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
Thiazole derivatives as potential hiv-1 protease inhibitors
- Authors: Hlongwe, Zola
- Date: 2021-04
- Subjects: Gqeberha (South Africa) , Eastern Cape (South Africa) , Enzyme kinetics
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10948/52116 , vital:43427
- Description: Series of compounds were screened using Schrodinger suite (Maestro). The DFT calculations were used for geometry optimization of the ligands using the B3YLYP functional and 6-31G basis set, and these structures were used for docking studies. Maestro was used to predict the activity of thiazole derivatives against HIV-1 protease. The range of estimated inhibition constants for these thiazole derivatives (65 nM-5 μM) indicate moderate to weak activity against HIV-1 protease, given that the activity of current protease inhibitors is typically found have experimental inhibition constants around 0.1-2.0 nM. Twenty compounds were selected based on the docking results and they were synthesized and characterized by NMR, FT-IR and elemental analysis. The cytotoxicity studies were done at two different concentrations (100 μM and 10 μM), using the brine shrimp bioassay. All compounds were highly toxic at 100 μM, with the percentage mortality between 20 to 75%. Eight compounds were selected for the enzyme bioassay based on the results obtained from lower concentration (10 μM). In the enzyme inhibition studies, the profile of HIV-1 activity was done at different inhibitor concentrations (800 μM – 10 μM) by measuring the cleavage of the synthetic substrate (Abz-Thr-lle-PNO2Phe=Gln-Arg-NH2) at excitation wavelength of 345/490 nm using fluorescence. Ligands 5 (unsubstituted derivative), 7 (4-nitro derivative) and 16 (4-methoxy derivative) gave percentage inhibition of 39, 45 and 42%, and this activity was very low compared to the activity of the positive control ritonavir (85% enzyme inhibition). Ligands 8 (4-methoxy derivative) and 12 (4-methoxy derivative) gave enzyme inhibition of 70% and 75%. These results suggest that the presence of the methoxy substituents ii increases activity of these compounds against HIV-1 protease. Most of the compounds gave good IC50 values between 12.5-42.7 nM. The bromo-substituted ligand 7 gave the lowest IC50 (12.5 nM). Ligand 11 also gave a good IC50 value of 14.86 nM. The bromo-substituted derivatives showed to be very active compared to other types of thiazole derivatives. Enzyme kinetics were carried out to compare the inhibition constants obtained via computational modelling. Ligand 7 (4-methoxy derivatives) binds better in the active site of HIV-1 protease than other compounds in Class B, with Ki = 50 nM, Km = 23.8 Nm and Vmax = 83.3 nM/min. The unsubstituted (L5), 4-bromo (L7) and 4-nitro (L8) substituted compounds gave inhibition constants of 100 to 112 nM. The in vitro testing yielded higher activity than that determined in silico. , Thesis (MSc) -- Faculty of Science, School of Biomolecular and Chemical Sciences, 2021
- Full Text: false
- Date Issued: 2021-04
- Authors: Hlongwe, Zola
- Date: 2021-04
- Subjects: Gqeberha (South Africa) , Eastern Cape (South Africa) , Enzyme kinetics
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10948/52116 , vital:43427
- Description: Series of compounds were screened using Schrodinger suite (Maestro). The DFT calculations were used for geometry optimization of the ligands using the B3YLYP functional and 6-31G basis set, and these structures were used for docking studies. Maestro was used to predict the activity of thiazole derivatives against HIV-1 protease. The range of estimated inhibition constants for these thiazole derivatives (65 nM-5 μM) indicate moderate to weak activity against HIV-1 protease, given that the activity of current protease inhibitors is typically found have experimental inhibition constants around 0.1-2.0 nM. Twenty compounds were selected based on the docking results and they were synthesized and characterized by NMR, FT-IR and elemental analysis. The cytotoxicity studies were done at two different concentrations (100 μM and 10 μM), using the brine shrimp bioassay. All compounds were highly toxic at 100 μM, with the percentage mortality between 20 to 75%. Eight compounds were selected for the enzyme bioassay based on the results obtained from lower concentration (10 μM). In the enzyme inhibition studies, the profile of HIV-1 activity was done at different inhibitor concentrations (800 μM – 10 μM) by measuring the cleavage of the synthetic substrate (Abz-Thr-lle-PNO2Phe=Gln-Arg-NH2) at excitation wavelength of 345/490 nm using fluorescence. Ligands 5 (unsubstituted derivative), 7 (4-nitro derivative) and 16 (4-methoxy derivative) gave percentage inhibition of 39, 45 and 42%, and this activity was very low compared to the activity of the positive control ritonavir (85% enzyme inhibition). Ligands 8 (4-methoxy derivative) and 12 (4-methoxy derivative) gave enzyme inhibition of 70% and 75%. These results suggest that the presence of the methoxy substituents ii increases activity of these compounds against HIV-1 protease. Most of the compounds gave good IC50 values between 12.5-42.7 nM. The bromo-substituted ligand 7 gave the lowest IC50 (12.5 nM). Ligand 11 also gave a good IC50 value of 14.86 nM. The bromo-substituted derivatives showed to be very active compared to other types of thiazole derivatives. Enzyme kinetics were carried out to compare the inhibition constants obtained via computational modelling. Ligand 7 (4-methoxy derivatives) binds better in the active site of HIV-1 protease than other compounds in Class B, with Ki = 50 nM, Km = 23.8 Nm and Vmax = 83.3 nM/min. The unsubstituted (L5), 4-bromo (L7) and 4-nitro (L8) substituted compounds gave inhibition constants of 100 to 112 nM. The in vitro testing yielded higher activity than that determined in silico. , Thesis (MSc) -- Faculty of Science, School of Biomolecular and Chemical Sciences, 2021
- Full Text: false
- Date Issued: 2021-04
Isolation and characterisation of a channel inhibitor from Bunodosoma capense
- Authors: Van Losenoord, Wynand
- Date: 2019
- Subjects: Bioactive compounds , Potassium channels Medical microbiology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/44134 , vital:37114
- Description: Voltage gated ion channels have recently become a subject of investigation as possible pharmaceutical targets. Research has linked the activity of ion channels directly to antiinflammatory pathways, energy homeostasis, cancer proliferation and painful diabetic neuropathy. Sea anemones secrete a diverse array of bioactive compounds including potassium and sodium channel inhibitors. A novel sodium channel inhibitor (molecular mass of 4619.7 ± 0.6 Da) with a predicted sequence: CLCNSDGPSV RGNTLSGILW LAGCPSGWHN CKKHKPTIGW CCK was isolated from Bunodosoma capense using a modified stimulation technique to induce the secretion of the neurotoxin rich mucus confirmed by an Artemia nauplii swimming assay. The peptide purification combined size-exclusion and reverse-phase high performance liquid chromatography. A thallium-based ion flux assay confirmed the presence of a sodium channel inhibitor and purity was determined using a modified tricine SDS-PAGE system. The peptide isolated indicated a tight conformation with the presence of multiple disulfide bonds in a cystine knot conformation. An IC50 value of 26 nM was determined for sodium channel inhibition on MCF-7 cells, indicating increased toxicity in comparison to sodium channel inhibitors previously isolated from Bunodosoma species.
