Pacific hagfish (Eptatretus stoutii) slime mucin vesicles are released by holocrine secretion with membranes that remain intact until the vesicle contacts seawater and ruptures. This thesis is an investigation of the mechanisms that drive mucin vesicle rupture for mucin release. Using isolated mucin vesicles collected from the slime glands of the hagfish, I tested the effects of a variety of solutions and drugs on vesicle rupture. I found that there are two categories of mucin vesicle that differ in their sensitivity to calcium ions, and that calcium-dependent vesicle rupture was inhibited with anion channel inhibitors. I also found that vesicle swelling rate was reduced by the aquaporin inhibitor mercuric chloride. Together, these data suggest that mucin vesicle rupture is partially dependent on the movement of chloride ions from seawater through calcium-activated anion channels and the rapid influx of water through aquaporin-like proteins in the vesicle membrane. / NSERC Discovery Grant, NSERC CGSM scholarship, Canada Foundation for Innovation, Ontario Ministry of Research and Innovation

This thesis tested the hypothesis that nitrogen use efficiency of subirrigated potted chrysanthemum (Chrysanthemum morifolium Ramat.) can be improved by managing the NO3- status of the plant. Replacement of NO3- with water one week prior to flower development was the most effective method of reducing tissue NO3- content and improving the nitrogen usage index, as compared to various combinations of NO3- and Cl-. Shoot or flower dry mass and N content were unaffected and the medium electrical conductivity was reduced. Supplying N in the NH4+ form in combination with NO3- did not affect nitrogen use indices. Reducing N supply improved various indices of nitrogen use efficiency, with a slight loss of quality. Elimination of NO3- prior to flower development and reduction in N supplied are effective strategies for improving nitrogen use efficiency in subirrigated potted chrysanthemums without sacrificing quality. / Cecil Delworth Foundation

No description available.

Cystic fibrosis is caused primarily by deletion of Phe508. An exciting discovery was that CFTR’s sister protein, the P-glycoprotein (P-gp) containing the equivalent mutation (ΔY490), could be repaired by a drug-rescue approach. Drug substrates showed specificity, and their mechanism involves direct binding to the transmembrane domains (TMDs) since arginine suppressor mutations were identified in TMDs that mimicked drug-rescue to promote maturation. We tested the possibility of rescuing CFTR processing mutants with a drug-rescue approach. 1) Arginine mutagenesis was performed on TM6, 8, and 12. 2) Correctors were tested for specificity. 3) Truncation mutants were used to map the VX-809 rescue site. Correctors 5a, 5c, and VX-809 were specific for CFTR. VX-809 appeared to specifically rescue CFTR by stabilizing TMD1. Therefore, the TMDs are potential targets to rescue CFTR. Rescue of P-gp and CFTR appeared to occur by different mechanisms since no arginine suppressor mutations were identified in CFTR.

Cystic fibrosis is caused primarily by deletion of Phe508. An exciting discovery was that CFTR’s sister protein, the P-glycoprotein (P-gp) containing the equivalent mutation (ΔY490), could be repaired by a drug-rescue approach. Drug substrates showed specificity, and their mechanism involves direct binding to the transmembrane domains (TMDs) since arginine suppressor mutations were identified in TMDs that mimicked drug-rescue to promote maturation. We tested the possibility of rescuing CFTR processing mutants with a drug-rescue approach. 1) Arginine mutagenesis was performed on TM6, 8, and 12. 2) Correctors were tested for specificity. 3) Truncation mutants were used to map the VX-809 rescue site. Correctors 5a, 5c, and VX-809 were specific for CFTR. VX-809 appeared to specifically rescue CFTR by stabilizing TMD1. Therefore, the TMDs are potential targets to rescue CFTR. Rescue of P-gp and CFTR appeared to occur by different mechanisms since no arginine suppressor mutations were identified in CFTR.

An examination of the seasonal and spatial distribution of Polar Stratospheric Clouds (PSCs) inferred from standard temperature profiles in the lower-middle atmosphere above Antarctica, as derived from the Earth Observing System (EOS) Aura Microwave Limb Sounder (MLS) satellite observations and NCEP/NCAR assimilations, is provided. Chemical volume mixing ratio (VMR) observations of EOS Aura MLS v2.2 hydrogen chloride (HCl) were used to show the interannual variability of PSC formation with respect to stratospheric chlorine partitioning during five Southern Hemisphere Antarctic seasons from 2005 to 2009. A remarkable first set of results, obtained from an algorithm developed for modelling HCl depletion areas in the Antarctic polar vortex region, and based on satellite observations, is presented. In particular, the analysis of HCl concentration data obtained from 2006 indicated that the area processed for HCl was larger than the area of PSC during some periods of Antarctic winter, and that this result was robust with respect to the various PSC formation and HCl depletion thresholds utilized. The results suggest that an underestimation in chlorine activation area can occur when temperature thresholds for PSC formation thresholds are employed. The work presented here also evaluated chlorine activation via sulfate aerosol (SA) in the Southern Hemisphere 2006 stratosphere, based on satellite measurements of water vapor (H2O) and constant values of SA, by implementing the TACL formula of Drdla and Müller [2010] in contrast to the TNAT formula of Hanson and Mauersberger [1988]. The results indicated that the former formula was not completely sufficient for accurately modeling areas of depleted HCl and chlorine deactivation for all pressure surfaces in the Antarctic stratosphere. Based on the results of this study, the role of SA in chlorine activation appears to be more important at lower altitudes than for areas higher in the stratosphere.

