Chemoenzymatically synthesized low molecular weight lignin polymers have been previously found to be potent inhibitors of a number of serine proteases via allosteric mechanisms targeting heparin binding sites. Herein, we describe the creation of synthetic sulfated β-O4 lignin (SbO4L) polymer, which is more homogenous compared to previous lignins with respect to its inter-monomeric linkage. SbO4L is a selective inhibitor of thrombin and plasmin. SbO4L was found to act via a unique mechanism targeting thrombin exosite 2 in a manner similar to platelet glycoprotein Ibα (GPIbα). Advanced hemostasis and thrombosis assays demonstrated that SbO4L acts via a dual mechanism: as an anticoagulant, by allosteric inhibition of thrombin catalysis; and as an antiplatelet agent, by competing with platelet GPIbα. These mechanisms are comparable in potency to low molecular weight heparins currently used in the market, indicating that targeting exosite 2 may yield clinically useful drugs in the future. Since the β-O4 type lignin was found to be selective for thrombin and plasmin, we hypothesized that other scaffolds from lignins could be potent inhibitors of other serine proteases. In particular, we screened a library of synthetic sulfated small molecules against factor XIa – an emerging target for prophylactic anticoagulation. Our search identified a sulfated benzofuran trimer (a mimic of β-5 type linkage found in lignins) as a potent inhibitor of factor XIa. Surprisingly, this inhibitor did not compete with heparin. A plausible binding site in the A3 domain of factor XIa was proposed by using molecular modeling techniques. The binding pose demonstrated good correlation with the structure activity data from in vitro studies. Further confirmation that the apple domains were required was proved by testing the trimer against recombinant catalytic domain. A 40-fold decrease in activity was observed. A temperature-dependant perrin plot demonstrated that factor XIa undergoes a large conformational change in the presence of the trimer, which is possibly converting the enzyme back into the zymogen-like shape. In general, the synthetic sulfated lignins can act as a useful foundation to develop anticoagulant, antiplatelet, and anti-inflammatory molecules in the future.

The herpes virus family consists of more than hundred members that infect organisms, of which eight, differing markedly in the biology are known to infect humans. HSV- I is the most common one, causing oral lesions and sporadic encephalitis. These infections are highly prevalent affecting at least one in three individuals in the United States.The entry of the herpes virus into the cell is a two-step process. The initial step involves the cell surface heparan sulfate and glycoproteins in the viral envelope which enables the virus to penetrate into the cell. The second step is the fusion step. Depending on the nature of interaction and size of HS chain, a single chain may bind multiple viral ligands on a virion. There is substantial evidence showing that HS plays an important role in viral binding.HS is a heterogeneous, linear sulfated oligosaccharide composed of alternating glucosamine and uronic acid residues, which could specify distinct receptor for various viral ligands. HS, present on most exposed cell surfaces, make an ideal snare for the capture of most herpes viruses and may facilitate subsequent interactions with other co-receptors required for entry. Number of viruses, including HSV- I, HSV- II, HIV- I and dengue virus use sites of HS as receptors for binding to cells. Recently 2000 Liu et.al have characterized a HS based octasaccharide that binds to HSV-I gD. The distinguished feature in the composition of the octasaccharide is the presence of 3-O-sulfate glucosamine residue, which is an uncommon structural modification in HS. Its presence in the HSV-I gD binding sequence may confer specificity of interaction and assist HSV-I entry into the cell.Numerous sulfated molecules have been explored as mimics of HS in the inhibition of HSV-1 entry into cells. To date, most of the sulfated molecules screened for anti-viral activity have been carbohydrates. So, we reasoned that it should be possible to mimic critical interactions of HS with one or more viral glycoprotein using synthetic, non-polysaccharide, sulfated compounds. Further, it may be possible to mimic specific sequence(s) in HS, which play a role in HSV infection, with small synthetic, sulfated, non-carbohydrate molecules. In a search for synthetic mimics of HS as inhibitors of HSV-I infection, we screened a small, synthetic, sulfated flavonoids to discover a potent inhibitory activity arising from sulfation of a macromolecule present as an impurity in a crude natural product.The active principle was identified through an array of biophysical and chemical analyses as lignin sulfate, a heterogeneous; polydisperse network polymer composed of substituted phenylpropanoid monomers. Further, LC-MS with APCI in negative ionization mode, which have been reported in here for the first time for analysis of lignin, has been successfully used to deduce oligomeric structures present in the precursor of the active macromolecule based on the spectrum of the depolymerized lignin. This corroborates well with the structural information obtained using other analytical techniques. We hypothesize that the structural heterogeneity and polydispersity of lignin coupled with optimal combination of sulfate charge and hydrophobicity result in high potency. Given that the native lignin is inactive, lignin sulfate discovered here provides a variety of organic scaffolds that with the critical sulfate groups in space can mimic the HSV-I gD binding sequence.

