactions. This resulted in a good î of almost 60% for Rif, despite the presence of bulky HSA in the lipid bilayer. These details were confirmed using proton nuclear magnetic resonance (1H NMR) and Fourier-transform infrared spectroscopy (FTIR). Furthermore, energy dispersive X-ray (EDX) and DLS data suggested the presence of HSA poking out on the surface of liposomes, which is encouraging for potential targeted delivery in the future. The in vitro release studies also depicted a substantial improvement in the diffusion of Rif in dual liposomes versus free Rif, from 65% after 12 hours for free Rif to 95% after only 5 hours for Rif- HSA-lip. Finally, stability studies conducted over 30 days at room temperature, showed that the freeze-dried formulations of Rif-HSA-lip exhibited good shelf stability over liposomes with no HSA. This study represents an illustrative example of co-loading of antibiotics and proteins into liposomes, which could encourage further development of novel nanoparticulate tools for the effective management of both drug-susceptible and -resistant infectious diseases such as TB.
- Full Text:
- Date Issued: 2020
- Authors: Bapolisi, Alain Murhimalika
- Date: 2020
- Subjects: Liposomes , Serum albumin , Rifampin , Mycobacterium tuberculosis
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/161780 , vital:40670
- Description: Tuberculosis (TB) is a devastating infectious disease caused by Mycobacterium tuberculosis and is the leading cause of death from a single infectious agent. The high morbidity and mortality rates of TB are partly due to factors such as the lengthy regimen (of 6–24 months), the development of drug resistance, and the pathogen location within the macrophages. These, with poor physiochemical properties of existing drugs hamper the effectiveness of the treatment despite the existence of potent antibiotics such as Rifampicin (Rif). Hydrophobicity plagues many drugs, including Rif, which are then particularly affected due to inherently poor intracellular availability. Novel drug delivery approaches are therefore needed in order to optimize the cytotoxic potential of said antitubercular drugs. To improve the bioavailability of hydrophobic drugs, numerous delivery strategies have been developed. Amongst these, the coordination of cytotoxic drugs to therapeutic proteins have shown some success for improved efficacy in the management of illnesses including infectious diseases. Of therapeutic proteins, Human Serum Albumin (HSA) is an attractive drug carrier with interestingbenefits such as low immunogenicity, antioxidant properties and improving cellular uptake ofdrugs through HSA-specific binding sites which are expressed on most cells including macrophages, where M. tuberculosis often resides. Hence, combination of Rif to HSA (Rif-HSA)seems a promising approach for improved intracellular delivery of Rif. However, the in vivo stability of colloidal protein-based therapeutics is mostly challenging and an effective vehicle is needed to control the biological fate of such conjugates.Liposomes seem to be appropriate carriers for the Rif-HSA complex due to their reputable applicability for encapsulating diverse materials (i.e., hydrophobic and hydrophilic compounds or small and complex molecules) and preventing chemical and biological degradation of the cargo. Therefore, the main objective of this study was to simultaneously encapsulate Rif and HSA in liposomes, which, to the best of our knowledge, has not been done before. The dual liposomes (Rif-HSA-lip) were made by a modified “Reverse Phase Evaporation” method (REV), following a Design of Experiments (DOE) approach to determine which factors impact the formulation. In addition, liposomes were made from crude soybean lecithin (CSL), rather than expensive and highly purified lipids. The liposomes were fully characterised, and the encapsulation efficiency (î) was monitored using high-performance liquid chromatography (HPLC). The results were correlated with factors such as organic and aqueous phase composition, as well as the in vitro release profile of Rif. Transmission electron microscopy (TEM) results confirmed the formation of spherical dual liposomes nanoparticles of roughly 200 nm. Dynamic light scattering (DLS) and Zeta potential measurements showed a negative charge (<–45 mV) and with satisfactory polydispersity (PDI<0.5). HSA dramatically improved the aqueous solubility of Rif (from1.9 mg/ml in water to around 4.3 mg/ml in HSA 10% solution) mainly due to Rif-HSA hydrophobic interactions. This resulted in a good î of almost 60% for Rif, despite the presence of bulky HSA in the lipid bilayer. These details were confirmed using proton nuclear magnetic resonance (1H NMR) and Fourier-transform infrared spectroscopy (FTIR). Furthermore, energy dispersive X-ray (EDX) and DLS data suggested the presence of HSA poking out on the surface of liposomes, which is encouraging for potential targeted delivery in the future. The in vitro release studies also depicted a substantial improvement in the diffusion of Rif in dual liposomes versus free Rif, from 65% after 12 hours for free Rif to 95% after only 5 hours for Rif- HSA-lip. Finally, stability studies conducted over 30 days at room temperature, showed that the freeze-dried formulations of Rif-HSA-lip exhibited good shelf stability over liposomes with no HSA. This study represents an illustrative example of co-loading of antibiotics and proteins into liposomes, which could encourage further development of novel nanoparticulate tools for the effective management of both drug-susceptible and -resistant infectious diseases such as TB.
- Full Text:
- Date Issued: 2020
Design, formulation and evaluation of liposomes co-loaded with human serum albumin and rifampicin
- Authors: Bapolisi, Alain Murhimalika
- Date: 2020
- Subjects: Liposomes , Rifampin , Antitubercular agents , Serum albumin , Albumins , Tuberculosis -- Treatment
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/163179 , vital:41016
- Description: Tuberculosis (TB) is a devastating infectious disease caused by Mycobacterium tuberculosis and is the leading cause of death from a single infectious agent. The high morbidity and mortality rates of TB are partly due to factors such as the lengthy regimen (of 6–24 months), the development of drug resistance, and the pathogen location within the macrophages. These, with poor physiochemical properties of existing drugs hamper the effectiveness of the treatment despite the existence of potent antibiotics such as Rifampicin (Rif). Hydrophobicity plagues many drugs, including Rif, which are then particularly affected due to inherently poor intracellular availability. Novel drug delivery approaches are therefore needed in order to optimize the cytotoxic potential of said antitubercular drugs. To improve the bioavailability of hydrophobic drugs, numerous delivery strategies have been developed. Amongst these, the coordination of cytotoxic drugs to therapeutic proteins have shown some success for improved efficacy in the management of illnesses including infectious diseases. Of therapeutic proteins, Human Serum Albumin (HSA) is an attractive drug carrier with interesting benefits such as low immunogenicity, antioxidant properties and improving cellular uptake of drugs through HSA-specific binding sites which are expressed on most cells including macrophages, where M. tuberculosis often resides. Hence, combination of Rif to HSA (Rif-HSA) seems a promising approach for improved intracellular delivery of Rif. However, the in vivo stability of colloidal protein-based therapeutics is mostly challenging and an effective vehicle is needed to control the biological fate of such conjugates. Liposomes seem to be appropriate carriers for the Rif-HSA complex due to their reputable applicability for encapsulating diverse materials (i.e., hydrophobic and hydrophilic compounds or small and complex molecules) and preventing chemical and biological degradation of the cargo. Therefore, the main objective of this study was to simultaneously encapsulate Rif and HSA in liposomes, which, to the best of our knowledge, has not been done before. The dual liposomes (Rif-HSA-lip) were made by a modified “Reverse Phase Evaporation” method (REV), following a Design of Experiments (DOE) approach to determine which factors impact the formulation. In addition, liposomes were made from crude soybean lecithin (CSL), rather than expensive and highly purified lipids. iv The liposomes were fully characterised, and the encapsulation efficiency (î) was monitored using high-performance liquid chromatography (HPLC). The results were correlated with factors such as organic and aqueous phase composition, as well as the in vitro release profile of Rif. Transmission electron microscopy (TEM) results confirmed the formation of spherical dual liposomes nanoparticles of roughly 200 nm. Dynamic light scattering (DLS) and Zeta potential measurements showed a negative charge (<–45 mV) and with satisfactory polydispersity (PDI<0.5). HSA dramatically improved the aqueous solubility of Rif (from1.9 mg/ml in water to around 4.3 mg/ml in HSA 10% solution) mainly due to Rif-HSA hydrophobic interactions. This resulted in a good î of almost 60% for Rif, despite the presence of bulky HSA in the lipid bilayer. These details were confirmed using proton nuclear magnetic resonance (1H NMR) and Fourier-transform infrared spectroscopy (FTIR). Furthermore, energy dispersive X-ray (EDX) and DLS data suggested the presence of HSA poking out on the surface of liposomes, which is encouraging for potential targeted delivery in the future. The in vitro release studies also depicted a substantial improvement in the diffusion of Rif in dual liposomes versus free Rif, from 65% after 12 hours for free Rif to 95% after only 5 hours for Rif- HSA-lip. Finally, stability studies conducted over 30 days at room temperature, showed that the freeze-dried formulations of Rif-HSA-lip exhibited good shelf stability over liposomes with no HSA. This study represents an illustrative example of co-loading of antibiotics and proteins into liposomes, which could encourage further development of novel nanoparticulate tools for the effective management of both drug-susceptible and -resistant infectious diseases such as TB.
