Prediction of mass spectra using an ab initio approach
- Authors: Novokoza, Yolanda
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Doctoral theses , text
- Identifier: http://hdl.handle.net/${Handle} , vital:72818
- Description: Access restricted. Expected release date in 2025. , Thesis (PhD) -- Faculty of Science, Chemistry, 2023
- Full Text:
- Date Issued: 2023-10-13
- Authors: Novokoza, Yolanda
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Doctoral theses , text
- Identifier: http://hdl.handle.net/${Handle} , vital:72818
- Description: Access restricted. Expected release date in 2025. , Thesis (PhD) -- Faculty of Science, Chemistry, 2023
- Full Text:
- Date Issued: 2023-10-13
Screening isolation and biological characterization of antibacterial secondary metabolites from macrofauna endemic to the southern African coast
- Authors: Njanje, Idris
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/431944 , vital:72817
- Description: Access restricted. Expected release date in 2025. , Thesis (PhD) -- Faculty of Science, Chemistry, 2023
- Full Text:
- Date Issued: 2023-10-13
- Authors: Njanje, Idris
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/431944 , vital:72817
- Description: Access restricted. Expected release date in 2025. , Thesis (PhD) -- Faculty of Science, Chemistry, 2023
- Full Text:
- Date Issued: 2023-10-13
The effect of Cannabis extract on the morphological and metabolic characteristics of various fat depots in diet-induced Obese and STZ-induced male wistar rats
- Authors: Ramlugon, Sonaal
- Date: 2023-04
- Subjects: Rats as laboratory animals , Diabetes in practice , Cannabis -- South Africa
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10948/61282 , vital:70044
- Description: To investigate the potential anti-diabetic/obesity properties of oral cannabis administration in an obese and streptozotocin (STZ)-induced diabetic rat model, as well as an obese rat model, and to determine the mechanism of action, with a focus on the peritoneal and intramuscular fat depots. Experimental Design: Obese and STZ-induced diabetic rats were allocated a high fat diet (HFD) and intraperitoneally injected with STZ to mimic an obese and diabetic state. The rats were then orally administered cannabis extract (CE) of 1.25, 2.5 and 5.0 mg/kg body weight (relative to tetrahydrocannabinol (THC) content) or metformin as a positive control. For the obese rat model, the rats were allocated either a high carbohydrate diet (HCD) or high fat diet (HFD) and orally administered with cannabis extract of 1.25 mg/kg body weight (relative to THC content). Weight, blood and insulin-resistant parameters of the rats were monitored. The mitochondrial to genomic DNA ratio (MT:18S DNA), average adipocyte area of the various adipose tissues, citrate synthase and carnitine palmitoyltransferase 1 (CPT1) enzyme activities of the peritoneal and intramuscular fat were measured. Gene expression levels of uncoupling protein 1 (UCP1), cell-death inducing DNA fragmentation factor alpha like effector-a (Cidea), perilipin, hormone-sensitive lipase (HSL) and mitochondrial transcription factor A (TFAM) were measured in peritoneal fat, intramuscular fat and brown adipose tissue (BAT). Main Findings: Obese and STZ-induced diabetic rat model: Due to the biphasic nature of cannabinoids, cannabis dosage plays an important role in the observed effects. CE1.25 was the only cannabis treatment effective in improving the insulinresistant parameters of the rats unlike the other higher cannabis concentrations (CE2.5 and CE5.0). In the peritoneal fat, CE1.25 increased MT:18S DNA, increased citrate synthase activity, and decreased the average adipocyte area when compared to the STZ group. CE1.25 also induced fat beigeing by upregulating gene expression levels of UCP1 and Cidea. XIX Furthermore, an increase in gene expression levels of perilipin, HSL, and TFAM showed increased fat mobilization and metabolic activity. In the intramuscular fat, CE1.25 also reduced the average adipocytes area. However, a different mechanism of action was observed where CE1.25 did not induce fat beigeing, but instead increased both citrate synthase and CPT1 enzyme activities and gene expression levels of HSL, thereby indicating increased fat oxidation and mitochondrial activity. , Thesis (PhD) -- Faculty of Science, School of Biomolecular and Chemical Sciences, 2023