Phenylephrine (PE) is the most commonly used over-the-counter nasal decongestant. The problem associated with phenylephrine is that it undergoes extensive first pass metabolism in the intestinal gut wall leading to its poor and variable oral bioavailability. This research project aims at developing strategies in order to increase the oral bioavailability of PE by co-administration of GRAS compounds. A HILIC assay method was developed to detect the parent drug, phenylephrine (PE) and its sulfate metabolite (PES).The enzyme kinetic studies were done with phenolic dietary or GRAS compounds using LS180 human intestinal cell model, recombinant SULT enzymes and human intestinal cytosol (HIC). From the screening studies done, one inhibitor was selected in order to study the mechanism of inhibition. In conclusion the studies done in vitro provided a basis in order to predict in vivo intrinsic clearance through the sulfation pathway.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Global environmental concerns arise when marine deposits with fine-grained iron sulfide-rich sediments (FeS and FeS2), now situated above sea level, oxidize from anthropogenic lowering of the groundwater table. The oxidation of iron sulfides decreases the soil pH and the acidic environment of these Acid Sulfate Soils (AS) soils increase weathering and mobilization of metals into adjacent watercourses, lakes and estuaries. Low pH and enhanced concentrations of metals are known to influence water quality negatively, causing fish kills and reduced aquatic diversity. Sulfide rich sediments were deposited in the Baltic Sea after the last glaciation and are now abundantly found along the coasts of for example the Bothnian Bay as a result of isostatic rebound. Recent studies from Finland have stated associations between leached (AS) soils and increased concentrations of metals in estuary sediments, thus the effects are likely to be similar in Sweden. With financial support from the Interreg Nord project “Ecological restoration in coastal river basins in the Bothnian Bay” in cooperation with the Geological Survey of Sweden (SGU), sediments from three lakes and two estuarine sites in the Rosån catchment in Norrbotten county, northern Sweden, were sampled and analyzed with fpXRF, ICP-MS and LOI methods. Elemental concentrations and organic contents were compared to establish accumulation trends over time and relationships between metal concentrations in recently deposited sediments and potential influence from previously sampled (AS) soils. Correlations in time, to anthropogenic activity, such as ditching were also considered. A primary allover trend with increasing concentrations of Aluminum (Al), Arsenic (As), Cadmium (Cd), Cobalt (Co), Cupper (Cu), Iron (Fe), Manganese (Mn), Nickel (Ni), Lead (Pb), Rare Earth Elements (REE) and Zinc (Zn), was noticed in a majority of the lake and estuary sediment samples. With some site variation, also two discrete peaks at different depth, were found in the upper 20 to 30 cm of the sampled sediments. Significant correlations to organic matter were also found for a considerable amount of the elements. Soil samples from (AS) soils in areas related to Rosån show pH values as low as 2.62 and oxidation depths down to 170 cm. Substantial elemental depletion in the oxidized zone suggest increased weathering, leaching and mobilization of Al, Cd, Co, Mn, Ni, REE, Zn and to some extent As, Cu, Fe and Pb from the soil, as a consequence of the acidic environment. A relationship between (AS) soils and increased metal concentrations is therefore likely. The sedimentation rate of roughly 0.2 cm/year was calculated from the separation age of the lakes and the sediment depth to an interpreted transition from more marine environment to lake settings. Consequently the distinct peaks of increased metal concentrations are thus suggested to be related to anthropogenic activities as for example improved drainage methods after the Second World War, but proper dating of the sediments is needed to establish any certain correlations. / På senare tid har man uppmärksammat de miljöproblem som uppstår när finkorniga sediment, innehållande järnsulfider, oxiderar på grund av mänsklig påverkan, t ex. dikning, eller annan typ av aktivitet som sänker grundvattennivån. När järnsulfiderna oxiderar bildas svavelsyra som gör att pH- värdet i marken blir väldigt lågt. Den sura miljön i dessa så kallade sura sulfatjordar, gör i sin tur att mineraler i marken vittrar fortare med påföljden att både surt vatten och ökade mängder metaller, sprids till närliggande vattendrag, sjöar och hav. Lågt pH och höga halter av metaller i vatten påverkar även vattenkvalitén negativt och har rapporterats orsaka fiskdöd och minskad