- Full Text:
- Date Issued: 2020
- Authors: Bapolisi, Alain Murhimalika
- Date: 2020
- Subjects: Liposomes , Rifampin , Antitubercular agents , Serum albumin , Albumins , Tuberculosis -- Treatment
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/163179 , vital:41016
- Description: Tuberculosis (TB) is a devastating infectious disease caused by Mycobacterium tuberculosis and is the leading cause of death from a single infectious agent. The high morbidity and mortality rates of TB are partly due to factors such as the lengthy regimen (of 6–24 months), the development of drug resistance, and the pathogen location within the macrophages. These, with poor physiochemical properties of existing drugs hamper the effectiveness of the treatment despite the existence of potent antibiotics such as Rifampicin (Rif). Hydrophobicity plagues many drugs, including Rif, which are then particularly affected due to inherently poor intracellular availability. Novel drug delivery approaches are therefore needed in order to optimize the cytotoxic potential of said antitubercular drugs. To improve the bioavailability of hydrophobic drugs, numerous delivery strategies have been developed. Amongst these, the coordination of cytotoxic drugs to therapeutic proteins have shown some success for improved efficacy in the management of illnesses including infectious diseases. Of therapeutic proteins, Human Serum Albumin (HSA) is an attractive drug carrier with interesting benefits such as low immunogenicity, antioxidant properties and improving cellular uptake of drugs through HSA-specific binding sites which are expressed on most cells including macrophages, where M. tuberculosis often resides. Hence, combination of Rif to HSA (Rif-HSA) seems a promising approach for improved intracellular delivery of Rif. However, the in vivo stability of colloidal protein-based therapeutics is mostly challenging and an effective vehicle is needed to control the biological fate of such conjugates. Liposomes seem to be appropriate carriers for the Rif-HSA complex due to their reputable applicability for encapsulating diverse materials (i.e., hydrophobic and hydrophilic compounds or small and complex molecules) and preventing chemical and biological degradation of the cargo. Therefore, the main objective of this study was to simultaneously encapsulate Rif and HSA in liposomes, which, to the best of our knowledge, has not been done before. The dual liposomes (Rif-HSA-lip) were made by a modified “Reverse Phase Evaporation” method (REV), following a Design of Experiments (DOE) approach to determine which factors impact the formulation. In addition, liposomes were made from crude soybean lecithin (CSL), rather than expensive and highly purified lipids. iv The liposomes were fully characterised, and the encapsulation efficiency (î) was monitored using high-performance liquid chromatography (HPLC). The results were correlated with factors such as organic and aqueous phase composition, as well as the in vitro release profile of Rif. Transmission electron microscopy (TEM) results confirmed the formation of spherical dual liposomes nanoparticles of roughly 200 nm. Dynamic light scattering (DLS) and Zeta potential measurements showed a negative charge (<–45 mV) and with satisfactory polydispersity (PDI<0.5). HSA dramatically improved the aqueous solubility of Rif (from1.9 mg/ml in water to around 4.3 mg/ml in HSA 10% solution) mainly due to Rif-HSA hydrophobic interactions. This resulted in a good î of almost 60% for Rif, despite the presence of bulky HSA in the lipid bilayer. These details were confirmed using proton nuclear magnetic resonance (1H NMR) and Fourier-transform infrared spectroscopy (FTIR). Furthermore, energy dispersive X-ray (EDX) and DLS data suggested the presence of HSA poking out on the surface of liposomes, which is encouraging for potential targeted delivery in the future. The in vitro release studies also depicted a substantial improvement in the diffusion of Rif in dual liposomes versus free Rif, from 65% after 12 hours for free Rif to 95% after only 5 hours for Rif- HSA-lip. Finally, stability studies conducted over 30 days at room temperature, showed that the freeze-dried formulations of Rif-HSA-lip exhibited good shelf stability over liposomes with no HSA. This study represents an illustrative example of co-loading of antibiotics and proteins into liposomes, which could encourage further development of novel nanoparticulate tools for the effective management of both drug-susceptible and -resistant infectious diseases such as TB.
- Full Text:
- Date Issued: 2020
Development and assessment of a fixed dose combination of perindopril arginine and indapamide loaded microparticles
- Authors: Mandava, Tavonga Tyomai
- Date: 2020
- Subjects: Uncatalogued
- Language: English
- Type: thesis , text , Masters , MSc
- Identifier: http://hdl.handle.net/10962/164413 , vital:41116
- Description: Thesis (MSc)--Rhodes University, Faculty of Pharmacy, Pharmaceutics, 2020
- Full Text:
- Date Issued: 2020
- Authors: Mandava, Tavonga Tyomai
- Date: 2020
- Subjects: Uncatalogued
- Language: English
- Type: thesis , text , Masters , MSc
- Identifier: http://hdl.handle.net/10962/164413 , vital:41116
- Description: Thesis (MSc)--Rhodes University, Faculty of Pharmacy, Pharmaceutics, 2020
- Full Text:
- Date Issued: 2020
Development of a low-cost bioprinting system for engineering of Human Tumour Models
- Authors: Fanucci, Sidne
- Date: 2020
- Subjects: Uncatalogued
- Language: English
- Type: thesis , text , Masters , MSc
- Identifier: http://hdl.handle.net/10962/163295 , vital:41026
- Description: Thesis (MSc)--Rhodes University, Faculty of Science, Biotechnology Innovation Centre, 2020.
- Full Text:
- Date Issued: 2020
- Authors: Fanucci, Sidne
- Date: 2020
- Subjects: Uncatalogued
- Language: English
- Type: thesis , text , Masters , MSc
- Identifier: http://hdl.handle.net/10962/163295 , vital:41026
- Description: Thesis (MSc)--Rhodes University, Faculty of Science, Biotechnology Innovation Centre, 2020.
