This project investigates the moisture barrier properties of bio-based polymers and ways of improving them. The first section addresses the effect of crystallinity on the water permeability of poly(lactic acid) (PLA). The second section investigates PLA/talc composites and PLA/ montmorillonite nanocomposites. The third section is focused on a new polymer, polybutylene succinate (PBS), and its nanocomposites with montmorillonite. In the first section, the water vapour transmission rates (WVTR) through samples of polylactic acid of different crystallinities have been measured. Three different grades of commercial PLA were used with different ratios of L-lactide and D-lactide to give a range of crystallinities from 0 to 50%. Sheets of PLA were prepared by melt compounding followed by compression moulding and annealing at different temperatures and for different times to give the range of crystallinities required. Crystallinity was measured by differential scanning calorimetry (DSC) and the morphology of the samples was observed under crossed polars in a transmitted light microscope. Water vapour transmission rates through the films were measured at 38°C and at a relative humidity of 90%. It was found that the measured values of WVTR decreased linearly with increasing crystallinity of the PLA from 0 to 50%. The results are discussed in terms of the effect of crystallinity on solubility and shown to fit the tortuous path model. The model was also successfully used to explain published data on water permeability of polyethylene terephthalate. In the second section, a series of PLA/talc composites and PLA/ montmorillonite nanocomposites were prepared by melt compounding followed by compression moulding. The morphologies of the composites were investigated using transmission electron microscopy (TEM) and wide-angle X-ray diffraction (WAXD) and it was found that the fillers were well dispersed in the polymer matrix. The average aspect ratio of the compounded talc was found to be 8, and that of the nanoclay was found to be 50. Water vapour transmission rates (WVTR) through the films were measured at 38°C and at a relative humidity of 90%. It was found that the measured values of WVTR decreased with increasing filler content and the results gave good agreement with predictions from the Nielsen tortuous path model. In the third section, PBS/ montmorillonite nanocomposites were prepared by melt compounding followed by compression moulding. The melting and crystallisation behaviour of the pure PBS samples were investigated using differential scanning calorimetry (DSC) and cross polarised optical microscopy. A slight decrease of the degree of crystallinity was found in PBS containing 5% nanoclay. The morphology of the composites was investigated using transmission electron microscopy (TEM) and wide-angle X-ray diffraction (WAXD) and it was confirmed that that composite structures were intercalated. Water vapour transmission rates (WVTR) through the PBS sheets were measured using a MOCON Permatran-W®398. The measured values of WVTR decreased with increasing nanoclay content. However, the experimental values were all higher than the values predicted by the Nielsen tortuosity model. This result shows that in the case of PBS, which is a highly crystalline polymer, the nanoclay is not as well dispersed and is not as effective in reducing water vapour permeability as in the case of PLA.

The poly(A)-binding proteins (PABPs) are a family of eukaryotic RNA-binding proteins with key roles in mRNA translation and stability. The molecular function of PABPs have been largely revealed through study of the prototypical cytoplasmic poly(A)-binding protein, PABP1. Thus, little is known regarding other PABP family members. PABP5 contains four RNA-recognition motifs characteristic of the cytoplasmic PABPs yet is structurally distinct as it lacks a portion of the C-terminus. This region contains a proline-rich section linked to a globular domain that facilitates a number of protein-protein interactions. To date, little information has been presented regarding the expression of PABP5 and there is no data pertaining to the function of this protein, despite being mapped to a region of the X-chromosome associated with human pathological conditions. In this thesis, I present the first data documenting the expression of PABP5 within mouse tissues, and find it to be expressed at the highest levels within the brain, ovary, and testis. The limited data available suggests that gonads may be the only tissue to contain all PABPs therefore I additionally describe the expression of PABP1 and PABP4 to ascertain their cellular distribution within these tissues. This revealed that PABPs have overlapping yet distinct expression patterns in mouse gonads. The distinct structure of PABP5 suggested that its function may vary from PABP1. Characterisation of its activities in translational regulation was therefore investigated. When tethered to a reporter mRNA PABP5 had limited translational stimulatory activity, and in addition could not be isolated via m7G cap chromatography and failed to interact with translation initiation factors including eIF4G and PAIP-1. These factors interact with PABP1 to positively promote translation, implying that PABP5 function in translational regulation differs from other PABPs investigated. Examining why PABP5 failed to display translational stimulatory activity also revealed an interaction with the negative regulator of translation, PAIP-2. In summary, I present the first description of PABP5 cellular localisation, and have gone some way towards elucidating the molecular function of this uncharacterised protein.