- Full Text:
- Date Issued: 2023-04
- Authors: Ramlugon, Sonaal
- Date: 2023-04
- Subjects: Rats as laboratory animals , Diabetes in practice , Cannabis -- South Africa
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10948/61282 , vital:70044
- Description: To investigate the potential anti-diabetic/obesity properties of oral cannabis administration in an obese and streptozotocin (STZ)-induced diabetic rat model, as well as an obese rat model, and to determine the mechanism of action, with a focus on the peritoneal and intramuscular fat depots. Experimental Design: Obese and STZ-induced diabetic rats were allocated a high fat diet (HFD) and intraperitoneally injected with STZ to mimic an obese and diabetic state. The rats were then orally administered cannabis extract (CE) of 1.25, 2.5 and 5.0 mg/kg body weight (relative to tetrahydrocannabinol (THC) content) or metformin as a positive control. For the obese rat model, the rats were allocated either a high carbohydrate diet (HCD) or high fat diet (HFD) and orally administered with cannabis extract of 1.25 mg/kg body weight (relative to THC content). Weight, blood and insulin-resistant parameters of the rats were monitored. The mitochondrial to genomic DNA ratio (MT:18S DNA), average adipocyte area of the various adipose tissues, citrate synthase and carnitine palmitoyltransferase 1 (CPT1) enzyme activities of the peritoneal and intramuscular fat were measured. Gene expression levels of uncoupling protein 1 (UCP1), cell-death inducing DNA fragmentation factor alpha like effector-a (Cidea), perilipin, hormone-sensitive lipase (HSL) and mitochondrial transcription factor A (TFAM) were measured in peritoneal fat, intramuscular fat and brown adipose tissue (BAT). Main Findings: Obese and STZ-induced diabetic rat model: Due to the biphasic nature of cannabinoids, cannabis dosage plays an important role in the observed effects. CE1.25 was the only cannabis treatment effective in improving the insulinresistant parameters of the rats unlike the other higher cannabis concentrations (CE2.5 and CE5.0). In the peritoneal fat, CE1.25 increased MT:18S DNA, increased citrate synthase activity, and decreased the average adipocyte area when compared to the STZ group. CE1.25 also induced fat beigeing by upregulating gene expression levels of UCP1 and Cidea. XIX Furthermore, an increase in gene expression levels of perilipin, HSL, and TFAM showed increased fat mobilization and metabolic activity. In the intramuscular fat, CE1.25 also reduced the average adipocytes area. However, a different mechanism of action was observed where CE1.25 did not induce fat beigeing, but instead increased both citrate synthase and CPT1 enzyme activities and gene expression levels of HSL, thereby indicating increased fat oxidation and mitochondrial activity. , Thesis (PhD) -- Faculty of Science, School of Biomolecular and Chemical Sciences, 2023
- Full Text:
- Date Issued: 2023-04
Interaction of catechol O-methyltransferase with gold and silver nanoparticles
- Authors: Usman, Aminu
- Date: 2018
- Subjects: Parkinson's disease , Methyltransferases , Catechol , Nanoparticles
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/61818 , vital:28063 , DOI https://doi.org/10.21504/10962/61818