- Full Text:
- Date Issued: 2020
Development of a protocol for extracting and quantifying the concentration of thiafentanil in blesbok (Damaliscus pygargus phillipsi) matrices 72-74 hours post administration
- Authors: Webber, Judith Tracy
- Date: 2020
- Subjects: Uncatalogued
- Language: English
- Type: thesis , text , Masters , MSc
- Identifier: http://hdl.handle.net/10962/164738 , vital:41159
- Description: Thesis (MSc)--Rhodes University, Faculty of Science, Chemistry, 2020
- Full Text:
- Date Issued: 2020
- Authors: Webber, Judith Tracy
- Date: 2020
- Subjects: Uncatalogued
- Language: English
- Type: thesis , text , Masters , MSc
- Identifier: http://hdl.handle.net/10962/164738 , vital:41159
- Description: Thesis (MSc)--Rhodes University, Faculty of Science, Chemistry, 2020
- Full Text:
- Date Issued: 2020
Development of graphene materials and phthalocyanines for application in dye-sensitized solar cells
- Authors: Chindeka, Francis
- Date: 2020
- Subjects: Dye-sensitized solar cells , Graphene , Phthalocyanines , Molecular orbitals , Impedance spectroscopy
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/166092 , vital:41328
- Description: Two sets of dye-sensitized solar cells (DSSCs) were fabricated. In the first set, dye-sensitized solar cells (DSSC) were fabricated by incorporating graphene materials as catalysts at the counter electrode. Platinum was also used as a catalyst for comparative purposes. Different phthalocyanines: hydroxyl indium tetracarboxyphenoxy phthalocyanine (1), chloro indium octacarboxy phthalocyanine (2) and dibenzoic acid silicon phthalocyanine (3) were used as dyes. Complex 3 gave the highest power conversion efficiency (η) of 3.19% when using nitrogen doped reduced graphene oxide nanosheets (NrGONS) as a catalyst at the counter electrode, and TiO2 containing rGONS at the anode. The value obtained is close to 3.8% obtained when using Pt catalyst instead of NrGONS at the cathode, thus confirming that NrGONS is a promising candidate to replace the more expensive Pt. The study also shows that placing rGONS on both the anode and cathode improves efficiency. In the second set, DSSCs were fabricated by using 2(3,5-biscarboxyphenoxy), 9(10), 16(17), 23(24)-tri(tertbutyl) phthalocyaninato Cu (4) and Zn (5) complexes as dyes on the ITO-TiO2 photoanodes containing reduced graphene oxide nanosheets (rGONS) or nitrogen-doped rGONS (NrGONS). The evaluation of the assembled DSSCs revealed that using ITO-TiO2-NrGONS-CuPc (4) photoanode had the highest fill factor (FF) and power conversion efficiency (ɳ) of 69 % and 4.36 % respectively. These results show that the asymmetrical phthalocyanine complexes (4) and (5) showed significant improvement on the performance of the DSSC compared to previous work on symmetrical carboxylated phthalocyanines with ɳ = 3.19%.
- Full Text:
- Date Issued: 2020
- Authors: Chindeka, Francis
- Date: 2020
- Subjects: Dye-sensitized solar cells , Graphene , Phthalocyanines , Molecular orbitals , Impedance spectroscopy
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/166092 , vital:41328
- Description: Two sets of dye-sensitized solar cells (DSSCs) were fabricated. In the first set, dye-sensitized solar cells (DSSC) were fabricated by incorporating graphene materials as catalysts at the counter electrode. Platinum was also used as a catalyst for comparative purposes. Different phthalocyanines: hydroxyl indium tetracarboxyphenoxy phthalocyanine (1), chloro indium octacarboxy phthalocyanine (2) and dibenzoic acid silicon phthalocyanine (3) were used as dyes. Complex 3 gave the highest power conversion efficiency (η) of 3.19% when using nitrogen doped reduced graphene oxide nanosheets (NrGONS) as a catalyst at the counter electrode, and TiO2 containing rGONS at the anode. The value obtained is close to 3.8% obtained when using Pt catalyst instead of NrGONS at the cathode, thus confirming that NrGONS is a promising candidate to replace the more expensive Pt. The study also shows that placing rGONS on both the anode and cathode improves efficiency. In the second set, DSSCs were fabricated by using 2(3,5-biscarboxyphenoxy), 9(10), 16(17), 23(24)-tri(tertbutyl) phthalocyaninato Cu (4) and Zn (5) complexes as dyes on the ITO-TiO2 photoanodes containing reduced graphene oxide nanosheets (rGONS) or nitrogen-doped rGONS (NrGONS). The evaluation of the assembled DSSCs revealed that using ITO-TiO2-NrGONS-CuPc (4) photoanode had the highest fill factor (FF) and power conversion efficiency (ɳ) of 69 % and 4.36 % respectively. These results show that the asymmetrical phthalocyanine complexes (4) and (5) showed significant improvement on the performance of the DSSC compared to previous work on symmetrical carboxylated phthalocyanines with ɳ = 3.19%.
- Full Text:
- Date Issued: 2020
Distribution, habitat associations and conservation of the Eastern Cape rocky Sandelia bainsii (Perciform: Anabantidae) in the Great Fish River system
- Authors: Sifundza, Delsy Sindy
- Date: 2020
- Subjects: Anabantidae -- South Africa -- Great Fish River , Perciformes -- South Africa -- Great Fish River , Labyrinth fishes -- South Africa -- Great Fish River , Fishes -- Effect of human beings on -- South Africa -- Great Fish River , Fish populations -- South Africa -- Great Fish River , Fish declines -- South Africa -- Great Fish River , Fish populations -- Monitoring -- South Africa -- Great Fish River , Anabantidae -- Evolution -- South Africa -- Great Fish River , Gene flow , Mitochondrial DNA , Microsatellites (Genetics) , Eastern Cape rocky Sandelia bainsii , Freshwater ecology -- South Africa -- Great Fish River
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/142692 , vital:38102
- Description: In South Africa, freshwater ecosystems are under pressure due to increasing anthropogenic factors degrading river systems. The Cape Fold, the lower Southern Temperate Highveld and the Amatolo – Winterberg Highlands freshwater ecoregions contain some of the heavily impacted river systems in the country. One such river system is the Great Fish River system, which has been affected by anthropogenic modification (hydrological modification, water pollution and invasion by non-native fish species). These modifications have raised conservation concerns on an imperilled anabantid fish, the Eastern Cape rocky Sandelia bainsii that is confined to the Kat and Koonap rivers, which are tributaries of the Great Fish River. Historically, the species was reportedly common and abundant, with a range spanning across seven river systems (Kowie, Great Fish, Keiskamma, Igoda, Gxulu, Buffalo and Nahoon rivers) in the Eastern Cape Province. However, surveys have indicated that the species has experienced decline in population size and distribution range, with localised extirpations being reported. This resulted in its listing on the IUCN Red List of threatened species as Endangered. The proposed shale gas exploration and potential infrastructure development in the Karoo basin, which encompasses the headwaters of the Kat River, represent a potential future threat to the remnant populations of this species. The aim of the present study was to review the past and present distribution range of S. bainsii, determine the habitat associations and assess the conservation of S. bainsii in the Great Fish River system. This information is essential for informing conservation and management decisions for this species in the Great Fish River system. This study undertook a comprehensive survey of the Kat and the Koonap rivers in 2017, and augmented this information with data from more recent surveys that were conducted from 2009 - 2014 to map the distributions freshwater fishes in the Great Fish River system. Data from these recent surveys (2009 – 2017) were compared with historical records (1961 – 2005) obtained from the South African Institute for Aquatic Biodiversity to evaluate changes in distribution patterns of S. bainsii. Historical records indicated that S. bainsii occurred at 11 localities in the Kat River and three localities in the Koonap River. Data from recent surveys indicated that the species has persisted at these historical localities, except one locality in the Koonap River where no individuals of S. bainsii were captured, despite the use of multiple sampling approaches and gears. Although the species still persists at two of the three historical localities in the Koonap River, the population sizes have considerably declined as the species was reportedly in high abundance at these localities in the 1980s. Factors threatening the continued existence of this species in the Koonap River likely include non-native species, extreme drought conditions and the associated excessive abstraction of water which has resulted in the drying of some of the refugia pools where the species was historically common and abundant. In contrast to the Koonap River, the 2017 survey indicated that in the Kat River, S. bainsii had a similar distribution range compared to historical records. The species occurred at 11 of the 41 localities that were sampled in the Kat River, and the sampled populations consisted of all size classes. Sandelia bainsii occurred with other native fish species in the system, including Labeo umbratus, Enteromius anoplus and Glossogobius callidus. A comparison of historical and recent data indicates that non-native fishes have likely not expanded their ranges in the Kat River. This is likely due to the presence of weirs and dams that have prevented upstream movement of non-native fish species L. aeneus, C. gariepinus and T. sparrmanii which are now abundant in the lower Kat and mainstem Great Fish River. Surveys from the present study indicated that although S. bainsii still persists within the Kat River, there are a number of factors that pose a considerable threat to the continued existence of this species in the Great Fish River system. Chapter 3 evaluated the habitat associations of S. bainsii based on 10 physical and chemical variables and distribution data collected from 30 localities in the Upper Kat River and seven of its tributaries. A non-metric multidimensional scaling model was used to assess the distribution patterns of S. bainsii in relation to environmental variables. Furthermore, a negative binomial generalised linear model (GLM) was used to assess the relationship between the abundance of S. bainsii and environmental variables. Results from the present study indicated that S. bainsii was a habitat generalist as the distribution of this species showed no specific association with any of the habitat variables that were considered in this study. The species occurred in a wide range of habitats, including rocky sections in the upper Kat River and its tributaries as well as the mainstem sections. The GLM however indicated that the abundance of S. bainsii was negatively associated with increasing conductivity. In addition, the Kat River contains multiple weirs of variable sizes. Future studies should therefore evaluate the effects of these weirs on population fragmentation, gene flow and long-term evolutionary potential of S. bainsii through the application of techniques such as mitochondrial DNA and microsatellites.
