Spatiotemporal ecology and potential displacement of white sharks in southern Africa
- Authors: Towner, Alison
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/432275 , vital:72857
- Description: Restricted access. Expected release date in 2025. , Thesis (PhD) -- Faculty of Science, Ichthyology and Fisheries Science, 2023
- Full Text:
- Date Issued: 2023-10-13
- Authors: Towner, Alison
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/432275 , vital:72857
- Description: Restricted access. Expected release date in 2025. , Thesis (PhD) -- Faculty of Science, Ichthyology and Fisheries Science, 2023
- Full Text:
- Date Issued: 2023-10-13
Fucoidans from South African brown seaweeds: establishing the link between their structure and biological properties (anti-diabetic and anti-cancer activities)
- Authors: Mabate, Blessing
- Date: 2022-10-14
- Subjects: Fucoidan , Diabetes Treatment , Cancer Treatment , Brown algae
- Language: English
- Type: Academic theses , Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/365677 , vital:65775 , DOI https://doi.org/10.21504/10962/365677
- Description: Type 2 diabetes mellitus (T2DM) and cancer are major non-communicable diseases causing a heavy morbidity-mortality and economic burden globally. The therapeutic efforts in managing these diseases are primarily chemotherapeutic and are associated with demerits, including side effects and toxicity, limiting the prescribed amounts. These dosage limits may cause drug resistance, another major challenge in maintaining quality global health. The pursuit of novel natural bioproducts is a reasonable strategy to add to the arsenal against T2DM and cancer. Fucoidans, sulphated fucose polysaccharides abundant in brown seaweeds, have recently become popular for their biological activities, including anti-diabetic and anti-cancer properties. However, endemic South African brown seaweeds have not been adequately explored. Therefore, this study sought to characterise fucoidans extracted from South African brown seaweeds and elucidate their structure to their biological activities. Also, this study highlighted carbohydrate and glucose metabolism as major target processes in the control efforts of T2DM and cancer using fucoidans. Harvested brown seaweeds were identified as Ecklonia radiata and Sargassum elegans. E. maxima was kindly donated by KelpX. The fucoidans were then extracted using hot water, EDTA assisted, and acid extraction protocols. The integrity of the extracted fucoidan was confirmed through structural analysis using FTIR, NMR and TGA. The fucoidan extracts were then chemically characterised to determine their carbohydrate and monosaccharide composition and sulphate content. The characterised fucoidans were profiled for inhibiting the major amylolytic enzymes, namely α-amylase and α-glucosidase. The mode of inhibition by fucoidans and synergy experiments with the commercial anti-diabetic drug acarbose were also investigated. Furthermore, the fucoidans were screened for potential anti-cancer activities on the human colorectal HCT116 cancer cell line. The cytotoxicity of fucoidans was quantified using the resazurin assay. The effect of fucoidan on HCT116 cell adhesion on the tissue culture plastic was also investigated using the crystal violet-based cell adhesion assay. In addition, cancer antimigration properties of fucoidans were also investigated using 2D wound healing and 3D spheroid-based assays. Furthermore, the long-term survival of HCT116 cells was investigated through the clonogenic assay after treatment with fucoidans. Lastly, glucose uptake and lactate export assays revealed the influence of fucoidan on glucose uptake and the glycolytic flux of HCT116 cells. Fucoidans were successfully extracted with a yield between 2.2% and 14.2% on a dry weight basis. EDTA extracts produced the highest yields than the water and the acid extracts. Ecklonia spp. fucoidans displayed the highest total carbohydrate content, with glucose and galactose being the major monosaccharides. S. elegans and commercial Fucus vesiculosus had lower carbohydrate contents but contained more sulphates than the Ecklonia spp. fucoidans. Furthermore, the extracted fucoidan contained little to no contaminants, including proteins, phenolics and uronic acids. In addition, the extracted fucoidans were determined to be >100 kDa through ultracentrifugation. Mass spectrometry also detected the most abundant peak for all fucoidans to be around 700 Da (m/z). Extracted fucoidans inhibited the activity of α-glucosidase more strongly than the commercial anti-diabetic agent acarbose but were inactive on α-amylase. Fucoidans were also shown to be mixed inhibitors of α-glucosidase. Compellingly, fucoidans synergistically inhibited α-glucosidase in combination with the