The incidences of diabetes are on the constant rise, and thus need an alternative dietary approach to improve glucose metabolism in type II diabetes patients. Fruits and vegetables have long been known for their beneficial fiber, phenolics and antioxidants, and by incorporating these dietary components into our everyday diet have proven beneficial in controlling the glucose metabolism. In this study, Prime-Jan and Kiowa blackberry cultivars were evaluated for their anti-diabetic properties using a variety of in vitro assays. Overall, the Kiowa blackberry reigned supreme over the Prime-Jan blackberry in the majority of functionality assays, more specifically alpha-amylase, alpha-glucosidase, total soluble phenolics and DPPH free radical scavenging activity. In an attempt to incorporate anti-hypertensive properties into this study, we chose to integrate the Bartlett pear, known as the most widely consumed pear in the United States and recognized for its moderate ACE inhibition. Our results indicated a combination of 70% pear to 30% blackberry as the most beneficial ratio for exhibiting high alpha-amylase (96.5%) and alpha-glucosidase inhibition (95.6%). Additionally, the 70/30 combination exhibited high DPPH free radical scavenging activity (80.7%), total soluble phenolics (1.9 mg/g FW), while also maintaining moderately high ACE inhibition (25.2%). Therefore, there is vast potential for a combination of 70% pear to 30% blackberry to serve as a beneficial alternative in the diet of patients suffering from type 2 diabetes.

Type 2 diabetes is a worldwide and costly disease that requires complex cellular metabolic breakdown for its development. The onset of this disease is indicated by an abnormal elevated blood glucose level (hyperglycemia) and coexistence with hypertension. Apple and blueberry are the two popular fruits in the U.S. that are high in bioactive phenolic compounds. The objectives of this thesis were to investigate the synergistic health benefits of apple cider and blueberry juice for potential management of early stages of type 2 diabetes using in vitro enzyme assay models and enhance their synergistic effects through fermentation using probiotic strains of lactic acid bacteria. Apple cider and whole blueberry juice were combined at 5 different ratios. Synergistic interaction of the phenolic bioactives between the two juice sources was observed in all assays. For maintenance of high α-glucosidase, α-amylase and ACE inhibition activities, a proportion of 80% apple cider and 20% whole blueberry juice was selected as the optimum combination reflecting maximum health benefits to potentially manage hyperglycemia and hypertension associated with type 2 diabetes. The 80/20 apple cider and blueberry juice combination was fermented at 24 and 48 h using Lactobacillus helveticus R0052 and Bifidobacterium longum isolate. High inhibition of α-glucosidase, α-amylase and ACE enzymes was observed in apple cider/blueberry juice combination. Strong antimicrobial activity against H. pylori was exerted by L. helveticus- and B. longum-fermented sample at 48 h. A possible mode of action through the synergy between lactic acid and proline oxidation pathway was suggested.

The incidence of diabetes mellitus and cardiovascular diseases is increasing at a worrisome rate globally. Diabetes mellitus is known to occur due to high blood glucose levels, caused by defects in insulin levels. Adult on-set type II diabetes, which is closely associated with obesity, is reported to be 90-95% of all diabetic cases and linked to diet and lifestyle factors. A large population of the developed and developing countries is now being effected by this epidemic. Natural sources of phenolic antioxidants and inhibitors of digestive enzymes from food sources have potential for low cost dietary management of type II diabetes. Therefore, the main focus of this study was to evaluate, develop and design effective dietary strategies based on a combination of Chilean potatoes and herb synergies for the management of hyperglycemia and hypertension linked to type II diabetes. Antioxidant, antihypertensive and anti-hyperglycemic potentials of Chilean potato (Solanum tuberosum ssp. tuberosum L.), herbs from the Apiaceae (Dill, Ajowan, Fennel, Caraway, Coriander and Anise) and Lamiaceae (Sage and Marjoram) families were evaluated, with a goal to target a new dietary management strategy for early stages of type II diabetes through lowering of hyperglycemia and related complications of hypertension The results indicated a high correlation between total phenolic content and total antioxidant activity in several Chilean potato varieties evaluated, which indicates that certain phenolic compounds may be responsible for high antioxidant activity. Also, certain varieties of Chilean potato had antihypertensive potentials, with ACE inhibition upto 88%. The -glucosidase inhibition relevant for hyperglycemia management for Apiaceae family ranged upto 50% (Dill) for aqueous extracts. A high correlation (r = 0.86) was observed between -glucosidase inhibition and total phenolic content for aqueous extracts of all species investigated in the Apiaceae family. A high rosmarinic acid activity was observed in aqueous extracts of Lamiaceae family, which ranged upto 39.7 mg/g of sample dry weight (DW). This suggests that high phenolic content and associated antioxidant activity found in sage and marjoram is dominated by rosmarinic acid. High enzyme inhibitory activities, reflecting in vitro anti-hyperglycemic and anti-hypertensive potentials indicates that consumption of these food sources in our diet would prove to be beneficial towards our health. Further in vivo studies for type II diabetes-linked functionalities of these natural sources of antioxidants and inhibitors would confirm the human health benefits achieved through dietary intake.