- Full Text:
- Date Issued: 2020
- Authors: Sifundza, Delsy Sindy
- Date: 2020
- Subjects: Anabantidae -- South Africa -- Great Fish River , Perciformes -- South Africa -- Great Fish River , Labyrinth fishes -- South Africa -- Great Fish River , Fishes -- Effect of human beings on -- South Africa -- Great Fish River , Fish populations -- South Africa -- Great Fish River , Fish declines -- South Africa -- Great Fish River , Fish populations -- Monitoring -- South Africa -- Great Fish River , Anabantidae -- Evolution -- South Africa -- Great Fish River , Gene flow , Mitochondrial DNA , Microsatellites (Genetics) , Eastern Cape rocky Sandelia bainsii , Freshwater ecology -- South Africa -- Great Fish River
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/142692 , vital:38102
- Description: In South Africa, freshwater ecosystems are under pressure due to increasing anthropogenic factors degrading river systems. The Cape Fold, the lower Southern Temperate Highveld and the Amatolo – Winterberg Highlands freshwater ecoregions contain some of the heavily impacted river systems in the country. One such river system is the Great Fish River system, which has been affected by anthropogenic modification (hydrological modification, water pollution and invasion by non-native fish species). These modifications have raised conservation concerns on an imperilled anabantid fish, the Eastern Cape rocky Sandelia bainsii that is confined to the Kat and Koonap rivers, which are tributaries of the Great Fish River. Historically, the species was reportedly common and abundant, with a range spanning across seven river systems (Kowie, Great Fish, Keiskamma, Igoda, Gxulu, Buffalo and Nahoon rivers) in the Eastern Cape Province. However, surveys have indicated that the species has experienced decline in population size and distribution range, with localised extirpations being reported. This resulted in its listing on the IUCN Red List of threatened species as Endangered. The proposed shale gas exploration and potential infrastructure development in the Karoo basin, which encompasses the headwaters of the Kat River, represent a potential future threat to the remnant populations of this species. The aim of the present study was to review the past and present distribution range of S. bainsii, determine the habitat associations and assess the conservation of S. bainsii in the Great Fish River system. This information is essential for informing conservation and management decisions for this species in the Great Fish River system. This study undertook a comprehensive survey of the Kat and the Koonap rivers in 2017, and augmented this information with data from more recent surveys that were conducted from 2009 - 2014 to map the distributions freshwater fishes in the Great Fish River system. Data from these recent surveys (2009 – 2017) were compared with historical records (1961 – 2005) obtained from the South African Institute for Aquatic Biodiversity to evaluate changes in distribution patterns of S. bainsii. Historical records indicated that S. bainsii occurred at 11 localities in the Kat River and three localities in the Koonap River. Data from recent surveys indicated that the species has persisted at these historical localities, except one locality in the Koonap River where no individuals of S. bainsii were captured, despite the use of multiple sampling approaches and gears. Although the species still persists at two of the three historical localities in the Koonap River, the population sizes have considerably declined as the species was reportedly in high abundance at these localities in the 1980s. Factors threatening the continued existence of this species in the Koonap River likely include non-native species, extreme drought conditions and the associated excessive abstraction of water which has resulted in the drying of some of the refugia pools where the species was historically common and abundant. In contrast to the Koonap River, the 2017 survey indicated that in the Kat River, S. bainsii had a similar distribution range compared to historical records. The species occurred at 11 of the 41 localities that were sampled in the Kat River, and the sampled populations consisted of all size classes. Sandelia bainsii occurred with other native fish species in the system, including Labeo umbratus, Enteromius anoplus and Glossogobius callidus. A comparison of historical and recent data indicates that non-native fishes have likely not expanded their ranges in the Kat River. This is likely due to the presence of weirs and dams that have prevented upstream movement of non-native fish species L. aeneus, C. gariepinus and T. sparrmanii which are now abundant in the lower Kat and mainstem Great Fish River. Surveys from the present study indicated that although S. bainsii still persists within the Kat River, there are a number of factors that pose a considerable threat to the continued existence of this species in the Great Fish River system. Chapter 3 evaluated the habitat associations of S. bainsii based on 10 physical and chemical variables and distribution data collected from 30 localities in the Upper Kat River and seven of its tributaries. A non-metric multidimensional scaling model was used to assess the distribution patterns of S. bainsii in relation to environmental variables. Furthermore, a negative binomial generalised linear model (GLM) was used to assess the relationship between the abundance of S. bainsii and environmental variables. Results from the present study indicated that S. bainsii was a habitat generalist as the distribution of this species showed no specific association with any of the habitat variables that were considered in this study. The species occurred in a wide range of habitats, including rocky sections in the upper Kat River and its tributaries as well as the mainstem sections. The GLM however indicated that the abundance of S. bainsii was negatively associated with increasing conductivity. In addition, the Kat River contains multiple weirs of variable sizes. Future studies should therefore evaluate the effects of these weirs on population fragmentation, gene flow and long-term evolutionary potential of S. bainsii through the application of techniques such as mitochondrial DNA and microsatellites.
- Full Text:
- Date Issued: 2020
Do submarine canyons influence the structure and diversity of benthic fish assemblages on the continental shelf edge?