The research presented in this thesis includes a detailed investigation into factors influencing ionic conductivity in the crystalline polymer electrolyte PEO₆:LiPF₆. It has previously been shown that preparing PEO₆:LiPF₆ with PEO modified with larger –OC₂H₅ end groups increases ionic conductivity by one order of magnitude [¹],primarily due to disruption of the crystal structure caused by the inclusion of the larger end groups. In this study it is shown that by reducing PEO molecular weight in crystalline PEO₆:LiPF₆ ionic conductivity is also increased. This was attributed to an increasing concentration of polymer chain end regions upon lowering molecular weight resulting in the creation of more defects, as well as possible increases in crystallite size resulting in longer continuous pathways for ion transport. Similar results were observed using both polydispersed and monodispersed PEO to prepare complexes. In addition, it is demonstrated here that ionic conductivity in crystalline polymerelectrolytes is not confined to PEO₆:LiXF₆ (X=P, As, Sb)[²][³] type materials. The structures and ionic conductivity data are reported for a series of new crystalline polymer complexes: the alkali metal electrolytes. They are composed of low molecular weight PEO and different alkali metal hexafluoro salts (Na⁺, K⁺ and Rb⁺), and include the best conductor poly(ethylene oxide)₈:NaAsF₆ discovered to date [⁴], with a conductivity 1.5 orders of magnitude higher than poly(ethylene oxide)₆:LiAsF₆. A new class of solid ion conductor is reported: the crystalline small-molecule electrolytes. Such materials consist of lithium salts dissolved in low molecular weight glyme molecules [CH₃O(CH₂CH₂O)[subscript(n)]CH₃, n=1-12], forming crystalline complexes [⁵][⁶]. These materials are soft solids unlike ceramic electrolytes and unlike polymer electrolytes they are highly crystalline, are of low molecular weight and have no polydispersity. By varying the number of repeat units in the glyme molecule, many complexes may be prepared with a wide variety of structures. Here, ionic conductivity and cation transference number (t₊) data for several such complexes is presented [⁷][⁸][⁹].These complexes have appreciable ionic conductivities for crystalline complexes and their t₊ values vary markedly depending on the glyme molecule utilized. The differences in t₊ values can be directly attributed to differences in their crystal structures. [¹] Staunton, E., Andreev, Y.G. & Bruce, P.G. Factors influencing the conductivity of crystalline polymer electrolytes. Faraday Discussions 134, 143-156 (2007). [²] Gadjourova, Z., Andreev, Y.G., Tunstall, D.P. & Bruce, P.G. Ionic conductivity in crystalline polymer electrolytes. Nature 412, 6846 (2001). [³] Stoeva, Z., Martin-Litas, I., Staunton, I., Andreev, Y.G. & Bruce, B.G. Ionic Conductivity in the Crystalline Polymer Electrolytes PEO₆:LiXF₆, X = P, As, Sb. J. Am. Chem. Soc. 125, 4619-4626(2003). [⁴] Zhang, C., Gamble, S., Ainsworth, D., Slawin, A.M.Z., Andreev, Y.G. & Bruce, P.G. Alkali metal crystalline polymer electrolytes. Nature Materials 8, 580-584 (2009). [⁵] Henderson, W.A., Brooks, N.R., Brennessel, W.W. & Young Jr, V.G. Triglyme-Li⁺ Cation Solvate Structures: Models for Amorphous Concentrated Liquid and Polymer Electrolytes (I). Chem. Mater. 