Detection of cellular messenger RNA is a useful diagnostic strategy for the detection of patho-gens. A rapid and sensitive method for on-site detection of specific pathogens would be of great use in a number of fields. For example, a simple and inexpensive method for the detection of harmful biological agents in train stations and airports is useful for national security. Rapid detection of pathogenic E. coli strains in food production would also be of great benefit in ensuring the safety and quality of our food supply. Here we present a method for the rapid de-tection of cellular mRNA. This system is based on the 3’-labeling approach in which targeted RNA is simultaneously extended and labeled with the use of biotin labeled-dNTPs and DNA po-lymerase on an immobilized nucleic acid probe. The biotin is subsequently converted to an enzymatic label, which produces a detectable chemiluminescent reaction in the presence of substrate. Detection time of this system is short (approximately 20 minutes) because there is no need for amplification by PCR, transcription, or fluorophore labeling. This novel methodology has been successfully demonstrated by selective detection of lac Z mRNA in a total RNA sample from E. coli.

Minerals, metals and mining are the cornerstone of technological development and play an essential role in achieving the United Nations Sustainable Development Goals. Mining, however, is not a wastefree process, with mine wastes being a source of a host of environmental problems. One of these is acid rock drainage (ARD), which forms through a complex series of acid formation (mainly via sulfide oxidation), neutralisation (primarily by carbonates) and gangue mineral dissolution reactions in waste rock and tailings storage piles. The resulting drainage waters are often acidic, highly saline and may contain elevated levels of deleterious elements. Effective ARD mitigation requires accurate ARD characterisation and prediction strategies. To date, standard guidelines recommend a suite of geochemical static (characterisation) and kinetic (prediction) tests. Characterisation tests such as acid-base accounting (ABA) and net acid generation (NAG) tests provide a quick and relatively inexpensive estimate of the "worst case" scenario for acid formation and neutralisation, while kinetic tests (commonly humidity or column leach tests) aim to predict the longterm weathering potential of waste material. The UCT biokinetic test (not currently industry standard practice) was developed to address the effect of microorganisms on ARD formation and allow for the collection of relative kinetic data on neutralisation and acidification within a shortened time period. None of these tests, however, account for the additional layer of complexity introduced by mineral texture, which describes the interrelationship of mineral grains to one another, their shapes and sizes, with some frequently studied textural parameters including mineral liberation, association, grain size distribution and particle size. Mineralogical and textural analyses are infrequently practiced in the context of ARD assessment due to the difficulty in obtaining statistically sound quantitative textural data, high costs of measurement, and standard ARD assessment protocols recommending (rather than necessitating) these assessments. An ARD assessment approach that includes static, kinetic, mineralogical and textural assessments has nonetheless been suggested by several researchers. This project assessed the dominating textural parameters on the scales of kinetic (humidity cell) test (HCT) feed material (meso-scale) and characterisation (static and UCT batch biokinetic) test (SCT) feed material (micro-scale) using four waste rock samples (A, B, C and D) from a greenstone belt gold deposit as a case study. More specifically, the study aimed to assess the role of mineralogy and texture in the ARD assessment "toolbox" and to investigate the role of coarse material sampling for ARD assessment. Data sets collected included the PSD of the micro- and meso-scale material, sample chemistry data obtained from XRF spectrometry and LECO total sulfur, bulk mineralogy data from QXRD and QEMSCAN, as well as textural and mineralogical data from QEMSCAN for sized and unsized micro- and meso-scale material. ARD-specific data sets included results of geochemical characterisation tests such as ANC and single-addition NAG tests, the UCT batch biokinetic test with and without pH control for samples C and D, as well as prediction test data from water-fed and modified humidity cell tests. The geochemical static tests performed on samples A, B, C and D classified them as PAF, PAF, uncertain and NAF, respectively. Non-pH-controlled UCT batch biokinetic tests remained