Green chemistry principles served as a guide for three industrially-relevant projects. In the first project, silylamines were applied as reversible ionic liquids for carbon dioxide capture from post-combustion flue gas streams. The effect of silylamine structure was thoroughly researched to develop a comprehensive library of silylamines and an accompanying set of structure-property relationships. The proposed solvent systems have the potential to present significant energy savings, as design has focused on their use in a non-aqueous, solvent-free environment. The second project also dealt extensively with carbon dioxide capture, as a reversible, in-situ protecting group for amines. Three strategies for the reversible protection of amines using carbon dioxide were developed and evaluated. Further, a chemoselective reaction was performed using carbon dioxide to protect a reactive amine and consequentially direct reactivity elsewhere within the same molecule. The carbon dioxide-protection technology developed has significant impact in multi-step industrial syntheses, as reversible, in-situ protection with carbon dioxide could eliminate the need for separate protection and deprotection unit operations. Lastly, a study was performed on the thermal degradation and stabilization of PVC in the presence of both plasticizers and thermal stabilizers. The study combined both model compound experiments as well as work with bulk PVC blends to gain a holistic understanding of the processes that take place during the degradation and stabilization of PVC. A bio-based plasticizer was investigated as a replacement for petroleum-based phthalate plasticizers. Additionally, two novel thermal stabilizers for PVC were presented and evaluated.