15, 4679-4684 (2003). [⁶] Henderson, W.A., Brooks, N.R. & Young Jr, V.G. Tetraglyme-Li⁺ Cation Solvate Structures: Models for Amorphous Concentrated Liquid and Polymer Electrolytes (II). Chem. Mater. 15, 4685-4690 (2003). [⁷] Zhang, C., Andreev, Y.G. & Bruce, P.G. Crystalline small-molecule electrolytes. Angewandte Chemie, International Edition 46, 2848-2850 (2007). [⁸] Zhang, C., Ainsworth, D., Andreev, Y.G. & Bruce, P.G. Ionic Conductivity in the Solid Glyme Complexes [CH₃O(CH₂CH₂O)[subscript(n)]CH₃]:LiAsF₆ (n = 3,4). J. Am. Chem. Soc. 129, 8700- 8701 (2007). [⁹] Zhang, C., Lilley, S.J., Ainsworth, D., Staunton, E., Andreev, Y.G., Slawin, A.M.Z. & Bruce, P.G. Structure and Conductivity of Small-Molecule Electrolytes [CH₃O(CH₂CH₂O)[subscript(n)]CH₃]:LiAsF₆ (n = 8-12). Chem. Mater. 20, 4039-4044 (2008).

Thesis (MSc (Chemistry and Polymer Science))--Stellenbosch University, 2008. / Reversible addition fragmentation chain transfer (RAFT)-mediated polymerization has emerged as a versatile method for preparing polymers with control over molecular weight and polydispersity. Inherent in its mechanism is the retention of the chain transfer agent, the RAFT agent, at the polymer chain ends. Typically RAFT agents are made up of two parts, the so called R (leaving) and Z (thiocarbonyl thio, stabilizing) groups. These are retained as the a-and the w-end groups of the final polymer, respectively. RAFT polymerization offers a ready method for preparing polymers with well defined end functionalities. The a-end functionality can easily be built into the R group. The Z group, however, is thermally unstable and can impart color and smell to the polymer. Hence, two new methods for Z end group removal were introduced. Both methods take advantage of the facile reaction between thiocarbonyl thio compounds and radicals. By matching the functionalities of the R group (a-end group) with that of the end modified w-chain end, both methods offer an easy route to accessing telechelic functional polymers. End functional polymers have many important uses in industry and in the biomedical field. An alcohol functional xanthate RAFT agent was synthesized and successfully used to conduct the RAFT-mediated polymerization of N-vinylpyrrolidone (NVP). Characterization by NMR and MALDI ToF MS confirmed that a-hydroxyl-w-xanthate-functional PVP was easily produced. In the first end group modification method radicals were generated as in atom transfer radical addition (ATRA). A hydroxyl functional a-haloester was used as the ATRA initiator with a Cu catalyst system. The alkyl radical produced by this ATRA initiator then replaced the Z group giving a telechelic hydroxyl functional polymer. NMR analysis showed that the thiocarbonyl thio end group was completely removed. The hydroxyl functionality was quantified by derivatizing with trichloro acetyl isocyanate and subsequent analysis by NMR. MALDI ToF MS analysis, however, was inconclusive. In the second method the thiocarbonyl thio end group was removed by simply heating the polymer with hydrogen peroxide, thereby replacing the Z group with a hydroxyl end group at the w-chain end, giving a telechelic functional polymer. The telechelic hydroxyl functional polymer was subsequently crosslinked with a trifunctional isocyanate to make a PVP hydrogel. This confirmed that the end-modified polymer was indeed telechelic. The swelling kinetics of this hydrogel were determined in water at 37 oC.