circumneutral for samples B, C and D over the duration of 90 days, while for sample A the pH became acidic over time. The pH-controlled tests demonstrated a steady depletion of neutralisation potential over the first 30 days. Humidity cell test results demonstrated no acidic leachate formation for waterfed tests over 40 weeks, while modified tests showed a decreasing pH over time as the neutralisation capacity was reduced. The mineralogy was important for the interpretation of test results on both the micro-and meso-scales and was assessed in terms of both discrete minerals and reactivity groupings (Fe-Sulfide, other sulfide, dissolving (carbonate), fast weathering, intermediate weathering, slow weathering, inert and other). For sample A the mineralogy was dominated by the inert (quartz), slow weathering (magnetite, plagioclasealbite) and intermediate weathering (Fe-amphibole) categories, with lesser contributions from the FeSulfide (pyrrhotite), dissolving (calcite) and fast weathering (epidote) groups. The main groups contributing to the sample B mineralogy were the slow weathering (plagioclase-albite, magnetite, Kfeldspar), inert (quartz) and intermediate weathering (Fe-mica, chlorite) groups, followed by Fe-Sulfide (pyrite), dissolving (calcite) and fast weathering (epidote) mineral groups. Sample C mineralogy comprised predominantly inert (quartz, titanite), Fe-Sulfide (pyrrhotite), dissolving (calcite) and intermediate weathering (Fe-mica, chlorite) minerals, with lesser contributions from slow weathering (K-feldspar) and fast weathering (epidote) minerals. Sample D comprised intermediate weathering (Feamphibole, chlorite, Fe-mica) and slow weathering minerals, with lesser contributions from slow weathering (magnetite), dissolving (calcite) and Fe-Sulfide (pyrrhotite) minerals. Textural parameters (liberation and association, grain size distribution and liberation spectrum) were evaluated for the FeSulfide and dissolving minerals. On the micro-scale, a large portion of the Fe-Sulfide and dissolving minerals in the samples was found in the liberated category (50%) of the texturally significant size fractions (>1mm), which comprised predominantly locked Fe-Sulfide and dissolving minerals. Evidence of a bimodal distribution was, however, found for sample C via the liberation spectrum and grain size distribution (early liberation size of 8mm), which accounted for the larger degree of liberation observed in the >1mm size fractions, and a larger degree of liberation for the sample overall. The association of Fe-Sulfide and dissolving minerals for all samples was found to be primarily to inert, intermediate weathering and slow weathering minerals, with a larger degree of association of Fe-Sulfide to dissolving minerals observed in sample C. On the micro-scale the mineralogy helped inform the placement of the samples on the geochemical classification plot based on the Fe-Sulfide, dissolving and intermediate weathering mineral contents. For the non-pH-controlled UCT batch biokinetic test, the presence and abundance of calcite was thought to dictate the PAF/NAF nature of the test, as even relatively low amounts of calcite rendered the pH circumneutral for the duration of the test (sample B). For pH-controlled tests, however, the calcite was depleted over time, which led to a favourable acidic environment for the acidophilic bacteria used in the batch biokinetic test. For both the geochemical characterisation and the pH-controlled UCT batch biokinetic tests there was evidence to suggest the contribution of intermediate weathering (Feamphibole, Fe-mica, chlorite) and slow weathering (magnetite) minerals to the neutralisation potential in the sample. On the meso-scale the effects of mineralogy were most prominent for the modified humidity cell tests, which showed some pH fluctuations and a steady depletion of the primary neutralisation potential. The pH fluctuation after the depletion of the dissolving minerals was attributed to the dissolution of intermediate weathering minerals over the 40 weeks of the tests. These effects were not observed during the 40 weeks of the water-fed experiments. Given sufficient time for the latter test, however, it would be expected that upon the onset of acidification, similar effects of the mineralogy on the leachate quality would be observed as in the modified tests. Knowledge of the Fe-sulfide and dissolving mineral texture yielded several insights. on the micro-scale, the liberation and grain size distribution data provided an indication that a sample-customised grinding size should be established to ensure adequate "worst case" scenario determination via characterisation tests, as material with fine Fe-Sulfide or dissolving mineral grains may not be fully liberated