anti-diabetic agent acarbose, highlighting prospects for combination therapy. Finally, fucoidans demonstrated some anti-proliferative characteristics on HCT116 cancer cells by inhibiting their ability to adhere to the tissue culture plate matrix. Furthermore, some fucoidan extracts inhibited the migration of HCT116 cancer cells from 3D spheroids. Some of our fucoidan extracts also inhibited HCT116 colony formation, demonstrating inhibition of long-term cell survival. The E. maxima water extract also inhibited glucose uptake by HCT116 cells, thereby influencing the glycolytic flux. In conclusion, biologically active fucoidans were successfully extracted from South African brown seaweeds. These fucoidans demonstrated anti-diabetic and anti-cancer properties, revealing their relevance as potential drugs for these diseases. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2022
- Full Text:
- Date Issued: 2022-10-14
- Authors: Mabate, Blessing
- Date: 2022-10-14
- Subjects: Fucoidan , Diabetes Treatment , Cancer Treatment , Brown algae
- Language: English
- Type: Academic theses , Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/365677 , vital:65775 , DOI https://doi.org/10.21504/10962/365677
- Description: Type 2 diabetes mellitus (T2DM) and cancer are major non-communicable diseases causing a heavy morbidity-mortality and economic burden globally. The therapeutic efforts in managing these diseases are primarily chemotherapeutic and are associated with demerits, including side effects and toxicity, limiting the prescribed amounts. These dosage limits may cause drug resistance, another major challenge in maintaining quality global health. The pursuit of novel natural bioproducts is a reasonable strategy to add to the arsenal against T2DM and cancer. Fucoidans, sulphated fucose polysaccharides abundant in brown seaweeds, have recently become popular for their biological activities, including anti-diabetic and anti-cancer properties. However, endemic South African brown seaweeds have not been adequately explored. Therefore, this study sought to characterise fucoidans extracted from South African brown seaweeds and elucidate their structure to their biological activities. Also, this study highlighted carbohydrate and glucose metabolism as major target processes in the control efforts of T2DM and cancer using fucoidans. Harvested brown seaweeds were identified as Ecklonia radiata and Sargassum elegans. E. maxima was kindly donated by KelpX. The fucoidans were then extracted using hot water, EDTA assisted, and acid extraction protocols. The integrity of the extracted fucoidan was confirmed through structural analysis using FTIR, NMR and TGA. The fucoidan extracts were then chemically characterised to determine their carbohydrate and monosaccharide composition and sulphate content. The characterised fucoidans were profiled for inhibiting the major amylolytic enzymes, namely α-amylase and α-glucosidase. The mode of inhibition by fucoidans and synergy experiments with the commercial anti-diabetic drug acarbose were also investigated. Furthermore, the fucoidans were screened for potential anti-cancer activities on the human colorectal HCT116 cancer cell line. The cytotoxicity of fucoidans was quantified using the resazurin assay. The effect of fucoidan on HCT116 cell adhesion on the tissue culture plastic was also investigated using the crystal violet-based cell adhesion assay. In addition, cancer antimigration properties of fucoidans were also investigated using 2D wound healing and 3D spheroid-based assays. Furthermore, the long-term survival of HCT116 cells was investigated through the clonogenic assay after treatment with fucoidans. Lastly, glucose uptake and lactate export assays revealed the influence of fucoidan on glucose uptake and the glycolytic flux of HCT116 cells. Fucoidans were successfully extracted with a yield between 2.2% and 14.2% on a dry weight basis. EDTA extracts produced the highest yields than the water and the acid extracts. Ecklonia spp. fucoidans displayed the highest total carbohydrate content, with glucose and galactose being the major monosaccharides. S. elegans and commercial Fucus vesiculosus had lower carbohydrate contents but contained more sulphates than the Ecklonia spp. fucoidans. Furthermore, the extracted fucoidan contained little to no contaminants, including proteins, phenolics and uronic acids. In addition, the extracted fucoidans were determined to be >100 kDa through ultracentrifugation. Mass spectrometry also detected the most abundant peak for all fucoidans to be around 700 Da (m/z). Extracted fucoidans inhibited the activity of α-glucosidase more strongly than the commercial anti-diabetic agent acarbose but were inactive on α-amylase. Fucoidans were also shown to be mixed inhibitors