Thesis (PhD)--Stellenbosch University, 2015. / ENGLISH ABSTRACT : In this thesis, a number of combinatorial objects are enumerated. Du and Yin as well as Shin and Zeng (by a different approach) proved an elegant formula for the number of labelled trees with respect to a given in degree sequence, where each edge is oriented from a vertex of lower label towards a vertex of higher label. We refine their result to also take the number of sources (vertices of in degree 0) or sinks (vertices of out degree 0) into account. We find formulas for the mean and variance of the number of sinks or sources in these trees. We also obtain a differential equation and a functional equation satisfied by the generating function for these trees. Analogous results for labelled trees with two marked vertices, related to functional digraphs, are also established. We extend the work to count reachable vertices, sinks and leaf sinks in these trees. Among other results, we obtain a counting formula for the number of labelled trees on n vertices in which exactly k vertices are reachable from a given vertex v and also the average number of vertices that are reachable from a specified vertex in labelled trees of order n. In this dissertation, we also enumerate certain families of set partitions and related tree-like structures. We provide a proof for a formula that counts connected cycle-free families of k set partitions of {1, . . . , n} satisfying a certain coherence condition and then establish a bijection between these families and the set of labelled free k-ary cacti with a given vertex-degree distribution. We then show that the formula also counts coloured Husimi graphs in which there are no blocks of the same colour that are incident to one another. We extend the work to count coloured oriented cacti and coloured cacti. Noncrossing trees and related tree-like structures are also considered in this thesis. Specifically, we establish formulas for locally oriented noncrossing trees with a given number of sources and sinks, and also with given indegree and outdegree sequences. The work is extended to obtain the average number of reachable vertices in these trees. We then generalise the concept of noncrossing trees to find formulas for the number of noncrossing Husimi graphs, cacti and oriented cacti. The study is further extended to find formulas for the number of bicoloured noncrossing Husimi graphs and the number of noncrossing connected cycle-free pairs of set partitions. / AFRIKAANSE OPSOMMING : In hierdie tesis word ’n aantal kombinatoriese objekte geenumereer. Du en Yin asook Shin en Zeng (deur middel van ’n ander benadering) het ’n elegante formule vir die aantal geëtiketteerde bome met betrekking tot ’n gegewe ingangsgraadry, waar elke lyn van die nodus met die kleiner etiket na die nodus met die groter etiket toe georiënteer word. Ons verfyn hul resultaat deur ook die aantal bronne (nodusse met ingangsgraad 0) en putte (nodusse met uitgangsgraad 0) in ag te neem. Ons vind formules vir die gemiddelde en variansie van die aantal putte of bronne in hierdie bome. Ons bepaal verder ’n differensiaalvergelyking en ’n funksionaalvergelyking wat deur die voortbringende funksie van hierdie bome bevredig word. Analoë resultate vir geëtiketteerde bome met twee gemerkte nodusse (wat verwant is aan funksionele digrafieke), is ook gevind. Ons gaan verder voort deur ook bereikbare nodusse, bronne en putte in hierdie bome at te tel. Onder andere verkry ons ’n formule vir die aantal geëtiketteerde bome met n nodusse waarin presies k nodusse vanaf ’n gegewe nodus v bereikbaar is asook die gemiddelde aantal nodusse wat bereikbaar is vanaf ’n gegewe nodus. Ons enumereer in hierdie tesis verder sekere families van versamelingsverdelings en soortgelyke boom-vormige strukture. Ons gee ’n bewys vir ’n formule wat die aantal van samehangende siklus-vrye families van k versamelingsverdelings op {1, . . . , n} wat ’n sekere koherensie-vereiste bevredig, en ons beskryf ’n bijeksie tussen hierdie familie en die versameling van geëtiketteerde vrye k-êre kaktusse met ’n gegewe nodus-graad-verdeling. Ons toon ook dat hierdie formule ook gekleurde Husimi-grafieke tel waar blokke van dieselfde kleur nie insident met mekaar mag wees nie. Ons tel verder ook gekleurde georiënteerde kaktusse en gekleurde kaktusse. Nie-kruisende bome en soortgelyke boom-vormige strukture word in hierdie tesis ook beskou. On bepaal spesifiek formules vir lokaal georiënteerde nie-kruisende bome wat ’n gegewe aantal bronne en putte het asook nie-kruisende bome met gegewe ingangs- en uitgangsgraadrye. Ons gaan voort deur die gemiddelde aantal bereikbare nodusse in hierdie bome te bepaal. Ons veralgemeen dan die konsep van nie-kruisende bome en vind formules vir die aantal nie-kruisende Husimi-grafieke, kaktusse en georiënteerde kaktusse. Laastens vind ons ’n formule vir die aantaal tweegekleurde nie-kruisende Husimi-grafieke en die aantal nie-kruisende samehangende siklus-vrye pare van versamelingsverdelings.

