Faculty of Applied Sciences
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Item AB initio studies of a pentacyclo-undecane cage lactam(2003) Singh, ThishanaThe purpose of this study is to utilize computational techniques in the determination of the mechanistic pathways for the one-pot conversion of a pentacyclo-undecane (PCU) dione 1.1 to a pentacyclo-undecane cage lactam 1.2.Item The accumulation of heavy metals by aquatic plants(2003) Maharaj, Saroja; Moodley, Kandasamy Govindsamy; Southway-Ajulu, F. A.; Baijnath, HimansuThe pollution of water bodies by heavy metals is a serious threat to humanity. The technique known as phytoremediation is used to clean up these polluted water bodies. The accumulation of heavy metals by aquatic plants is a safer, . cheaper and friendlier manner of cleaning the environment. The aquatic plants -studied in this project are A.sessilis, P.stratiotes, R.steudelii and T.capensis. The accumulation of heavy metals in aquatic plants growing in waste water treatment ponds was investigated. The water, sludge and plants were collected from five maturation ponds at the Northern Waste Water Treatment Works, Sea Cow Lake, Durban. The samples were analysed for Zn, Mn, Cr, Ni, Pb and Cu using ICP-MS. In general it was found that the concentrations of the targeted metals were much lower in the water (0.002 to 0.109 mg/I) compared to sediment/sludge (44 to 1543mg/kg dry wt) and plants (0.4 to 2246 mg/kg dry wt). These results show that water released into the river from the final maturation pond has metal concentrations well below the maximum limits set by international environmental control bodies. It also shows that sediments act as good sinks for metals and that plants do uptake metals to a significant extent. Of the four plants investigated it was found that }t.sessi[ir (leaves, roots and stems) and }A.sessilis (roots and stems) are relatively good collectors of Mn and Cu respectively. These findings are described in the thesis. The concentration of heavy metals in the stems, leaves and roots of the three plants were compared to ascertain if there were differences in the ability of the plant at different parts of the plant to bioaccumulate the six heavy metals studied.Item Active targeting of cancer cells using gemcitabine conjugated platinum nanoparticles(2017) Odayar, Kriya; Odhav, Bharti; Mohanlall, VireshNanotechnology is explained as the science of engineered materials and systems on a molecular scale. This innovation is currently used in a wide variety of applications which include using these nanoparticles as drug delivery vehicles. Such nanocarriers are relatively smaller than 100 nm in size with the ability to convey therapeutic drugs to a number of disease sites. Platinum-based nanoparticles have been extensively used in a number of applications namely catalysts, gas sensors, glucose sensors and cancer therapy. The properties of platinum nanoparticles (PtNP’s) typically depend on characteristics such as shape, particle size, elemental composition and structure, all of which can be manipulated and controlled in the fabrication process. Their unique size in the nanometer scale makes platinum nanoparticles an ideal candidate as targeted drug delivery vehicles. To target an anticancer drug to a diseased site is a distinctive feature of most studies, which aim to transfer an adequate dosage of the drug to cancer cells. Transport systems used as carriers of anticancer drugs offer numerous advantages, which include improved efficacy and a decrease in toxicity towards healthy cells when compared to standard drugs. The aim of this study was to determine the effect of platinum nanoparticles, gemcitabine and gemcitabine conjugated platinum nanoparticles (Hybrids) against cancer cells and healthy cells and to determine the mode of cell death and cell death pathways using flow cytometry. Platinum nanoparticles were synthesized via the reduction of hexachloroplatinic acid using sodium borohydride in the presence of capping agents. Synthesized platinum nanoparticles and the hybrids were characterized by observing peaks at 301 nm and 379 nm respectively using UV-visible spectroscopy. TEM images revealed that the PtNP’s and the conjugate compounds were spherical in shape with core sizes of 1.14 nm - 1.65 nm and 1.53 - 2.66 nm respectively. The bioactivity platinum nanoparticles, gemcitabine and the hybrids were investigated using MCF7 and Melanoma cancer cells at different concentrations from 0.10 to 100 µg/ml. Results indicated that conjugated nanoparticles induced the highest cell inhibition against both cell lines compared to gemcitabine and platinum nanoparticles. Bioactivity against PBMC (peripheral blood mononuclear) cells indicated that all three compounds show little or no effect towards the healthy cell line compared to the control. Melanoma cell line was used to determine the mode of cell death. Apoptosis was detected using Annexin V-FITC to detect membrane changes, JC-1 to detect a loss in mitochondrial membrane potential and caspase-3 assay kits. Results indicated that a significant amount of cell death was caused by cleavage of caspase-3. Nanoparticle drug delivery is an area that has shown significant promise in cancer treatment. Interaction of nanoparticles with human cells is an interesting topic for understanding toxicity and developing potential drug candidates. Imagine, something that is atleast or more than 80,000 times smaller than the edge of the ridge on a fingertip and unlocks a new wilderness into cancer research. Nanotechnology, known as the science of minute, is changing the approach to cancer and especially future diagnosis and treatment. Nanotechnology permits scientists to fabricate new apparatuses that are definitely smaller than cells, giving them the chance to attack tumor diseased cells. This innovation not just empowers practitioners to recognize malignancies prior but additionally holds the guarantee of halting cancer growth before it further develops. This progressive approach is so exact, specialists will in future be able to outline a unique treatment for an individual’s own restorative and hereditary profile. Researchers are designing nanoparticles that detect and destroy diseased cells and this optimistic innovation could be personalized for targeted drug delivery, enhanced imaging and ongoing affirmation of cancer cell death. The National Cancer Institute remains hopeful that facilitated development, nanotechnology will drastically change cancer treatment.Item Activity coefficients at infinite dilution for ILs : methyltrioctylammonium cation and bis(trifluoromethylsulfonyl)imide or thiosalicylate anions using glc(2009) Gwala, Nobuhle V.; Deenadayalu, NirmalaThe activity coefficients at infinite dilution ( γ13 ) were calculated for alkanes, alkenes, cycloalkanes, alkynes, ketones, alcohols and aromatic compounds from gas liquid chromatography (glc) measurements at three temperatures (303.15 and 313.15 and 323.15) K. The γ13 values were calculated from the retention data for two ionic liquids (ILs) with the same cation but different anions. The ionic liquids: [methyltrioctylammonium bis(trifluoromethylsulfonyl)imide and methyltrioctylammonium thiosalicylate] were used as the stationary phase. The γ13 data for methyltrioctylammonium thiosalicylate were higher than for methyltrioctylammonium bis(trifluoromethylsulfonyl)imide. For each temperature and each ionic liquid, γ13 values were determined for two columns and the average γ13 values were used for the calculation of the partial molar excess enthalpies at infinite dilution, H E 1 Δ , using the Gibbs-Helmholtz equation. Graphs of In γ13 vs 1/T were plotted to determine H E 1 Δ at T = 303.15 K. The selectivity, S12 , value for the hexane/benzene separation was calculated from the ratio of the avarage γ13 values to determine the suitability of the ILs as an entrainer for extractive distillation in the separation of aromatic and aliphatic compounds.The S12 values were also compared to the literature values for other ionic liquids. It was found that the longer chain alkyl group attached to the cation or anion gave lower S12 values. Both ILs have S12 values greater than one and in theory can be used as an entrainer for the hexane/ benzene separation. The capacities at infinite dilutions, k1 , were also calculated from the inverse of the avarage γ13 values for each IL at each temperature.Item Adressing food and nutrition insecurity through the development and implementation of an agricultural and nutrition education programme on legumes for older farmers living in Mariannhill, KwaZulu-Natal, South Africa(2020) Mkhize, Nkumbulo Xolile; Napier, Carin E.; Oldewage-Theron, Wilna; Duffy, Kevin JanIntroduction Food and nutrition security can be influenced by various external and internal factors that contribute to a compromised wellbeing status amongst older farmers who are considered as a vulnerable group. This group faces diverse socio-economic and health challenges within an evolving food system. The complexity of this challenge persistently requires multisectoral intervention programmes, as reflected in the nature of this study. Aim and Objectives The aim of the study was to determine the levels of and address the food intake and nutrition insecurity status of