- Full Text:
- Date Issued: 2019
- Authors: Van Losenoord, Wynand
- Date: 2019
- Subjects: Bioactive compounds , Potassium channels Medical microbiology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/44134 , vital:37114
- Description: Voltage gated ion channels have recently become a subject of investigation as possible pharmaceutical targets. Research has linked the activity of ion channels directly to antiinflammatory pathways, energy homeostasis, cancer proliferation and painful diabetic neuropathy. Sea anemones secrete a diverse array of bioactive compounds including potassium and sodium channel inhibitors. A novel sodium channel inhibitor (molecular mass of 4619.7 ± 0.6 Da) with a predicted sequence: CLCNSDGPSV RGNTLSGILW LAGCPSGWHN CKKHKPTIGW CCK was isolated from Bunodosoma capense using a modified stimulation technique to induce the secretion of the neurotoxin rich mucus confirmed by an Artemia nauplii swimming assay. The peptide purification combined size-exclusion and reverse-phase high performance liquid chromatography. A thallium-based ion flux assay confirmed the presence of a sodium channel inhibitor and purity was determined using a modified tricine SDS-PAGE system. The peptide isolated indicated a tight conformation with the presence of multiple disulfide bonds in a cystine knot conformation. An IC50 value of 26 nM was determined for sodium channel inhibition on MCF-7 cells, indicating increased toxicity in comparison to sodium channel inhibitors previously isolated from Bunodosoma species.
- Full Text:
- Date Issued: 2019
The synthesis, characterization, and application of peptide-capped magnetite nanoparticles for the targeting of cancer cells
- Authors: Hickson, Matthew Victor
- Date: 2019
- Subjects: Nanomedicine -- Research , Nanostructured materials Cancer -- Alternative treatment
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/40172 , vital:35965
- Description: In this study, a comparative analysis was performed upon three co-precipitation techniques for the synthesis of capped magnetite nanoparticles as to optimize the approach to the highest quality nanoparticles. Three techniques were applied whereby the capping agent either introduced before the stage of nanoparticle precipitation, simultaneously to the stage of precipitation, or after the stage of precipitation. The resultant nanoparticles were tested in terms of their size, dispersity, crystallinity, and magnetic characteristics. The three techniques gave nanoparticles of varying sizes and characteristics. Out of the three synthetic techniques, the post precipitation introduction method gave the highest quality nanoparticles in terms of size distribution, crystallinity and magnetic character. Three novel peptides were synthesized, incorporating amino acids to varying degrees. Structure was confirmed via IR and NMR spectroscopy. The peptides were studied potentiometrically to explore their acid nature and were explored computationally as to discern possible modes of interaction with the nanoparticles. These three peptides were further used in the capping of magnetite nanoparticles. For this set of nanoparticles, a higher synthesis temperature and larger iron content were used as to obtain larger nanoparticles. For the capping procedure, the post precipitation technique was used due to its previous positive results, once again yielding high quality nanoparticles with low size dispersity, high crystallinity, and high magnetic saturations. The nanoparticles were also seen to display positive zeta potentials, which are beneficial for cellular interactions. The peptides and peptide-capped nanoparticles were tested for biological activity against the healthy MCF-10A and cancerous MCF-7 cell lines. The MTT assay displayed increased proliferation for both the cell lines treated with the nanoparticles, while the peptide treatments decreased the MCF-10A cell lines proliferation and increased the MCF-7 proliferation. TEM analysis displayed nanoparticles in the cellular sections. An ICP-OES analysis on the cells showed that the capped nanoparticles of similar zeta potentials were seen to be taken up excessively by cells as compared to the uncapped. The nanoparticles of lower zeta potentials but higher L-glutamine content were taken up to a lesser degree.
- Full Text:
- Date Issued: 2019
- Authors: Hickson, Matthew Victor
- Date: 2019
- Subjects: Nanomedicine -- Research , Nanostructured materials Cancer -- Alternative treatment
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/40172 , vital:35965
- Description: In this study, a comparative analysis was performed upon three co-precipitation techniques for the synthesis of capped magnetite nanoparticles as to optimize the approach to the highest quality nanoparticles. Three techniques were applied whereby the capping agent either introduced before the stage of nanoparticle precipitation, simultaneously to the stage of precipitation, or after the stage of precipitation. The resultant nanoparticles were tested in terms of their size, dispersity, crystallinity, and magnetic characteristics. The three techniques gave nanoparticles of varying sizes and characteristics. Out of the three synthetic techniques, the post precipitation introduction method gave the highest quality nanoparticles in terms of size distribution, crystallinity and magnetic character. Three novel peptides were synthesized, incorporating amino acids to varying degrees. Structure was confirmed via IR and NMR spectroscopy. The peptides were studied potentiometrically to explore their acid nature and were explored computationally as to discern possible modes of interaction with the nanoparticles. These three peptides were further used in the capping of magnetite nanoparticles. For this set of nanoparticles, a higher synthesis temperature and larger iron content were used as to obtain larger nanoparticles. For the capping procedure, the post precipitation technique was used due to its previous positive results, once again yielding high quality nanoparticles with low size dispersity, high crystallinity, and high magnetic saturations. The nanoparticles were also seen to display positive zeta potentials, which are beneficial for cellular interactions. The peptides and peptide-capped nanoparticles were tested for biological activity against the healthy MCF-10A and cancerous MCF-7 cell lines. The MTT assay displayed increased proliferation for both the cell lines treated with the nanoparticles, while the peptide treatments decreased the MCF-10A cell lines proliferation and increased the MCF-7 proliferation. TEM analysis displayed nanoparticles in the cellular sections. An ICP-OES analysis on the cells showed that the capped nanoparticles of similar zeta potentials were seen to be taken up excessively by cells as compared to the uncapped. The nanoparticles of lower zeta potentials but higher L-glutamine content were taken up to a lesser degree.