The major objective of this research is a systematic experimental exploration of hydrophilic materials that can be applied as coating materials for conventional ultrafiltration (UF) membranes to improve their fouling resistance against organic components. This objective is achieved by developing new, fouling-reducing membrane coatings and applying these coatings to conventional UF membranes, which can provide unprecedented reduction in membrane fouling and marked improvements in membrane lifetime. Novel polymeric materials are synthesized via free-radical photopolymerization of mixtures containing poly(ethylene glycol) diacrylate (PEGDA), photoinitiator, and water. PEGDA chain length (n=10-45, where n is the average number of ethylene oxide units in the PEGDA molecule) and water content in the prepolymerization mixture (0-80 wt.%) were varied. Crosslinked PEGDA (XLPEGDA) exhibited high water permeability and good fouling resistance to oil/water mixtures. Water permeability increased strongly with increasing the water content in the prepolymerization mixture. Specifically, for XLPEGDA prepared with PEGDA (n=13), water permeability increased from 0.6 to 150 L um/(m2 h bar) as prepolymerization water content increased from 0 to 80 wt.%. Water permeability also increased with increasing PEGDA chain length. Moreover, water permeability exhibits a strong correlation with equilibrium water uptake. However, solute rejection, probed using poly(ethylene glycol)s of well defined molar mass, decreased with increasing prepolymerization water content and increasing PEGDA chain length. That is, there is a tradeoff between water permeability and separation properties: Materials with high water permeability typically exhibit low solute rejections, and vice versa. The fouling resistance of XLPEGDA materials was characterized via contact angle measurements and static protein adhesion experiments. From these results, XLPEGDA surfaces are more hydrophilic in samples prepared at higher prepolymerization water content or with longer PEGDA chains, and the more hydrophilic surfaces generally exhibit less BSA accumulation. These materials were applied to polysulfone (PSF) UF membranes to form coatings on the surface of the PSF membranes. Oil/water crossflow filtration experiments showed that the coated PSF membranes had water flux values 400% higher than that of an uncoated PSF membrane after 24 h of operation, and the coated membranes had higher organic rejection than the uncoated membranes. / text

<p>Degradation of mRNA is a highly regulated step important for proper gene expression. Degradation of eukaryotic mRNA is initiated by shortening of the 3’ end located poly(A) tail. Poly(A)-specific ribonuclease (PARN) is an oligomeric enzyme that degrades the poly(A) tail with high processivity. A unique property of PARN is its ability to interact not only with the poly(A) tail but also with the 5’ end located mRNA cap structure. A regulatory role in protein synthesis has been proposed for PARN based on its ability to bind the cap that is required for efficient initiation of eukaryotic mRNA translation. Here we have investigated how the cap structure influences PARN activity and how PARN binds the cap. We show that the cap activates PARN and enhances the processivity of PARN. Further we show that the cap binding complex (CBC) inhibits PARN activity through a protein-protein interaction. To investigate the cap binding property of PARN, we identified the cap binding site at the molecular level using site-directed mutagenesis and fluorescence spectroscopy. We identified tryptophan 475, located within the RNA recognition motif (RRM) of PARN, as crucial for cap binding. A crystal structure of PARN bound to cap revealed that cap binding is mediated by the nuclease domain and the RRM of PARN. Tryptophan 475 binds the inverted 7-Me-guanosine residue through a stacking interaction. Involvement of the nuclease domain in cap binding suggests that the cap site and the active site overlap. Mutational analysis showed that indeed amino acids involved in cap binding are crucial for hydrolytic activity of PARN. Taken together, we show that PARN is an allosteric enzyme that is activated by the cap structure and that the allosteric cap binding site in one PARN subunit corresponds to the active site in the other PARN subunit.</p>