elderly farmers. The main objective was to address the challenges that compromised the health of selected previously non-legume producing farmers by encouraging legume production and consumption amongst them. The findings of the study informed the proposal of appropriate strategies to develop an intervention programme intended to educate the identified elderly farmers and to promote their dietary diversification by means of legume production and consumption. Methodology The study employed a cross-sectional design. A baseline study was conducted that aimed at examining variable relationships using a mixed methods approach involving both qualitative and quantitative data collection techniques. A sample of 112 elderly farmers participated in the baseline study, followed by a case-controlled intervention programme that included preand post-intervention surveys. The measuring instruments included socio-demographic as well as health and medical surveys to elicit data such as blood pressure, blood indices for glucose and cholesterol due to dietary intake, household food security, legume knowledge, legume acceptability and agricultural practices. The data informed the findings of both the baseline and the intervention phases of the study. The baseline study involved a sample of 112 elderly farmers, whereas the intervention study involved two case controls (n=103) that comprised an experimental group (EG: n=53) and a control group (CG: n=50). Data were obtained for both groups by analysing blood indices, dietary intake, yields of legume production (EG only) and legume acceptability levels using statistical analyses of variance, significance and trends of pre- and post-intervention surveys over two years. A follow-up survey (FU) was conducted six months after the conclusion of the intervention phase and involved only the EG (n=32), to assess the impact of the intervention programme on their dietary intake and farming practices. The follow-up study thus compared the intervention data of the farmers’ consumption and production patterns with data obtained postintervention. During this latter study, only dietary intake and production levels were measured. All the data were analysed using IBM Statistical Package for Social Sciences (SPSS), Food Finder software, the Mann-Whitney U test, Analysis of Variance (ANOVA) and T-tests for statistical frequencies, variances and correlations. Results In the baseline study, 79.5% of the participants were women. The mean age of the participants was 63.3 ± 8.2 years. A diverse legume consumption was not observed as indicated by a low legume food group diversity score (FGDS) for 0-9 varieties with a mean of 3.5 (SD ± 1.70). Legumes were the least (19.0%) preferred crop planted by the farmers prior to the study. In the intervention study, the majority of participants were also women (73.6% in the EG and 66.0% in the CG). Of the six varieties of legumes under study, red kidney beans was the most preferred (141.60 kg/ha-1 SD ± 162.11). There was a high significance (p=0.001) amongst the six legume varieties produced, which implies that some legumes had a higher yield than others. A comparison between the genders indicated that the women produced 81.70 kg/ha-1 (SD ± 77.95), while the men produced 33.63 kg/ha-1 (SD ± 166.40). The higher yields produced by the women were significant (p=0.011). Legume consumption improved for the EG because the intervention programme resulted in a significantly (p=0.000) increased dietary diversity score (DDS) of legume intake with a mean (±SD) FGDS from 2.4 (SD ± 1.35) to 5.7 (SD ± 2.56) and a mean (±SD) Dietary Diversity Score (DDS) from 8.59 ± 0.74 to 8.23 ± 1.11, while that of the CG was 3.0 (SD ± 1.82). According to the post-intervention survey, the blood results of the EG indicated nutrient improvement at the 5.0% significance level for blood glucose (p=0.038) and cholesterol (p=0.008). The blood glucose levels of the EG were statistically significant (p=0.037) when this group was compared with the CG in both the pre-intervention and post-intervention surveys. Trend analyses were conducted and compared between those men and women who participated throughout the study (i.e. participation from the baseline to the post-intervention surveys). It was found that cholesterol (p=0.033) and Systolic Blood Pressure (SBP) (p=0.013) were statistically significant when the genders were compared across all phases of the study. Data obtained for the EG in the Follow Up (FU) study six months after the intervention study indicated that the production of legumes had declined (71.8%). This was because the legume intake with a mean (±SD) FGDS of 3.0 ± 2.2 indicated a significant (p=0.000) drop from 5.7 (SD ± 2.56) with DDS from 8.23 (± 1.11) to 8.81 (± 0.47) at p=0.005 significance as determined by the FU survey. It was also found that limited marketing of legumes had occurred, which suggests that the sustainability of legume production and consumption post-intervention by the elderly farmers was a challenge. However, 100.0% of the farmers indicated that they intended to continue their production and consumption of legumes, provided that the constant supply of legumes did not cease. Conclusions Evidence was obtained that the intervention project positively impacted the production and consumption of legumes and that the elderly farmers thus diversified their dietary intake for the better during this phase of the study. However, further investigations need to be conducted to explain the decline in consumption and production in the post-intervention period if support for a healthier lifestyle amongst elderly farmers is to be sustained. The technology to produce legumes with high yields to address food insecurity through more appropriate consumption patterns also needs to be explored in greater depth. Because the two streams of knowledge transfer that involved hands-on training of elderly farmers improved legume production and consumption patterns, this study established a sound foundation on which future studies and intervention strategies for food security amongst vulnerable groups may be built.Item Analysis of nanoscale ingredients in commercial food and cosmetic products by field-flow fractionation and single particle ICP-MS(2020-09) Naidoo, Lyndon; Bisetty, Krishna; Kanchi, Suvardhan; Sabela, Myalowenkosi InnocentThere is a growing need to disclose the possible presence of nanostructures on the labels of commercial food and cosmetic products in South Africa. Synthetic amorphous silica (SiO2) and titanium dioxide (TiO2) are the two widely used nanoparticles (NPs) selected for this study. This work was undertaken in two stages. The first part deals with an analytical method developed for the separation and characterization of TiO2 NPs capped with poly(ethylene glycol) (PEG) as a potential reference material for sunscreen analysis. The second aspect of this work focuses on SiO2, a highly attractive biomaterial widely used as a food additive (E551) to improve the flow properties of powdered food ingredients. Also, modern computational methods were implemented in both case studies, to better understand the nanoparticulate interactions with the organic substrates at an atomistic and molecular level. For the cosmetics study, asymmetric flow field-flow fractionation (AF4) coupled online to multi-angle light scattering (MALS) and dynamic light scattering (DLS) detectors were employed to assess the geometry and size of the PEGylated TiO2 NPs in terms of the evaluated radius of gyration (rg) and hydrodynamic radius (rh). Single particle inductively coupled plasma mass spectrometry (spICP-MS) and transmission electron microscopy (TEM) were implemented to provide information on the core diameter (d) of the TiO2 particle. To overcome agglomeration, Monte Carlo simulations were employed in this study to assess the effectiveness of PEG capped onto spherically constructed TiO2 anatase nanoclusters by systematically performing a series of adsorption studies. For PEG-TiO2 NPs, AF4-MALS-DLS reported rg and rh values of 28.7 nm and 40.3 nm respectively, with the shape factor (rg/rh) values generally reported in the range of 0.7 to 0.8, indicative of spherical particle geometry. SpICP-MS and TEM obtained complementary measurements of d = 32.0 nm and d = 38.4 nm, respectively. The computational modelling results demonstrated the strong binding affinity of PEG as a capping agent to TiO2, exhibiting stabilisation of TiO2 NPs in aqueous medium. Finally, the developed AF4-MALS-DLS method was applied to two commercial SPF 50 sunscreens, exhibiting promising separation and detection efficiency. These findings can contribute to regulatory measures in line with the South African National Nanotechnology Strategy for the cosmetics industry. With regards to the food application study, a multivariate method was developed for the detection and characterization of SiO2 particles also based on AF4-MALS-DLS. This analytical approach attempts to address the fate and the presence of nanoparticulate SiO2 additives (E551) in food products. The experimental design using SiO2 NP standards resulted in the following optimum conditions of the system: crossflow, 0.8 ml/min; injection time, 5 min and sonication time, 60 min. It was observed that the average geometric diameters (Dgeo) for SiO2 NPs in three selected commercial coffee creamers (A, B and C) detected by AF4-MALS were 286.7 nm, 129.8 nm and 190.7 nm respectively. Similar trends for the coffee creamers were observed for the hydrodynamic diameter (Dh) measurements using AF4-DLS i.e. 301.5 nm, 141.3 nm and 197.8 nm respectively. The rg/rh values were reported ranging from 0.7 to 0.8, indicative of spherical particle geometry. Also, the electrostatic interactions between SiO2 NPs and glucose/water mixtures, as evaluated by Monte Carlo simulations, revealed O-interactions dominating over the flat amorphous SiO2 surface. The strongest interaction observed (around - 239 kcal/mol) for the SiO2-water/glucose mixture demonstrates the hydrophilic nature of SiO2 NPs. The findings in both case studies provide fundamental information to improve the understanding of nanoparticulate interactions with additives and paves the way for the labelling of cosmetic and food products that potentially exhibit nanomaterials in complex matrices.Item Analysis of selected organic pollutants in water using various concentration techniques(2014-08-08) Ramphal, Sayjil Rohith; Moodley, Kandasamy Govindsamy; Chetty, Deenadayalan KistenAmong persistent organic pollutants, chlorobenzenes are some of the most frequently encountered compounds in aqueous systems. These compounds can enter the environment via natural and anthropogenic sources, and are ubiquitous due to their extensive use over the past several decades. Several chlorobenzene compounds, once in the environment, can biologically accumulate, and are reputed to be carcinogens and extremely hazardous to health. Several chlorobenzenes are listed as priority pollutants by the United States Environmental Protection Agency. Excessive exposure to these compounds affects the central nervous system, irritates skin and upper respiratory tract, hardens skin and leads to haematological disorders including anaemia. In spite of these harmful effects, chlorobenzenes are still used widely as process solvents and raw materials in the manufacture of pesticides, chlorinated phenols, lubricants, disinfectants, pigments and dyes. In the light of the above, it is imperative to monitor the levels of chlorinated benzenes in all types of surface waters, using low-cost but sensitive methods of preconcentration and detection. In this study, a simple and relatively cheap preconcentration method using direct immersion solid phase microextraction (DI-SPME) followed by gas chromatography equipped with a flame ionisation detector (GC-FID) was developed for the analysis of 7 chlorinated benzenes in dam water. Experimental parameters affecting the extraction efficiency of the selected chlorobenzenes, such as fibre type, sample size, rate of agitation, salting-out effect and extraction time, were optimised and applied to the Grootdraai Dam water samples. The optimised method comprises the use of a 100 µm polydimethylsiloxane (PDMS) fibre coating; 5 ml sample size; 700 revolutions per minute rate of agitation and an extraction time of 30 minutes. The calibration curves were linear with correlation coefficients ranging from 0.9957–0.9995 for a concentration range of 1–100 ng/ml. The respective limits of detection and quantification for each analyte was as follows: 1,3-dichlorobenzene, 0.02 and 0.2 ng/ml; 1,4-dichlorobenzene, 0.04 and 0.4 ng/ml; 1,2-dichlorobenzene, 0.02 and 0.2 ng/ml; 1,2,4-trichlorobenzene, 0.3 and 2.7 ng/ml; 1,2,4,5-tetrachlorobenzene, 0.09 and 0.9 ng/ml; 1,2,3,4-tetrachlorobenzene, 0.07 and 0.7 ng/ml; pentachlorobenzene, 0.07 and 0.7 ng/ml. Recoveries ranged from 83.6–107.2% with relative standard deviation of less than 9%, indicating that the method has good precision, is reliable and free of matrix interferences. Water samples collected from the Grootdraai Dam were analysed using the optimised conditions to assess the potential of the method for trace level screening and quantification of chlorobenzenes. The method proved to be efficient, as 1,3 dichlorobenzene, 1,4-dichlorobenzene and pentachlorobenzene were detected at concentrations of 0.4 ng/ml, 1.7 ng/ml and 1.4 ng/ml, respectively.Item Analysis of the nutritional status and dietary intake data of a group of elderly at a day and frail care centre in Verulam(2011) Govender, Theloshni; Oldewage-Theron, Wilna; Napier, Carin E.Background: South Africa, a richly diverse developing country has been faced by the consequences of transition attributed to urbanisation and acculturation. A Westernised lifestyle has, therefore, resulted in increasing disease patterns that are characterized by a combination of poverty-related diseases together with the emerging chronic diseases. The shift to a Westernised lifestyle has resulted to a shift in the composition of dietary staples leading to dietary factors related to an increase in lifestyle diseases. These include a high fat, low fibre diet, as well as an inadequate intake of fruits and vegetables. However, this in turn has led to higher energy intakes with insufficient and imbalanced micronutrient intake. Research conducted amongst the elderly in South Africa has clearly indicated that the elderly live within a limited financial budget leading to extreme levels of food insecurity and the social burden of being the head of the household, in addition to being the caregiver to grandchildren and sick children. Due to the current living status the elderly encounter reduced food intake in addition to a reduced variety in their diet, therefore, micronutrient deficiencies are common amongst this age group. Therefore, a consumption of energy-dense foods, particularly staple foods, to stretch the food budget which are more affordable and thus allow for an increased consumption is evident. Aim: To determine the socio-demographic profile, health and nutritional status in relation to the dietary intake patterns to reflect malnutrition among free living elderly (60yrs+) in Verulam. Methodology: Fifty nine randomly selected men and 191 women aged 60+ participated on a voluntary basis in this study. A descriptive survey method was used for this cross sectional study. Trained fieldworkers assisted with the administration of all questionnaires and a registered nurse measured blood pressure. Socio-demographic questionnaires were administered to determine the socio-economic characteristics of the elderly within this community. Anthropometric measurements determined the Body Mass Index according to the World Health Organisation and Asian cut-off points to indentify the risk factors. The Health questionnaire identified the health status correlated to the respondent’s profundity of disease and deficiencies associated to dietary patterns. Blood pressure measurements were taken to determine the hypertension prevalence related to the dietary intake. Two 24-Hour Recall questionnaires were completed by the 250 respondents to identify actual vii food intake and measured against the Dietary Recommended Intake (DRIs). A food frequency questionnaire (FFQ) determined the respondent’s food variety score over a period of one week. The socio-demographic questionnaire, health questionnaire, food frequency questionnaire and anthropometric measurements were captured on an Excel® spread sheet by the researcher and analyzed for descriptive statistics using the Statistical Package for the Social Sciences (SPSS) version 17.0 with the assistance of a statistician. The 24-Hour Recall data were captured and analyzed by a nutrition professional using the MRC Food Finder® version 3.0 software, based on the South African Food Composition tables. Results: The majority of the respondent’s role in the family was mothers (70.8%) and lived in an urban area (68.8%). In addition, 73.2 percent (n=183) of the respondents shared the house with one to five people, and lived in a brick house (74.4%, n=186) with more than 3 rooms (74.0%, n=185). The elderly in this sample were pensioners and, therefore, 76.0 percent (n=190) received state grants of which the total household income ranged between R1001-R1500 (35.2%, n=88). Food insecurity is prevalent as reported by 28.4 percent. Primary school was the highest level of education completed by 52.4 percent (n=131) and English is the most spoken language amongst this group (74.0%, n=185). Women had higher BMI values particularly in the overweight (18.32%, n=35) and obese I and II (58.6%, n=112) categories when compared with men. Blood pressure measurements indicated that 60.0 percent (n=150) of the respondents suffered from hypertension. The use of chronic medication was reported by 84.4 percent (n=212). The total range of individual food items consumed by an individual during the seven-day data collection period measured by the (FFQ) was between 4-66 foods. However, the highest consumption was four food items by 23.2 percent (n=58) of the respondents. The summary of the food variety within food groups indicated a high dietary diversity, of which the other vegetable group reported the highest individual mean FVS (±SD) of 10.86 (±5.82), followed by other fruit, cereal, flesh and Vitamin A rich groups with 5.73 (±4.41), 5.03 (±1.85), 4.08 (±2.23) and 2.43 (±1.09) respectively. The nutrient analysis indicated a deficient intake by both men and women of all the nutrients, except for the mean (±SD) total