akvatisk mångfald. Sura sulfatjordar är globalt förekommande och återfinns bland annat längs Bottenvikens kuster. De har kunnat bildas genom att sulfidhaltiga sediment, som avsattes i Östersjön efter den senaste istiden, nu befinner sig ovan havsnivån på grund av landhöjningen. I Finland har man i flera studier sett ett samband mellan sura sulfatjordar och ökade metallhalter i nyligen avsatta kustsediment och man kan anta att liknande förhållande gäller även i Sverige. Med finansiellt stöd från EU-projektet “Kustmynnande Vattendrag i Bottenviken-Metodutveckling och Ekologisk Restaurering” (Interreg Nord) genom Sveriges geologiska undersökning (SGU) och i samarbete med Länsstyrelsen i Norrbotten, har bottensediment från tre sjöar och två fjärdar i Rosåns avrinningsområde i Norrbotten provtagits och analyserats. För att fastställa hur koncentrationerna har förändrats över tiden har metallhalterna i de nyligen avsatta sedimenten jämförts med koncentrationerna i äldre sediment. Även jordprofiler från sura sulfatjordar i området har studerats för att kunna utvärdera ett ev. samband mellan urlakade ämnen i jordarna och ökade halter i sedimenten. Granskning av hur långt metallerna transporteras i systemet har gjorts, liksom försök att hitta kopplingar i tiden till mänsklig påverkan som t.ex. dikning. I de översta sedimenten kan man, förutom en generellt ökande trend av aluminium (Al), arsenik (As), kadmium (Cd), kobolt (Co), koppar (Cu), järn (Fe), mangan (Mn), nickel, (Ni) bly (Pb), sällsynta jordartsmetaller (REE) och zink (Zn), också se tydliga toppar med ökade halter av dessa ämnen på minst två specifika djup. De sura sulfatjordarna, som uppmätte pH-värden ner till 2,62, visade tecken på urlakning av Al, Cd, Co, Ni, Mn, REE, Zn och till viss del också As, Cu, Fe och Pb, varpå en trolig relation mellan sura sulfatjordar och ökade metallhalter i nyligen avsatta sediment kan fastställas. En uppskattning av ackumulationshastigheten, som gjordes utifrån när de provtagna sjöarna skiljdes från havet, och sedimentdjupet som visar övergången från hav till sjö, visar att de observerade topparna av ökade metallkoncentrationer i sedimenten skulle kunna vara förknippade med dikning i början av 1900-talet och efter andra världskriget. För en säker bestämning av sambandet till specifika händelser behövs dock en riktig datering.

Master of Science / Department of Animal Sciences and Industry / Jim Nelssen / Seven experiments using a total of 3,251 preweaned pigs, nursery pigs, and sows were used to determine the effects of: 1) supplemental vitamin D[subscript]3 on suckling and nursery pig growth, and maternal performance, and 2) high sulfate water, dietary zeolite and humic substance on nursery pig performance. Also, a web-based survey was developed to question pork producers and advisors of the swine industry on their knowledge of feed efficiency. Experiment 1 tested an oral dose of either; none, 40,000 or 80,000 IU vitamin D[subscript]3 given to pigs 24 to 48 h after farrowing. No differences in growth performance or bone mineralization were observed, but vitamin D[subscript]3 supplementation increased serum 25(OH)D[subscript]3 on d 10, 20, and 30, but returned to control values by d 52. Experiments 2 and 3 evaluated an oral dose of vitamin D[subscript]3 to pigs just before weaning, as well as added D[subscript]3 in nursery diets and in drinking water. There were no effects on growth performance; however, serum 25(OH)D[subscript]3 increased with all sources of vitamin D[subscript]3 supplementation. Experiment 4 evaluated if pigs had a preference to 1 of 3 dietary concentrations of vitamin D[subscript]3. Pigs ate less feed from diets containing very high levels of vitamin D[subscript]3 compared to commonly supplemented levels. Experiment 5 evaluated 3 levels of vitamin D[subscript]3 in sow diets. There were no effects on sow productivity, subsequent pig performance, or piglet bone ash content. However, increasing vitamin D[subscript]3 increased sow serum 25(OH)D[subscript]3, milk vitamin D, and pig serum 25(OH)D[subscript]3. Experiment 6 and 7 evaluated the effects of dietary zeolite and humic substances in nursery pigs drinking high sulfate water. Ultimately, pigs drinking high sulfate water had increased fecal moisture content and decreased growth performance, and feed additives evaluated were ineffective in ameliorating these negative effects. Finally, data collected from the feed efficiency survey suggest that there are knowledge gaps about practices that effect feed efficiency. Results from this survey will help extension educators better target specific industry segments with current information and provide more specific areas of future research where lack of information has been identified.