Besides exerting cytostatic or cytotoxic effects on tumor cells, some anti-cancer therapies (anthracyclines, oxaliplatin, X-Rays) could trigger an immunogenic cell death modality, releasing danger signals to alert immune system. We have shown that tumor-specific IFN- producing CD8+ T cells (Tc1) are mandatory for the success of chemotherapy to prevent tumor outgrowth. Priming of Tc1 response depends on IL-1β secretion by DC confronted with anthracycline-treated tumor cells releasing ATP. To identify the inflammatory components which link innate and cognate immune responses, we analyzed the influence of immunogenic chemotherapy on tumor microenvironment. We found an upregulated Th1- and Th17-related gene expression pattern in growth-retarded tumor after anthracycline treatment. By interfering with IFN- or IL-17A pathways, therapeutic effect of doxorubicin and oxaliplatin was abolished and dying tumor cell-based vaccine lost its efficacy to protect mice from live tumor cell rechallenge. Interestingly, we discovered that distinct subsets of  T lymphocytes (V4+ and V6+) colonized tumors shortly after chemotherapy, where they proliferated and became the dominant IL-17 producers within tumor beds. In three tumor models treated with chemotherapy or radiotherapy, a strong correlation between the presence of IL-17-producing  T ( T17) and IFN--producing CD8+ TIL (Tc1) was discovered. IL-17A signaling acts as upstream of IFN- since defect in IL-17RA led to complete loss of antigen specific Tc1 priming. The contribution of  T17 cells (V4+ and V6+) to chemotherapy is critical as V4/6-/- mice showed reduced sensitivity to chemotherapy and vaccination. Also, tumor infiltrating  T17 and Tc1 cells were reduced to basal level in this strain. IL-1β/IL-1R, but not IL-23/IL-23R, is pivotal for IL-17 production by  T cells and the success of chemotherapy. Importantly, adoptive transfer of  T cells could restore the efficacy of chemotherapy in IL-17A-/- mice and ameliorate the effect of chemotherapy in wild type host, provided that they retain the expression of IL-1R and IL-17A. Our research suggest a DC (IL-1β) →  T cells (IL-17) → Tc1 (IFN-) immune axis triggered by chemotherapy-induced dying tumor cells, which is critical for the favorable therapeutic response. To boost the immune system, we try to combine immunogenic chemotherapy with tumor vaccine in the presence of TLR3 agonist Poly (A:U). This sequential combined therapy, which we named VCT, could significantly retard tumor growth or even completely eradicate tumor and establish long-term protection against rechallenge in highly tumorigenic models. To dissect the effect of Poly (A:U) on immune system and that on TLR3 expressing-tumor cells, we performed VCT treatment in nude mice, TRIF-/- mice and with TRIF-silencing tumors. Interestingly, our results suggested that anti-tumor effect of VCT required T cells and intact TRIF signaling pathway at the level of the host and that of tumor cells. Poly (A:U) treatment could induce high level of CCL5 and CXCL10 production from tumor cells both in vitro and in vivo, which could negatively and positively influence the therapeutic outcome. By uncoupling the effect of CCL5 from that of CXCL10, the VCT treatment can be ameliorated. Our study emphasizes that both tumor and host derived inflammatory factors participate in regulating anti-tumor response. We also highlight that therapeutic application of TLR agonists can be optimized through regulating the profile of chemokines and their downstream signaling events.