- Full Text:
- Date Issued: 2019
The development of rhenium(III) oxide nanoradiopharmaceuticals
- Authors: Joseph, Sinelizwi Veronica
- Date: 2018
- Subjects: Radiopharmaceuticals , Radiopharmaceuticals Rhenium
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/30507 , vital:30955
- Description: The study details the experimental work on the development of rhenium(III) oxide nanoradiopharmaceuticals for imaging and therapy of disease states. The nanoparticles (NPs) were capped with covalently linked tetraaminophthalocyanine-folate and ethylenediamine-folate to enhance their targeting ability. The capping agents were successfully synthesised and structurally characterised using Ultraviolet-Visible Spectroscopy (UV-Vis), Fourier Transform-Infrared Spectroscopy (FT-IR), Proton Nuclear Magnetic Resonance (1H-NMR), and Liquid Chromatography-Mass Spectroscopy (LC-MS). The nanoparticles were characterised using UV-Vis, spectrofluorimetry, Transmission Electron Microscopy (TEM), dynamic light scattering (DLS) and Zeta potential. Nanoparticles of sizes between 10 and 100 nm size were envisaged to be suitable for applications in biological systems. The preferred surface charge for the uptake of NPs must be between -30 and +30 mV, Re2O3 NPs capped with ethylenediamine were found to have a surface charge of -49 mV as compared with NPs capped with ethylenediamine-folate which gave -18.6 mV. The cytotoxicity studies of the nanoparticles were tested against four different cell lines: MDA-MB-468, MDA-MB-231, MCF-7, and MCF-10A. The cell survival rate after treatment was done with different capped rhenium(III) oxide nanoparticles obtained at a 10 μM concentration showed more than 80% cell viability. A comparison was conducted based on different nanoparticle sizes of capping agents across the four cell lines of varying folate receptor. All the cell lines were compared, and it was observed that MCF-7 had high percentage of cell viability especially with the cells treated with folate conjugated nanoparticles. Further investigation was done on the effects of folate conjugates and the effects of size. It was observed that the tetraaminophthalocyanine-folate favoured the MCF-7, for large-sized nanoparticles. However, further work is required to test the cancer cell internalisation of the nanoparticles using TEM as well as the correct size for endocytosis. Thereafter, the mice model study will be carried out for investigation of biodistribution of particles in tumour tissue using hot isotopes (186/188Re) and this will be done in a radiophamarceutical laboratory.
- Full Text:
- Date Issued: 2018
- Authors: Joseph, Sinelizwi Veronica
- Date: 2018
- Subjects: Radiopharmaceuticals , Radiopharmaceuticals Rhenium
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/30507 , vital:30955
- Description: The study details the experimental work on the development of rhenium(III) oxide nanoradiopharmaceuticals for imaging and therapy of disease states. The nanoparticles (NPs) were capped with covalently linked tetraaminophthalocyanine-folate and ethylenediamine-folate to enhance their targeting ability. The capping agents were successfully synthesised and structurally characterised using Ultraviolet-Visible Spectroscopy (UV-Vis), Fourier Transform-Infrared Spectroscopy (FT-IR), Proton Nuclear Magnetic Resonance (1H-NMR), and Liquid Chromatography-Mass Spectroscopy (LC-MS). The nanoparticles were characterised using UV-Vis, spectrofluorimetry, Transmission Electron Microscopy (TEM), dynamic light scattering (DLS) and Zeta potential. Nanoparticles of sizes between 10 and 100 nm size were envisaged to be suitable for applications in biological systems. The preferred surface charge for the uptake of NPs must be between -30 and +30 mV, Re2O3 NPs capped with ethylenediamine were found to have a surface charge of -49 mV as compared with NPs capped with ethylenediamine-folate which gave -18.6 mV. The cytotoxicity studies of the nanoparticles were tested against four different cell lines: MDA-MB-468, MDA-MB-231, MCF-7, and MCF-10A. The cell survival rate after treatment was done with different capped rhenium(III) oxide nanoparticles obtained at a 10 μM concentration showed more than 80% cell viability. A comparison was conducted based on different nanoparticle sizes of capping agents across the four cell lines of varying folate receptor. All the cell lines were compared, and it was observed that MCF-7 had high percentage of cell viability especially with the cells treated with folate conjugated nanoparticles. Further investigation was done on the effects of folate conjugates and the effects of size. It was observed that the tetraaminophthalocyanine-folate favoured the MCF-7, for large-sized nanoparticles. However, further work is required to test the cancer cell internalisation of the nanoparticles using TEM as well as the correct size for endocytosis. Thereafter, the mice model study will be carried out for investigation of biodistribution of particles in tumour tissue using hot isotopes (186/188Re) and this will be done in a radiophamarceutical laboratory.
- Full Text:
- Date Issued: 2018
The effect of ER stress in INS-1E cells using IL-1β under hyperglycaemic conditions
- Authors: Jackson, Simon
- Date: 2018
- Subjects: Diabetes -- Pathogenesis , Endoplasmic reticulum -- Pathophysiology Endoplasmic reticulum Diabetes
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/30397 , vital:30938
- Description: Diabetes afflicts millions of individuals worldwide, and the statistics rise each year. It is associated with various ailments which reduce the quality of life and has been shown to be associated with seemingly unrelated diseases or conditions. One aspect of the pathogenesis of diabetes is endoplasmic reticulum (ER) stress, which is an imbalance in the protein loading and protein folding capacities of the ER of a cell. Under chronic hyperglycaemic conditions associated with the development of diabetes, excessive insulin production disrupts the ER homeostasis, leading to ER stress. If the ER stress is severe or chronic, cell death of pancreatic β-cells may occur, leading to the onset of diabetes. There is currently a gap in available models that closely resemble the pathogenesis of diabetes in insulinoma-1E (INS-1E) pancreatic β-cells to study ER stress under hyperglycaemic conditions. This study optimised various ER stress-inducing models and tested terpenoid treatments to investigate their potential in modulating ER stress. Using various cell viability assays, five models to induce ER stress were optimised (hyperglycaemic, interleukin-1β (IL-1β), tunicamycin (Tm), brefeldin-A (BFA) and thapsigargin (Tg)). The five models were shown to induce ER stress through the expression of the downstream ER stress marker CCAAT-enhancer binding protein (C/EBP) homologous protein (CHOP). The various models induced ER stress under different mechanisms. Insulin secretion analysis using enzyme-linked immunosorbent assays (ELISAs) demonstrated that low concentrations of IL-1β promoted insulin secretion. Several of the terpenoid treatments showed potential in alleviating different aspects of either ER stress or inflammation.