protein in the women 45.10 (±12.55) and carbohydrate 212.83 (±36.97) in the men. The energy contribution indicated 98.3 percent (n=58) men and 85.72 percent (n=158) women consumed <100 percent of the EER for viii energy. However, the findings from the Top 20 food items measured by the 24-Hour Recall indicated that this community’s diet is largely carbohydrate-based, containing primarily starchy staple foods, sufficient intake of animal products, and insufficient intakes of dairy foods, fruit and vegetables, possibly resulting in the micronutrient deficiencies. The energy distribution of the macronutrients from the average of both 24-Hour Recalls indicates that both men and women are in range of 15-30 percent total fat intake, 10-15 percent protein and 55-75 percent carbohydrate. Conclusion: The results of the study reflect that the elderly in this community are faced with poverty, food insecurity as well as social factors thus contributing to a compromised nutritional status. The progression of malnutrition in particular overnutrition is experienced by the majority of the respondents in this study, however, an increased BMI and the prevalence of hypertension is a risk marker for noncommunicable diseases. However, the high prevalence of inadequate food and nutrient intake amongst elderly discloses the need for nutrition interventions and should be aimed at modifying the elderly food choices when purchasing food, healthier food preparation methods, increasing fruit and vegetable portions and improving daily physical activity to attain a better quality of life.Item Anthropometric profile and food consumption patterns of an emerging middle income community in Umlazi(2017) Memela, Sinenhlanhla Ntokozo; Napier, Carin E.Introduction: The nutrition transition is a global phenomenon affected by developing economic prosperity. ‘Nutrition transition’ may be defined as changes in dietary patterns and nutrient intakes occurring in individuals, families, groups of people or whole populations when the food environment and other circumstances change. Post-1994 democratic elections in South Africa, economic laws called for the de-racialization of the economy. As a result of the annulment of the apartheid laws in the 1990s entrepreneurial opportunities became available for black people. Based on the 2011 Census, the middle class constituted 8.3 million (17%) out of the 51 million population in South Africa. Blacks made up 51%, whites 34%, coloureds 9% and Indians 6%. This is a vast difference from two decades prior to this when blacks made up only 3.3% of the population, with coloureds making up 9.1% and Indians 18.5%. In a number of Sub-Saharan African studies obesity has been shown to have a significant link to socio-economic status determined by access to water and electricity, smaller numbers of members residing in one household and increased amounts of money spent on food among other factors, which all promote a tendency to leading a more Westernised lifestyle. The main objective of the study was to determine the anthropometric status and analyse the food consumption of a community of emerging middle income status in Umlazi Township (Ward 84) outside Durban in the province of KwaZulu-Natal. Methods: This study had a sample size of n=250 adult men (n=56) and women (n=194) who were heads of households or primary caregivers above the age of 19 years old with households receiving a total household income of more than R100 000.00 per annum/ more than R8000.00 per month. The study was quantitative and descriptive in nature with different tools used to measure the sample population. These included waist circumference and height and weight measurement to determine BMI and WHtR. A socio-demographic questionnaire was completed where the multi-dimensional poverty index (MPI) was established; a food frequency questionnaire (FFQ) and three 24-hour food recall were also administered to all participants; one weekend day recall and two weekday recalls. Data were analysed on SPSS and nutrient analysis done through Food Finder Version 3 Software and compared to the World Health Organisation standards recommendations for optimal health. Results: The mean age of the group was 52 years old. Slightly more than half (51.6%; n=129) of the population was unemployed. Retired participants made up 27.2% (n=68) of the population. The majority (80%) of the households indicated that between one to three members contributed to the total household income. The MPI of the Ward 84 community amounted to 9.09. The prevalence of obesity was 37.59% (n=21) among men and 76.80% (n=149) for women. Women had a higher prevalence of risk factors linked to non-communicable diseases than men. Mineral and fat soluble vitamin intake was deficient; however protein and carbohydrates were consumed in excess with means of 187.70% and 111.43% respectively. Top 20 foods consumed lists indicated that little fruit or vegetables were consumed. The average food variety score was medium indicating the consumption of between >30 but <60 different foods across the total population. A positive relationship of statistical significance was found between food variety score and income (p=0.007). Conclusion: The Ward 84 community consisted of an ageing population with households mostly supported by another member’s income from within or outside the household. The adults had sufficient education to have secured promotions in the employment arena and could afford a lifestyle similar to that of those who had always been more privileged. Although the Umlazi residents benefited from good housing and infrastructure and appeared to have averted poverty, the prevalence of overnutrition malnutrition was high and linked with an excessive intake of macronutrients. Majority (70%) of the community of Ward 84 is ‘food secure’ as indicated by the number of participants’ households indicating never having a shortage of money to buy food. This correlates negatively with the high prevalence of overweight and obesity (BMI) (p=-0.029). According to literature a persistent issue in black communities is the acceptance of an overweight body image being perceived as ‘healthy’ and an indicator of affluence. This community appears to be in the 4th stage/ pattern of the nutrition transition. It appears that a ‘westernised diet’ and lifestyle is more accessible and is embraced by this urban community as described in various literature and seen in the top 20 foods lists compiled from the 24 hour recalls. There is some oversight regarding the nutrient quality of a varied diet and achieving energy balance with macronutrient intake linked to informed nutritional choices. Suggested interventions could include the dissemination of information regarding nutrition and practical suggestions to achieve energy balance. In addition, future research could investigate the prospects of introducing participation in urban agriculture, investigate the issue of physical activity, and foster an environment for more physical activity and increase availability and access to micronutrient dense foods and investigate the development of a body mass index specifically for the black African population.Item Anti-carcinogenic activity of Centella asiatica and Elytropappus rhinocerotis on a human colon cancer cell line(2012) Dwarka, Depika; Odhav, Bharti; Sewram, VikashRecently our understanding of cancer has advanced in the realization that apoptosis and the genes that control it have a profound effect on the malignant phenotype. It is now clear that some oncogenic mutations disrupt apoptosis, leading to tumor initiation, progression or metastasis. Conversely, compelling evidence indicates that other oncogenic changes promote apoptosis, thereby producing selective pressure to override apoptosis during multistage carcinogenesis. Finally, it is now well documented that most cytotoxic anti-cancer agents induce apoptosis, raising the intriguing possibility that defects in apoptotic programs contribute to treatment failure. Because the same mutations that suppress apoptosis during tumor development also reduce treatment sensitivity, apoptosis provides a conceptual framework to link cancer genetics with cancer therapy. An intense research effort is uncovering the underlying mechanisms of apoptosis, such that, in the next decade, one envisions that this information will produce new strategies to exploit apoptosis for therapeutic benefit. Plants have a long history in cancer treatment. More than 3000 species have been known for their anti-cancer potential. Over 60% of currently used anti-cancer agents are derived in one way or another from higher plants. Indeed, compounds derived from natural sources, including plants, have played, and continue to play, a dominant role in the discovery of leads for the development of conventional drugs for the treatment of most human diseases especially cancer. Thus the aim of this study was to investigate if Centella asiatica and Elytropappus rhinocerotis possess anti-cancer potential and determine the effect on the modulation of apoptosis. In South Africa C. asiatica is known anecdotally to treat various forms of cancers and E. rhinocerotis is known to treat colic and diarrhoea. The anti-cancer activity of C. asiatica has been studied in some parts India but E. rhinocerotis