Previous evidence has shown that the simple sequences microsatellites and poly-purine/poly-pyrimidine tracts (PPTs) could be both a cause, and an effect, of meiotic recombination. The causal link between simple sequences and recombination has not been much explored, however, probably because other evidence has cast doubt on its generality, though this evidence has never been conclusive. Several questions have remained unanswered in the literature, and I have addressed aspects of three of them in my thesis. First, what is the scale and magnitude of the association between simple sequences and recombination? I found that microsatellites and PPTs are strongly associated with meiotic double-strand break (DSB) hotspots in yeast, and that PPTs are generally more common in human recombination hotspots, particularly in close proximity to hotspot central regions, in which recombination events are markedly more frequent. I also showed that these associations can't be explained by coincidental mutual associations between simple sequences, recombination and other factors previously shown to correlate with both. A second question not conclusively answered in the literature is whether simple sequences, or their high levels of polymorphism, are an effect of recombination. I used three methods to address this question. Firstly, I investigated the distributions of two-copy tandem repeats and short PPTs in relation to yeast DSB hotspots in order to look for evidence of an involvement of recombination in simple sequence formation. I found no significant associations. Secondly, I compared the fraction of simple sequences containing polymorphic sites between human recombination hotspots and coldspots. The third method I used was generalized linear model analysis, with which I investigated the correlation between simple sequence variation and recombination rate, and the influence on the correlation of additional factors with potential relevance including GC-content and gene density. Both the direct comparison and correlation methods showed a very weak and inconsistent effect of recombination on simple sequence polymorphism in the human genome.Whether simple sequences are an important cause of recombination events is a third question that has received relatively little previous attention, and I have explored one aspect of it. Simple sequences of the types I studied have previously been shown to form non-B-DNA structures, which can be recombinagenic in model systems. Using a previously described sodium bisulphite modification assay, I tested for the presence of these structures in sequences amplified from the central regions of hotspots and cloned into supercoiled plasmids. I found significantly higher sensitivity to sodium bisulphite in humans in than in chimpanzees in three out of six genomic regions in which there is a hotspot in humans but none in chimpanzees. In the DNA2 hotspot, this correlated with a clear difference in numbers of molecules showing long contiguous strings of converted cytosines, which are present in previously described intramolecular quadruplex and triplex structures. Two out of the five other hotspots tested show evidence for secondary structure comparable to a known intramolecular triplex, though with similar patterns in humans and chimpanzees. In conclusion, my results clearly motivate further investigation of a functional link between simple sequences and meiotic recombination, including the putative role of non-B-DNA structures.

Electrospinning is a method of producing nano structured material from a polymer solution or melt using high strength electric fields. It is a process that has yet to find extensive industrial application yet shows promise if obstacles such as low rate of production overcome perhaps by more complete theoretical modelling. This work examines the effects of adding an ionic salt to a solution of poly(vinyl alcohol) in water. The direct effect was an increase the charge density and electric current. It was found that an increase in charge density decreases the mass deposition rate and forms a thinner initial jet. When the sign of the charge on the polymer solution was changed from positive to negative the charge density increased and the initial jet diameter and mass deposition rate also decreased. It was proposed that a smaller radius of curvature is formed by the Taylor cone at higher charge densities resulting in a smaller “virtual orifice”. The extent of the bending instability was explored and it was found that adding ionic salt results in a decrease in the bending instability resulting in thicker fibres. Changing the sign of the charge on the polymer solution from positive to negative resulted in an increase in the bending instability and resulted in thinner fibres. The charge transfer mechanisms used in different electrospinning models are explored and some assumptions not explicitly stated are discussed. From this discussion a generalized equation describing the charge transport mechanisms is proposed.