Submitted in the fulfillment of the requirements for the degree of Master of Technology, Durban University of Technology, Durban, South Africa, 2011. / The main objective of this study was to evaluate the feasibility of conversion of crude tall oil and tall oil fatty acids into biodiesel. During the Kraft pulping process, Crude Tall Oil originates as tall oil soap, which is separated from recovered black liquor. The soap is then converted to Crude Tall Oil by acidulation with sulphuric acid. The Crude Tall Oil is then fractionated by distillation to produce tall oil fatty acids (TOFA), rosin and pitch. There were a number of conversional methods that were considered but proved to be inappropriate. A base-catalyzed method was inappropriate with due to the high free fatty acid content on the feedstock, and the acid-base catalyzed method was inappropriate due to the long reaction times and large excess of methanol required. An enzyme based conversion method was also found to be inappropriate because of the high price attached to the purchasing of the enzymes and the stability of the enzyme. A procedure of choice was the supercritical methanol treatment, due to the fact that it requires no separate catalyst. A procedure was developed for both the feedstocks (i.e. crude tall oil and tall oil fatty acids) using the supercritical methanol treatment. In supercritical methanol treatment, feedstock and methanol were charged to a reactor and were subjected to temperatures and pressures beyond the critical point of methanol (Tc = 240 °C, Pc = 35 bar). The maximum biodiesel yield obtained from Crude tall oil was 66% and was 81% for the tall oil fatty acids that was produced in a single stage process. The temperature and methanol to feedstock ratio effects was also found to yield a maximum biodiesel yield at 325°C and 40:1 respectively. A 20 minutes reaction time was found to be appropriate for the maximum yield of biodiesel. The final biodiesel produced was also evaluated against a commercial biodiesel product and its parameters measured. The biodiesel resulting from the tall oil fatty acid yielded parameters that were acceptable according to ASTM D6751 specifications for biodiesel. The biodiesel produced from the crude tall oil did not meet the ASTM D6751 specification, and this was mostly attributed to the presence of unsaponifiables which hindered the conversion of oil into biodiesel. / M

Ce projet de thèse vise à étudier les processus géochimiques de l'aluminium dans les sols acides à sulfates. Les résultats obtenus avec le dispositif de dialyse de Donnan montrent que l'aluminium dans la solution du sol est presque exclusivement (> 98%) présent sous forme de complexes de charge négative, vraisemblablement des organo-complexes d'aluminium. Les concentrations d'aluminium isotopiquement échangeables (valeurs-E) et les concentrations obtenues par extractions chimiques révèlent qu'une solution de 1 M KCl sous-estime systématiquement les valeurs E. Une méthode d'extraction séquentielle a révélé qu'une quantité importante de minéraux primaires (initialement présents dans les sols) a déjà été dissoute, comme en témoignent les concentrations élevées d'aluminium présentes en tant que minéraux secondaires d'aluminium. Ces études permettent de mieux comprendre la géochimie de l'aluminium dans les sols CLASS et de pouvoir intégrer ces connaissances dans la gestion des sols. / The aim of this thesis was to further understanding on Al geochemistry in coastal lowland acid sulfate soils (CLASS). It was observed that Al was present almost solely (> 98%) as negatively charged complexes in CLASS pore-waters, presumably with natural organic matter. Isotopically exchangeable concentrations (E-values) of Al and extraction solutions used to estimate the exchangeable pool showed that 1 M KCl always underestimated isotopically exchangeable Al concentrations in these soils and that 0.2 M CuCl2 improved agreement between both methodologies but sometimes overestimated corresponding E values. A sequential extraction procedure showed that substantial amounts of Al have already been dissolved from primary aluminosilicates initially present in the soils and remain in the soils mostly as reactive secondary Al minerals. The outcomes of this thesis significantly further our understanding of Al geochemistry in CLASS environments and how this knowledge can be incorporated into land management practices.

