Thesis (M.Sc. Agric. (Crop Science)) --University of Limpopo, 2013 / The study aims to provide insights into the nature and extent of development funding provided to various poverty eradication projects by the National Development Agency (NDA) and its subsequent impact on reducing poverty in the predominantly rural communities of the Makhuduthamaga Municipality in the Limpopo Province. It examines in detail the impact made by the NDA on community development through the disbursement of funds to poverty eradication projects. It also provides insights into the total number of the NDA-funded projects and the total proportion of the NDA-funds allocated to them in the Makhuduthamaga Municipality. The study further examines the nature of the NDA support regarding the design of the interventions, relevance, participation of communities, delivery modalities and sustainability. The effectiveness of the NDA-funded projects in community development is determined using employment opportunities created, income generated, skills transferred, assets accumulated, sustainability mechanisms and community empowerment indicators. Although all of these indicators are found to be tightly linked to the NDA’s mandate of poverty eradication, the extent to which the NDA has achieved its objectives in disbursing development funding earmarked for poverty eradication and strengthening of CSOs was yet to be determined, hence the relevance of this study. The study, therefore, highlights key issues regarding the types of employment opportunities created and levels of income emanating from the NDA-funded projects. The study further highlights various areas of community empowerment, financial and sustainability measures put in place for the sustainability of the NDA-funded projects. Using a combined method of research, that is the qualitative and quantitative case study approach, the study highlights in detail insights into the impact made by the NDA on community development, particularly on Makhuduthamaga Municipality. The study highlights that while the NDA made some strides in the creation of employment opportunities, income generation, food security and community empowerment, both financial and institutional sustainability proved to be a daunting challenge for the NDA-funded projects Tailor-made and accredited training interventions coupled with the introduction of market-driven products to the NDA-funded projects as opposed to heavy reliance on donor funding will go a long way in bringing about productivity and, most probably, positive balance sheets and the maximum impact on the NDA funded projects.

Human hydroxysteroid sulfotransferase (hSULT2A1) catalyzes the sulfation of a broad range of endogenous (e.g., hormones, neurotransmitters, bile acids) as well as xenobiotic (e.g, drugs, environmental pollutants) compounds. Alteration in the catalytic activity of hSULT2A1 can lead to outcomes like endocrine disruptions or aberrant drug metabolism and xenobiotic toxicity. Oxidative and electrophilic stresses are known to cause physiological damage and be implicated as possible underlying pathologic mechanisms of a wide range of diseases. To examine the oxidative as well as electrophilic regulation of hSULT2A1, model oxidants (glutathione disulfide (GSSG), 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), diamide, tert-butyl hydroperoxide (TBHP)) and electrophiles such as quinone metabolites of polychlorinated biphenyls (PCB-quinones) and phenyl-p- benzoquinone were chosen for this study. Mechanistic studies correlating the enzyme structural modifications with alteration in the catalytic properties were performed to elucidate the catalytic regulative mechanism of an individual oxidant or electrophile. Thiol oxidants including GSSG, DTNB, and diamide showed catalytic regulation of hSULT2A1. Changes in protein intrinsic fluorescence indicated conformational alterations in hSULT2A1 following the reaction with diamide. Binding properties of hSULT2A1 for its substrates were also altered after reaction with these thiol oxidants, which could be one major reason for the kinetic alteration due to oxidative modification. Formation of mixed disulfides with cysteines in hSULT2A1 was also identified as a result of reaction with GSSG and DTNB. TBHP was chosen as a model for lipid peroxides, and reaction with this hydroperoxide decreased the catalytic function of hSULT2A1. The dissociation constant for binding of dehydroepiandrosterone (DHEA) was significantly altered with TBHP-pretreatment, but this did not affect the binding of 3',5'-adenosine diphosphate (PAP) to the enzyme. Structural analysis identified cysteine sulfonic acids and methionine sulfoxide formation after reaction of hSULT2A1 with TBHP, which could account for the alterations in the binding properties and the catalytic activity. Both PCB-quinones and PBQ could regulate the catalytic activity of hSULT2A1. Although PCB-quinones only caused decreases in the catalytic activity at all concentrations tested, pretreatment with PBQ indicated that lower concentrations resulted in an increase in the catalytic activity of hSULT2A1 that was followed by a decrease in the catalytic activity of hSULT2A1 upon increasing the concentration of PBQ in the pretreatment. Differences in the dissociation constants of PAP after PBQ-pretreatment were also observed, indicating the key role played by these PCB-quinones in altering the binding of either PAP or the sulfuryl donors, PAPS. Adducts at cysteines in hSULT2A1 were formed following reactions with PCB-quinones and PBQ. Small amounts of cysteine sulfonic acids and methionine sulfoxides were also formed following reaction of the protein with PCB-quinones and PBQ. Therefore, alterations in both the catalytic function as well as the structural properties of hSULT2A1 by interaction with oxidants and electrophiles may lead to changes in the metabolism of xenobiotics, as well as alterations in the endogenous balance of various steroid hormones. Such changes may be an important component in physiological damage that occurs under oxidative and electrophilic stress.