at the recommended 75µm top size. On the meso-scale, the texture yielded insight into the circumneutral behaviour of the water-fed HCT, as most of the acid-forming minerals were contained in size fractions where the liberation was either limited or negligible, with predominant association to slow weathering, intermediate weathering and inert minerals. These findings highlighted the importance of considering mineralogy, texture and the PSD of the material for HCT result interpretation. When considering texture as a parameter for ARD assessment, the potential for sampling and mineralogical errors arose due to the coarse material size (specifically on the meso-scale) and the limitations on the number of particles that could be assessed. Quantitative mineralogy and texture data allowed for the quantitative assessment of the sampling and mineralogical errors, which were investigated through Pierre Gy's fundamental sampling error (FSE) equation, the binomial distribution approximation and the plotting of confidence intervals over the Fe-Sulfide liberation data. The results showed that although tools such as Gy's "safety line" provide a useful quick means of sampling error assessment, this approach may yield excessively large sample mass requirements for coarse material. Calculating the sampling error from the textural and mineralogical data provided a useful tool to estimate sample representativeness. Additionally, the estimation of sampling errors may help in the planning of an appropriate sampling approach, which may ultimately provide a means to relate data sets to one another across scales based on how representative samples are of one another, and therefore of the parent lot. The current study showed how mineralogy and texture are not simply "tools" in the ARD assessment "toolbox", but rather a key means for interpreting characterisation and prediction test data. Additionally, the quantitative assessment of mineralogy and texture provided the opportunity to assess the materialspecific sampling error, which, in turn, may allow for the correlation of data sets across various scales and for the planning of appropriate sampling strategies. Recommendations for future work include: the quantitative assessment of the ARDI for meso-scale material; the assessment of detailed characterisation and prediction test leachate chemistry; trace element assessment and deportment throughout UCT batch biokinetic and humidity cell testing; mineralogical and textural assessment on characterisation and prediction test residues during and after tests; an in-depth analysis of the minimal/optimal sample block/sub-sample mass required for minimal error; the assessment of samples using X-ray microcomputed tomography to assess and decrease the effects of stereological bias prevalent in 2D measurements; and the application of a similar texture and mineralogy assessment to additional waste types (such as coal wastes, or waste material containing non-Fe-bearing sulfides).

No description available.

Herein are presented collective studies of amino acid-based surfactants, also known as lipoamino acids, at liquid interfaces. Chapter III describes an investigation of domain morphology of N-Stearoylglutamic acid (N-SGA) Langmuir monolayers at the air/water interface by epifluorescence microscopy. Anisotropic feather-like domains were observed in L-enantiomeric monolayers while symmetric circular domains were found in racemic N-SGA monolayers. At a surface pressure of 30 mN/m the enantiomeric domains melted at 31 ??C while the racemic domains melted at 27 ??C. This result is exactly opposite to the behavior found in bulk crystals where the racemate melts at a higher temperature. These results were explained in terms of different molecular packing and hydrogen bonding between bulk crystals and two-dimensional thin films for enantiomeric and racemic compounds. Chapter IV summarizes the investigations of hydrogen bonding in N-acyl amino acid monolayers by vibrational sum-frequency spectroscopy (VSFS). The intermolecular hydrogen bonding interaction between the adjacent molecules through amide-amide groups in N-stearoylalanine (N-SA) is characterized by an NH stretch peak at 3311 cm-1. This is the first time that the amide NH stretching signals have been detected with the VSFS technique. A similar peak was detected at 3341 cm-1on N-SGA monolayer. The higher frequency indicates that the H-bond strength is weaker due to the larger size of the glutamic acid residue. The NH stretch mode can thus be used as a fingerprint of hydrogen bonding among amide-amide groups. A peak at 3050 cm-1 due to hydrogen bonding among carboxyl groups was also resolved from the VSFS spectra. Molecular models of intermolecular hydrogen bonding were proposed.