- Description: Catechol O-methyltransferase (S-adenosyl-Z-methionine: catechol O-methyltransferase; COMT; EC 2.1.1.6) is a ubiquitous enzyme that catalyses the transfer of a methyl group from the cofactor, S-adenosyl-Z-methionine (SAM) to a hydroxyl group of endogenous and exogenous catechol-containing moieties. The physiological role of this enzyme is the methylation and thereby inactivation of the catechol-containing bio-active and bio-toxic compounds, including catechol-neurotransmitters, catechol-estrogens and catechol-containing drugs. Activity of this enzyme is implicated in the treatment of Parkinson’s disease and is associated with other diseases including breast cancer and an array neuropsychological disorders, such as schizophrenia. This thesis explores the use of gold and silver nanoparticles (NPs) (AuNPs and AgNPs) to inhibit the catalytic activity of mammalian COMT. Because of its accessibility and availability, we initially investigated bovine soluble COMT (BSCOMT) from liver tissue. Bioinformatic analyses and structural modeling revealed high (>90%) sequence similarity between BSCOMT and human soluble COMT (HSCOMT). BSCOMT was partially purified to 7.78 fold, 1.65% yield and had a specific activity of 0.052 U/mg. It had pH and temperature optima of 8.5 and 40oC, respectively. The Km, Vmax, Kcat and Kcat/Km towards esculetin methylation were respectively 1.475±0.130 pM, 0.0353±0.001 pmol/ml/min, 1.748 x 10-2±5.0x10-4 min-1 and 1.18x10-2 M-1. min-1. HSCOMT was expressed in Escherichia coli BL21(DE3) which showed optimal activity for esculetin methylation at pH and temperature of 7.0 and 30°C, respectively. It was purified to 5.62 fold, 22.6% yield with a specific activity of 3.85 U/mg. HSCOMT kinetic plots, upon incubation of the reaction mixture at 30°C for 5 min before addition of SAM was hyperbolic with Km, Vmax, Kcat and Kcat/Km values of 1.79 pM, 0.412 pmol/ml/min, 2.08 min-1 and 1.165 M-1. min-1, respectively. AuNPs and AgNPs showed a concentration dependent inhibition of HSCOMT activity upon increasing the 5 min incubation time to 1 h. Interestingly, HSCOMT kinetics, with 1 h incubation at 30°C, showed a sigmoidal curve, as well as increased activity. Incubation of the reaction mixture in the presence of 60 pM AuNPs and/or AgNPs for 1 hreversed the observed sigmoidal to a hyperbolic curve, with kinetic parameters comparable to those of 5 min incubation. SDS-PAGE analyses of HSCOMT after the kinetic experiments showed the enzyme incubated for 5 min as a monomer, while that which was incubated for 1 h migrated substantially as dimer. However, the HSCOMT incubated for 1 h in the presence of 60 pM AuNPs and/or AgNPs migrated as a monomer. This indicated that the extension of the incubation period allowed the dimerization of HSCOMT, which exhibited sigmoidal kinetics and higher activity. The presence of NPs impeded the HSCOMT dimerization which decreased the activity. Varying the concentration of SAM suggested that SAM had an allosteric modulatory effect on HSCOMT. Absorption spectroscopy indicated adsorption of HSCOMT on the gold and silver NP surfaces and the formation of NPs-HSCOMT corona. Fluorescence spectroscopy showed that the interaction of HSCOMT with both gold and silver NPs was governed by a static quenching mechanism, implying the formation of a non-fluorescent fluorophore-NP complex at the ground state. Further fluorometric analyses indicated that both gold and silver NPs had contact with Trp143; that the interactions were spontaneous and were driven by electrostatic interactions. Fourier transform infrared spectroscopic studies showed the adsorption of HSCOMT of the NPs surfaces to cause relaxation of the enzyme’s B-sheet structures. Molecular docking studies indicated involvement of largely hydrophilic amino acids, with the interacting distances of less than 3.5A. These findings signify the potential of nanotechnology in the control of COMT catalytic activity for the management of the COMT-related disorders. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2018
- Full Text:
- Date Issued: 2018
- Authors: Usman, Aminu
- Date: 2018
- Subjects: Parkinson's disease , Methyltransferases , Catechol , Nanoparticles
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/61818 , vital:28063 , DOI https://doi.org/10.21504/10962/61818
