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Background: Low-grade chronic inflammation has been implicated as a risk factor in prostate-related pathologies including benign hyperplasia and cancer. Sweet cherry containing the bioactive anthocyanin (ACN), has demonstrated tumor inhibitory action in model systems, specifically inhibition of inflammatory molecules and prostaglandin biosynthesis. Objective: To assess the urinary and plasma concentrations of ACN from the daily consumption of 3 cups of sweet cherries for 4 weeks and test the relationship of ACN levels and cherry consumption to inflammatory biomarkers in an at risk population. Results: Prostaglandin E2 Metabolite (PGEM) levels were reduced with cherry consumption in men with elevated baseline values. Conclusion: We conclude that 1c (142g) of sweet cherries 3 times daily for 4 weeks significantly reduced the COX-2 metabolite, PGEM, in men with elevated baseline levels. This was the first study to examine the chronic effects of daily sweet cherries on COX-2 inhibition in an at risk population.

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Doctor of Philosophy / Department of Biological & Agricultural Engineering / Donghai Wang / Sweet sorghum accumulates high concentrations of fermentable sugars in the stem, produces significant amount of starch in the grain (panicle) and has shown to be a promising energy feedstock. Sweet sorghum has a short growing season so adding it to the sugar cane system would be good. The overall goal of this dissertation is to enhance the attractiveness of biofuel production from sweet sorghum to fully utilize fermentable sugars in the juice, starch in the panicle and structural carbohydrates in the stalk for high efficiency and low-cost ethanol production. Sweet sorghum juice was incorporated into the dry-grind process which increased ethanol yield by 28% increase of ethanol yield compared to the conventional ethanol method and decreased enzymatic hydrolysis time by 30 minutes. A very high gravity fermentation technique was applied using sweet sorghum juice and sorghum grain yielded 20.25% (v/v) of ethanol and 96% fermentation efficiency. Response surface methodology was applied in order to optimize diffusion conditions and to explore effects of diffusion time, diffusion temperature, and ratio of sweet sorghum biomass to grain on starch-to-sugar efficiency and total sugar recovery from sweet sorghum. Starch hydrolysis efficiency and sugar recovery efficiency of 96 and 98.5% were achieved, respectively, at an optimized diffusion condition of 115 minutes, 95 °C, and 22% grain loading. Extraction kinetics based on the optimized diffusion parameters were developed to describe the mass transfer of sugars in sweet sorghum biomass during the diffusion process. Ethanol obtained from fermented extracted sugars treated with granular starch hydrolyzing enzyme and those with traditional enzymes were comparable (14.5 – 14.6% v/v). Ethanol efficiencies also ranged from 88.92 –92.02%.

Doctor of Philosophy / Department of Biological & Agricultural Engineering / Donghai Wang / Sweet sorghum accumulates high concentrations of fermentable sugars in the stem, produces significant amount of starch in the grain (panicle) and has shown to be a promising energy feedstock. Sweet sorghum has a short growing season so adding it to the sugar cane system would be good. The overall goal of this dissertation is to enhance the attractiveness of biofuel production from sweet sorghum to fully utilize fermentable sugars in the juice, starch in the panicle and structural carbohydrates in the stalk for high efficiency and low-cost ethanol production. Sweet sorghum juice was incorporated into the dry-grind process which achieved 28% increase of ethanol yield compared to the conventional ethanol method and decreased enzymatic hydrolysis time by 30 minutes. A very high gravity fermentation technique was applied using sweet sorghum juice and sorghum grain yielded 20.25% (v/v) of ethanol and 96% fermentation efficiency. Response surface methodology was applied in order to optimize diffusion conditions and to explore effects of diffusion time, diffusion temperature, and ratio of sweet sorghum biomass to grain on starch-to-sugar efficiency and total sugar recovery from sweet sorghum. Starch hydrolysis efficiency and sugar recovery efficiency of 96 and 98.5% were achieved, respectively, at an optimized diffusion condition of 115 minutes, 95 °C, and 22% grain loading. Extraction kinetics based on the optimized diffusion parameters were developed to describe the mass transfer of sugars in sweet sorghum biomass during the diffusion process. Ethanol obtained from fermented extracted sugars treated with granular starch hydrolyzing enzyme and those with traditional enzymes were comparable (14.5 – 14.6% v/v). Ethanol efficiencies also ranged from 88.92 –92.02%.