Herein are presented collective studies of amino acid-based surfactants, also known as lipoamino acids, at liquid interfaces. Chapter III describes an investigation of domain morphology of N-Stearoylglutamic acid (N-SGA) Langmuir monolayers at the air/water interface by epifluorescence microscopy. Anisotropic feather-like domains were observed in L-enantiomeric monolayers while symmetric circular domains were found in racemic N-SGA monolayers. At a surface pressure of 30 mN/m the enantiomeric domains melted at 31 ??C while the racemic domains melted at 27 ??C. This result is exactly opposite to the behavior found in bulk crystals where the racemate melts at a higher temperature. These results were explained in terms of different molecular packing and hydrogen bonding between bulk crystals and two-dimensional thin films for enantiomeric and racemic compounds. Chapter IV summarizes the investigations of hydrogen bonding in N-acyl amino acid monolayers by vibrational sum-frequency spectroscopy (VSFS). The intermolecular hydrogen bonding interaction between the adjacent molecules through amide-amide groups in N-stearoylalanine (N-SA) is characterized by an NH stretch peak at 3311 cm-1. This is the first time that the amide NH stretching signals have been detected with the VSFS technique. A similar peak was detected at 3341 cm-1on N-SGA monolayer. The higher frequency indicates that the H-bond strength is weaker due to the larger size of the glutamic acid residue. The NH stretch mode can thus be used as a fingerprint of hydrogen bonding among amide-amide groups. A peak at 3050 cm-1 due to hydrogen bonding among carboxyl groups was also resolved from the VSFS spectra. Molecular models of intermolecular hydrogen bonding were proposed.

The overall aim of this thesis was to establish strategies to minimize secondary damage to the injured spinal cord. Secondary damage that follows spinal cord injury (SCI) involves inflammatory and excitotoxic pathways. Regulation of these pathways using immunomodulatory and neuroprotective substances potentially protects the injured spinal cord from further damage. We also developed and studied resorbable biomaterials to be used as carriers for potential neuroprotectants to the injured spinal cord. We used transversal spinal cord slice cultures (SCSCs) derived from postnatal mice as a model. SCSCs were maintained on different biomaterials and were studied after treatment with immunomodulatory and/or neurotrophic factors. They were further excitotoxically injured and subsequently treated with interleukin-1 receptor antagonist (IL1RA) or by neural crest stem cell (NCSC)-transplantation. The results show that biocompatible and resorbable hydrogels based on hyaluronic acid (HA) preserved neurons in SCSCs to a much higher extent than a conventional collagen-based biomaterial or standard polyethylene terephthalate (PET) membrane inserts. Glial activation was limited in the cultures maintained on HA-based hydrogel. The anti-inflammatory factor IL1RA protected SCSCs from degenerative mechanisms that occur during in vitro incubation, and IL1RA also protected SCSCs from excitotoxic injury induced by N-Methyl-d-Aspartate (NMDA). IL1RA specifically protected neurons that resided in the ventral horn, while other neuronal populations such as dorsal horn neurons and Renshaw cells did not respond to treatment. Finally, transplantation of NCSCs onto excitotoxically injured SCSCs protected from neuronal loss, apoptosis and glial activation, while NCSCs remained undifferentiated. The results presented in this thesis indicate that carriers based on HA seem to be more suitable than conventional collagen-based biomaterials since they enhance neuronal survival per se. The observed neuroprotection is likely due to biomechanical properties of HA. IL1RA protects SCSCs from spontaneous degeneration and from NMDA-induced injury, suggesting that excitotoxic mechanisms can be modulated through anti-inflammatory pathways. Different neuronal populations are affected by IL1RA to various degrees, suggesting that a combination of different neuroprotectants should be used in treatment strategies after SCI. Finally, NCSCs seem to protect SCSCs from excitotoxic injury through paracrine actions, since they remain undifferentiated and do not migrate into the tissue during in vitro incubation. It seems that combinations of neuroprotectants and carrier substances should be considered rather than one single strategy when designing future treatments for SCI. Incorporation of neuroprotectants such as IL1RA combined with stem cells in injectable biocompatible carriers based on HA is the final goal of our group in the treatment of SCI.