- Full Text:
- Date Issued: 2018
- Authors: Jackson, Simon
- Date: 2018
- Subjects: Diabetes -- Pathogenesis , Endoplasmic reticulum -- Pathophysiology Endoplasmic reticulum Diabetes
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/30397 , vital:30938
- Description: Diabetes afflicts millions of individuals worldwide, and the statistics rise each year. It is associated with various ailments which reduce the quality of life and has been shown to be associated with seemingly unrelated diseases or conditions. One aspect of the pathogenesis of diabetes is endoplasmic reticulum (ER) stress, which is an imbalance in the protein loading and protein folding capacities of the ER of a cell. Under chronic hyperglycaemic conditions associated with the development of diabetes, excessive insulin production disrupts the ER homeostasis, leading to ER stress. If the ER stress is severe or chronic, cell death of pancreatic β-cells may occur, leading to the onset of diabetes. There is currently a gap in available models that closely resemble the pathogenesis of diabetes in insulinoma-1E (INS-1E) pancreatic β-cells to study ER stress under hyperglycaemic conditions. This study optimised various ER stress-inducing models and tested terpenoid treatments to investigate their potential in modulating ER stress. Using various cell viability assays, five models to induce ER stress were optimised (hyperglycaemic, interleukin-1β (IL-1β), tunicamycin (Tm), brefeldin-A (BFA) and thapsigargin (Tg)). The five models were shown to induce ER stress through the expression of the downstream ER stress marker CCAAT-enhancer binding protein (C/EBP) homologous protein (CHOP). The various models induced ER stress under different mechanisms. Insulin secretion analysis using enzyme-linked immunosorbent assays (ELISAs) demonstrated that low concentrations of IL-1β promoted insulin secretion. Several of the terpenoid treatments showed potential in alleviating different aspects of either ER stress or inflammation.
- Full Text:
- Date Issued: 2018
The effects of terpenoids on the expression and function of cytokines and adipokines in pre-adipocytes and differentiated adipocytes
- Authors: Bloom, Carri-Ann
- Date: 2017
- Subjects: Terpenes , Cytokines , Fat cells
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/9208 , vital:26479
- Description: CURRENTLY UNDER EMBARGO UNTIL THE 26/4/2019: Type 2 diabetes is a metabolic disorder characterised by inflammation, insulin resistance and the inability of pancreatic β-cells to secrete enough insulin to produce a physiological effect. Obesity and high levels of triacylglycerol’s are associated with the development of Type 2 diabetes. Adipose tissue is an active endocrine organ that secretes various protein and peptide hormones, known as adipokines, which mediate important metabolic functions. In an insulin resistant and hyperglycaemic state, levels of anti-inflammatory adipokines, adiponectin, are reduced, whereas levels of pro-inflammatory cytokines, interleukin-6 and interleukin-1β, are elevated; this results in a shift from an anti- to a pro-inflammatory state that is accompanied by dysfunction and apoptosis of the pancreatic β-cells. Cannabis sativa L. has been traditionally used as an anti-inflammatory agent in Southern Africa, specifically treating snakebites, fever and malaria. Δ9-tetrahydrocannabinol is the main psychoactive compound derived from C. sativa, whereas the other major cannabinoids, cannabinol and cannabidiol, have shown anti-inflammatory and sedative properties respectively. Marrubiin is a compound derived from the plant Leonotis leonurus L. and has been traditionally used as an anti-inflammatory and anti-diabetic agent. To determine the effects of these compounds in a hyperglycaemic state, pre- and differentiated mouse adipocytes (3T3-L1 cells) were exposed for seven and fourteen days to the following treatments: Δ9-tetrahydrocannabinol, cannabidiol, cannabinol, marrubiin, anandamide (an endogenous endocannabinoid) and cannabis extract, individually and in combination, under normal glucose and hyperglycaemic conditions. Levels of adiponectin, interleukin-6, leptin, tumour-necrosis factor-α and interleukin-1β were quantified using mouse enzyme-linked immunosorbent assay kits and Oil Red O staining was carried out to determine lipid distribution and lipid droplet characteristics. Results indicate that various cannabinoids, in combination, mediate an anti-inflammatory effect by decreasing the expression of various pro-inflammatory cytokines, which may have allowed for a shift from a pro- to an anti-inflammatory state by these compounds, and may also contribute to the reduction of lipid, which may be used as a supplementary option to current diabetic treatment regimes.
- Full Text: false
- Date Issued: 2017
- Authors: Bloom, Carri-Ann
- Date: 2017
- Subjects: Terpenes , Cytokines , Fat cells
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/9208 , vital:26479
- Description: CURRENTLY UNDER EMBARGO UNTIL THE 26/4/2019: Type 2 diabetes is a metabolic disorder characterised by inflammation, insulin resistance and the inability of pancreatic β-cells to secrete enough insulin to produce a physiological effect. Obesity and high levels of triacylglycerol’s are associated with the development of Type 2 diabetes. Adipose tissue is an active endocrine organ that secretes various protein and peptide hormones, known as adipokines, which mediate important metabolic functions. In an insulin resistant and hyperglycaemic state, levels of anti-inflammatory adipokines, adiponectin, are reduced, whereas levels of pro-inflammatory cytokines, interleukin-6 and interleukin-1β, are elevated; this results in a shift from an anti- to a pro-inflammatory state that is accompanied by dysfunction and apoptosis of the pancreatic β-cells. Cannabis sativa L. has been traditionally used as an anti-inflammatory agent in Southern Africa, specifically treating snakebites, fever and malaria. Δ9-tetrahydrocannabinol is the main psychoactive compound derived from C. sativa, whereas the other major cannabinoids, cannabinol and cannabidiol, have shown anti-inflammatory and sedative properties respectively. Marrubiin is a compound derived from the plant Leonotis leonurus L. and has been traditionally used as an anti-inflammatory and anti-diabetic agent. To determine the effects of these compounds in a hyperglycaemic state, pre- and differentiated mouse adipocytes (3T3-L1 cells) were exposed for seven and fourteen days to the following treatments: Δ9-tetrahydrocannabinol, cannabidiol, cannabinol, marrubiin, anandamide (an endogenous endocannabinoid) and cannabis extract, individually and in combination, under normal glucose and hyperglycaemic conditions. Levels of adiponectin, interleukin-6, leptin, tumour-necrosis factor-α and interleukin-1β were quantified using mouse enzyme-linked immunosorbent assay kits and Oil Red O staining was carried out to determine lipid distribution and lipid droplet characteristics. Results indicate that various cannabinoids, in combination, mediate an anti-inflammatory effect by decreasing the expression of various pro-inflammatory cytokines, which may have allowed for a shift from a pro- to an anti-inflammatory state by these compounds, and may also contribute to the reduction of lipid, which may be used as a supplementary option to current diabetic treatment regimes.