Con la finalidad de insertar los genes cry3Ca1 y cry7Aa1 en Ipomoea batatas (L.) Lam. cv. “Huachano” para conferir resistencia a Cylas puncticollis y C. brunneus “gorgojos del camote”, se desarrolló la transformación genética mediada por Agrobacterium tumefaciens, siguiendo dos protocolos de regeneración y transformación. Partiendo de 46 meristemos y 282 hojas con peciolo, se obtuvieron 8 eventos transgénicos de inserción completa del ADN - T (4 por cada protocolo de transformación) y un evento de inserción incompleta. Con una eficiencia de transformación de 8.70%, el protocolo a partir de meristemos demostró ser más eficaz en la obtención de eventos transgénicos de inserciones completas que el protocolo a partir de hojas con peciolo, con el cual se obtuvo una eficiencia de transformación de 1.42%. A su vez, los regenerantes fueron evaluados mediante pruebas in vitro (resistencia a kanamicina) y moleculares (PCR), con los análisis de PCR se confirmó que los 8 regenerantes callo positivos (resistentes a la prueba de kanamicina) también presentaron la inserción de los transgenes de interés “cry3Ca1 y cry7Aa1” y del gen marcador selector “nptII”. Asimismo, se determinó que 4 de los eventos transgénicos integraron secuencias externas al ADN - T “backbone” en el genoma de la planta, uno de los cuales, presentó la inserción del gen bacteriano “virD2”.With the purpose of inserting the genes cry3Ca1 and cry7Aa1 in Ipomoea batatas (L.) Lam. cv. "Huachano" to develop resistance to Cylas puncticollis and C. brunneus “sweet potato weevils”, the genetic transformation was developed using Agrobacterium tumefaciens, following two protocols of regeneration and transformation. Starting from 46 meristems and 282 leaves with petiole, 8 transgenic events were obtained with complete insertion of T - DNA (4 for each transformation protocol) and one event with incomplete insertion. With an efficiency of transformation of 8.70 %, the protocol of meristems proved to be more effective in obtaining transgenic events of complete inserts than the protocol of leaves with petiole, which obtained a efficiency of transformation of 1.42 %. The regenerants were evaluated by testing in vitro (resistance to kanamycin) and molecular (PCR), the PCR analysis confirmed that the 8 regenerants positive callus (resistant to kanamycin test) also presented the insertion of transgenes of interest “cry3Ca1 and cry7Aa1” and selection marker gene “nptII”. Likewise, 4 of the transgenic events integrated sequences outside the T - DNA “backbone” into the plant genome, one of whom, showed the insertion of the bacterial gene “virD2”.

Master of Science / Department of Animal Sciences and Industry / Teresa L. Slough / Teresa L. Slough / There is increasing interest in the use of point-of-care glucometers to monitor glucose concentrations in horses with metabolic disorders. The first study reported herein compared equine glucose concentrations obtained by a handheld glucometer using whole blood or plasma, a YSI 2300 bench top glucose analyzer using whole blood or plasma, and a SEVEN continuous glucose monitoring (CGM) device that measured glucose in interstitial fluid to readings obtained by a standard laboratory glucose analyzer utilizing plasma. In addition, glucose concentrations obtained by the CGM were compared to those obtained by the handheld glucometer using whole blood or plasma. Post-prandial increases and decreases in glucose concentrations were detected utilizing all glucometers tested. When glucose measurements obtained with the CGM in interstitial fluid were compared to glucose measured using the handheld glucometer in plasma or whole blood, glucose measurements from plasma had better reproducibility. Although the CGM could be a useful instrument for collecting nearly continuous data for the researcher and clinician, there are technical difficulties related to the CGM that must first be overcome. The second study was designed to compare the effects of consuming a twice-daily meal of sweet feed (SF) to ad libitum access to a molasses-based block (BL) supplement on patterns of interstitial glucose concentrations in horses. A novelty effect of the BL was observed, in which horses consumed increased quantities in the first 12 h. Treatments had no effect on intake of forage in this study. The range and means of glucose values were similar between treatments, and significant glucose responses to treatments had lag times that were indirectly similar, even though molasses intake was greater for horses on BL. Variability between horses was noted in quantity of BL consumed as well as timing and magnitude of glucose responses. Based on the results of this experiment, there does not appear to be a clear advantage to either treatment, SF or BL, in attenuating post-prandial glucose increases or in minimizing glucose fluctuations in the horse.