More than 16,000 tons of olive cake was produced in the United States in 2017. Olive cake is a by-product of olive oil extraction, which has limited animal feed potential, and poses an environmental threat when landfilled due to its high organic load and polyphenol content. This residue has potential for biofuel (bioethanol and biomethane) production because it is rich in polysaccharides such as pectin, hemicellulose, and cellulose. Yet, olive cake contains olive stones that can impede its conversion to biofuel. Therefore, two methods of destoning, centrifugation and screening by horizontal screw press, were first compared. Both methods removed an equal percentage of stones (95%), but centrifugation partitioned the majority (57 – 79%) of digestible solids (olive pulp) with the stones. Then, two strategies were compared to maximize both biomethane and bioethanol production; enzymatic conversion of insoluble to soluble carbohydrates and destoning by screening. After 30 days of anaerobic digestion at 35 °C, both the enzymatically pretreated and the destoned olive cakes produced similar amounts of methane (~295 mL CH4/g VS), 42% more than the control (209.5 mL CH4/g VS). The biogas produced was composed of 60-70% methane. A comparison of biomethane yields with a broad range of agricultural residues demonstrated olive cake’s suitability for biomethane production. The digestate, residue from the anaerobic digestion, have high Kjeldahl nitrogen content (3.6%, db) and low polyphenol concentration (0.02 mg GAE/g), which then qualify it as an ingredient for soil amendment. Ethanol production investigations showed that after 3 days of fermentation at 32 °C, only the destoned and enzymatically pretreated olive cake produced ethanol (1.3 mg/mL). Acetic acid, an inhibitor of ethanol production, was present in all samples broth, suggesting microbial contamination was present. These results provide evidence that olive cake can be diverted from landfills to be converted into a biofuel. Sustainable pretreatments such as destoning and enzymatic pretreatment increase biomethane yield. The digestate created from the anaerobic digestion of olive cake can be used as a soil amendment, adding further value to olive cake.

For the past 40 years, antiangiogenic approaches have been of major interest in the development of methods to cure and prevent cancer. Angiogenesis, the development of blood vessels from pre-existing vascularization, is essential for cancer growth and spread of metastasis through the delivery of nutrients and oxygen essential to sustain the metabolic activity of these malignant cells. Blocking access to blood will cause cancerous cells to assume a dormant state creating inactive micro-tumors innocuous to the host. Angiostatin, the internal fragment of the fibrinolytic zymogen plasminogen, has shown great potential in reducing cancer size and number of metastatic colonies in animal models. Owing to the success of these preliminary results angiostatin is currently on clinical trials. Plasminogen is known to be transferred from blood to milk during lactation. The objectives of this research were to: 1) investigate the ability of various proteases in cleaving plasminogen, both from human and bovine sources, and consequently release the angiostatin like fragment; 2) determine the anticancer activity of bovine angiostatin; 3) examine ability of the antiangiogenic fragment to survive digestion; 4) purify the fragment of interest through column chromatography. Production of angiostatin was tested through hydrolysis of plasminogen via Bacillus Polymyxa protease (or dispase I), elastase, lactic acid bacteria and Bacilli originated enzymes. Once proteases capable of angiostatin like peptide production were identified, and sequence analysis of the fragments obtained conducted to confirm that bovine angiostatin was indeed produced, ability of angiostatin, both human and bovine, in inhibiting malignant melanoma as well as colon cancer cells was evaluated in vitro. From the results obtained we can confirm that bovine angiostatin inhibitory activity on cancerous cells is similar to that observed for human angiostatin. Analysis of bovine angiostatin survival through in vitro human digestion model was also examined. Results show good possibility of angiostatin surviving digestion, even if confirmation of these results is required through further in vivo studies. Additionally, digestive enzymes such as trypsin and α-chymotrypsin showed ability in cleaving plasminogen directly to release a 25kDa fragment. Knowing that each kringle has some degree of anticancer activity it would be of interest to further study the possibility of angiostatin related fragments to be produced during milk digestion. Finally, affinity chromatography through L-lysine used to purify human angiostatin resulted to be an adequate method for bovine angiostatin purification. Preliminary results obtained from this study open a new area worth investigating to uncover the potential of using bovine angiostatin in the development of novel food products capable of cancer prevention.