- Full Text: false
- Date Issued: 2017
Leonotis leonurus: understanding the mechanism of anti-diabetic action and investigating a nano drug delivery system
- Odei-Addo, Frank, Levendal, Ruby-Ann
- Authors: Odei-Addo, Frank , Levendal, Ruby-Ann
- Date: 2016
- Subjects: Diabetes Plant extracts
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10948/13116 , vital:27153
- Description: Diabetes mellitus is a metabolic disease characterised by hyperglycaemia resulting from defects in insulin secretion, insulin action, or both. The leaf extract of Leonotis leonurus and its active compound marrubiin, have been shown to possess anti-diabetic, antiplatelet, anti-inflammatory and anti-coagulation activity. In the present study, the mechanism by which L. leonurus and marrubiin exert their anti-diabetic properties, the cross-talk between the peripheral tissues and a nano drug delivery system were investigated. Marrubiin in the plant extract was effectively quantified by an optimised reversed phase highperformance liquid chromatography (HPLC) protocol using a pentafluorophenyl (PFP) column with water and acetonitrile (50:50) as mobile phase, and a flow rate of 1ml/min. The chemical structure was determined using liquid chromatography-tandem mass spectroscopy LC-MS/MS. Real-time quantitative polymerase chain reaction (RT-qPCR) gene expression of selected adipokines and proteins implicated in Type-2 diabetes (T2D) were investigated in specific peripheral tissues isolated from an in vivo obese rat model. An in vitro cell culture model was used to determine the crosstalk between the peripheral tissues and pancreatic (INS-1E) β-cells. Various nanoformulations of L. leonurus extract were prepared and their effect on cytotoxicity (in Chang liver and INS-1 cells), insulin-mediated glucose uptake (Change liver cells) and insulin secretion (INS-1) were investigated. The average yield of marrubiin from the plant extract was 10% (n=3), with a molecular mass of 333.20Da and a molecular formula of C20H29O4 +. Results from the in vivo study showed that the L. leonurus extract significantly (p<0.05) enhanced the gene expression of adiponectin, peroxisome proliferator-activated receptor gamma (PPAR-γ), glucokinase (GK), uncoupling protein-2 (UCP-2) and reduced leptin in adipose tissue, but resistin, glucose transporters (GLUT), fatty acid synthase (FAS), insulin receptor substrate -1 (IRS-1) and phosphoenolpyruvate carboxykinase (PEPCK) gene expression were not affected. Marrubiin decreased gene expression of leptin and resistin, and increased IRS-1 and glucokinase in adipose tissue. In liver and muscle tissues, marrubiin and the L. leonurus extract reduced gene expression of PPAR-γ, IRS-1, glucokinase and PEPCK. In the in vitro crosstalk study (under normoglycaemic and hyperglycaemic conditions), conditioned medium from 3T3-L1 cells significantly (p<0.01) enhanced insulin secretion. This was not observed in INS-1E cells exposed to muscle- and liver-conditioned medium, respectively. The in vitro studies using a nanostructured lipid formulation (NLC) of the plant extract was not cytotoxic to either INS-1 and Chang liver cells. The NLC formulation significantly (p<0.05) enhanced glucose uptake in Chang liver cells and improved chronic insulin release in INS-1 cells (p<0.05). Based on the above findings from the in vivo and in vitro studies, both L. leonurus and marrubiin exerted an insulinotropic effect via adipose tissue on pancreatic β-cells. The findings in the in vivo study showed that marrubiin and the L. leonurus extract were employing their major anti-diabetic action via the adipose tissue.
- Full Text:
- Date Issued: 2016
- Authors: Odei-Addo, Frank , Levendal, Ruby-Ann
- Date: 2016
- Subjects: Diabetes Plant extracts
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10948/13116 , vital:27153
- Description: Diabetes mellitus is a metabolic disease characterised by hyperglycaemia resulting from defects in insulin secretion, insulin action, or both. The leaf extract of Leonotis leonurus and its active compound marrubiin, have been shown to possess anti-diabetic, antiplatelet, anti-inflammatory and anti-coagulation activity. In the present study, the mechanism by which L. leonurus and marrubiin exert their anti-diabetic properties, the cross-talk between the peripheral tissues and a nano drug delivery system were investigated. Marrubiin in the plant extract was effectively quantified by an optimised reversed phase highperformance liquid chromatography (HPLC) protocol using a pentafluorophenyl (PFP) column with water and acetonitrile (50:50) as mobile phase, and a flow rate of 1ml/min. The chemical structure was determined using liquid chromatography-tandem mass spectroscopy LC-MS/MS. Real-time quantitative polymerase chain reaction (RT-qPCR) gene expression of selected adipokines and proteins implicated in Type-2 diabetes (T2D) were investigated in specific peripheral tissues isolated from an in vivo obese rat model. An in vitro cell culture model was used to determine the crosstalk between the peripheral tissues and pancreatic (INS-1E) β-cells. Various nanoformulations of L. leonurus extract were prepared and their effect on cytotoxicity (in Chang liver and INS-1 cells), insulin-mediated glucose uptake (Change liver cells) and insulin secretion (INS-1) were investigated. The average yield of marrubiin from the plant extract was 10% (n=3), with a molecular mass of 333.20Da and a molecular formula of C20H29O4 +. Results from the in vivo study showed that the L. leonurus extract significantly (p<0.05) enhanced the gene expression of adiponectin, peroxisome proliferator-activated receptor gamma (PPAR-γ), glucokinase (GK), uncoupling protein-2 (UCP-2) and reduced leptin in adipose tissue, but resistin, glucose transporters (GLUT), fatty acid synthase (FAS), insulin receptor substrate -1 (IRS-1) and phosphoenolpyruvate carboxykinase (PEPCK) gene expression were not affected. Marrubiin decreased gene expression of leptin and resistin, and increased IRS-1 and glucokinase in adipose tissue. In liver and muscle tissues, marrubiin and the L. leonurus extract reduced gene expression of PPAR-γ, IRS-1, glucokinase and PEPCK. In the in vitro crosstalk study (under normoglycaemic and hyperglycaemic conditions), conditioned medium from 3T3-L1 cells significantly (p<0.01) enhanced insulin secretion. This was not observed in INS-1E cells exposed to muscle- and liver-conditioned medium, respectively. The in vitro studies using a nanostructured lipid formulation (NLC) of the plant extract was not cytotoxic to either INS-1 and Chang liver cells. The NLC formulation significantly (p<0.05) enhanced glucose uptake in Chang liver cells and improved chronic insulin release in INS-1 cells (p<0.05). Based on the above findings from the in vivo and in vitro studies, both L. leonurus and marrubiin exerted an insulinotropic effect via adipose tissue on pancreatic β-cells. The findings in the in vivo study showed that marrubiin and the L. leonurus extract were employing their major anti-diabetic action via the adipose tissue.