In this thesis the effect of mechanical stimuli on A549 lung cancer cells is studied. Modifications of polydimethylsiloxane (PDMS) surfaces are employed to alter the mechanical stimuli applied to the cells. Flat substrates are first studied and then micropillared substrates are designed, fabricated, and tested as a method to alter the mechanical properties of the PDMS surfaces. Molds with micro-pillars are designed then fabricated from silicon using deep reactive ion etching. From these molds, a negative then a positive replicate is made using PDMS. The pillared PDMS substrates are fabricated in 10 geometries and used for experiments. A549 cells are cultured on these surfaces then analyzed using fluorescence microscopy and atomic force microscopy (AFM). Fluorescence microscopy images processed by ImageJ software measure the cell spreading area (m2) while AFM quantifies the cell stiffness (kPa). For flat substrates, the cell stiffness and spreading area increase with increasing substrate stiffness. Further, results on pillared substrates show a similar trend based on pillar geometry changes. For pillared substrates, the A549 cell stiffness and spreading area increase as the height decreases, yet there is decreased cell stiffness and spreading area as the diameter and spacing decreases. The experiments show that changes in surface properties and only mechanical stimuli alter cellular morphology and biomechanics

Currently, extensive research in using bio‐derived polymers is being done, highlighting the importance of sustainable, green polymeric materials. Some sustainable alternatives to synthetic polymers include lignin, starch, cellulose or blends of these with petroleum‐based polymers. In New Zealand, large quantities of animal derived proteins are available at very low cost, making it ideal as a sustainable alternative to petroleum‐derived polymers. However, the processability of most proteins is very difficult, but can be improved by blending with synthetic polymers, such as polyolefins. To improve, the compatibility between these substances, a functional monomer could be grafted onto the polyolefin chain. Using an appropriate functional group, the polyolefin could then react with certain amino acids residues in the protein. Lysine and cystein are the two most appropriate amino acid residues because of their reactivity and stability at a wide pH range. In this study, free radical grafting of itaconic anhydride (IA) onto polyethylene was investigated. IA was selected because it is capable of reacting with polyethylene and amino acid residues, such as lysine. The objective of the research was to identify and investigate the effect of reaction parameters on grafting. These were: residence time, temperature, initial monomer concentration as well as peroxide concentration and type. Grafting was characterized in terms of the degree of grafting (DOG), percentage reacted and the extent of side reactions. The reaction temperature was taken above the melting point of the polyethylene, monomer and decomposition temperature of the initiator. It was found that above 160 C polymer degradation occurred, evident from sample discolouration. A higher degree of grafting can be achieved by increasing the initial monomer concentration up to a limiting concentration. The highest DOG achieved was about 1.2 mol IA per mol PE, using 2 wt% DCP. When using 2 wt % peroxide, the limiting concentration was found to be 6 wt% IA, above which no improvement in DOG was achieved. It was found that DCP is much more effective at grafting, compared to DTBP because DTBP is more prone to lead to side reactions than DCP. iv It was found that a residence time of 168 seconds resulted in the highest DOG, corresponding to 4 extrusions in series. However, it was also found that an increase in residence time resulted in an increase in polymer degradation. The tensile strength of PE decreased after two extrusions when using DTBP, and three extrusions, when using DCP. Young's modulus decreased only slightly, while all samples showed a dramatic decrease in ductility, even after one extrusion. It was concluded that degradation had a more pronounced effect on mechanical properties than cross‐linking, and residence time should therefore not exceed three extrusions in series, which corresponded to about 126 seconds. It can be concluded that a high reaction temperature and high initiator concentration lead to a low degree of grafting, accompanied by high cross‐linking and increased degradation. On the other hand, high monomer concentration and high residence time lead to a high degree of grafting. Optimising grafting is therefore a trade off between maximal DOG and minimising side reactions such as cross‐linking and degradation and optimal conditions do not necessarily correspond to a maximum DOG. Other factors, such as the use of additives to prevent degradation should also be investigated and may lead to different optimum conditions.