- Nyawo, Mpilonhle Aura June-Rose
- Authors: Nyawo, Mpilonhle Aura June-Rose
- Date: 2020
- Subjects: Submarine valleys , iSimangaliso Wetland Park (South Africa) , Biotic communities -- South Africa -- iSimangaliso Wetland Park , Groundfishes -- South Africa -- iSimangaliso Wetland Park , Marine ecology -- South Africa -- iSimangaliso Wetland Park
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/167264 , vital:41462
- Description: Marine ecosystems play an important role in human welfare, such as providing food security and providing social, economic and environmental benefits to an increasing human population. However, due to the deleterious impact of overfishing as well as habitat degradation, pollution and climate change, many marine ecosystems and the substantial biodiversity they support, are under increasing threats. Submarine canyon ecosystems cut into the continental margin in all our oceans and are considered regions of enhanced diversity, abundance and biomass for many marine communities, including fish. In South Africa, the tropical Delagoa Ecoregion is characterised by numerous submarine canyons that are home to coelacanths Latimeria chalumnae, and a variety of commercially important line fish species. However, there is currently a lack of information on the ecological structure and functioning of these canyons, in relation to nearby non-canyon ecosystems. The aim of this thesis was to generate knowledge on the role of submarine canyons in influencing the benthic fish assemblage structure and diversity on the continental shelf and shelf edge within the high-latitude coral reef ecosystems in the tropical Delagoa Ecoregion of South Africa. First, a systematic literature review was conducted to examine the current knowledge on the role of submarine canyons on the benthic fish assemblages occurring on continental margins, as well as outline the trends in canyon research. For the fieldwork component of this thesis, baited remote underwater stereo-video systems were utilised to gather data. The first research chapter, chapter 3, investigated the effect of ecosystem and depth on the diversity and structure of benthic fish assemblages within iSimangaliso Wetland Park in the Delagoa Ecoregion. Data were collected in 2017 from Wright Canyon (canyon ecosystem) and adjacent slope ecosystems around the canyon (non-canyon ecosystem) at depths ranging between 70–240 m. The second research chapter, chapter 4, data were collected in 2018, from 60–100 m depths in three ubmarine canyons (Wright, Diepgat,Levens) and their adjacent habitat within iSimangaliso Wetland Park to investigate the diversity patterns and community structure of benthic fish and cons ider location as a factor. The systematic review showed that canyon research is still very much in its infancy, however more papers are expected to be published in the future. There were clear location associated biases detected, with more than 90% of the canyon research being concentrated in the Mediterranean Sea, while only one study was published from South Africa. Most of the reviewed literature focused on invertebrates, with benthic fish (including cartilaginous fish) being the second most studied taxa. A wide range of methods from both fishery- dependent and fishery-independent categories were used to gather the data of benthic fish from the canyons and adjacent areas. With over 8500 submarine canyons mapped globally, the number of submarine canyon research published and the detected location bias is of great concern. More studies are needed in order to understand the role of submarine canyons on adjacent slope habitats Overall, the results from the research chapters indicated that there were higher a bundances in the canyon ecosystem compared to the non-canyon ecosystem. In contrast, evenness and beta-diversity were higher in the non-canyon ecosystem. Relief, bottom type as well as depth played a major role in explaining the observed abundance and diversity patterns. Both evenness and beta-diversity decreased with increase in depth. The canyon and non-canyon also differed in terms of habitat structure. The canyon ecosystem was characterised by patched reef and reef habitats with high relief whereas the non-canyon ecosystem was characterised by flats and habitats, especially at shallower depths. Additionally, there were clear differences in terms of the fish community structure between the canyon and the non-canyon ecosystem. The heterogeneous and complex habitats of the canyon ecosystem were typified by species such as Epinephelus poecilonotus, Polysteganus coeruleopunctatus and Chrysoblephus puniceus in comparison to the less complex habitats outside the canyons that were preferred by species such as Lagocephalus sceleratus, and Rhinobatos leucospilus. The different locations sampled demonstrated that the canyon effect was not onsistent at all locations, with varying results detected in the different sample locations. This research demonstrates that clea r differences exists between the benthic fish assemblages associated with canyon and non-canyon ecosystems and provides important information on the role of submarine canyons in the iSimangaliso Wetland Park. From this thesis, it is clear that a ‘canyon effect’ exists in the continental shelf and shelf edge. The results suggested that heterogeneity of habitats was the most important environmental factor that differed between the canyon and non-canyon ecosystems and these differences in the habitat were driving the observed patterns. This thesis has provided a reference point, with regards to investigating the role and influence of these canyons on benthic fish assemblages occurring on the continental shelf edge. The information provided by this thesis can ultimately be used in other studies investigating benthic fish assemblages in other submarine canyons within iSimangaliso Wetland Park and the rest of South Africa./
- Full Text:
- Date Issued: 2020
- Authors: Nyawo, Mpilonhle Aura June-Rose
- Date: 2020
- Subjects: Submarine valleys , iSimangaliso Wetland Park (South Africa) , Biotic communities -- South Africa -- iSimangaliso Wetland Park , Groundfishes -- South Africa -- iSimangaliso Wetland Park , Marine ecology -- South Africa -- iSimangaliso Wetland Park
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/167264 , vital:41462
- Description: Marine ecosystems play an important role in human welfare, such as providing food security and providing social, economic and environmental benefits to an increasing human population. However, due to the deleterious impact of overfishing as well as habitat degradation, pollution and climate change, many marine ecosystems and the substantial biodiversity they support, are under increasing threats. Submarine canyon ecosystems cut into the continental margin in all our oceans and are considered regions of enhanced diversity, abundance and biomass for many marine communities, including fish. In South Africa, the tropical Delagoa Ecoregion is characterised by numerous submarine canyons that are home to coelacanths Latimeria chalumnae, and a variety of commercially important line fish species. However, there is currently a lack of information on the ecological structure and functioning of these canyons, in relation to nearby non-canyon ecosystems. The aim of this thesis was to generate knowledge on the role of submarine canyons in influencing the benthic fish assemblage structure and diversity on the continental shelf and shelf edge within the high-latitude coral reef ecosystems in the tropical Delagoa Ecoregion of South Africa. First, a systematic literature review was conducted to examine the current knowledge on the role of submarine canyons on the benthic fish assemblages occurring on continental margins, as well as outline the trends in canyon research. For the fieldwork component of this thesis, baited remote underwater stereo-video systems were utilised to gather data. The first research chapter, chapter 3, investigated the effect of ecosystem and depth on the diversity and structure of benthic fish assemblages within iSimangaliso Wetland Park in the Delagoa Ecoregion. Data were collected in 2017 from Wright Canyon (canyon ecosystem) and adjacent slope ecosystems around the canyon (non-canyon ecosystem) at depths ranging between 70–240 m. The second research chapter, chapter 4, data were collected in 2018, from 60–100 m depths in three ubmarine canyons (Wright, Diepgat,Levens) and their adjacent habitat within iSimangaliso Wetland Park to investigate the diversity patterns and community structure of benthic fish and cons ider location as a factor. The systematic review showed that canyon research is still very much in its infancy, however more papers are expected to be published in the future. There were clear location associated biases detected, with more than 90% of the canyon research being concentrated in the Mediterranean Sea, while only one study was published from South Africa. Most of the reviewed literature focused on invertebrates, with benthic fish (including cartilaginous fish) being the second most studied taxa. A wide range of methods from both fishery- dependent and fishery-independent categories were used to gather the data of benthic fish from the canyons and adjacent areas. With over 8500 submarine canyons mapped globally, the number of submarine canyon research published and the detected location bias is of great concern. More studies are needed in order to understand the role of submarine canyons on adjacent slope habitats Overall, the results from the research chapters indicated that there were higher a bundances in the canyon ecosystem compared to the non-canyon ecosystem. In contrast, evenness and beta-diversity were higher in the non-canyon ecosystem. Relief, bottom type as well as depth played a major role in explaining the observed abundance and diversity patterns. Both evenness and beta-diversity decreased with increase in depth. The canyon and non-canyon also differed in terms of habitat structure. The canyon ecosystem was characterised by patched reef and reef habitats with high relief whereas the non-canyon ecosystem was characterised by flats and habitats, especially at shallower depths. Additionally, there were clear differences in terms of the fish community structure between the canyon and the non-canyon ecosystem. The heterogeneous and complex habitats of the canyon ecosystem were typified by species such as Epinephelus poecilonotus, Polysteganus coeruleopunctatus and Chrysoblephus puniceus in comparison to the less complex habitats outside the canyons that were preferred by species such as Lagocephalus sceleratus, and Rhinobatos leucospilus. The different locations sampled demonstrated that the canyon effect was not onsistent at all locations, with varying results detected in the different sample locations. This research demonstrates that clea r differences exists between the benthic fish assemblages associated with canyon and non-canyon ecosystems and provides important information on the role of submarine canyons in the iSimangaliso Wetland Park. From this thesis, it is clear that a ‘canyon effect’ exists in the continental shelf and shelf edge. The results suggested that heterogeneity of habitats was the most important environmental factor that differed between the canyon and non-canyon ecosystems and these differences in the habitat were driving the observed patterns. This thesis has provided a reference point, with regards to investigating the role and influence of these canyons on benthic fish assemblages occurring on the continental shelf edge. The information provided by this thesis can ultimately be used in other studies investigating benthic fish assemblages in other submarine canyons within iSimangaliso Wetland Park and the rest of South Africa./