- Full Text:
- Date Issued: 2016
The role of cancer procoagulant on the MTOR pathway
- Authors: Chiuswa, Chengetanai
- Date: 2016
- Subjects: Cancer -- Research , Neovascularization , Biochemistry , Blood coagulation factors -- Biotechnology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/7303 , vital:21316
- Description: Cancer procoagulant (CP) is a cysteine protease found in tumour cells and amnion chorion membranes. The main function of CP is not yet known, but it has potential roles in tumour growth and metastasis. Initially, CP was believed to increase coagulation in cancer patients; however, research has shown that increase in CP concentration does not correlate with an increase in coagulation. The location of CP in amnion chorion membranes and tumour cells only led to the hypothesis that CP might be involved in inducing blood vessel and or lymph vessel formation. CP was shown to induce lymphangiogenesis (lymph vessel formation) in human telomerase reverse transcriptase-human dermal endothelial cells (hTERT-HDLEC) (Tshaka, 2011). CP-induced tube formation was inhibited by rapamycin; indicating that CP may be signalling via the mammalian target of rapamycin (mTOR) pathway. The aim of this study was to investigate the effect of CP on the mTOR signalling pathway using human umbilical endothelial vein cells (HUVECs) as a model. CP was isolated from amnion chorion membranes and purified using two anion exchange chromatography steps. Purified CP (2 μg/ml) was used to induce tube formation in endothelial cells (HUVECs) seeded on growth factor reduced (GFR) Matrigel. In addition, the 2 μg/ml CP was used to treat cultured HUVECs. Sodium dodecyl sulphate polyacrylamide electrophoresis (SDS-PAGE) and western blotting were used to determine phosphorylation levels of protein kinase B (Akt) and ribosomal protein S6 kinase (S6K). CP was successfully isolated and purified using anion exchange chromatography. The effect of CP on tube formation was not significant relative to the control in the HUVEC cell line. The role of CP on Akt and S6K phosphorylation still needs to be verified by using sensitive methods of quantification such as enhanced chemiluminescence (ECL) and enzyme linked immunosorbent assay (ELISA). The levels of CP activity were shown to be higher in early tumour growth than in advanced cancer suggesting that certain physiological factors could be increasing CP activity during early tumour growth. This study investigated the effect of cobalt chloride on CP activity in breast cancer cell lines. Cobalt chloride reduced CP activity in MCF-7 and promoted CP activity in MDA-MB-231.
- Full Text:
- Date Issued: 2016
- Authors: Chiuswa, Chengetanai
- Date: 2016
- Subjects: Cancer -- Research , Neovascularization , Biochemistry , Blood coagulation factors -- Biotechnology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/7303 , vital:21316
- Description: Cancer procoagulant (CP) is a cysteine protease found in tumour cells and amnion chorion membranes. The main function of CP is not yet known, but it has potential roles in tumour growth and metastasis. Initially, CP was believed to increase coagulation in cancer patients; however, research has shown that increase in CP concentration does not correlate with an increase in coagulation. The location of CP in amnion chorion membranes and tumour cells only led to the hypothesis that CP might be involved in inducing blood vessel and or lymph vessel formation. CP was shown to induce lymphangiogenesis (lymph vessel formation) in human telomerase reverse transcriptase-human dermal endothelial cells (hTERT-HDLEC) (Tshaka, 2011). CP-induced tube formation was inhibited by rapamycin; indicating that CP may be signalling via the mammalian target of rapamycin (mTOR) pathway. The aim of this study was to investigate the effect of CP on the mTOR signalling pathway using human umbilical endothelial vein cells (HUVECs) as a model. CP was isolated from amnion chorion membranes and purified using two anion exchange chromatography steps. Purified CP (2 μg/ml) was used to induce tube formation in endothelial cells (HUVECs) seeded on growth factor reduced (GFR) Matrigel. In addition, the 2 μg/ml CP was used to treat cultured HUVECs. Sodium dodecyl sulphate polyacrylamide electrophoresis (SDS-PAGE) and western blotting were used to determine phosphorylation levels of protein kinase B (Akt) and ribosomal protein S6 kinase (S6K). CP was successfully isolated and purified using anion exchange chromatography. The effect of CP on tube formation was not significant relative to the control in the HUVEC cell line. The role of CP on Akt and S6K phosphorylation still needs to be verified by using sensitive methods of quantification such as enhanced chemiluminescence (ECL) and enzyme linked immunosorbent assay (ELISA). The levels of CP activity were shown to be higher in early tumour growth than in advanced cancer suggesting that certain physiological factors could be increasing CP activity during early tumour growth. This study investigated the effect of cobalt chloride on CP activity in breast cancer cell lines. Cobalt chloride reduced CP activity in MCF-7 and promoted CP activity in MDA-MB-231.