Vaccinia virus encodes at least eight proteins that are modified post-translationally by the addition of a 16-carbon saturated fatty acid through thioester linkage to cysteine residues. This is referred to as palmitylation of proteins. The purpose of this work was to gain an understanding of palmitylation, focusing on what defined the substrate for the modification, and the biological relevance of protein palmitylation in the vaccinia virus life cycle. A systematic approach was taken to identify the genes in vaccinia virus that encode these proteins. We found that vaccinia virus palmitylproteins are of the "late" temporal class, associate with intracellular membranes, and are specific for a particular form of the infectious virion. These criteria were used to narrow the number of genes expressed by vaccinia virus that potentially encode palmitylproteins. The "candidate" palmitylprotein genes were cloned and transiently expressed in mammalian tissue culture cells and analyzed for incorporation of palmitic acid. In addition to three previously identified vaccinia virus palmitylproteins, three new palmitylproteins were identified. The six known palmitylprotein genes were mutated to determine the site of modification, leading to the identification of the modification site for four of the six proteins. One of the proteins, p37, was analyzed further for biological significance of the palmitate modification. A recombinant vaccinia virus was constructed that did not express the wild-type palmitylated form of p37, but expressed a nonpalmitylated form of the protein instead. This virus was severely inhibited from proceeding past a particular morphogenetic stage, leading to an attenuated phenotype in tissue culture systems. Although the expression of the nonpalmitylated protein appeared normal compared to the wild-type protein, the lack of the palmityl moiety resulted in the loss of a targeting signal that directed the protein to its normal intracellular location. By this work, significant contributions have been made toward understanding the process of protein palmitylation. We have identified, at least for vaccinia virus, primary structural determinants specifying the modification, leading to the identification of a palmitylation motif. Considering the attenuated phenotype of the mutant virus, our conclusion is that palmitylation is necessary for biological function, at least for p37. / Graduation date: 1999

Resident bull trout (Salvelinus confluentus) may be particularly vulnerable to human related disturbance, however very few studies have focused on resident bull trout populations. The abundance of bull trout is one measure of the strength and potential for persistence of a population. Habitat characteristics may influence resident bull trout abundance to differing degrees and by varying means at multiple spatial scales. We used day and night snorkel counts to assess relative bull trout abundance. A modification of the Forest Service R1/R4 Fish and Fish Habitat Inventory was used to assess habitat characteristics associated with resident bull trout. Logistic and multiple linear regression were used to assess the relationships between resident bull trout abundance and habitat characteristics at the patch (1 to 5 km), reach (0.5 to 1 km) and habitat unit (1 to 100 m) scales. Site categorical variables were used along with quantitative habitat variables to explain among-site and across-site variation in the data. The significance of both quantitative habitat variables and categorical site variables at various spatial scales suggest that relationships between bull trout abundance and habitat characteristics are complex and in part dependent on scale. The characteristics of individual habitat units explained little of the variation in bull trout presence/absence (logistic regression; Somers' D=0.44) and density (multiple linear regression; adjusted R��=0.08) in habitat units, however there were habitat characteristics that were significantly (P���0.05) correlated to bull trout presence/absence and density in habitat units. The relationships between habitat characteristics and bull trout presence/absence and density varied between habitat unit types. There was a strong quadratic relationship between bull trout abundance and mean summer water temperature at the reach (P=0.004) and patch scales (P=0.001). The mean temperature of patches appears to explain some of the variation in bull trout density at smaller spatial scales, such as reaches and habitat units. An appreciation of the complex nature of scale dependent interactions between bull trout abundance and habitat characteristics may help resource managers make wiser decisions regarding conservation of resident bull trout populations. / Graduation date: 2000