has not been investigated. This study was conducted using polarity guided fractionation (aqueous, ethanolic, methanolic and hexane), thereafter these extracts were tested for their toxicity on a colon cancer cell line (CaCO-2) and on normal cells vi (PBMC). Subsequently, the most active extract was used to isolate the active fraction. The fraction that displayed toxicity on the CaCO-2 cells were further investigated for their ability to induce apoptosis by observing the morphological effects and DNA changes using acridine orange-ethidium bromide staining. Apoptosis was confirmed using Annexin V- PI staining. Nuclear effects were studied by DNA fragmentation and by agarose gel electrophoresis. Nuclear fragmentation was studied by flow cytometry using bromodeoxyuridine (BrDU). Pro-apoptotic changes were determined with Caspase III enzyme levels using flow cytometry. The results were compared to the effect of a known anti-carinogen - Taxol. The anti-oxidant activity was also evaluated for the different extracts. The ethanolic extracts of both C. asiatica and E. rhinocerotis showed more than 100% radical scavenging activity. The methanolic extract (125 μg/ml -500 μg/ml) showed cytotoxicity on the CaCO-2 cells and a proliferative effect on the PBMC. Apoptosis was confirmed in the methanolic extract for both plants and was therefore used to carry forth this study. This included early apoptotic changes observed by the morphological study i.e., membrane blebbing, nuclear condensation and the presence of apoptotic bodies, in both C. asiatica and E. rhinocerotis fractions demonstrated more non-viable apoptotic cells than the methanolic extracts. Late changes of apoptosis were also found as indicated by DNA laddering and a positive outcome with BrDU. Both the active fractions from C. asiatica and E. rhinocerotis showed more DNA laddering and active caspase III than the methanolic extract. These features indicate that C. asiatica and E. rhinocerotis cause apoptotic death of colon cancer cells CaC0-2. In conclusion, there was a significant increase in apoptosis of CaCO-2 cells with little alteration of PBMC in the presence of the methanolic extract of C. asiatica and E. rhinocerotis. The semipure fractions resulted in changes related to late apoptosis. The results suggest that C. asiatica and E. rhinocerotis induces apoptosis in CaCO-2 cells which is an important step in elucidating the underlying molecular mechanism for anti-tumour activity.Item Anti-HIV activity of selected South African medicinal plants(2013-09-17) Hurinanthan, Vashka; Odhav, Bharti; Parboosing, RaveenSouth Africa has the largest number of people infected with HIV/AIDS. It also has more than 30 000 species of plants and many of these have a long tradition of medicinal use. It is highly likely that the treatment for HIV will come from this traditional knowledge. The need for effective preventative and therapeutic agents for HIV remains an urgent global priority. The aim of this study was to screen selected South African medicinal plants for anti-HIV activity and to identify and characterise an active compound from a plant that can be used for HIV treatment. The aqueous and methanolic extracts of the roots, leaves, flowers and stems of thirty eight plant species (108 extracts) were screened for anti-HIV activity. The plants which had anti-HIV activity were further screened for anti-reverse transcriptase activity. Thirty-two extracts exhibited varying degrees of anti-HIV activity. Cleome monophylla, Dichrostachys cinerea and Leonotis leonurus aqueous leaf extracts had anti-HIV-1 reverse transcriptase activity. The aqueous extracts of D. cinerea showed the best anti-HIV activity with a Selectivity Index of 43.5 and significant anti-HIV-1 reverse transcriptase activity. Crude phytochemical screening of D. cinerea showed that it had tannins, saponins, flavonoids and alkaloids but did not contain any phlobatannins, terpenoids, steroids or phenols. D. cinerea displayed a high degree of free radical scavenging activity with an IC50 of 25 μg/ml, therefore the anti-HIV activity could be attributed to the flavonoids present in the plant. Bio-guided fractionation was used to isolate and purify the active compound from the D. cinerea extract. Compounds were isolated by thin layer chromatography and were tested for anti-HIV-1 and anti-reverse transcriptase activity. From these results the active compound was identified, and purified using preparative TLC. The active compound was characterised by High Performance Liquid Chromatography, Ultraviolet-visible spectrophotometry, and Ultra Performance liquid chromatography coupled to MS/MS. Structural elucidation was performed using Nuclear Magnetic Resonance. From these results, it was deduced that the compound isolated from D. cinerea was a catechin. In this study we show that the catechins present in D. cinerea are responsible for the anti- HIV-I activity and inhibits the reverse transcriptase activity which is a key factor in the progression of HIV. Potentially, these results can be used to develop a new drug for the treatment of HIV or as a cost effective therapeutic agent in treating HIV-infected individuals with oxidative stress.Item Anti-inflammatory, anti-oxidant and wound-healing properties of selected South Africa medicinal plants(2017) Mzindle, Nonkululeko Betty; Odhav, BhartiSouth Africa has a wide range of medicinal plants that are used traditionally by black Zulu South Africans for the treatment of a range of illnesses, including inflammatory ailments; disease conditions caused by oxidative stress and wound healing. It has been indicated that bioactive compounds isolated from plants contribute to their anti-inflammatory, antioxidant and wound healing properties; hence, herbal remedies have been widely used traditionally in many countries in the management and treatment of wounds. Inflammation is the main condition that relates to a variety of diseases affecting most of the world’s population. It is the body’s immune response to infection and injury and is induced by the release of pro-inflammatory mediator’s —prostaglandins and leukotrienes—following wound occurrence. Wounds result in disruption of living tissue caused by oxidative stress. Anti-inflammatory agents, antioxidants, and antimicrobials play an important role in the wound healing process and they prevent aggravated wound conditions.Controlling inflammation during wound repair is important to minimize any additional complications that may result; hence, chemical agents such as non-steroidal anti-inflammatory drugs (NSAIDS), synthetic antioxidantsand steroids are frequently used. These drugs block the enzymes that are responsible for prostaglandin synthesis in inflammation, react with free radicals thereby interfering with oxidation process as a result affect one or more phases of wound healing. The use of these drugs, however, has been limited as they can cause detrimental side effects when used over long periods of time.There is, consequently, a need to find alternative natural therapeutic drugs. Studies on medicinal plants confirmed that herbal drugs exhibit fewer side effects in comparison with chemical agents and are more cost-effective.Thus the aim of this study was to investigate South African medicinal plants, for anti-inflammatory, antioxidant and wound healing properties. Dissolved extracts of thirty-eight medicinal plants were evaluated for theiranti-inflammatory activity using the 5-lipoxygenase assay as well as free radical scavenging activity using the 1; 1-diphenyl-2-picrylhydrazyl (DPPH) assay.Their safety was evaluated using brine shrimp lethality assay. Proliferation and viability of fibroblast cells was determined by the3-(4, 5-dimethylthiazolyl)-2, 5-diphenyltetrazolium bromide(MTT) assay furthermore a scratch wound assay was used to study the properties of wound healing in vitro and to confirm the anti-inflammatory activities of the dissolved extracts. Migration rate was evaluated quantitatively by an image analyzer. Methanol was chosen for extraction because it completely dissolves extracts. Anova was used for statistical analysis. Almost all aqueous extracts were found to be effective in inhibiting lipoxygenase enzyme when compared to nordihydroguaiaretic acid (NDGA). Aqueous extracts exhibited remarkably high percentage inhibition of lipoxygenase with most above 100% when compared to methanolic extracts. Amaranthus dubius and Portulaca oleracea were found to have good biological activities in the inhibition of 5-lipoxygenase enzymes when compared to the other plants. However, Galinsoga parviflora and Syzygium cordatumwere least effective in inhibiting enzyme activity with percentages as low as -2% and 34% respectively. Percentage inhibitions for methanolic extracts were lower than that of aqueous extracts. Amaranthus spinosus had the highest percentage inhibition among all the methanolic extracts andGalinsoga parviflorahad the lowest. The methanolic plant extracts were found to be more effective in scavenging DPPH free radicals than the corresponding aqueous extracts. All the methanolic extracts exhibited free radical scavenging ability in the range of 60%–104%. Asystasia gangetica, Ficus sur, Heteropyxis natalensis, Hibiscus sabdariffa, Pelargonium sp. showed notably higher