- Full Text:
- Date Issued: 2020
Ecological engineering: an assessment of the ecological impact of Reno mattress structures used in erosion control in the Keurbooms Estuary, South Africa
- Authors: De Villiers, Nina
- Date: 2020
- Subjects: Sediments (Geology) -- Management , Sediments (Geology) -- South Africa -- Keurboomstrand , Coast changes -- South Africa -- Keurboomstrand , Shore protection -- South Africa -- Keurboomstrand , Coastal engineering-- South Africa -- Keurboomstrand , Coastal zone management-- South Africa -- Keurboomstrand , Estuarine ecology-- South Africa -- Keurboomstrand , Eelgrass -- South Africa -- Keurboomstrand , Reno Mattresses
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/166138 , vital:41332
- Description: Global climate changes have been associated with ocean warming and sea-level rise. Armouring of coastlines has become common practice with the increasing threat of coastal erosion. The transformation of soft sediment habitats to hard, artificial habitats because of coastline armouring can lead to changes in species diversity, composition and distribution. It is, therefore, essential to assess changes to habitats from coastal development as well as the ecological impact erosion control structures have within coastal systems. Ecological engineering attempts to combine engineering principals and ecological processes to reduce environmental impacts from coastal development and the implementation of artificial structures. Estuaries are particularly vulnerable to anthropogenic impacts through development, and are extremely important systems offering nursery and foraging grounds for many species. These systems are, however, particularly vulnerable to anthropogenic impacts from urbanisation and development. Within South Africa many estuaries are being transformed by the addition of artificial structures to combat erosion, one such structure is the Reno mattress (a flattened wire box filled with rocks). This study compared the fish diversity and abundances of existing Reno mattress structures and natural eelgrass (Zostera capensis) habitat in the Keurbooms Estuary, South Africa. Benthic invertebrates were sampled using standard core sampling and an adapted suction sampling approach within the two habitats. The non-destructive method of mini Baited Remote Underwater Video Systems (BRUVs) was used to sample fish. Seasonal benthic invertebrate and fish abundances and assemblages were assessed from winter 2018 to spring 2019 with greater abundances of both recorded in summer. Significantly greater abundances, diversity, and richness of fish were found in the Reno mattress habitat compared to Z. capensis. Invertebrate taxa displayed some overlap between habitats, however, three higher taxonomic groups were only recorded within Reno mattress habitat and one only within the eelgrass habitat. Fish assemblages differed significantly between the two established habitats. A Before-After-Control-Impact (BACI) investigation was used to assess the ecological impact of newly installed Reno mattresses in the Keurbooms Estuary. Zostera capensis extent was sampled by determining the percentage cover of 0.5 m X 0.5 m quadrats and measurements of eelgrass blades. Percentage cover and blade length decreased during the installation of Reno mattress, but then recovered shortly after completion of the installation. Abundances, richness and diversity of invertebrates and fish were found to be similar before and after the installation which suggests that the installation had no net negative impact on the site. The Reno mattresses were found to attract fauna typical of rocky shore environments as well as a few invasive alien invertebrate species. This study noted that a hybrid habitat of Reno mattress and eelgrass was created and may in fact provide the positives of both to a system. In any coastal development it will be important to balance the demands of a growing population and the protection of natural habitats. The results of this study suggest that complex artificial structures such as Reno mattresses do provide habitat for fish and invertebrates. However, the use of these structures should be in combination with natural vegetation (e.g. as a hybrid habitat) and not one that replaces intertidal and subtidal natural habitat especially eelgrass. There is limited information regarding the ecological impacts of using Reno mattresses in estuaries and this study provides new information on their ecological efficacy that should be valuable for future coastal erosion control practices.
- Full Text:
- Date Issued: 2020
- Authors: De Villiers, Nina
- Date: 2020
- Subjects: Sediments (Geology) -- Management , Sediments (Geology) -- South Africa -- Keurboomstrand , Coast changes -- South Africa -- Keurboomstrand , Shore protection -- South Africa -- Keurboomstrand , Coastal engineering-- South Africa -- Keurboomstrand , Coastal zone management-- South Africa -- Keurboomstrand , Estuarine ecology-- South Africa -- Keurboomstrand , Eelgrass -- South Africa -- Keurboomstrand , Reno Mattresses
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/166138 , vital:41332
- Description: Global climate changes have been associated with ocean warming and sea-level rise. Armouring of coastlines has become common practice with the increasing threat of coastal erosion. The transformation of soft sediment habitats to hard, artificial habitats because of coastline armouring can lead to changes in species diversity, composition and distribution. It is, therefore, essential to assess changes to habitats from coastal development as well as the ecological impact erosion control structures have within coastal systems. Ecological engineering attempts to combine engineering principals and ecological processes to reduce environmental impacts from coastal development and the implementation of artificial structures. Estuaries are particularly vulnerable to anthropogenic impacts through development, and are extremely important systems offering nursery and foraging grounds for many species. These systems are, however, particularly vulnerable to anthropogenic impacts from urbanisation and development. Within South Africa many estuaries are being transformed by the addition of artificial structures to combat erosion, one such structure is the Reno mattress (a flattened wire box filled with rocks). This study compared the fish diversity and abundances of existing Reno mattress structures and natural eelgrass (Zostera capensis) habitat in the Keurbooms Estuary, South Africa. Benthic invertebrates were sampled using standard core sampling and an adapted suction sampling approach within the two habitats. The non-destructive method of mini Baited Remote Underwater Video Systems (BRUVs) was used to sample fish. Seasonal benthic invertebrate and fish abundances and assemblages were assessed from winter 2018 to spring 2019 with greater abundances of both recorded in summer. Significantly greater abundances, diversity, and richness of fish were found in the Reno mattress habitat compared to Z. capensis. Invertebrate taxa displayed some overlap between habitats, however, three higher taxonomic groups were only recorded within Reno mattress habitat and one only within the eelgrass habitat. Fish assemblages differed significantly between the two established habitats. A Before-After-Control-Impact (BACI) investigation was used to assess the ecological impact of newly installed Reno mattresses in the Keurbooms Estuary. Zostera capensis extent was sampled by determining the percentage cover of 0.5 m X 0.5 m quadrats and measurements of eelgrass blades. Percentage cover and blade length decreased during the installation of Reno mattress, but then recovered shortly after completion of the installation. Abundances, richness and diversity of invertebrates and fish were found to be similar before and after the installation which suggests that the installation had no net negative impact on the site. The Reno mattresses were found to attract fauna typical of rocky shore environments as well as a few invasive alien invertebrate species. This study noted that a hybrid habitat of Reno mattress and eelgrass was created and may in fact provide the positives of both to a system. In any coastal development it will be important to balance the demands of a growing population and the protection of natural habitats. The results of this study suggest that complex artificial structures such as Reno mattresses do provide habitat for fish and invertebrates. However, the use of these structures should be in combination with natural vegetation (e.g. as a hybrid habitat) and not one that replaces intertidal and subtidal natural habitat especially eelgrass. There is limited information regarding the ecological impacts of using Reno mattresses in estuaries and this study provides new information on their ecological efficacy that should be valuable for future coastal erosion control practices.