- Full Text:
- Date Issued: 2016
The antifungal activity of an aqueous Tulbaghia violacea plant extract against Aspergillus flavus
- Authors: Belewa, Xoliswa Vuyokazi
- Date: 2015
- Subjects: Medicinal plants , Antifungal agents , Fungi -- Biotechnology
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10948/5858 , vital:21001
- Description: Phytochemical analysis of both HEA1 and the crude plant extract showed the presence of phenolics, tannins and saponins. Saponins were the predominant secondary metabolites and were mostly abundant in the plant extract and to a lesser extent in the active compound. Steroidal saponins, tannins and phenolics were also detected in the plant extract, but only the phenolics were detected in the active compound. The results of the phytochemical analysis showed that those compounds that were not present in the active compound could be removed from the crude extract during the TLC purification process. Investigation on the mechanism of action of the crude plant extract on the sterol production by A. flavus showed that the plant extract affected ergosterol biosynthesis by causing an accumulation of oxidosqualene in the ergosterol biosynthetic pathway resulting in a decline in ergosterol production. An oscillatory response in lanosterol production was observed in the presence of the plant extract, which may be an adaptation mechanism of A. flavus to unfavourable conditions and compensation for the loss of enzyme activity which may have occurred as a result of the accumulation of oxidosqualene. The antifungal activity of the plant extract on ergosterol production by A. flavus may also be due to saponins which target the cell membrane and ergosterol production in fungi. The effect of the plant extract on the fungal cell wall of A. flavus also showed that the plant extract caused a decline in β-(1, 3) glucan production by inhibiting β-glucan synthase. The plant extract also affected the chitin synthesis pathway of A. flavus, by causing a decline in chitin production, which was due to the inhibition of chitin synthase. Investigation of chitinase production using 4MU substrates showed that the plant extract caused an accumulation of chitobioses, by activating chitobiosidases and endochitinases. A decline in N-acetylglucosaminidase activity in the presence of the plant extract was observed and this prevented the formation of N-acetylglucosamine. The accumulation of chitobiosidase and endochitinase may be as a result of autolysis that may be triggered by A. flavus as a survival mechanism in the presence of the plant extract and as a compensatory mechanism for the loss of β-glucans and chitin. The antifungal effect of the plant extract on various components of the cell wall of A. flavus, makes T. violacea aqueous plant extract an ideal chemotherapeutic agent against both human and plant pathogens of Aspergillus. The broad spectrum of antifungal activity of T. violacea against A. flavus also eliminates any chances of the fungus developing resistance towards it and would make it a candidate for use as a potential antifungal agent. Further identification and possible chemical synthesis is needed to shed light on the safety and efficacy of the active compound for further development as a chemotherapeutic agent.
- Full Text:
- Date Issued: 2015
- Authors: Belewa, Xoliswa Vuyokazi
- Date: 2015
- Subjects: Medicinal plants , Antifungal agents , Fungi -- Biotechnology
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10948/5858 , vital:21001
- Description: Phytochemical analysis of both HEA1 and the crude plant extract showed the presence of phenolics, tannins and saponins. Saponins were the predominant secondary metabolites and were mostly abundant in the plant extract and to a lesser extent in the active compound. Steroidal saponins, tannins and phenolics were also detected in the plant extract, but only the phenolics were detected in the active compound. The results of the phytochemical analysis showed that those compounds that were not present in the active compound could be removed from the crude extract during the TLC purification process. Investigation on the mechanism of action of the crude plant extract on the sterol production by A. flavus showed that the plant extract affected ergosterol biosynthesis by causing an accumulation of oxidosqualene in the ergosterol biosynthetic pathway resulting in a decline in ergosterol production. An oscillatory response in lanosterol production was observed in the presence of the plant extract, which may be an adaptation mechanism of A. flavus to unfavourable conditions and compensation for the loss of enzyme activity which may have occurred as a result of the accumulation of oxidosqualene. The antifungal activity of the plant extract on ergosterol production by A. flavus may also be due to saponins which target the cell membrane and ergosterol production in fungi. The effect of the plant extract on the fungal cell wall of A. flavus also showed that the plant extract caused a decline in β-(1, 3) glucan production by inhibiting β-glucan synthase. The plant extract also affected the chitin synthesis pathway of A. flavus, by causing a decline in chitin production, which was due to the inhibition of chitin synthase. Investigation of chitinase production using 4MU substrates showed that the plant extract caused an accumulation of chitobioses, by activating chitobiosidases and endochitinases. A decline in N-acetylglucosaminidase activity in the presence of the plant extract was observed and this prevented the formation of N-acetylglucosamine. The accumulation of chitobiosidase and endochitinase may be as a result of autolysis that may be triggered by A. flavus as a survival mechanism in the presence of the plant extract and as a compensatory mechanism for the loss of β-glucans and chitin. The antifungal effect of the plant extract on various components of the cell wall of A. flavus, makes T. violacea aqueous plant extract an ideal chemotherapeutic agent against both human and plant pathogens of Aspergillus. The broad spectrum of antifungal activity of T. violacea against A. flavus also eliminates any chances of the fungus developing resistance towards it and would make it a candidate for use as a potential antifungal agent. Further identification and possible chemical synthesis is needed to shed light on the safety and efficacy of the active compound for further development as a chemotherapeutic agent.
- Full Text:
- Date Issued: 2015
The detection of glyphosate and glyphosate-based herbicides in water, using nanotechnology
- De Almeida, Louise Kashiyavala Sophia
- Authors: De Almeida, Louise Kashiyavala Sophia
- Date: 2015
- Subjects: Water -- Glyphosate content , Aquatic herbicides -- South Africa , Aquatic herbicides -- Physiological effect , Nanotechnology , Invasive plants -- South Africa , Genetic toxicology , Thiazoles , Tetrazolium , Immunotoxicology , Colorimetry , Nanofibers
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4163 , http://hdl.handle.net/10962/d1019755
- Description: Glyphosate (N-phosphonomethylglycine) is an organophosphate compound which was developed by the Monsanto Company in 1971 and is the active ingredient found in several herbicide formulations. The use of glyphosate-based herbicides in South Africa for the control of alien invasive plants and weeds is well established, extensive and currently unregulated, which vastly increases the likelihood of glyphosate contamination in environmental water systems. Although the use of glyphosate-based herbicides is required for economic enhancement in industries such as agriculture, the presence of this compound in natural water systems presents a potential risk to human health. Glyphosate and glyphosate formulations were previously considered safe, however their toxicity has become a major focal point of research over recent years. The lack of monitoring protocols for pesticides in South Africa is primarily due to limited financial capacity and the lack of analytical techniques.
- Full Text:
- Date Issued: 2015
- Authors: De Almeida, Louise Kashiyavala Sophia
- Date: 2015
- Subjects: Water -- Glyphosate content , Aquatic herbicides -- South Africa , Aquatic herbicides -- Physiological effect , Nanotechnology , Invasive plants -- South Africa , Genetic toxicology , Thiazoles , Tetrazolium , Immunotoxicology , Colorimetry , Nanofibers
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4163 , http://hdl.handle.net/10962/d1019755
- Description: Glyphosate (N-phosphonomethylglycine) is an organophosphate compound which was developed by the Monsanto Company in 1971 and is the active ingredient found in several herbicide formulations. The use of glyphosate-based herbicides in South Africa for the control of alien invasive plants and weeds is well established, extensive and currently unregulated, which vastly increases the likelihood of glyphosate contamination in environmental water systems. Although the use of glyphosate-based herbicides is required for economic enhancement in industries such as agriculture, the presence of this compound in natural water systems presents a potential risk to human health. Glyphosate and glyphosate formulations were previously considered safe, however their toxicity has become a major focal point of research over recent years. The lack of monitoring protocols for pesticides in South Africa is primarily due to limited financial capacity and the lack of analytical techniques.