scavenging abilities, ranging from 101%–104% compared to Rutin. Methanolic extracts of Heteropyxis natalensis and Hibiscus sabdariffa exhibited scavenging ability even at the lowest concentration of 10μg/ml. Furthermore, aqueous extracts displayed remarkably lower activities than methanolic extracts with thirty-one extracts having a scavenging capacity ranging from 22%—59%. None of the extracts were found to be detrimental to brine shrimp. Almost all the extracts were shown to stimulate the growth of fibroblast cells except the methanolic extract of Solanum nodiflorum, which was shown to be killing the cells at high concentrations with a percentage viability of 46%.As the concentration decreased, however, the viability of cells with this extract increased to 143%. An increase in the number of fibroblast cells was observed in the scratched area of the treated cells and a significant migration rate was also noted with some of the extracts. Aqueous extracts of Sonchus oleraceus (86%), Justicia flava (85%) and Dichrostachys cinerea (85%) and methanolic extracts of Senna occidentalis and Hibiscus sabdariffa were found to have the highest migration rate compared to untreated cells that served as a control. No cell migration was observed with methanolic extract of Solanum nodiflorum.Instead, the extract was found to be toxic to the cells. Some of the plants evaluated in this study have been studied for either anti-inflammatory, antioxidantand wound healing properties in vivo, however, no work has been conducted to demonstrate a correlation between anti-inflammatory, antioxidant and wound healing properties of plant species in vitro. The current study was, therefore, conducted to review medicinal herbs considered as anti-inflammatory, antioxidants and wound healing agents as well as collecting evidence for their effectiveness and pharmacological mechanisms in modern science. In the plant species investigated Amaranthus dubius, Asystasia gangetica, Bidens pilosa, Buddleja saligna, Carpobrotus dimidiatus, Chenopodium album, Dichrostachys cinerea, Emex australis, Ficus sur, Guilleminea densa, Hibiscus sabdariffa, Physalis viscose, Syzygium cordatum, Taraxacum officinale and Tulbaghia violacea demonstrated good anti-inflammatory and wound healing properties.In conclusion the results from this study demonstrated promising anti-inflammatory and antioxidantactivities as well as wound healing properties,furthermoreit was aslo shown that the plant extracts were not toxic to the cells hencethis suggested that the plants investigated, can be used as substitutes or to formulate wound healing agents that are safe to use in primary healthcare.Item Anti-microbial activity of phenolic extracts from Virgilia oroboides and Chlorophora excelsa(2000) Padayachee, Thiriloshani; Odhav, BhartiThis study focussed on the anti-bacterial, anti-fungal and anti-protozoal activity of four plant extracts, maackiain and formononetin from Virgilia oroboides and chlorophorin and lroko from Chlorophora excelsaItem Anticancer activity of ceratotheca triloba(2016) Naicker, Leeann; Odhav, Bharti; Matsabisa, M.G.; Mohanlall, VireshPlants have provided a source of medicine from the beginning of human history and are the core of modern medicine. Moreover, plant based drug discovery has led to the development of various anticancer drugs (such as vincristine, vinblastine, etoposide, paclitaxel, camptothecin, topotecan and irinotecan). The use of botanical, phytochemical, biological and molecular techniques have facilitated the discovery of anthraquinones from Ceratotheca triloba that can inhibit the human topoisomerase II enzyme (target for anticancer drugs) and kill cancer cells. However, the C. triloba plant has not been extensively studied for its anticancer activity. Therefore, the aim of this study was to further investigate the anticancer activity of C. triloba and determine the classes of compounds that contributed towards its activity. In this study the leaf and root extracts were prepared by using hexane, DCM, hexane: DCM (1:1), methanol and/or water. These extracts were examined for their growth inhibitory potential on three cancer cell lines (A375 [melanoma], MDA-MB-231[breast] and WHCO1 [esophageal]) by using the MTT assay. Then, different mobile phases were prepared for optimizing the separation of the compounds of the active extract by TLC. Column chromatography was performed with the active extract by using five mobile phases (hexane : DCM [60 : 40, 40 : 60], DCM, DCM : ethyl acetate [90 : 10, 70 : 30, 60 : 40, 50 : 50, 50 : 60, 30: 60, 20 : 80], ethyl acetate and ethyl acetate: methanol [80 : 20, 70 : 30, 50 : 50]). The fractions collected from the column were examined for their growth inhibitory potential on two melanoma cell lines (A375 and UACC-62). The IC50 and TGI (total growth inhibition) values of the active fractions were determined. Also, the apoptosis inducing effects of the active fractions and standards (camptothecin and doxorubicin) were determined by using flow cytometer based assays (FITC annexin assay, PE active caspase 3 assay and BD MitoScreen assay). Subsequently, the chemical structures of the compounds that contributed towards the activity of these fractions were obtained by EI-LC-MS analysis. The results demonstrated that the hexane root extract exhibited the best percentage of growth inhibition (%GI) on all three cancer cell lines. The separation of the compounds of the hexane root extract was optimized on TLC plates by using different ratios of hexane and DCM. Column chromatography allowed for fractionation of this extract. Purified compounds were not obtained due to co-elution. Further research would have to be conducted to obtain purified compounds. This may involve the use of mini-column chromatography and PTLC. Overall a total of ten combined fractions were collected from the column. Four of these fractions (F2, F4, F5 and F8) displayed a high %GI on the A375 and UACC-62 cell lines. Moreover, fraction F4 was the most active fraction as it had the lowest IC50 (0.70 µg.ml-1 [A375] and 0.39 µg.ml-1 [UACC-62]) and TGI (12.50 µg.ml-1[A375] and 25 µg.ml-1 [UACC-62]) values in comparison to the other fractions. All four fractions induced depolarization of the mitochondria membrane potential (ΔΨ), caspase 3 activation, early apoptosis (phospholipid phosphatidylserine exposure) and/or late apoptosis in the melanoma cells. The results also revealed that fraction F4 (25 µg.ml-1) induced depolarization of the ΔΨ in a higher percentage of A375 (78.11%) and UACC-62 (87.4%) cells than the other fractions and standards. This fraction also induced caspase 3 activation in a high percentage of A375 (90.56%) and UACC-62 (96.78%) cells. Therefore fraction F4 was also the most active fraction in terms of apoptosis activity. Based on our results and literature findings we can deduce that the active fractions induced the intrinsic or extrinsic (type II) apoptosis pathway in the melanoma cells. Six classes of compounds were identified from the four active fractions. These were: benzothiophenones, benzopyranones, naphthoquinones, anthraquinones, androstanes and quinazolines. In conclusion, this is the first study that evaluated the growth inhibition potential of the leaf and root extracts of C. triloba on a panel of cancer cells. This research indicated that the hexane root extract displayed the best levels of cell growth inhibition. The active constituents of this extract were isolated into four fractions which elicited apoptosis inducing effects that promoted the extrinsic (type II) or intrinsic apoptosis pathway in the melanoma cells. Furthermore, fraction F4 contained the most active compounds from C. triloba as it had the lowest IC50 and TGI values (in comparison to the other fractions) and induced depolarization of the ΔΨ in the highest percentage of melanoma cells. It was confirmed that six classes of compounds were accountable for the anticancer activity of these fractions. Thus, the C. triloba plant is a rich source of anticancer compounds.Item Anticancer activity of silver nanoparticles embedded in porous starch as a potential delivery system(2024-05) Mohan, Naaznee; Mellem, John JasonSilver nanoparticles have been proven to have anticancer abilities but they have been known to agglomerate and become toxic. Therefore, various studies have been conducted to explore ways of preventing aggregation using biopolymers such as starch. This study makes use of Lablab purpureus (hyacinth bean) porous starch to biosynthesize and encapsulate silver nanoparticles and then test its anticancer potential. Porous starches were produced from hyacinth bean using three different techniques. These were compared against the native starch with silver nanoparticles, then synthesized and encapsulated using the porous starch. In comparison to the native starch, the porous starches made through solvent exchange and enzyme hydrolysis had similar outcomes with granules exhibiting pores, as shown by the structural and chemical characteristics. The lack of pasting properties and extremely distinct chemical and structural graphs of the porous starch, produced by freeze-thaw procedures, may be related to the presence of mercaptosuccinic acid. It was decided to employ porous starch made by solvent-exchange (SE) for the