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Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnology, Durban University of Technology, 2007 xi, 114 leaves / Eighteen traditionally leafy vegetables consumed as food or medicinal compounds by a majority of people in the KwaZulu Natal province of South Africa were analysed for the presence of potentially harmful chemicals (antinutrients) and for their toxicity and mutagenicity. The purpose of the study was to determine whether leafy vegetables were safe for human consumption. Chemical analysis showed that none of the vegetables contained cyanogenic glycosides, however all the vegetables contained oxalic acid ranging from 24.1 mg/ml to 798.2 mg/ml with Solanum nigrum, Portulaca oleracea and Mormodica balsamina showing the highest concentrations. Most of the vegetables contained negligible amounts of phytic acid and saponins, except for Momordica balsamina (3.01 mg/ml and 1.83 mg/ml, respectively). Fourteen of the plants contained alkaloids with Portulaca oleracea having the highest content (1.53 g total alkaloids/5 g leaf material). Eight of the plants were found to inhibit trypsin activity. These chemical analyses were carried out in duplicate and the mean and standard deviation were used. The Ames test revealed that none of the leafy vegetables produced a mutagenic frequency above 1, except 10 000 µg/ml organic extract of Senna occidentalis (mutagenecity considered at mutagenic frequency above 2), thus none were considered mutagenic. All 18 organic extracts did not kill off more than 50% brine shrimp and were thus considered non-toxic. On the other hand the aqueous extracts of seven vegetables, namely, Physalis viscosa, Amaranthus dubius, Justicia flava, Bidens pilosa, Senna occidentalis, Chenopodium album and Ceratotheca triloba, killed more than 50% of the shrimp and are thus considered toxic above 100 µg/ml. The MTT assay carried out on the organic extracts indicated that 17 vegetables did not kill off more than 50% of HepG2 cells and were thus considered non-cytotoxic. The aqueous extracts of four vegetables, namely, Justicia flava, Asystasia gangetica, Momordica balsamin and Senna occidentalis, however killed more than 50% of the shrimp and were thus considered cytotoxic above 1 000 µg/ml. It may be concluded from the antinutrient analyses and the bioassays on the 18 vegetables that caution needs to be maintained with the consumption of certain leafy vegetables included in this study, especially Senna occidentalis.

UDP-sugar metabolizing pyrophosphorylases provide the primary mechanism for de novo synthesis of UDP-sugars, which can then be used for myriads of glycosyltranferase reactions, producing cell wall carbohydrates, sucrose, glycoproteins and glycolipids, as well as many other glycosylated compounds. The pyrophosphorylases can be divided into three families: UDP-Glc pyrophosphorylase (UGPase), UDP-sugar pyrophosphorylase (USPase) and UDP-N-acety lglucosamine pyrophosphorylase (UAGPase), which can be discriminated both by differences in accepted substrate range and amino acid sequences. This thesis focuses both on experimental examination (and re-examination) of some enzymatic/ biochemical properties of selected members of the UGPases and USPases and UAGPase families and on the design and implementation of a strategy to study in vivo roles of these pyrophosphorylases using specific inhibitors. In the first part, substrate specificities of members of the Arabidopsis UGPase, USPase and UAGPase families were comprehensively surveyed and kinetically analyzed, with barley UGPase also further studied with regard to itspH dependency, regulation by oligomerization, etc. Whereas all the enzymes preferentially used UTP as nucleotide donor, they differed in their specificity for sugar-1-P. UGPases had high activity with D-Glc-1-P, but could also react with Frc-1-P, whereas USPase reacted with arange of sugar-1-phosphates, including D-Glc-1-P, D-Gal-1-P, D-GalA-1-P, β-L-Ara-1-P and α-D-Fuc-1-P. In contrast, UAGPase2 reacted only with D-GlcNAc-1-P, D-GalNAc-1-P and, to some extent, with D-Glc-1-P. A structure activity relationship was established to connect enzyme activity, the examined sugar-1-phosphates and the three pyrophosphorylases. The UGPase/USPase/UAGPase active sites were subsequently compared in an attempt to identify amino acids which may contribute to the experimentally determined differences in substrate specificities. The second part of the thesis deals with identification and characterization of inhibitors of the pyrophosphorylases and with studies on in vivo effects of those inhibitors in Arabidopsis-based systems. A novel luminescence-based high-throughput assay system was designed, which allowed for quantitative measurement of UGPase and USPase activities, down to a pmol per min level. The assay was then used to screen a chemical library (which contained 17,500 potential inhibitors) to identify several compounds affecting UGPase and USPase. Hit-optimization on one of the compounds revealed even stronger inhibitors of UGPase and USPase which also strongly inhibited Arabidopsis pollen germination, by disturbing UDP-sugar metabolism. The inhibitors may represent useful tools to study in vivo roles of the pyrophosphorylases, as a complement to previous genetics-based studies. The thesis also includes two review papers on mechanisms of synthesis of NDP-sugars. The first review covered the characterization of USPase from both prokaryotic and eukaryotic organisms, whereas the second review was a comprehensive survey of NDP-sugar producing enzymes (not only UDP-sugar producing and not only pyrophosphorylases). All these enzymes were discussed with respect to their substrate specificities and structural features (if known) and their proposed in vivo functions.