Doctor of Philosophy / Department of Agronomy / P. V. Vara Prasad / Sweet sorghum (Sorghum bicolor L. Moench) is an important bioenergy crop. There is a wide array of genetic diversity in sweet sorghum germplasm collections. However, information on traits associated with sugar yield, optimum harvesting time for maximum sugar yield, effects of abiotic stresses on sugar yield is scarce. The objectives of the present study were: to identify traits that are associated with sugar yield, to determine the optimum harvesting time for maximum sugar yield and to understand the physiological responses of different sweet sorghum genotypes to drought and high temperature. In order to meet these objectives, five independent field and greenhouse studies were conducted. Field experiments were conducted using 280 sweet sorghum germplasm and were evaluated for 2 years. From this study, 30 genotypes representing high and low sugar yielders were selected for the subsequent experiment. We observed a significant variation in physiological, morphological and sugar yield traits associated with biofuel production. In the selection experiment, investigations on the morphological, physiological attributes helped to identify those characters which influence or limit sugar yield in the sweet sorghum. Another field study was conducted to optimize the harvesting time for obtaining highest sugar and juice yields in sweet sorghum. Sweet sorghum variety M81E was harvested at ten growth stages. Our results suggest that the optimum time for harvesting of sweet sorghum cultivar M81E is between milk and hard dough stages when highest sugar yield was observed. Studies on different levels of water stress were studied under greenhouse conditions. Four sweet sorghum genotypes (Awanlek, Smith, Tracy and Wray) were subjected to three water stress treatments (100% pot capacity (PC); 70% PC and 30% PC) for 20 days at early seed filling (Milk) stage. The results showed that genotypes differed significantly for all growth and yield, biochemical and physiological traits. Severe water stress significantly decreased juice and sugar yields by decreasing net photosynthetic rate, transpiration rate, stomatal conductance and sucrose content in the stem juice. Genotypes Tracy and Wray produced significantly highest brix, stem fresh weight, juice and sugar yield under both irrigated and water stress conditions. In another greenhouse study, we quantified the effects of drought, high temperature, and their combinations on growth, physiology and yield of sweet sorghum genotypes. The same four genotypes above were subjected to four treatments, T1 - control, T2 - drought stress, T3 - high temperature stress and T4 - combination of drought and high temperature for 16 days after anthesis. The result showed that significant difference was observed for growth and yield traits, physiological traits and non-reducing and total sugar content in juice for genotypes and treatments. Among the genotypes Tracy recorded higher juice and sugar yield. Among the various treatments, combination of drought and high temperature was found to be more deleterious in reducing most of the biofuel traits followed by drought and high temperature stress. The above studies gave significant findings with regards to the identification of superior sweet sorghum germplasm, their tolerance capacity to different abiotic stresses, which allows better selection for the use of bioenergy production.