The attachment of myristic acid to the N-terminal glycine residue of many eukaryotic and viral proteins is often essential for the acquisition of the protein's biological activity. Vaccinia virus (VV), the prototype member of the Poxviridae, expresses several myristylated proteins during the course of infection. Only one of these proteins, L1R, has been identified and characterized. Experiments were performed to identify and analyze four additional VV myristylproteins. These proteins were identified as the A-type inclusion protein (92 kDa), G9R (39 kDa), A16L (36 kDa), and E7R (17 kDa). The latter three proteins were shown to be myristylated on an N-terminal glycine residue. Additional studies demonstrated that both A16L and E7R are soluble proteins, unlike L1R, which is a constituent of the viral envelope. Furthermore, A16L could not be detected in either purified extracellular enveloped virus (EEV) or in intracellular mature virus (IMV). These are the two predominant forms of infectious virions produced during a VV infection. E7R was detected in EEV and, to a lesser extent, in IMV. Unlike the other proteins, the amino terminal sequence of the A-type inclusion protein did not fit the consensus sequence for N-myristylation (M-G-X-X-X-S/T/A/C/N), suggesting that it was internally myristylated. A combination of studies revealed that the protein is both myristylated and palmitylated. Addition of each acyl group could be separated temporally: myristylation occured co-translationally, while palmitylation occurred post-translationally. Genetic analyses of lysine doublets and arginine/lysine doublets within the A-type inclusion protein indicated that these sites are not utilized for myristylation. This is in contrast to the precursors of TNFoc and Ilia which are internally-myristylated on a lysine doublet. It is not clear why this protein would be both myristylated and palmitylated. Only class four palmitylproteins, such as the Src family of proteins, have been shown to be both myristylated and palmitylated. The A-type inclusion protein expressed by cowpox virus forms a large symmetrical matix in the cytoplasm of infected cells and generally contains mature virions. It is possible, therefore, that the function of acylation may be to stabilize the protein matrix or to assist in occlusion of enveloped virus particles. / Graduation date: 1998

Thesis (Ph. D)--Ohio State University, 2006. / Available online via OhioLINK's ETD Center; full text release delayed at author's request until 2007 Aug 16

The Roosevelt elk (Cervus elephus roosevelti) is a National Forest management indicator species on the westside of the Cascade mountains, Western Oregon. A Habitat Effectiveness model is used by State and Federal agencies to evaluate elk habitat in the region. Concerns about the model's lack of differentiation between winter and summer ranges in the analyses and assumptions that burning will increase forage quality on winter range prompted this study. I investigated the effect of broadcast burning, plant association, and time since disturbance on the quality of trailing blackberry (Rubus ulna), red huckleberry (Vaccinium parviflorum), willow (Salix spp.), vine maple (Acer circinatum), salal (Gaultheria shallon), and red elderberry (Sambucus racemosa). Crude protein, neutral detergent fiber, acid detergent fiber, acid insoluble ash, lignin, astringency, condensed tannin, and hydrolyzable tannin contents were measured. No significant effects of burning, plant association, or age were observed for crude protein, hydrolyzable tannins, or neutral detergent fiber. Crude protein varied among taxa: trailing blackberry contained 9.65%, and the other taxa ranged from 5.21-7.24%. Neutral detergent fiber was highly variable: trailing blackberry contained 30.90%, and the other taxa ranged from 52.20%-65.06%. Acid detergent fiber content ranged from 44.88%-49.49% for all taxa except trailing blackberry (17.78%). Lignin varied among taxa: trailing blackberry had the lowest content (6.37%) and salal had the highest (30.25%). Lignin content in salal was greater on recently disturbed sites. Astringency ranged from 0.0015 mg protein precipitated per mg plant tissue in vine maple to 0.6737 in trailing blackberry. Salal and willow had intermediate astringencies: elderberry, huckleberry, and vine maple had the lowest. Hydrolyzable tannins were present in all species except red elderberry. Burning and plant association effected astringency and condensed tannin content in trailing blackberry and huckleberry. Samples from burned, very dry and resource-poor sites had higher astringencies than on similar unburned units and non-resource-limiting sites. Condensed tannin contents increased with unit age in salal, huckleberry, and trailing blackberry, possibly accumulating during peak years of re-establishment after disturbance. Vine maple and red huckleberry were the only taxa with positive digestible protein levels. Digestible protein content may be higher in winter forage on less severe sites. Elk forage enhancement in winter range should be evaluated on a site-specific basis. Burning did not promote a detectable increase in quality for these forage taxa, and it decreased the quality of species sensitive to site conditions. / Graduation date: 1991

Occupational therapists assess and make recommendations for modifying home environments to facilitate caregiving of children in their homes. Children who are profoundly and permanently physically disabled remain dependent for their care throughout their lifespan. As they mature, the physical demands on family members to provide care become more challenging requiring major modifications to ease caregiving. Qualitative interview data of ten educationally influential occupational therapists (EIOTs) were collected to examine what information the EIOTs collect and how they prioritize and organize these data to make modification recommendations. The data were analyzed using grounded theory methodology. The findings detail myriad information required by the EIOT to make recommendations for modifications. Findings suggest a future oriented and highly contextualized information gathering process. The themes were integrated into a process model that suggests how therapists may be developing recommendations. The findings highlight important implications for research, education, clinical practice and service delivery policy.