[pt] Íons de potássio (K+) são essenciais na nutrição das plantas e são, normalmente, fornecidos na forma de KCl ou K2SO4. Este cloreto de potássio é responsável por 90 por cento da quantidade de potássio oferecido pela indústria de fertilizantes. No entanto, alguns cultivos são sensíveis à presença de íons cloreto (Cl) ou exigem maior teor de enxofre, o que explica o interesse em fertilizantes contendo íons sulfato (SO42-). No contexto de um mercado de fertilizantes competitivo, a indústria está constantemente à procura de novos métodos para obtenção dos produtos desejados, assim como novas matérias-primas. Portanto, o concentrado mineral de glauconita pode ser visto como uma fonte potencial de matéria-prima para obtenção de Sulfato de Potássio, por ser este um mineral essencialmente constituído por óxidos e hidróxidos de Mg, Al, Si, Fe e K. Em geral, pode-se dizer que a glauconita tem um teor de potássio variando de 5 a 8 por cento de K2O, dando suporte à sua utilização em um processo de obtenção de K2SO4, para uso no enriquecimento de fertilizantes. Dentro dessa perspectiva, o trabalho proposto tem como objetivo apreciar o processamento químico de um concentrado de glauconita, baseado em uma rota que tem início numa digestão com H2SO4 seguida de separações seletivas. O trabalho também contempla a caracterização dos materiais envolvidos, tanto o concentrado inicial de glauconita bem como os produtos obtidos ao longo do processamento químico. Para isso foram utilizadas as técnicas de Microscopia Eletrônica de Varredura (MEV / EDS), Espectrometria de Infravermelho por Transformada de Fourier (FTIR), Difração de Raios X (XRD) e Fluorescência (FRX). Estudos preliminares indicam que a digestão com H2SO4 tem um comportamento seletivo em relação ao SiO2, que não é sensível ao ataque ácido. Desta forma, o produto resultante pode ser lavado com água e filtrado consecutivas vezes formando soluções ricas em Mg, Al, Fe e K, bem como os ânions de sulfato. Tais soluções foram, em seguida, submetidos a evaporações controladas, a fim de determinar as condições em que a precipitação seletiva de um precursor rico em K e Al (por exemplo, alúmen de potássio) seria viável. O material obtido foi calcinado a 800 graus C, de modo a estimular sua decomposição em K2SO4 e Al2O3. A separação final entre estes componentes foi levada a efeito por meio de solubilização seletiva com água, permitindo a separação do óxido insolúvel. Os resultados sugerem que a rota de processamento proposto caracteriza-se como um método em potencial para obtenção do K2SO4, de interesse para a indústria de fertilizantes, podendo ainda viabilizar a formação do Al2O3 que, preferencialmente, deve conter níveis baixos de ferro para ter uma eventual utilização, com atrativos ambientais, na indústria do alumínio. / [en] Potassium ions (K+) are essentials for plant nutrition and are, usually, supplied in the form of KCl or K2SO4. The former is responsible of 90 percent of the potassium amount offered by the fertilizer industry. However, some crops are sensitive to presence of chloride ions (Cl-) or require higher sulfur content, thereby keeping the interest in fertilizers with sulfate ions (SO42-). In the context of a competitive fertilizer market, the industry is constantly searching for new methods to obtain their desired products as well as new raw materials. Therefore, glauconite bearing ores can be seen as a potential source of raw material as this hydrate silicate mineral is essentially constituted by oxides and hydroxides of Mg, Al, Si, K and Fe. In general, it can be said that glauconita has a potassium content varying from 5 to 8 percent of K2O, encouraging its use in a process to obtain K2SO4 with fertilizers purposes. Within this perspective, the proposed work aims to appreciate the chemical processing of a glauconite concentrate based on a combined aqueous and pyro metallurgical route. The present work also incorporates the materials characterization of the received concentrate as well as of the obtained products by means of Scanning Electron Microscopy (SEM/EDS), Fourier Transform Infrared Spectroscopy (FTIR), X-ray Diffraction (XRD) and Fluorescence (XRF). Preliminary studies have shown that H2SO4 digestion has a selective behavior towards SiO2 which is not sensible to the acid attack. It was also found that the resulting product can be washed with water forming solutions rich in Mg, Al, K and Fe as well as sulfate ions. These were then submitted to controlled evaporations in order to establish conditions in which the selective precipitation of a precursor rich in K and Al (e.g. potassium alúmens) is feasible. The obtained material was calcined at 800 degree C in order to stimulate decomposition in K2SO4 and Al2O3. The separation between these constituents was taken into effect through selective solubilization with water allowing the separation of the insoluble oxide. The results suggest that the proposed processing route characterize itself as a potential method to obtain K2SO4, of interest for the fertilizer industry, and Al2O3 which, in containing a small amount of iron, could be used, with environmental attractiveness, in the aluminum industry.