Doctor of Philosophy / Department of Human Nutrition / Weiqun Wang / Anthocyanins are flavonoid pigments that account for the purple color in many plant foods. It has been investigated that anthocyanins’ predominant occurrences in human diet and their health beneficial activities such as antioxidant, anti-inflammatory, and anti-carcinogenetic effects. Based on those scientific evidences, anthocyanins are now recognized as potential therapeutic compounds. Particularly, the chemopreventive effect of anthocyanins has been widely studied by many researchers in nutrition. However, their bioactivities are diverse due to different chemical structures of anthocyanins from different sources. In this study, we discuss the chemopreventive activity of anthocyanins from purple sweet potato. Previously, we selected a purple-fleshed sweetpotato clone, P40, crossbred seeds obtained from the International Potato Center in Lima, Peru. We hypothesized that anthocyanins enriched P40 may provide health beneficial activities in cancer prevention. For the first part of this study, we analyzed nutrient compositions, dietary fiber content, anthocyanins contents, total phenolics contents and total antioxidant activity. Even thought P40 presents similar composition and amount of nutrients with the control cultivars, white-fleshed O’Henry and yellow-fleshed NC Japanese, HPLC-MS analysis confirmed that it possesses much higher anthocyanin content even up to 7.5g/kg dry matter. Also, dietary fiber, particularly soluble dietary fiber content, total phenolics content, and total antioxidant capacity of P40 were significantly higher. For the second part of the study, we tested the potential anticancer characteristic of P40 cultivar in human colonic SW480 cancer cells and in azoxymethane-induced aberrant crypt foci in mice. Treatment with 0 – 40 μM of peonidin-3-glucoside or P40 extract containing corresponding amount of anthocyanins resulted in inhibition of cell growth in a dose-dependent manner. Interestingly, even though the patterns of growth inhibition were similar in the two treatment groups, the cells treated with P40 extract tend to survive significantly less than those treated with peonidin-3-glucoside. Cell cycle analysis confirmed that the growth inhibition was not due to cytotoxicity, but cytostatic mechanism with increased number at the G1 phase of the cell cycle. The cell cycle arrest was also significantly correlated with the anthocyanin contents in P40 cultivar when compared with the white-fleshed O’Henry and yellow-fleshed NC Japanese controls. After Azoxymethane (AOM) or saline injected mice were fed basal AIN-93M diet or diets containing 10~30% of P40, 20% O’Henry or 20% NC Japanese for 6 weeks, aberrant crypt foci (ACF) multiplicity was significantly inhibited by 10~30% P40 diet. Imunohistochemistry results of colonic mucosa showed that the expression level of apoptosis marker, caspase-3, was significantly induced in the mice treated with 10~20% P40 diet. Also, PCNA expression level, which is proliferation marker, was significantly inhibited by the 30% P40 diet. These findings indicated that consuming a purple sweet potato, P40, may prevent colon cancer by modulating antioxidant status, inducing apoptosis, and reducing cell proliferation.

M. Tech. (Biotechnology, Department of Biotechnology, Faculty of applied and computer sciences) Vaal University of Technolog / Sweetpotato (Ipomoea batatas) is an important food crop that is grown in many countries. A number of viral and fungal sweetpotato diseases have been reported worldwide. One of the major and most economic diseases of the sweetpotato is Alternaria blight which is caused by the fungal pathogen Alternaria bataticola. This disease can be managed in a short term using fungicides and cultural practices. However, a long term and inexpensive approach is through the development of resistant cultivars. A prerequisite to this approach is the knowledge of the genetic diversity of this fungal pathogen. This study assessed the genetic diversity of 25 South African isolates of A. bataticola from naturally infected leaves and stems collected from different sweetpotato growing regions in South Africa by (i) characterising the isolates based on their morphology (ii) pathogenicity tests (iii) random amplified polymorphic DNA (RAPD) (iv) variation of the ITS2 sequences and (v) prediction of the ITS2 secondary structures. The isolates revealed some variation in colony colour pigments after culturing but Koch’s postulates were confirmed by their pathogenicity tests. The analysis of RAPD and variation of the ITS2 sequences showed high levels of variation (100%) among the isolates. Dendrograms generated from these analyses had many subclusters and did not cluster the isolates according to their geographic origins. The ITS2 secondary structures were predicted and can be used to identify and distinguish the isolates. This information in addition to the genetic diversity of the A. bataticola isolates will aid plant breeders in the development of resistant sweetpotato cultivars and early management of blight disease in South Africa.