D. Tech. (Department of Chemistry, Faculty of Applied and Computer Sciences)|, Vaal University of Technology. / Medicinal plants provide valuable alternative sources of drugs and drug discovery because many have been used in traditional practices for centuries to manage or treat various forms of ailments. The aim of this study was to evaluate the biological activities of nine medicinal plants used by Zulus in Mabandla village, KwaZulu-Natal province, South Africa to treat inflammation and to isolate selected active compounds against studied pathogens from Alepidea amatymbica and Pomaria sandersonii. The plants were selected on the basis of an ethnobotanical survey based on questionnaire response and verbal interviews that were conducted in Mabandla village with the local traditional healers and herbalists. The isolation of compounds from Alepidea amatymbica and Pomaria sandersonii was based on the bioassay based study which was carried out in this study. Bioassay guided study involving in vitro anti-inflammatory measurement using soya bean derived 15 Lipoxygenase, free radical scavenging capacity against the ABTS●+ radical cation and DPPH● radicals; antimicrobial and bioautography assays against Staphylococcus aureus, ATCC 29213, Pseudomonas aeruginosa ATCC 27853, Enterococcus faecalis ATCC 29212, Escherichia coli, ATCC25922, Candida albicans, Cryptococcus neoformans and Aspergillus fumigatus were carried out using the plants extracts, fractions and pure compounds. Isolation of compounds displaying biological activity was carried out by using open column chromatography and preparative thin layer chromatography (PTLC). The compounds were characterised by use of Nuclear Magnetic resonance, (NMR) and Mass Spectrometry (MS). The DPPH sprayed TLC showed that all the nine plants contained antioxidants. Most of which were contained in polar fractions of acetone and methanol. Results of the assays displayed a range of biological activities comparable to the positive controls used for each assay. DPPH● scavenging displayed EC50 values ranging between 1.008 and 467 Kg/ml. The highest activity was observed with the methanol fraction of Berkheya setifera with an EC50 value of 1.008 Kg/ml followed by the crude extract of Gunnera perpensa with EC50 value of 1.069 Kg/ml. Carissa bispinosa hexane fraction had the lowest activity of 467.7 Kg/ml. The Pomaria sandersonii DCM extract had the highest ABTS●+ radical scavenging activity by Pomaria sandersonii DCM extract, (1.273 Kg/ml) for the ethyl acetate, (5.973 Kg/ml) while the hexane fraction from Eucomis autumnalis had the lowest activity (929.4 Kg/ml). The activity of Pomaria sandersonii extracts and fractions demonstrated that the plant contains antioxidants that react with both DPPH and ABTS radicals although higher activities were shown by ABTS as displayed by the lower EC50 values. All the crude fractions and extracts had high to moderate antibacterial activities (20-625 Kg/ml) and anti-fungal activities (20-2500 Kg /ml). Pomaria sandersonii crude and fractions had the highest antimicrobial activity compared to other plants. Some MIC values for P. sandersonii dichloromethane and ethyl acetate fractions (80 Kg/ml in each case) compared well with gentamycin (4 Kg/ml) since they showed same values against Staphylococcus aureus, Enterococcus faecalis, Escherichia coli and Pseudonomus aeruginosa. The dichloromethane, acetone and methanol fractions were also active (20 Kg/ml) against both Candida albicans and Aspergillus fumigatus. Inhibition of pathogen growth demonstrated by the polar fractions of the studied plants suggested that some of the active compounds would be soluble in water. A total of seven compounds were isolated from Alepidea amatymbica and Pomaria sandersonii. We propose three were new compounds after considering literature search involving closely related research to this investigation. These were two diterpenes from Alepidea amatymbica, namely, 14-acetoxo-12-oxokaur-16-en-19-oic acid labelled as 0657 and 16-hydroxy-kaur-6-en-19-oic acid given the label 06-2 in this study. The third suspected new compound is the chalcone dimer, which is referred to as EM86 in this study from Pomaria sandersonii. EM80-2 was obtained as a mixture of the cis and trans of 2’, 4, 4,’-trihydroxychalcone or 1-(2,4-dihydroxyphenyl)-3-(4-hydroxyphenyl)-2-propen-1-one, from Pomaria sandersonii. The three diterpenes, 14-acetoxokaur-16-en-19-oic acid (0652), 13-hydroxy-16-kauren-19-oic acid (06B) and 14-oxokaur-16-en-19-oic acid (06431) were isolated from Alepidea amatymbica for the first time. Isolated compounds were further tested as individual compounds and results showed that 16-hydroxy-kaur-6-en-19-oic acid (06-2) had weak activity against tested bacteria and fungi with the MIC: Staphylococcus aureus (320 Kg/ml) and Candida albicans, (320 Kg/ml). On the other hand 13-hydroxy-kaur-16-en-19-oic acid (06B) was more active against, Staphylococcus aureus (160 Kg/ml) and Aspergillus fumigatus (40 Kg/ml). The yellow compound that was isolated from Pomaria sandersonii, 1-(2, 4-ihydroxyphenyl)-3-(4-hydroxyphenyl)-2-propen-1-one was antimicrobial with the following MICs: Candida albicans: 80 Kg/ml; Pseudomonas aeruginosa, Escherichia coli, Enterococcus faecalis, Staphylococcus aureus: 160 Kg/ml and Aspergillus fumigatus: 625 Kg/ml. There were two mixtures referred to as EM 49 and EM 77 from Pomaria sandersonii which were difficult to purify but had anti-microbial inhibitory activities worth reporting. EM49 had MIC against Candida albicans of: 160μg/ml; Pseudomonas aeruginosa: 320 Kg/ml, Escherichia coli: 80μg/ml, Enterococcus faecalis 80μg/ml, and Staphylococcus aureus: 80μg/ml and Aspergillus fumigatus: 320μg/ml. EM 77 had MIC against Escherichia coli: 80 Kg/ml and Cryptococcus neoformans: 80μg/ml. Further work on their purification need to be done since in this research we are just reporting on their high MIC activities. The medicinal plants used to treat inflammation under different disease conditions in the Zulu community of Mabandla village, Kwa-Zulu Natal, South Africa have some relevant biological activities. The various antimicrobial, antioxidant and anti-inflammatory activities support the validity of their healing capacities that the traditional healers of the community claim to possess. Although there is evidence of good antimicrobial, antioxidant and anti-inflammatory activities by the crude extracts, the high levels of sucrose in P. prunelloides and glucose in G. perpensa should be borne in mind when using their decoctions in traditional medicine particularly by diabetic patients. In vitro results for the antioxidant, antinflammtory and antimicrobial activities carried out in this investigation illustrate that the plants can be a source of treatment and management for inflammation related conditions. These therefore justify their use in Zulu traditional medicine. However, in vivo assays should be carried out in order to completely validate claims by the traditional healers that they treat inflammation related conditions. / Vaal University of Technology

Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnology, Durban University of Technology, 2007 xi, 114 leaves / Eighteen traditionally leafy vegetables consumed as food or medicinal compounds by a majority of people in the KwaZulu Natal province of South Africa were analysed for the presence of potentially harmful chemicals (antinutrients) and for their toxicity and mutagenicity. The purpose of the study was to determine whether leafy vegetables were safe for human consumption. Chemical analysis showed that none of the vegetables contained cyanogenic glycosides, however all the vegetables contained oxalic acid ranging from 24.1 mg/ml to 798.2 mg/ml with Solanum nigrum, Portulaca oleracea and Mormodica balsamina showing the highest concentrations. Most of the vegetables contained negligible amounts of phytic acid and saponins, except for Momordica balsamina (3.01 mg/ml and 1.83 mg/ml, respectively). Fourteen of the plants contained alkaloids with Portulaca oleracea having the highest content (1.53 g total alkaloids/5 g leaf material). Eight of the plants were found to inhibit trypsin activity. These chemical analyses were carried out in duplicate and the mean and standard deviation were used. The Ames test revealed that none of the leafy vegetables produced a mutagenic frequency above 1, except 10 000 µg/ml organic extract of Senna occidentalis (mutagenecity considered at mutagenic frequency above 2), thus none were considered mutagenic. All 18 organic extracts did not kill off more than 50% brine shrimp and were thus considered non-toxic. On the other hand the aqueous extracts of seven vegetables, namely, Physalis viscosa, Amaranthus dubius, Justicia flava, Bidens pilosa, Senna occidentalis, Chenopodium album and Ceratotheca triloba, killed more than 50% of the shrimp and are thus considered toxic above 100 µg/ml. The MTT assay carried out on the organic extracts indicated that 17 vegetables did not kill off more than 50% of HepG2 cells and were thus considered non-cytotoxic. The aqueous extracts of four vegetables, namely, Justicia flava, Asystasia gangetica, Momordica balsamin and Senna occidentalis, however killed more than 50% of the shrimp and were thus considered cytotoxic above 1 000 µg/ml. It may be concluded from the antinutrient analyses and the bioassays on the 18 vegetables that caution needs to be maintained with the consumption of certain leafy vegetables included in this study, especially Senna occidentalis.