of α-glucosidase. Compellingly, fucoidans synergistically inhibited α-glucosidase in combination with the anti-diabetic agent acarbose, highlighting prospects for combination therapy. Finally, fucoidans demonstrated some anti-proliferative characteristics on HCT116 cancer cells by inhibiting their ability to adhere to the tissue culture plate matrix. Furthermore, some fucoidan extracts inhibited the migration of HCT116 cancer cells from 3D spheroids. Some of our fucoidan extracts also inhibited HCT116 colony formation, demonstrating inhibition of long-term cell survival. The E. maxima water extract also inhibited glucose uptake by HCT116 cells, thereby influencing the glycolytic flux. In conclusion, biologically active fucoidans were successfully extracted from South African brown seaweeds. These fucoidans demonstrated anti-diabetic and anti-cancer properties, revealing their relevance as potential drugs for these diseases. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2022
- Full Text:
- Date Issued: 2022-10-14
Synthesis, In-Silico molecular modelling and biological studies of 1,4-Dihydroxyanthraquinone and its derivatives
- Authors: Kisula, Lydia Mboje
- Date: 2022-10-14
- Subjects: Computer simulation , Molecules Models , Dihydroxyanthraquinone , Trypanosomiasis , Leishmaniasis , Docking
- Language: English
- Type: Academic theses , Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/365828 , vital:65793 , DOI https://doi.org/10.21504/10962/365828
- Description: This current study of investigation reports on the synthesis of 1,4-dihydroxyanthraquinone and its derivatives on explorations of their medicinal potential. The study initially aimed to synthesize an analogue of a natural anthraquinone, 1,3,6-trihydroxy-7-((S)-1- hydroxyethyl)anthracene-9,10-dione 5 using Friedel-Crafts acylation of phthalic anhydride and a benzene derivative. Synthetic transformation of anacardic acid 63, obtained as a by- product of the cashew industry successfully afforded 4-ethoxyisobenzofuran-1,3-dione 89. However, when attempted to couple 4-ethoxyisobenzofuran-1,3-dione 89 with 2- hydroxyacetophenone 91 in a Friedel-Crafts acylation manner to form 2-acetyl-1,8- dihydroxyanthracene-9,10-dione 87 the reaction did not work efficiently. A simple derivative of benzene which is; benzene-1,4-diol 102 was reacted instead with 3-ethoxyphthalic acid 71 and isobenzofuran-1,3-dione 96 to form 1,4,5-trihydroxy anthraquinone 72 and 1,4- dihydroxyanthraquinone 42, respectively. A modified Marschalk reaction was then used to introduce the hydroxyl alkyl group to 1,4-dihydroxy anthraquinone 42, which allowed further elaboration of the hydroxyl-substituent in moderate to good yields (22-80%). A molecular docking study was performed using Schrödinger software to predict the binding affinity of the test compounds to the target protein trypanothione reductase (PDB ID: 6BU7). An in-vitro screening of 1,4-dihydroxyanthraquinone derivatives and some selected precursors for antitrypanosomal, antiplasmodial, antibacterial, and cytotoxicity activities produced encouraging results. Derivatives of anacardic acid and cardanol from CNSL were found to have moderate activity against trypanosomes with no activity against Plasmodium falciparum. Almost 63% of synthesized 1,4-dihydroxyanthraquinone derivatives displayed activity against trypanosomes. The in-vitro evaluation and the in silico molecular docking studies revealed that 1,4-dihydroxyanthraquinone derivatives can be potential drug-like candidates active against T.brucei parasites (IC50 = 0.70-1.20 μM). Only four 1,4- iv dihydroxyanthraquinone derivatives with thiosemicarbazone, chloride, pyrrole, and diethanolamine functionality displayed activity against Plasmodium falciparum (IC50 = 3.17- 14.36 μM). In-vitro evaluated of test compounds against antibacterial screen and cytotoxicity effects significantly showed that 2-hydroxy-6-pentadecylbenzoic acid 63a and 2-((2- chlorophenyl)(piperazin-1-yl) methyl)-1,4-dihydroxyanthracene-9,10-dione 78 have potency against Staphylococcus aureus and reduced the viability of the cells below 20% at an initial concentration of 50 μg/mL. Only 1,4-dihydroxyanthraquinone derivatives with thiosemicarbazone 76, piperazine 78, and diethanolamine 80 motifs were active against HeLa cells and reduced the viability of cells below 20% at a concentration of 50 μg/mL. In conclusion, this current reported study has generated useful knowlege on the applicability of the agro-waste CNSL as an agent active against trypanosomiasis but also as a low-cost starting material to synthesize hydroxy anthraquinones. The study has further given an overview to the understanding of the medicinal value 1,4-dihydroxyanthraquinone derivatives as promising candidates towards developing drugs suitable for treating neglected tropical diseases particularly trypanosomiasis. , Thesis (PhD) -- Faculty of Science, Chemistry, 2022