- Full Text:
- Date Issued: 2015
The effect of phytocannabinoid treatment on adipogenesis and lipolysis in 3T3-L1 cells
- Authors: Ramlugon, Sonaal
- Date: 2014
- Subjects: Cannabinoids , Adipose tissues , Cannabis -- Therapeutic use
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10361 , http://hdl.handle.net/10948/d1021072
- Description: During the 1800’s cannabis use was described as a treatment for a variety of metabolic disorders but its recreational use in the twentieth century resulted in laws which made the usage of cannabis illegal despite its medicinal properties. Cannabis usage has been reported to be useful in the treatment of Type 2 diabetes but unfortunately conflicting results are often published and its mechanism of action is still unknown. The aim of this project was to investigate the effect of phytocannabinoid treatment on adipogenesis and lipolysis in 3T3-L1 cells, to unravel their mechanism of action and also to test for potential anti-diabetic properties. The studies showed that phytocannabinoid treatment promoted higher glucose uptake and significantly less fat accumulation when compared to Rosiglitazone. Rosiglitazone is an anti-diabetic drug that has recently been withdrawn from the market since its usage has been associated with severe side effects. It was also found that during the 1800’s cannabis use was described as a treatment for a variety of metabolic disorders but its recreational use in the twentieth century resulted in laws which made the usage of cannabis illegal despite its medicinal properties. Cannabis usage has been reported to be useful in the treatment of Type 2 diabetes but unfortunately conflicting results are often published and its mechanism of action is still unknown. The aim of this project was to investigate the effect of phytocannabinoid treatment on adipogenesis and lipolysis in 3T3-L1 cells, to unravel their mechanism of action and also to test for potential anti-diabetic properties. The studies showed that phytocannabinoid treatment promoted higher glucose uptake and significantly less fat accumulation when compared to Rosiglitazone. Rosiglitazone is an anti-diabetic drug that has recently been withdrawn from the market since its usage has been associated with severe side effects. It was also found that phytocannabinoid treatment was able to reverse the insulin-resistant state of 3T3-L1 cells. The study indicates that the mechanism of action occurs at the mitochondrial level where enzymes such as succinate dehydrogenase and glycerol-3-phosphate dehydrogenase are modulated thereby affecting oxidative phosphorylation involved in the respiratory chain. In addition the effect observed with phytocannabinoid treatment is time dependent and affects the cells differently at different developmental stages. Therefore it can be concluded that phytocannabinoid treatment not only helps to maintain the balance between adipogenesis and lipolysis in 3T3-L1 cells but its use may also be helpful in the treatment of Type 2 diabetes and/or obesity-related insulin resistance.phytocannabinoid treatment was able to reverse the insulin-resistant state of 3T3-L1 cells. The study indicates that the mechanism of action occurs at the mitochondrial level where enzymes such as succinate dehydrogenase and glycerol-3-phosphate dehydrogenase are modulated thereby affecting oxidative phosphorylation involved in the respiratory chain. In addition the effect observed with phytocannabinoid treatment is time dependent and affects the cells differently at different developmental stages. Therefore it can be concluded that phytocannabinoid treatment not only helps to maintain the balance between adipogenesis and lipolysis in 3T3-L1 cells but its use may also be helpful in the treatment of Type 2 diabetes and/or obesity-related insulin resistance.
- Full Text:
- Date Issued: 2014
- Authors: Ramlugon, Sonaal
- Date: 2014
- Subjects: Cannabinoids , Adipose tissues , Cannabis -- Therapeutic use
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10361 , http://hdl.handle.net/10948/d1021072
- Description: During the 1800’s cannabis use was described as a treatment for a variety of metabolic disorders but its recreational use in the twentieth century resulted in laws which made the usage of cannabis illegal despite its medicinal properties. Cannabis usage has been reported to be useful in the treatment of Type 2 diabetes but unfortunately conflicting results are often published and its mechanism of action is still unknown. The aim of this project was to investigate the effect of phytocannabinoid treatment on adipogenesis and lipolysis in 3T3-L1 cells, to unravel their mechanism of action and also to test for potential anti-diabetic properties. The studies showed that phytocannabinoid treatment promoted higher glucose uptake and significantly less fat accumulation when compared to Rosiglitazone. Rosiglitazone is an anti-diabetic drug that has recently been withdrawn from the market since its usage has been associated with severe side effects. It was also found that during the 1800’s cannabis use was described as a treatment for a variety of metabolic disorders but its recreational use in the twentieth century resulted in laws which made the usage of cannabis illegal despite its medicinal properties. Cannabis usage has been reported to be useful in the treatment of Type 2 diabetes but unfortunately conflicting results are often published and its mechanism of action is still unknown. The aim of this project was to investigate the effect of phytocannabinoid treatment on adipogenesis and lipolysis in 3T3-L1 cells, to unravel their mechanism of action and also to test for potential anti-diabetic properties. The studies showed that phytocannabinoid treatment promoted higher glucose uptake and significantly less fat accumulation when compared to Rosiglitazone. Rosiglitazone is an anti-diabetic drug that has recently been withdrawn from the market since its usage has been associated with severe side effects. It was also found that phytocannabinoid treatment was able to reverse the insulin-resistant state of 3T3-L1 cells. The study indicates that the mechanism of action occurs at the mitochondrial level where enzymes such as succinate dehydrogenase and glycerol-3-phosphate dehydrogenase are modulated thereby affecting oxidative phosphorylation involved in the respiratory chain. In addition the effect observed with phytocannabinoid treatment is time dependent and affects the cells differently at different developmental stages. Therefore it can be concluded that phytocannabinoid treatment not only helps to maintain the balance between adipogenesis and lipolysis in 3T3-L1 cells but its use may also be helpful in the treatment of Type 2 diabetes and/or obesity-related insulin resistance.phytocannabinoid treatment was able to reverse the insulin-resistant state of 3T3-L1 cells. The study indicates that the mechanism of action occurs at the mitochondrial level where enzymes such as succinate dehydrogenase and glycerol-3-phosphate dehydrogenase are modulated thereby affecting oxidative phosphorylation involved in the respiratory chain. In addition the effect observed with phytocannabinoid treatment is time dependent and affects the cells differently at different developmental stages. Therefore it can be concluded that phytocannabinoid treatment not only helps to maintain the balance between adipogenesis and lipolysis in 3T3-L1 cells but its use may also be helpful in the treatment of Type 2 diabetes and/or obesity-related insulin resistance.
- Full Text:
- Date Issued: 2014
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