Perfluoroalkylated substances (PFASs) are a group of synthetic compounds that have gained growing attention due to their environmental persistence, toxicity and their potential to bioaccumulate. Even though PFASs are not occurring naturally in our environment, they are globally distributed and can be found ubiquitously in air, water, soil, wildlife as well as in humans. PFASs have primarily been used, due to their unique properties of being both hydrophilic and hydrophobic, as surfactants in numerous products such as firefighting foams, paint, leather and textile coating. The occurrence of PFASs in drinking water as well as in wastewater makes it important to develop effective techniques to remove these compounds from drinking water sources and wastewater. To be able to effectively remove PFASs from drinking water and wastewater it is important to understand which treatment process is most efficient and how the removal efficiency is affected by the physicochemical properties of PFASs and characteristics of water. In this study, the removal efficiency of PFASs was investigated using six different water types with varying dissolved organic carbon (DOC) character. Four different treatment techniques were evaluated including anion exchange using MIEX® resins, coagulation with iron (III) chloride (FeCl3), adsorption using powdered activated carbon (PAC) and nanofiltration (NF) membrane. The batch experiments were performed in laboratory-scale for 14 individual PFASs including C3-11, C13 perfluoroalkyl carboxylic acids (PFCAs), C4, C6, C8 perfluoroalkyl sulfonic acids (PFSAs) and perfluorooctane sulfonamide (FOSA). The results showed that the removal efficiency of PFASs was dependent on both perfluorocarbon chain length as well as functional group, with an increase in removal efficiency with increased perfluorocarbon chain length. Short-chained PFASs (C!6) were removed in less extent than the long chained PFASs for all treatment techniques. Amongst the four treatment techniques investigated, NF membrane exhibited the best removal efficiency for both short- and long chained PFASs (on average, 51%). Lower removal efficiencies for PFASs were observed for MIEX (33%) < FeCl3 (16%) < PAC (14%). However, all tested treatment techniques used in this study exhibited generally low removal efficiency (< 78%), in particular for the short-chained PFASs (C!6, < 41%) Results using three different doses of PAC (i.e. 20, 50, 100 mg L-1) showed an increase in removal (i.e. 2.2-41%, 8.0-78% and 12-92% respectively) with increasing dose. No significant trends were found between PFAS removal and DOC removal for any of the treatments (p<0.05, student t-test). However, the removal efficiency was different of the six different water types, which indicates that the DOC characteristics (i.e. Freshness, humification index, pH and absorbance) have an influence on the removal efficiency of PFASs in water. / Perfluoroalkylerade ämnen (PFASer) är en grupp syntetiska ämnen som har fått allt större uppmärksamhet den senaste tiden då de har visat sig vara persistenta, toxiska och bioackumulerande. Även om PFASer inte förekommer naturligt i vår miljö är de globalt fördelade och kan återfinnas i luft, vatten, mark, djur och hos människor. PFASer har främst använts, på grund av sina unika egenskaper att vara både hydrofila och hydrofoba, som tensider i många produkter såsom brandsläckningsskum, färg, läder och textil. Förekomsten av PFASer i dricksvattentäkter och i många reningsverk gör det viktigt att utveckla effektiva metoder för att ta bort dessa föreningar i vattenreningsverk. För att effektivt kunna avlägsna PFASer från dricks- och avloppsvatten är det viktigt att ha kunskap om vilken behandlingsmetod som är effektivast och hur reningseffektiviteten påverkas av ämnenas fysikalisk-kemiska egenskaper och vattnets karaktär.   Syftet med denna studie var att undersökta reningseffektiviteten för PFASer i sex olika vatten innehållande olika typer av löst organiskt kol (DOC). Detta undersöktes för fyra olika behandlingsteknikert; jonbyte med MIEX®, koagulering med järnklorid (FeCl3), adsorption med hjälp av pulveriserat aktivt kol (PAC) och nanofiltrering. Försöken gjordes små skaligt i laboratorie och 14 olika PFASer undersöktes; C3-11,13  perfluoralkyl karboxylsyror (PFCAer), C4, C6, C8, perfluoralkyl sulfonsyror (PFSAer) och perfluoroktan sulfonamid (FOSA). Resultaten visar att reningseffektiviteten för PFASer var beroende av både den perfluorerade kolkedjans längd och funktionell grupp, med en ökning av reningseffektivitet med längre perfluorerad kolkedja. PFASer med kort perfluorerad kolkedja (C≤6) renades i mindre utsträckning än PFASer med lång perfluorerad kolkedjade; detta gällde för alla behandlingstekniker. Bland de fyra behandlingstekniker som undersöktes uppvisade nanofiltreringen den bästa reningseffektiviteten för PFASer med både korta och långa kolkedjor (i genomsnitt, 51%.). Lägre reningseffektivitet för PFASer observerades för MIEX®(33%), < FeCl3(16%) < PAC (14%). Totalt sett erhölls en relativt låg reningseffektivitet (<78%) för samtliga reningstekniker, speciellt för de kortkedjade PFASer (C£6, < 41%). Resultat från försök med tre olika doser PAC (e.g. 20, 50, 100 mg L-1) visade på en ökad reningseffektivitet (2,2-41%, 8,0-78% och 12-92%) med ökad dos PAC. Inga signifikanta trender kunde urskiljas vad gäller reningseffektivitet av PFASer och rening av DOC (p<0.05, student t-test), detta gällde för samtliga behandlingstekniker. Det fanns dock tydliga skillnader i reningseffektivitet mellan de sex olika vattentyperna vilket indikerar på att DOC egenskaperna (Freshnessindex, humifieringsindex, pH, absorbans) har en påverkan på reningseffektiviteten för PFASer i vatten.

The purpose of this research project was to develop and characterise matrix type microspheres prepared from Eudragit® L100, containing insulin as model peptide drug as well as an absorption enhancer, N-trimethyl chitosan chloride (TMC), to improve intestinal absorption via the paracellular route. Insulin loaded microspheres were prepared using a single water in oil emulsification/evaporation method in accordance with a fractional factorial design (23) and subsequently characterised in terms of morphology as well as internal structure. Also, insulin and TMC loading were determined using a high pressure liquid chromatography analysis (HPLC) and colorimetric assay, respectively. Scanning electron microscopic characterisation revealed that most microsphere formulations showed a spherical shape and smooth surface with a sponge-like internal structure as well as relatively good homogeneity in terms of size distribution. Insulin loading ranged from 27.9 ± 14.25 – 52.4 ± 2.72% between the different formulations. TMC loading was lower than for insulin and ranged from 29.1 ± 3.3 - 37.7 ± 2.3% between the different formulations. The pronounced difference in insulin and TMC loading between the microsphere formulations is probably the result of the multitude parameters involved as well as the complex physicochemical processes which govern emulsification/solvent evaporation. Based on the microsphere characterisation results, two formulations were selected (i.e. B and F) for further characterisation (i.e. particle size distribution, dissolution behaviour, and enteric nature) and for in vitro evaluation of insulin transport across excised Fischer (FSR) rat intestinal tissue using a Sweetana-Grass diffusion chamber. Particle size analysis by means of laser light diffraction of the two selected microsphere formulations revealed that the mean particle size (based on volume) ranged from 135.7 ± 41.05 to 157.3 ± 31.74 m. Dissolution results for microsphere Formulations B and F revealed that both insulin and TMC were released from the microsphere formulations in an alkaline environment (pH 7.4). The mean dissolution time (MDT) for insulin ranged from 34.5 ± 4.01 to 42.6 ± 9.06 min, while the MDT for TMC ranged from 1.2 ± 1.73 to 6.8 ± 6.42 min. Statistical analysis revealed no significant differences in the MDT of either insulin or TMC (p-value > 0.05) between the two formulations, although the difference between insulin and TMC of each formulation was significant (p-value < 0.05). Microsphere formulations B and F released 36.92 and 48.21% of their total drug content over a period of 1 h in 0.1 M HCl. Microsphere Formulation B showed 8.3 ± 0.52% and formulation F 8.9 ± 2.26% transport of the initial insulin dose after a period of 120 min across excised rat intestinal tissue. The increase in insulin transport by the microsphere formulations compared to that of the control group (i.e. insulin alone) correlated well with the decrease in transepithelial electrical resistance (TEER) caused by the microsphere formulations. The transport of insulin from Formulations B and F represented transport enhancement ratios of 10.67 and 9.68, respectively. Insulin loaded EudragitL100 microspheres containing TMC were successfully prepared by emulsification/solvent evaporation that demonstrated promising potential to serve as oral drug delivery systems for insulin. The microspheres exhibited improved insulin permeability across intestinal epithelial tissue; however, its enteric properties should be improved and clinical effectiveness need to be confirmed by future in vivo studies. / Thesis (MSc (Pharmaceutics))--North-West University, Potchefstroom Campus, 2013.