- Full Text:
- Date Issued: 2020
Effect of the nature of nanoparticles on the photophysicochemical properties and photodynamic antimicrobial chemotherapy of phthalocyanines
- Authors: Magadla, Aviwe
- Date: 2020
- Subjects: Nanoparticles , Phthalocyanines , Anti-infective agents -- Therapeutic use , Photochemotherapy , Photochemistry
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/123107 , vital:35406
- Description: In this work, the syntheses and characterisation of Zn monocaffeic acid tri–tert–butyl phthalocyanine (1), Zn monocarboxyphenoxy tri– tert–butylphenoxyl phthalocyanine (2), tetrakis phenoxy N,N-dimethyl-4-(methylimino) phthalocyanine indium (III) chloride (3) and tetrakis N,N-dimethyl-4-(methylimino) phthalocyanine indium (III) chloride (5) are presented. Complexes 3 and 5 were further quartenised with 1,3- propanesultone to form corresponding complexes (4) and (6), respectively. Complexes 1 and 2 were covalently linked to amino functionalised nanoparticles (NPs). Complexes 3, 4, 5 and 6 where linked to oleic acid/oleylamine capped (OLA/OLM) silver-iron dimers (Ag-Fe3O4 OLA/OLM) and silver-iron core shell (Ag@Fe3O4 OLA/OLM) NPs via interaction between the nanoparticles and the imino group on the phthalocyanines. The phthalocyanine-NP conjugates afforded an increase in triplet quantum yields with a corresponding decrease in fluorescence quantum yield as compared to the phthalocyanine complexes alone. Complexes 3, 4 and their conjugates were then used for photodynamic antimicrobial chemotherapy on E. coli. The zwitterionic photosensitiser 4 and its conjugates showed better efficiency for photodynamic antimicrobial chemotherapy compared to their neutral counterparts.
- Full Text:
- Date Issued: 2020
- Authors: Magadla, Aviwe
- Date: 2020
- Subjects: Nanoparticles , Phthalocyanines , Anti-infective agents -- Therapeutic use , Photochemotherapy , Photochemistry
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/123107 , vital:35406
- Description: In this work, the syntheses and characterisation of Zn monocaffeic acid tri–tert–butyl phthalocyanine (1), Zn monocarboxyphenoxy tri– tert–butylphenoxyl phthalocyanine (2), tetrakis phenoxy N,N-dimethyl-4-(methylimino) phthalocyanine indium (III) chloride (3) and tetrakis N,N-dimethyl-4-(methylimino) phthalocyanine indium (III) chloride (5) are presented. Complexes 3 and 5 were further quartenised with 1,3- propanesultone to form corresponding complexes (4) and (6), respectively. Complexes 1 and 2 were covalently linked to amino functionalised nanoparticles (NPs). Complexes 3, 4, 5 and 6 where linked to oleic acid/oleylamine capped (OLA/OLM) silver-iron dimers (Ag-Fe3O4 OLA/OLM) and silver-iron core shell (Ag@Fe3O4 OLA/OLM) NPs via interaction between the nanoparticles and the imino group on the phthalocyanines. The phthalocyanine-NP conjugates afforded an increase in triplet quantum yields with a corresponding decrease in fluorescence quantum yield as compared to the phthalocyanine complexes alone. Complexes 3, 4 and their conjugates were then used for photodynamic antimicrobial chemotherapy on E. coli. The zwitterionic photosensitiser 4 and its conjugates showed better efficiency for photodynamic antimicrobial chemotherapy compared to their neutral counterparts.
- Full Text:
- Date Issued: 2020
Effectiveness of different interventions to reduce occupational sitting among office administrators at Rhodes University
- Authors: Malesa, Thato
- Date: 2020
- Subjects: Standing position , Sedentary behavior , Work environment , Employee health promotion , Office furniture -- Design , Industrial hygiene , Employees -- Health risk assessment , Human engineering , Compliance
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/166403 , vital:41360
- Description: Background: Sitting for prolonged periods is common in the working environment. Office workers are exposed to long periods of sitting time at work. Research has reported associations between prolonged sitting and negative health implications. As such studies have proposed different interventions aimed at reducing sitting times in the workplace. The aim of the current study was to evaluate the effectiveness of two different interventions, physical intervention (sit-stand worktables) and personal intervention (instruction to stretch) on compliance and reducing occupational sitting behaviour. The study also aimed to investigate the effectiveness of interventions in improving or impairing mood. Methods: The study took the form of a fieldbased study atRhodes University, Makhanda, South Africa. Fifteen full–time office workers in different divisions participated in the study. Participants were randomly allocated to either the physical or personal intervention group. In both intervention groups, partic ipants were prompted to stand for five minutes every hour during the workday to either work in a standing position or perform stretches. Over the course of the experiment, desk occupancy, sitting/standing time and mood effect were recorded in both intervention groups Both intervention groups were monitored for a period of three months (58 days). Results: The study found that the physical intervention group was an effective approach to reduce prolonged sitting in the workplace in comparison to the personal intervention group. Over the course of the experiment, there was sustain usage of interventions in both groups, however, in the physical intervention group desk usage decreased overtime. The findings of the study also show that some participants were more compliant with the study procedures than others. It was also found that mood improved upon introducing interventions in the workplace. However, with the outcomes of the results, the study acknowledges that several factors emerged which are likely to impact compliance, which future studies may investigate. Conclusion: Although sit-stand worktables are expensive, it seems like introducing them was successful in reducing sitting time and changing sitting behaviour in comparison to an intervention that instruct s individuals to stand up and stretch.
- Full Text:
- Date Issued: 2020
- Authors: Malesa, Thato
- Date: 2020
- Subjects: Standing position , Sedentary behavior , Work environment , Employee health promotion , Office furniture -- Design , Industrial hygiene , Employees -- Health risk assessment , Human engineering , Compliance
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/166403 , vital:41360
- Description: Background: Sitting for prolonged periods is common in the working environment. Office workers are exposed to long periods of sitting time at work. Research has reported associations between prolonged sitting and negative health implications. As such studies have proposed different interventions aimed at reducing sitting times in the workplace. The aim of the current study was to evaluate the effectiveness of two different interventions, physical intervention (sit-stand worktables) and personal intervention (instruction to stretch) on compliance and reducing occupational sitting behaviour. The study also aimed to investigate the effectiveness of interventions in improving or impairing mood. Methods: The study took the form of a fieldbased study atRhodes University, Makhanda, South Africa. Fifteen full–time office workers in different divisions participated in the study. Participants were randomly allocated to either the physical or personal intervention group. In both intervention groups, partic ipants were prompted to stand for five minutes every hour during the workday to either work in a standing position or perform stretches. Over the course of the experiment, desk occupancy, sitting/standing time and mood effect were recorded in both intervention groups Both intervention groups were monitored for a period of three months (58 days). Results: The study found that the physical intervention group was an effective approach to reduce prolonged sitting in the workplace in comparison to the personal intervention group. Over the course of the experiment, there was sustain usage of interventions in both groups, however, in the physical intervention group desk usage decreased overtime. The findings of the study also show that some participants were more compliant with the study procedures than others. It was also found that mood improved upon introducing interventions in the workplace. However, with the outcomes of the results, the study acknowledges that several factors emerged which are likely to impact compliance, which future studies may investigate. Conclusion: Although sit-stand worktables are expensive, it seems like introducing them was successful in reducing sitting time and changing sitting behaviour in comparison to an intervention that instruct s individuals to stand up and stretch.