- Full Text:
- Date Issued: 2022-10-14
- Authors: Kisula, Lydia Mboje
- Date: 2022-10-14
- Subjects: Computer simulation , Molecules Models , Dihydroxyanthraquinone , Trypanosomiasis , Leishmaniasis , Docking
- Language: English
- Type: Academic theses , Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/365828 , vital:65793 , DOI https://doi.org/10.21504/10962/365828
- Description: This current study of investigation reports on the synthesis of 1,4-dihydroxyanthraquinone and its derivatives on explorations of their medicinal potential. The study initially aimed to synthesize an analogue of a natural anthraquinone, 1,3,6-trihydroxy-7-((S)-1- hydroxyethyl)anthracene-9,10-dione 5 using Friedel-Crafts acylation of phthalic anhydride and a benzene derivative. Synthetic transformation of anacardic acid 63, obtained as a by- product of the cashew industry successfully afforded 4-ethoxyisobenzofuran-1,3-dione 89. However, when attempted to couple 4-ethoxyisobenzofuran-1,3-dione 89 with 2- hydroxyacetophenone 91 in a Friedel-Crafts acylation manner to form 2-acetyl-1,8- dihydroxyanthracene-9,10-dione 87 the reaction did not work efficiently. A simple derivative of benzene which is; benzene-1,4-diol 102 was reacted instead with 3-ethoxyphthalic acid 71 and isobenzofuran-1,3-dione 96 to form 1,4,5-trihydroxy anthraquinone 72 and 1,4- dihydroxyanthraquinone 42, respectively. A modified Marschalk reaction was then used to introduce the hydroxyl alkyl group to 1,4-dihydroxy anthraquinone 42, which allowed further elaboration of the hydroxyl-substituent in moderate to good yields (22-80%). A molecular docking study was performed using Schrödinger software to predict the binding affinity of the test compounds to the target protein trypanothione reductase (PDB ID: 6BU7). An in-vitro screening of 1,4-dihydroxyanthraquinone derivatives and some selected precursors for antitrypanosomal, antiplasmodial, antibacterial, and cytotoxicity activities produced encouraging results. Derivatives of anacardic acid and cardanol from CNSL were found to have moderate activity against trypanosomes with no activity against Plasmodium falciparum. Almost 63% of synthesized 1,4-dihydroxyanthraquinone derivatives displayed activity against trypanosomes. The in-vitro evaluation and the in silico molecular docking studies revealed that 1,4-dihydroxyanthraquinone derivatives can be potential drug-like candidates active against T.brucei parasites (IC50 = 0.70-1.20 μM). Only four 1,4- iv dihydroxyanthraquinone derivatives with thiosemicarbazone, chloride, pyrrole, and diethanolamine functionality displayed activity against Plasmodium falciparum (IC50 = 3.17- 14.36 μM). In-vitro evaluated of test compounds against antibacterial screen and cytotoxicity effects significantly showed that 2-hydroxy-6-pentadecylbenzoic acid 63a and 2-((2- chlorophenyl)(piperazin-1-yl) methyl)-1,4-dihydroxyanthracene-9,10-dione 78 have potency against Staphylococcus aureus and reduced the viability of the cells below 20% at an initial concentration of 50 μg/mL. Only 1,4-dihydroxyanthraquinone derivatives with thiosemicarbazone 76, piperazine 78, and diethanolamine 80 motifs were active against HeLa cells and reduced the viability of cells below 20% at a concentration of 50 μg/mL. In conclusion, this current reported study has generated useful knowlege on the applicability of the agro-waste CNSL as an agent active against trypanosomiasis but also as a low-cost starting material to synthesize hydroxy anthraquinones. The study has further given an overview to the understanding of the medicinal value 1,4-dihydroxyanthraquinone derivatives as promising candidates towards developing drugs suitable for treating neglected tropical diseases particularly trypanosomiasis. , Thesis (PhD) -- Faculty of Science, Chemistry, 2022
- Full Text:
- Date Issued: 2022-10-14
Understanding the replication biology of Providence virus: elucidating the function of non-structural proteins
- Authors: Nakayinga, Ritah
- Date: 2014
- Subjects: Insects Viruses , Viruses Reproduction , Tombusviridae , RNA viruses , RNA polymerases
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/193930 , vital:45408