Adjuvants can enhance the performance of the pesticide active ingredients in many ways including decreasing surface tension and reducing evaporation. Understanding how adjuvants effect pesticide behavior (e.g., surface persistence) is crucial for developing effective pesticide formulations, as well as facilitating the development of effective approaches to reduce pesticide residues from the surface of fresh produce post-harvest. The objective of this study is to investigate the effect of a non-ionic surfactant, Surf-Ac 910, on the persistence of two model pesticides, thiabendazole and phosmet on apple surfaces. The result shows that the addition of Surf-Ac 910 increased both the maximum wetted area and evaporation rate of thiabendazole, a systemic pesticide, and phosmet, a non-systemic pesticide. Utilizing surface-enhanced Raman spectroscopy to explore the surface and penetrative behaviors of thiabendazole and phosmet revealed that the addition of Surf-Ac 910 influenced the Raman signal of pesticides as well. The addition of Surf-Ac 910 decreased the Raman signal intensity when added to phosmet but did not affect the Raman signal intensity when added to thiabendazole. In terms of penetration, the addition of Surf-Ac 910 did not affect the penetration depth of phosmet but slightly increased the penetration depth of thiabendazole. These findings were true for both short-term, 40 minutes, and long-term, 3 days, exposure. Next, the effects of adjuvants on the removal of pesticide residues were investigated. Common household materials, such as baking soda, were effective at removing surface pesticide residues. After testing a variety of baking soda concentrations and starch granules, 2% baking soda and 2% corn starch were found to be the most effective baking soda concentration and starch granule respectively. 2% corn starch was the most effective removal method overall, with 99% of pesticide with/without adjuvant removed in just 5 minutes of wash time. Overall, this study demonstrated that although adjuvant Surf-Ac 910 could affect the surface persistence of pesticides, washing with common household materials such as 2% corn starch can be used as an effective, safe, and economic way to reduce pesticide exposure through fresh produce.

Methylglyoxal is a highly reactive dicarbonyl compound, which is widely distributed in food products and beverages, and is particularly high in Manuka honey. In addition to its antibacterial effects, methylglyoxal is also known as a major precursor of advanced glycation end products (AGEs), that produces altered macromolecules (such as proteins and DNA), leading to abnormal physiological changes. However, the effects of methylglyoxal on development is unclear. Thus, this study aimed to determine the role of methylglyoxal in this aspect using Caenorhabditis elegans (C. elegans). Treatment of methylglyoxal at 0.1 mM and 1 mM for 48 h significantly inhibited development of C. elegans and reduced pumping rate. Activity, measured by moving speed, was increased with 0.1 mM methylglyoxal, but reduced with 1 mM methylglyoxal. Lifespan of C. elegans was not influenced by methylglyoxal at 0.1 mM, but was shortened at 1 mM. Treatment methylglyoxal on the mutant, lin-41, which has a precocious phenotype, could alleviate the implication on wild-type worms. These results suggested that methylglyoxal significantly influenced the development of C. elegans through the heterochronic pathway.