- Full Text:
- Date Issued: 2020
Evaluation of an NADPH-dependent assay for inhibition screening of Salmonella enterica DOXP Reguctoisomerase for identification of novel drug hit compounds
- Authors: Ngcongco, Khanyisile
- Date: 2020
- Subjects: 1-Deoxy-D-xylulose 5-phosphate , Antibiotics , Drug development , Salmonella , Enterobacteriaceae , Vaccines , Plasmodium falciparum , Mycobacterium tuberculosis
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/167132 , vital:41440
- Description: Invasive non-typhoidal Salmonella, caused by the intracellular pathogen Salmonella enterica, has emerged as a major cause of bloodstream infections. It remains a neglected infection responsible for many deaths in Africa, as it fails to receive the level of support that is given to most better known infections. There are currently no vaccines against invasive non-typhoidal Salmonella. First-line antibiotics have been used for treatment, however, the rise in the resistance of the bacteria against these antibiotics has made treatment of invasive salmonellosis into a clinical problem. Therefore, the discovery of new compounds for the development of antibiotic drugs is required. Central metabolic pathways can be a useful source for identifying drug targets and among these is the non-mevalonate pathway, one of the pathways used for the biosynthesis of isoprenoid precursors. The second step of the non-mevalonate pathway involves the NADPH-dependent reduction of 1-deoxy-D-xylulose 5-phosphate (DOXP) into 2-C-methyl-D-erythritol 4-phosphate (MEP). 1-Deoxy-D-xylulose 5-phosphate (DOXP) reductoisomerase plays a vital role in the catalysis of this reaction and requires NADPH and divalent metal cations as co-factors for its activity. In this investigation recombinant DOXP reductoisomerase from Salmonella enterica, Plasmodium falciparum and Mycobacterium tuberculosis were biochemically characterized as potential targets for developing drugs that could be used as treatment of the disease. The expression and nickel-chelate affinity purification of S. enterica DOXP reductoisomerase in a fully functional native state was successfully achieved. However, the expression and purification of P. falciparum DXR and M. tuberculosis DXR was unsuccessful due to the formation of insoluble inclusion bodies. Although alternative purification strategies were explored, including dialysis and slow dilution, these proteins remained insoluble, making their functional analysis not possible. An NADPH-dependent enzyme assay was used to determine the activity of S. enterica DXR. This assay monitors the reduction of DOXP to MEP by measuring the absorbance at 340 nm, which reflects the concentration of NADPH. An alternative assay, resazurin reduction, which monitors the NADPH-dependent reduction of resazurin to resorufin, was explored for detecting enzyme activity. The recombinant S. enterica DOXP reductoisomerase had a specific activity of 0.126 ± 0.0014 μmol/min/mg protein and a Km and Vmax of 881 μM and 0.249 μmol/min/mg respectively. FR900098, a derivative of fosmidomycin, is a well-known inhibitor of DXR, however, the sensitivity of S. enterica DXR towards FR900098 has not yet been reported. The NADPH dependent enzyme and resazurin reduction assays were used to determine whether FR900098 has enzyme inhibitory effects against S. enterica DXR. Upon confirming that FR900098 is able to inhibit S. enterica DXR, FR900098 was used as a control compound in the screening of novel compounds. The S. enterica DXR enzyme was screened for inhibition by the collection of compounds from the Pathogen Box. Compounds that exhibited the desired inhibitory activity, referred to as ‘hits’ were selected for further investigation. These hits were confirmed using the NADPH-dependent enzyme assay, resulting in the identification of two different DXR inhibitor compounds, MMV002816, also known as diethylcarbamazine, and MMV228911. The inhibitory concentration (IC50) values of FR900098, MMV002816 and MMV228911 against S. enterica DXR were 1.038 μM, 2.173 μM and 6.861 μM respectively. The binding mode of these compounds to S. enterica DXR could lead to the discovery of novel druggable sites on the enzyme and stimulate the development of new antibiotics that can be used for treating Salmonella infections.
- Full Text:
- Date Issued: 2020
- Authors: Ngcongco, Khanyisile
- Date: 2020
- Subjects: 1-Deoxy-D-xylulose 5-phosphate , Antibiotics , Drug development , Salmonella , Enterobacteriaceae , Vaccines , Plasmodium falciparum , Mycobacterium tuberculosis
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/167132 , vital:41440
- Description: Invasive non-typhoidal Salmonella, caused by the intracellular pathogen Salmonella enterica, has emerged as a major cause of bloodstream infections. It remains a neglected infection responsible for many deaths in Africa, as it fails to receive the level of support that is given to most better known infections. There are currently no vaccines against invasive non-typhoidal Salmonella. First-line antibiotics have been used for treatment, however, the rise in the resistance of the bacteria against these antibiotics has made treatment of invasive salmonellosis into a clinical problem. Therefore, the discovery of new compounds for the development of antibiotic drugs is required. Central metabolic pathways can be a useful source for identifying drug targets and among these is the non-mevalonate pathway, one of the pathways used for the biosynthesis of isoprenoid precursors. The second step of the non-mevalonate pathway involves the NADPH-dependent reduction of 1-deoxy-D-xylulose 5-phosphate (DOXP) into 2-C-methyl-D-erythritol 4-phosphate (MEP). 1-Deoxy-D-xylulose 5-phosphate (DOXP) reductoisomerase plays a vital role in the catalysis of this reaction and requires NADPH and divalent metal cations as co-factors for its activity. In this investigation recombinant DOXP reductoisomerase from Salmonella enterica, Plasmodium falciparum and Mycobacterium tuberculosis were biochemically characterized as potential targets for developing drugs that could be used as treatment of the disease. The expression and nickel-chelate affinity purification of S. enterica DOXP reductoisomerase in a fully functional native state was successfully achieved. However, the expression and purification of P. falciparum DXR and M. tuberculosis DXR was unsuccessful due to the formation of insoluble inclusion bodies. Although alternative purification strategies were explored, including dialysis and slow dilution, these proteins remained insoluble, making their functional analysis not possible. An NADPH-dependent enzyme assay was used to determine the activity of S. enterica DXR. This assay monitors the reduction of DOXP to MEP by measuring the absorbance at 340 nm, which reflects the concentration of NADPH. An alternative assay, resazurin reduction, which monitors the NADPH-dependent reduction of resazurin to resorufin, was explored for detecting enzyme activity. The recombinant S. enterica DOXP reductoisomerase had a specific activity of 0.126 ± 0.0014 μmol/min/mg protein and a Km and Vmax of 881 μM and 0.249 μmol/min/mg respectively. FR900098, a derivative of fosmidomycin, is a well-known inhibitor of DXR, however, the sensitivity of S. enterica DXR towards FR900098 has not yet been reported. The NADPH dependent enzyme and resazurin reduction assays were used to determine whether FR900098 has enzyme inhibitory effects against S. enterica DXR. Upon confirming that FR900098 is able to inhibit S. enterica DXR, FR900098 was used as a control compound in the screening of novel compounds. The S. enterica DXR enzyme was screened for inhibition by the collection of compounds from the Pathogen Box. Compounds that exhibited the desired inhibitory activity, referred to as ‘hits’ were selected for further investigation. These hits were confirmed using the NADPH-dependent enzyme assay, resulting in the identification of two different DXR inhibitor compounds, MMV002816, also known as diethylcarbamazine, and MMV228911. The inhibitory concentration (IC50) values of FR900098, MMV002816 and MMV228911 against S. enterica DXR were 1.038 μM, 2.173 μM and 6.861 μM respectively. The binding mode of these compounds to S. enterica DXR could lead to the discovery of novel druggable sites on the enzyme and stimulate the development of new antibiotics that can be used for treating Salmonella infections.
- Full Text:
- Date Issued: 2020