- Description: Catechol O-methyltransferase (S-adenosyl-Z-methionine: catechol O-methyltransferase; COMT; EC 2.1.1.6) is a ubiquitous enzyme that catalyses the transfer of a methyl group from the cofactor, S-adenosyl-Z-methionine (SAM) to a hydroxyl group of endogenous and exogenous catechol-containing moieties. The physiological role of this enzyme is the methylation and thereby inactivation of the catechol-containing bio-active and bio-toxic compounds, including catechol-neurotransmitters, catechol-estrogens and catechol-containing drugs. Activity of this enzyme is implicated in the treatment of Parkinson’s disease and is associated with other diseases including breast cancer and an array neuropsychological disorders, such as schizophrenia. This thesis explores the use of gold and silver nanoparticles (NPs) (AuNPs and AgNPs) to inhibit the catalytic activity of mammalian COMT. Because of its accessibility and availability, we initially investigated bovine soluble COMT (BSCOMT) from liver tissue. Bioinformatic analyses and structural modeling revealed high (>90%) sequence similarity between BSCOMT and human soluble COMT (HSCOMT). BSCOMT was partially purified to 7.78 fold, 1.65% yield and had a specific activity of 0.052 U/mg. It had pH and temperature optima of 8.5 and 40oC, respectively. The Km, Vmax, Kcat and Kcat/Km towards esculetin methylation were respectively 1.475±0.130 pM, 0.0353±0.001 pmol/ml/min, 1.748 x 10-2±5.0x10-4 min-1 and 1.18x10-2 M-1. min-1. HSCOMT was expressed in Escherichia coli BL21(DE3) which showed optimal activity for esculetin methylation at pH and temperature of 7.0 and 30°C, respectively. It was purified to 5.62 fold, 22.6% yield with a specific activity of 3.85 U/mg. HSCOMT kinetic plots, upon incubation of the reaction mixture at 30°C for 5 min before addition of SAM was hyperbolic with Km, Vmax, Kcat and Kcat/Km values of 1.79 pM, 0.412 pmol/ml/min, 2.08 min-1 and 1.165 M-1. min-1, respectively. AuNPs and AgNPs showed a concentration dependent inhibition of HSCOMT activity upon increasing the 5 min incubation time to 1 h. Interestingly, HSCOMT kinetics, with 1 h incubation at 30°C, showed a sigmoidal curve, as well as increased activity. Incubation of the reaction mixture in the presence of 60 pM AuNPs and/or AgNPs for 1 hreversed the observed sigmoidal to a hyperbolic curve, with kinetic parameters comparable to those of 5 min incubation. SDS-PAGE analyses of HSCOMT after the kinetic experiments showed the enzyme incubated for 5 min as a monomer, while that which was incubated for 1 h migrated substantially as dimer. However, the HSCOMT incubated for 1 h in the presence of 60 pM AuNPs and/or AgNPs migrated as a monomer. This indicated that the extension of the incubation period allowed the dimerization of HSCOMT, which exhibited sigmoidal kinetics and higher activity. The presence of NPs impeded the HSCOMT dimerization which decreased the activity. Varying the concentration of SAM suggested that SAM had an allosteric modulatory effect on HSCOMT. Absorption spectroscopy indicated adsorption of HSCOMT on the gold and silver NP surfaces and the formation of NPs-HSCOMT corona. Fluorescence spectroscopy showed that the interaction of HSCOMT with both gold and silver NPs was governed by a static quenching mechanism, implying the formation of a non-fluorescent fluorophore-NP complex at the ground state. Further fluorometric analyses indicated that both gold and silver NPs had contact with Trp143; that the interactions were spontaneous and were driven by electrostatic interactions. Fourier transform infrared spectroscopic studies showed the adsorption of HSCOMT of the NPs surfaces to cause relaxation of the enzyme’s B-sheet structures. Molecular docking studies indicated involvement of largely hydrophilic amino acids, with the interacting distances of less than 3.5A. These findings signify the potential of nanotechnology in the control of COMT catalytic activity for the management of the COMT-related disorders. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2018
- Full Text:
- Date Issued: 2018
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