During the COVID pandemic of 2019, Sweden saw a huge increase in food prices, as many of the country's grocery chains import a large amount of all foods. This research is motivated by the Swedish Parliament's motion "Self-Sufficiency of Food in Sweden" and the need for innovative solutions to enhance Sweden's food supply. This study focuses on alternative urban agriculture methods, with a focus on aeroponics, aquaponics, hydroponics, and vertical farming, and their potential for self-sufficiency in Sweden. This study aims to fill the gap in Swedish research on soilless urban farming methods and contribute to the understanding and acceptance of the alternative methods above. The study seeks to determine whether the yield of alternative cultivation methods is comparable to traditional methods, soil-based and greenhouse-based farming, in terms of resources invested and to identify the alternative method with the highest yield. Resources in this thesis will be defined as the amount of water that is used. Our thesis aims to support businesses and local governance in decision-making regarding implementation of alternative urban farming in Sweden. A literature search was conducted using keywords such as hydroponic, vertical farming and sustainability. Statistical analysis of collected data indicates significant differences in yield and water usage among the collected examples which resulted in vertical hydroponic farming offering the highest yield in terms of space utilization, with significant statistical significance (p < 0.05). Additionally, hydroponic farming requires the least amount of water compared to other methods although this is not a certainty as there was no significant statistical difference between the different methods. The study demonstrates that hydroponic farming can achieve higher yields compared to traditional soil-based farming and other alternative farming methods. The study emphasizes organizational and governmental solutions to improve self-sufficiency at the local and national levels. Alternative urban farming methods offer a sustainable contribution to increasing self-sufficiency while minimizing resource usage. / Denna studie undersöker potentialen hos alternativa urbana odlingsmetoder för självförsörjning i Sverige. Den svenska riksdagen stödjer motion om "Självförsörjning av mat i Sverige" och behovet av innovativa lösningar för matförsörjning i landet. Denna forskning fyller en kunskapsbrist i den svenska litteraturen om jordlösa odlingssystem såsom aeroponiska, akvaponiska, hydroponiska och vertikala odlingar. Syftet med studien är att sammanfatta den befintliga kunskapen om dessa alternativa odlingsmetoder och klargöra deras fördelar i en urban miljö. Denna studie strävar att stödja företag och lokala myndigheter vid implementering av alternativa odlingsmetoder, genom att studera de alternativa odlingsmetodernas avkastning i förhållande till resursanvändning. Studien har samlat exempel från forskningsartiklar och företags-hemsidor som använde de ovan nämnda alternativa odlingsmetoder som sedan analyserats och jämfört deras avkastning med traditionella jordbaserade odlingar såsom åker odlingar och växthusbaserade odlingar. Med den statistisk analys som gjorts har vi funnit att vertikal hydroponisk odling visar den högsta avkastningen per yta jämfört med de andra alternativa odlingsmetoderna och traditionella odlingar. Vi har dessutom fastställt att hydroponiska odlingar använder minst vatten per kg skörd i jämförelse med andra metoder. Studien betonar vikten av att fokusera på organisatoriska och statliga lösningar för att öka självförsörjningsgraden på lokal och nationell nivå. Medan privatpersoner kan vara självförsörjande med grönsaker och frukt på en normalstor tomt, kan det vara svårt för alla privatpersoner att upprätthålla en egen grönsaksträdgård och undvika skador på marken och ekosystemtjänsterna i staden. Därmed riktar vi denna studie till kommunen och staten som ett stöd för att säkerställa självförsörjning på en lokal och regional nivå. För att uppnå hållbar samhällsutveckling och stödja självförsörjning av mat i Sverige, är integreringen av alternativa urbana odlingsmetoder på lokal, regional och nationell nivå ett innovativt steg framåt. Metoderna nämnda i denna rapport erbjuder fördelar såsom effektiv minskad vatten- och markanvändning samt möjlighet att integrera odling i stadsområden. Genom att använda dessa odlingsmetoder kan Sverige öka sin självförsörjningsgrad och minska sin miljöpåverkan från livsmedelsproduktionen. Framtida forskning bör utforska och utveckla dessa innovativa lösningar för att möta Sveriges matförsörjnings- och hållbarhetsmål.