manufacture of silver nanoparticles as it contained resistant starch. Nanoparticles were produced using the porous starch from solvent-exchange, characterised and tested for their anticancer potential. Silver nanoparticles were indicative of a colour change from clear to brown, as well as, the characteristic peak at 425 nm for silver nanoparticle formation. Silver nanoparticles were implanted into porous starch at a size of around 50 nm, as further evidenced by the particle size distribution and TEM images of spherical granules with dark spots within. The zeta potential for the silver nanoparticles was -34 mV, thereby indicating that aggregation was minimized and particles were stable. The nanoparticles demonstrated less cytotoxicity in the human colon (CACO) and cervical (HELA) cancer cell lines, but more inhibition in the human breast (MCF-7) cancer cell line than the positive control camptothecin. The human muscle (C2C12), normal cell line's capacity to sustain cell viability for silver nanoparticles demonstrated that AgNP were not toxic. However, to maximize the potential of the silver nanoparticles implanted in porous starch, more research is necessary.Item Anticancer and anti-reactive oxygen species activity of bioactive peptides isolated from vigna unguiculata(2024-05) Ramsookmohan, Sonaal; Mellem, John Jason; Dwarka, DepikaCancer is a major cause of death globally and continues to escalate with current anticancer drugs associated with severe side effects and resistance driving the need for safer alternative therapeutics. Food proteins, from legumes, are a source of bioactive peptides and studies revealed that they are associated with various therapeutic properties. Cowpea (Vigna unguiculata) is an underutilized nutritious legume crop with promising potential due to its documented protein profile. Therefore, this study evaluated the in vitro anticancer effect of V. unguiculata peptides derived from alcalase and flavourzyme. Physicochemical properties such as water and oil absorption capacities, emulsifying properties, sub-unit composition, amino acid composition among others, were also assessed. Peptides were also evaluated for their antioxidant activity using superoxide radical scavenging, 1,1-diphenyl-2-picrylhydrazil (DPPH) and 2,2’-azino-bis (3 ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays as well as for their apoptotic potential using 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), reactive oxygen species (ROS) and caspase 3/7 assays on cancerous (Caco-2 and MCF 7) and healthy (C2C12) cell lines. From results obtained it was observed that the foam capacity for the peptides derived from alcalase and flavourzyme were 78.34 and 82.39%, respectively, which was noted to be significantly different. The physicochemical properties determined their potential application in food industries. Glutamic acid was the most abundant amino acid in all samples while methionine was noted to be the least concentrated amino acid in the flour and alcalase derived sample while proline was the least concentrated in the flavourzyme sample. Results from this study suggest that cowpea samples have antioxidant capabilities with enzymatic hydrolysis contributing to a higher capacity compared to that of raw flour flour samples. From the cowpea flour, the peptide sample derived from alcalase demonstrated the highest DPPH free radical scavenging activity (70.88-80.47%), followed by flavourzyme (67.27-75.84%), while the raw flour sample showed the lowest activity (24.28-66.17%). The ABTS scavenging capacity of the alcalase peptide was in the range of 35.26-85.92%. The MTT cytotoxicity assay revealed that the cowpea peptides and camptothecin showed different sensitivities on the MCF-7 cell lines. The IC50 values of flavourzyme peptide, alcalase peptide and camptothecin were 0.07, 0.09 and 0.07 µg/mL respectively. Cell viability of the cowpea peptides and camptothecin (control) on the Caco-2 cells varied with the different concentrations. Alcalase and flavourzyme samples had IC50 values of 0.15 and 0.11 µg/mL respectively. The apoptotic potential of the peptides was further shown by the caspase 3/7 activity. From the results in this study, it can be ascertained that the cowpea peptides have potential as an anticancer therapeutic agent. Further research is necessary to determine mechanism of action and to conduct in vivo evaluations of these peptides using animal modelsItem Anticancer and antioxidant potential of Amaranthus cruentus protein and its hydrolysates(2020) Ramkisson, Shanece; Mellem, John J.; Venter, SonjaTraditionally, amaranth has been acknowledged to possess vital pharmacological properties with anticancer peptides having been found in Amaranthus cultivars. However, limited knowledge is available over the use of pepsin and alcalase enzymes to form hydrolysates. Thus, this study was aimed at comparing the in vitro anticancer effect of Amaranthus cruentus (grain) protein isolate and hydrolysates using alcalase, trypsin and pepsin. The safety of hydrolysates was investigated using the Ames mutagenicity and Brine Shrimp Lethality assay. Protein hydrolysates were thereafter investigated for their antioxidant potential using the FRAP, ABTS and DPPH assays. Subsequently, the protein hydrolysates were tested for their anticancer and apoptotic potential. The MTT assay was conducted to evaluate the cytotoxic potential of the protein hydrolysates using the HEK 293 (non-cancerous), A549 (cancerous) and MCF-7 (cancerous) cell lines. After that, morphological alterations were examined using the acridine orange and ethidium bromide double stain. Following this, the Annexin V apoptotic detection kit was used to quantify apoptosis together with the Glomax Caspase 3/7 kit to detect changes in the cell cycle Results show A. cruentus isolate and hydrolysates had no mutagenic response against Salmonella typhimurium TA 98 and TA 100 strains. The tested samples did not induce any significant increase in the death percentage of Artemia spp. in comparison to potassium dichromate (control). DPPH assay revealed that the hydrolysed samples had an enhanced scavenging activity compared to the unhydrolyzed sample, with pepsin having the greatest IC50 of 23.06 µg/ml. Amaranthus cruentus isolate (IC50 17.57 µg/ml) was a greater scavenger of the Fe+ ions compared to the control glutathione (IC50 79.81 µg/ml). For ABTS, all hydrolysates had a greater antioxidant scavenging potential compared to the isolate. The MTT cytotoxicity assay revealed that the isolate produced a greater cytotoxic effect on the MCF-7 and A549 cell line when compared to the control (camptothecin). For the non-cancerous cell line (HEK 293), trypsin hydrolysate had the highest toxicity. Apoptotic results revealed that trypsin hydrolysate was the most effective compared to the isolate, which was confirmed from morphological and Caspase 3/7 results. It may be concluded from the findings of this research that hydrolysates from food protein isolates have the potential for use as possible anticancer therapeutics. However, more research needs to be conducted to determine the peptides responsible for anticancer activity as well as the possible mechanism of action.Item Antimicrobial activity of ciprofloxacin-coated gold nanoparticles on selected pathogens(2014-08-08) Moodley, Nivrithi; Odhav, BhartiAntibiotic resistance amongst bacterial pathogens is a crisis that has been worsening over recent decades, resulting in serious and often fatal infections that cannot be treated by conventional means. Diseases caused by these drug resistant agents result in protracted illnesses, greater mortality rates and increases in treatment costs. Improvements to existing therapies and the development of novel treatments are urgently required to deal with this escalating threat to human health. One of the more promising strategies to combat antibiotic resistance is the use of metallic nanoparticles. Research into this area has shown that the binding of antibiotics to nanoparticles enhances their antimicrobial effects, reduces side-effects due to requirement of lower dosages of the drug, concentrates the drug at the interaction site with bacterial cells and in certain cases, has re-introduced susceptibility into bacterial strains that have developed drug resistance. Furthermore, these nanoparticles can be used in cancer treatment in similar drug delivery roles. Based on the promising data that demonstrated the synergistic effects of antimicrobial agents with nanoparticles, the aim of our research is to determine the effect of ciprofloxacin-conjugated gold nanoparticles as antimicrobial agents. To achieve this aim our objectives were: (i) to synthesize citrate-capped and ciprofloxacin-conjugated gold nanoparticles; (ii) to determine the physical and chemical characteristics of the ciprofloxacin-nanoparticle hybrid molecule; (iii) to investigate the antimicrobial activity of the conjugated nanoparticles against various species of common pathogens and (iv) to investigate the anti-cancer potential of the citrate-capped nanoparticles against a Caco-2 cell line. In this study, citrate-capped gold nanoparticles were conjugated to the antibiotic, ciprofloxacin, and their antibacterial and anti-cancer activity was evaluated. Initial experiments involved the