Thesis (D. Tech.: Biotechnology)-Dept. of Biotechnology, Durban University of Technology, 2006 xii, 274 leaves / In Sub-Saharan Africa there is an urgent need to sustain and improve the quality of its food resources. Poverty eradication features high on the agenda of a number of world health organisations, while the number of underweight children in Africa continues to increase (Pellet, 1996). Providing nutritionally enhanced foods to the poor will help towards achieving this objective. Protein-energy malnutrition has been identified as one of the most important problems facing Africa, with maize as the staple diet (Nkama et al., 1995). However, a combination of several factors limits availability and the nutritional quality of maize. During starvation, energy and protein intakes decrease by 20-30%, with most of the children in Africa having an average protein intake of only 20 g per day (Igbedioh, 1996). Energy availability also affects protein utilization because of interrelationships of protein and energy metabolism (Elwyn, 1993). The diets of inhabitants in developing regions depend mainly on cereals (maize) for both protein and dietary energy which lacks indispensable amino acids, minerals, vitamins and carbohydrates. In light of these growing concerns an attempt was made to devise a scientific strategy to combat the nutritional shortfalls of maize meal. A multidisciplinary and concerted approach was followed within this project aimed at designing an improved thermostable amylase and applying the enzyme to nutritionally enhance maize meal. It was envisaged that the manipulation of maize meal, by the application of enzyme technology will improve the nutritional status of this staple food. The consequences is that an alternate solution for the eradication of an ailing, poverty stricken and malnourished African population is achievable. It is possible that the boundaries defining the limits of life will extend to even greater extremes through the application of novel technologies.

Thesis ( PhD. (Biotechnology )) --University of Limpopo, 2005 / The marula tree (Sclerocarya birrea subsp. caffra), an indigenous, multipurpose, drought tolerant tree of Africa harbors great economic potential. Acceptance of marula-derived products internationally will directly increase the demand for marula resource. Rapid multiplication of marula trees of superior quality forms the basis of sustainable export growth. In vitro propagation and genetic improvement offer the opportunity for accelerated multiplication of selected tree material as well as to dramatically increase production, quality and efficiencies. The objectives of the study were therefore to develop a protocol for in vitro multiplication of marula and to determine the feasibility of Agrobacterium-mediated transformation of the marula tree. Nodal sections with axillary bud (s) were cultured on Murashige and Skoog (MS) medium supplemented with 4.8μM BA and 2.4μM KN and 0.1% polyvinylpyrrolidone (PVP) to obtain on average 2.5 microshoots per responding explant. The proliferated microshoots were elongated on MS medium supplemented with 1.2μM BA and 1.0μM KN. Elongated microshoots were rooted in MS medium at half salts strength supplemented with 10μM IBA and 0.3% activated charcoal (AC). On average 82% of the shoots rooted. Survival of acclimatized plantlets was 90%. RAPD analysis confirmed intraclonal genetic stability between parent plants and their clones within the limits of the technique.Nodal sections cocultivated with Agrobacterium tumefaciens for 3 days on MS multiplication medium supplemented with 100μM acetosyringone resulted on average in transient expression of 52.5% of the explants with 1.6 blue stained zones per explant. Cocultivated explants on MS selection medium containing 300mgl-1 kanamycin resulted in 1.5% chimeric putative transgenic shoots. This is the first report on the micropropagation and genetic transformation of marula, Sclerocarya birrea subsp caffra. / South Africa’s National Research Foundation Institutional Research Development Program (NRF-IRDP)

Thesis (D. Tech.: Biotechnology)-Dept. of Biotechnology, Durban University of Technology, 2006 xii, 274 leaves / In Sub-Saharan Africa there is an urgent need to sustain and improve the quality of its food resources. Poverty eradication features high on the agenda of a number of world health organisations, while the number of underweight children in Africa continues to increase (Pellet, 1996). Providing nutritionally enhanced foods to the poor will help towards achieving this objective. Protein-energy malnutrition has been identified as one of the most important problems facing Africa, with maize as the staple diet (Nkama et al., 1995). However, a combination of several factors limits availability and the nutritional quality of maize. During starvation, energy and protein intakes decrease by 20-30%, with most of the children in Africa having an average protein intake of only 20 g per day (Igbedioh, 1996). Energy availability also affects protein utilization because of interrelationships of protein and energy metabolism (Elwyn, 1993). The diets of inhabitants in developing regions depend mainly on cereals (maize) for both protein and dietary energy which lacks indispensable amino acids, minerals, vitamins and carbohydrates. In light of these growing concerns an attempt was made to devise a scientific strategy to combat the nutritional shortfalls of maize meal. A multidisciplinary and concerted approach was followed within this project aimed at designing an improved thermostable amylase and applying the enzyme to nutritionally enhance maize meal. It was envisaged that the manipulation of maize meal, by the application of enzyme technology will improve the nutritional status of this staple food. The consequences is that an alternate solution for the eradication of an ailing, poverty stricken and malnourished African population is achievable. It is possible that the boundaries defining the limits of life will extend to even greater extremes through the application of novel technologies.