The purpose of this research project was to develop and characterise matrix type microspheres prepared from Eudragit® L100, containing insulin as model peptide drug as well as an absorption enhancer, N-trimethyl chitosan chloride (TMC), to improve intestinal absorption via the paracellular route. Insulin loaded microspheres were prepared using a single water in oil emulsification/evaporation method in accordance with a fractional factorial design (23) and subsequently characterised in terms of morphology as well as internal structure. Also, insulin and TMC loading were determined using a high pressure liquid chromatography analysis (HPLC) and colorimetric assay, respectively. Scanning electron microscopic characterisation revealed that most microsphere formulations showed a spherical shape and smooth surface with a sponge-like internal structure as well as relatively good homogeneity in terms of size distribution. Insulin loading ranged from 27.9 ± 14.25 – 52.4 ± 2.72% between the different formulations. TMC loading was lower than for insulin and ranged from 29.1 ± 3.3 - 37.7 ± 2.3% between the different formulations. The pronounced difference in insulin and TMC loading between the microsphere formulations is probably the result of the multitude parameters involved as well as the complex physicochemical processes which govern emulsification/solvent evaporation. Based on the microsphere characterisation results, two formulations were selected (i.e. B and F) for further characterisation (i.e. particle size distribution, dissolution behaviour, and enteric nature) and for in vitro evaluation of insulin transport across excised Fischer (FSR) rat intestinal tissue using a Sweetana-Grass diffusion chamber. Particle size analysis by means of laser light diffraction of the two selected microsphere formulations revealed that the mean particle size (based on volume) ranged from 135.7 ± 41.05 to 157.3 ± 31.74 m. Dissolution results for microsphere Formulations B and F revealed that both insulin and TMC were released from the microsphere formulations in an alkaline environment (pH 7.4). The mean dissolution time (MDT) for insulin ranged from 34.5 ± 4.01 to 42.6 ± 9.06 min, while the MDT for TMC ranged from 1.2 ± 1.73 to 6.8 ± 6.42 min. Statistical analysis revealed no significant differences in the MDT of either insulin or TMC (p-value > 0.05) between the two formulations, although the difference between insulin and TMC of each formulation was significant (p-value < 0.05). Microsphere formulations B and F released 36.92 and 48.21% of their total drug content over a period of 1 h in 0.1 M HCl. Microsphere Formulation B showed 8.3 ± 0.52% and formulation F 8.9 ± 2.26% transport of the initial insulin dose after a period of 120 min across excised rat intestinal tissue. The increase in insulin transport by the microsphere formulations compared to that of the control group (i.e. insulin alone) correlated well with the decrease in transepithelial electrical resistance (TEER) caused by the microsphere formulations. The transport of insulin from Formulations B and F represented transport enhancement ratios of 10.67 and 9.68, respectively. Insulin loaded EudragitL100 microspheres containing TMC were successfully prepared by emulsification/solvent evaporation that demonstrated promising potential to serve as oral drug delivery systems for insulin. The microspheres exhibited improved insulin permeability across intestinal epithelial tissue; however, its enteric properties should be improved and clinical effectiveness need to be confirmed by future in vivo studies. / Thesis (MSc (Pharmaceutics))--North-West University, Potchefstroom Campus, 2013.