synthesis and characterization of gold nanoparticles and ciprofloxacin conjugated nanoparticles. The gold nanoparticles were synthesized using the Turkevich citrate reduction technique which has been extensively used in studies thus far. The synthesized nanoparticles were characterized for specific absorbance using a UV-Spectrophotometer. The bond between the nanoparticles and ciprofloxacin was characterized by FTIR. Ultra structural details of the gold nanoparticles were established by TEM. The colloidal stability of the nanoparticles was determined by spectroscopic analysis. The antibacterial activity of the ciprofloxacin-conjugated gold nanoparticles was studied by exposure to pathogenic bacteria (Staphyloccocus aureus, E. coli, Klebsiella pneumoniae, Enterocococcus spp., Enterobacter spp., and Psuedomonas spp.). MIC values were measured to give indication of antimicrobial effect. These bactericidal properties of the conjugate nanoparticles were further investigated by electron microscopy. To evaluate the action of the citrate capped gold nanoparticles on cancer cells, we exposed Caco-2 cells to various concentrations of the nanoparticles and its effect was evaluated by measuring the viability of the cells. The results showed that 0.5 mM trisodium citrate reduced gold chloride to yield gold nanoparticles, which were spherical and 15 to 30 nm (by TEM characterization) and had an absorption maxima of 530 nm. The ciprofloxacin conjugated nanoparticles had an absorption maxima of 667nm. The colloidal stability, which is used to assess whether the synthesized particles will retain their integrity in solution showed that citrate-capped GNPs were most stable at 37°C over a 14 day storage period while ciprofloxacin-conjugated GNPs were found to be most stable at 4°C over a 14 day period. The FTIR results showed that chemical bonding in the conjugated nanoparticles occurs between the pyridone moiety of ciprofloxacin and the nanoparticle surface. The antimicrobial results of ciprofloxacin-conjugated GNPs had a significantly improved killing response compared to ciprofloxacin on both Gram positive and Gram negative bacteria. The citrate-capped GNPs are shown to exert a similar cytotoxic effect to gemcitabine on the Caco-2 cell line at a concentration of 0.5 mM. These results indicate that combining gold nanoparticles and ciprofloxacin enhances the antimicrobial effect of the antibiotic. The conjugate nanoparticles increase the concentration of antibiotics at the site of bacterium-antibiotic interaction, and thus enhance the binding and entry of antibiotics into bacteria. This has great implications for treatment of infection, as these antibiotic-conjugated nanoparticles can be incorporated into wound dressings, be administered intravenously as drug delivery agents, be engineered to possess multiple functionalities in addition to antibacterial activity and act as dual infection tracking and antimicrobial agents. Likewise, in this study, gemcitabine, an anticancer drug and gold nanoparticles were shown to kill cancer cells. In addition to their use in photothermal therapy and as drug delivery agents, the nanoparticles themselves possess anti-cancer activity against the Caco-2 cells. Thus, they have potential to act alone as a form of cancer treatment if functionalized with certain targeting agents that are specific to cancer cells, reducing the side-effects that come with regular chemotherapeutic drugs. It can be concluded that ciprofloxacin-conjugated gold nanoparticles enhance antibacterial effects of the antibiotic ciprofloxacin against bacterial cells and citrate-capped gold nanoparticles have anti-cancer activity against the Caco-2 cell line.Item Antimicrobial efficacy of nisin, oregano and ultrasound treatments against foodborne pathogens in ready-to-eat vegetables(2020) Takundwa, Brianmax Aubrey; Ijabadeniyi, Oluwatosin Ademola; Pillai, Santhosh Kumar KuttanA marked increase in the prevalence of foodborne illnesses resulting in foodborne outbreaks globally have been noticed, primarily due to the contamination of foods with bacterial pathogens. Recently, there has been a growing interest in employing alternative decontamination methods against bacterial pathogens, as a result of problems associated with the commonly used chemical and thermal methods. Therefore, the current impetus is on employing decontamination techniques involving green technology (physical, biological) and hurdle technology. In this study, oregano, nisin and ultrasound treatments were employed on lettuce and cabbage at varying levels to elucidate their antimicrobial efficacy and subsequent effect on the physical and sensory properties of the vegetables. Specifically, fresh cut lettuce and cabbage samples were inoculated with Escherichia coli O157:H7 and Listeria monocytogenes and were subjected to a series of combined treatment applications, of nisin, oregano essential oil and ultrasound. The Box Behnken, response surface methodology (RSM) technique was used in formulating the various combination treatments that would, in turn, demonstrate the synergistic capabilities of the three factors, nisin, oregano and ultrasound when combined. Results from the RSM were then used in the optimization of the combined treatment parameters by maximizing the microbial log reduction in both ready-to-eat vegetables. The physical properties studied on lettuce and cabbage subjected to combined treatments were colour and texture, and their structural damage was investigated using electrolyte leakage. Sensory properties were also analyzed on non-inoculated cabbage samples previously subjected to combined antimicrobial treatments. The efficacy of nisin, oregano and ultrasound on the reduction of E. coli O157:H7 and L. monocytogenes on lettuce studied using RSM/Box-Behnken model design, was found to be reliable (p<0.05). The most effective treatment on both pathogens was a combination of 771.2 IU/g nisin, 0.185% v/v oregano and 14.65 min ultrasound which showed log reductions of 3.43 and 9.20 CFU/mL for E. coli O157:H7 and L. monocytogenes, respectively. Lettuce treatment with the combined antimicrobial treatments resulted in no significant differences in textural properties, specifically hardness. However, mild colour changes and a slight increase in the electrolyte leakage rate was observed, though they were within the permissible limits. The reduction of E. coli O157:H7 on cabbage was increased with the use of combined treatments of nisin, oregano and ultrasound. Hence, the combination (comprising 607.85 IU/g nisin, 0.20% v/v oregano and 14.98 min ultrasound) exhibited the highest log reduction of 3.66 CFU/mL. In the samples inoculated with L. monocytogenes, 731.25 IU/g nisin, 0.12% v/v oregano and 13.21 min ultrasound treatments were found to be the best combination exhibiting the highest log reduction of 8.27 CFU/mL. No significant colour and textural changes were observed between untreated and treated cabbage. However, a slight increase in the electrolyte leakage rate was observed after the application of the combined treatment. Sensory evaluation scores also had some factors that were slightly below par. Overall, results from the study demonstrated that a combination of nisin, oregano and ultrasound, is a promising alternative to chemical treatments for the reduction of E. coliO157:H7 and L. monocytogenes as well as retaining the quality characteristics of fresh produce. Prospectively, other studies could explore the frontier field of artificial intelligence and machine learning, in the form of predictive microbiology and mathematical modelling, in the fresh produce industry. This would present a better understanding of risk assessment that is powered by the technological advantage of data analytics.Item Antimycobacterial, docking and molecular dynamic studies of pentacyclic triterpenes from Buddleja saligna leaves(Taylor and Fancis Online, 2017) Singh, Alveera; Venugopala, Katharigatta Narayanaswamy; Khedr, Mohammed A.; Pillay, Mellendran; Nwaeze, Kenneth U.; Coovadia, Yacoob; Shode, Francis; Odhav, BhartiBuddleja saligna (family Buddlejaceae) is a medicinal plant endemic to South Africa. Two isomeric pentacyclic triterpenes, oleanolic acid and ursolic acid, were isolated from the leaves of B. saligna using silica gel column chromatography. Compounds oleanolic acid and ursolic acid were subjected to derivatization with acetic anhydride in the presence of pyridine to obtain oleanolic acid-3-acetate and ursolic acid-3-acetate, respectively. The structures of these compounds were fully characterized by detailed nuclear magnetic resonance (NMR) investigations, which included 1H and 13C NMR. Molecular docking studies predicted the free binding energy of the four triterpenes inside the steroid binding pocket of Mycobacterium tuberculosis fadA5 thiolase compared to a reported inhibitor. Thus, their ability to inhibit the growth of M. tuberculosis was predicted and was confirmed to possess significant antimycobacterial activity when tested against Mycobacterium smegmatis, M. tuberculosis H37Rv (ATCC 25177), clinical isolates of multi-drug-resistant M. tuberculosis (MDR-TB) and extensively drug-resistant M. tuberculosis (XDR-TB) using the Micro Alamar Blue Assay. Ursolic acid was isolated from this plant for the first time.