- Description: Tetraviruses are non-enveloped, small insect RNA viruses with a single stranded positive RNA genome that is either monopartite or bipartite. Providence virus (PrV) is the only member of the three tetravirus families with a viral replicase similar to the replicases of tombusviruses and umbraviruses. The principle aim of this thesis was to study PrV replication, focusing on subcellular localization and potential interactions between PrV replication proteins. The first objective of this study was to generate an anti-p104 antibody that does not cross-react with p40. Expression of the C-terminal portion of p104 in E. coli resulted in no detectable protein. Further expression in an insect cell based expression system resulted in the production of an insoluble protein. Attempts to improve protein solubility with a range of solubilization treatments were unsuccessful. Bioinformatic analysis was used to detect an antigenic region at the C-terminus of p104 and the peptide was used to raise anti-p104 antibodies. These antibodies did not detect native protein by western blot detection however they were used for immunoprecipitation. The establishment of the subcellular localization of PrV required two approaches; immunofluorescence in persistently infected Helicoverpa zea MG8 cells using antip40 and anti-dsRNA antibodies and the expression of EGFP-replicase fusion protein in Spodoptera frugiperda Sf9 cells. Replication of PrV was found to take place in cytosolic punctate structures. Co-immunoprecipitation experiments revealed that p40 self-interacts and interacts with p104. Bioinformatic analysis of PrV p104 suggests that the RdRp is similar to viral RdRps of the carmo-like supergroup II. Potential RNA binding regions are present within p104. A potential p40 interaction domain that shares hydrophilic and surface exposed properties with the TBSV p33 interaction domain is present. A putative arginine-rich region and disordered C-terminal region is present in p130. In conclusion, PrV p104 is the viral replicase. The resemblance of the expression strategy and putative functional domains with tombusviruses and umbraviruses suggest that PrV replication is related to the replication system of the tombusviruses and umbraviruses. This has led to propose that tetravirus replication strategies are diverse and raises questions on the origin and evolution of PrV. , Thesis (PhD) -- Faculty of Science, Biochemistry, Microbiology and Biotechnology, 2014
- Full Text:
- Date Issued: 2014
- Authors: Nakayinga, Ritah
- Date: 2014
- Subjects: Insects Viruses , Viruses Reproduction , Tombusviridae , RNA viruses , RNA polymerases
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/193930 , vital:45408
- Description: Tetraviruses are non-enveloped, small insect RNA viruses with a single stranded positive RNA genome that is either monopartite or bipartite. Providence virus (PrV) is the only member of the three tetravirus families with a viral replicase similar to the replicases of tombusviruses and umbraviruses. The principle aim of this thesis was to study PrV replication, focusing on subcellular localization and potential interactions between PrV replication proteins. The first objective of this study was to generate an anti-p104 antibody that does not cross-react with p40. Expression of the C-terminal portion of p104 in E. coli resulted in no detectable protein. Further expression in an insect cell based expression system resulted in the production of an insoluble protein. Attempts to improve protein solubility with a range of solubilization treatments were unsuccessful. Bioinformatic analysis was used to detect an antigenic region at the C-terminus of p104 and the peptide was used to raise anti-p104 antibodies. These antibodies did not detect native protein by western blot detection however they were used for immunoprecipitation. The establishment of the subcellular localization of PrV required two approaches; immunofluorescence in persistently infected Helicoverpa zea MG8 cells using antip40 and anti-dsRNA antibodies and the expression of EGFP-replicase fusion protein in Spodoptera frugiperda Sf9 cells. Replication of PrV was found to take place in cytosolic punctate structures. Co-immunoprecipitation experiments revealed that p40 self-interacts and interacts with p104. Bioinformatic analysis of PrV p104 suggests that the RdRp is similar to viral RdRps of the carmo-like supergroup II. Potential RNA binding regions are present within p104. A potential p40 interaction domain that shares hydrophilic and surface exposed properties with the TBSV p33 interaction domain is present. A putative arginine-rich region and disordered C-terminal region is present in p130. In conclusion, PrV p104 is the viral replicase. The resemblance of the expression strategy and putative functional domains with tombusviruses and umbraviruses suggest that PrV replication is related to the replication system of the tombusviruses and umbraviruses. This has led to propose that tetravirus replication strategies are diverse and raises questions on the origin and evolution of PrV. , Thesis (PhD) -- Faculty of Science, Biochemistry, Microbiology and Biotechnology, 2014
- Full Text:
- Date Issued: 2014
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