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L’objectif de ces travaux de thèse a été de développer des polyesters issus de ressources oléagineuses pour les utiliser comme additifs pour moduler la viscosité d’huiles lubrifiantes. Pour ce faire, l’approche par polycondensation de monomères de type hydroxy-acide a été privilégiée. Dans un premier temps, le poly(ricinoléate de méthyle) et son homologue saturé, le poly(12-hydroxystéarate de méthyle), ont été synthétisés dans une large gamme de masses molaires et leur utilisation comme épaississant d’huiles lubrifiantes a été démontrée. Dans un second temps, des polyesters dérivés du poly(ricinoléate de méthyle) et présentant des architectures de polymère en peigne ont été synthétisés par addition thiol-ène et polycondensation. Une étude de l’impact de l’architecture de ces polyesters sur leur comportement en solution a permis de prouver que les structures en peigne étaient les plus adaptées pour des applications visant, à la fois, un épaississement et une diminution du point d’écoulement de l’huile lubrifiante. Par la suite, des copoly(9-alkyl 12-hydroxystéarate)s en peigne possédant différentes chaînes pendantes ont été synthétisés afin de contrôler leur solubilité dans une huile minérale, la Yubase 4+, et ont permis de réduire la diminution de viscosité de cette huile avec la température. Finalement, l’étude dans le dodécane de deux copoly(9-alkyl 12-hydroxystéarate)s en peigne a révélé un phénomène d’agrégation des chaînes polymères lesquelles se désagrègent avec l’augmentation de la température, ce qui est en accord avec un des mécanismes d’action des additifs modulant la viscosité des huiles lubrifiantes décrit dans la littérature. / The aim of this thesis was to promote the use of polyesters from oleaginous resources as viscosity control additives for lubricants. The hydroxyl-acid type monomers were polymerized through polycondensation route. First, poly(methyl ricinoleate) and its homologous poly(methyl-12-hydroxystearate) were synthesized in a large range of molecular weights and their use as thickeners of lubricant oils was demonstrated. Secondly, comb polyesters derived from poly(methyl ricinoleate) were designed via thiol-ene addition and polycondensation process. The effect of the polyester architecture on their behavior in solution was investigated and revealed that comb polymers are the most suitable for applications that required a thickening efficiency and a pour point depressant effect. Then, comb (co)poly(9-alkyl 12-hydroxystearate)s with various pendant alkyl chains were designed in order to control their solubility in a mineral oil, the Yubase 4+, and to limit the oil viscosity decrease of these oils with temperature. Finally, the behavior in dodecane of two comb (co)poly(9-alkyl 12-hydroxystearate)s revealed that the polymer chains tend to aggregate at low temperature and to disaggregate with the temperature increase. This phenomenon is in accordance with one of the oil Viscosity Index Improver behaviors, described in literature.

Hyaluronic acid (HA) is one of the primary chemical building blocks of the extracellular matrix and thus is an attractive material for biomedical applications. FDA approved HA-based materials are available as dermal fillers, joint viscosupplements, vitreous substitutes, and abdominal adhesion barriers. The engineering of new HA-based materials and applications is an active area of research. Here we develop several new types of HA-based hydrogels with unique and useful properties. To address the challenge of delivering hydrophobic drugs from hydrophilic hydrogel matrices we have grafted HA hydrogels with [Beta]-cyclodextrin to create hydrogels capable of binding poorly water soluble drugs. To create HA hydrogels with unique anisotropic swelling behavior we have developed a dual-crosslinking technique in which a super-swelling chemically crosslinked hydrogel is patterned with low-swelling photocrosslinked domains. When this dual-crosslinked hydrogel is swelled it contorts into a new shape because of differential swelling among photopatterned regions. To address the challenge of creating hydrogel scaffolds with biomimetic branched porosity we have invented a "crystal templating" technique. This technique grows dendritic crystals throughout a biopolymer solution, crosslinks the biopolymer around the crystals, and washes the crystals away to yield a hydrogel with a dendritic macroporous network. Lastly, we invented a method for patterning a substrate with a microarray of hydrogel compartments. A microarray of living cells is obtained when cells are seeded on the hydrogel patterned substrate. This method addresses the need for an inexpensive, simple method for obtaining living cell microarrays that does not require clean room labs and lithographic expertise. Each of these new materials were based on hyaluronic acid hydrogels but the methods are generalizable to hydrogels of other polymers too. In conclusion, the novel methods in this dissertation are a significant contribution to the engineering of HA-based materials. / text

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