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In Escherichia coli, the utilization of glycerol and sn-glycerol 3-phosphate is mediated by gene products of the glp regulon. The regulon encompasses five operons, including the glpFKX operon. Although glpF and glpK encode glycerol diffusion facilitator and glycerol kinase,respectively, the function of glpX was unknown. In the present work, we show that glpX encodes a fructose 1,6-bisphosphatase (FBPase), which catalyzes the hydrolysis of fructose 1,6-bisphosphate to fructose 6-phosphate and phosphate. The purified FBPase was dimeric, dependent on Mn2+ for activity and exhibited an apparent Km of 35 μM for fructose 1,6-bisphosphate. The enzyme was inhibited by ADP, ATP and phosphate and activated by PEP. The attributes of the glpX-encoded FBPase were different from those of the previously characterized E. coli FBPase encoded by fbp. Mutants deleted in fbp (Δfbp) display a growthnegative phenotype on gluconeogenic carbon sources such as glycerol, indicating the inability of chromosomal glpX+ to complement Δfbp. However, a Δfbp mutation was complemented by overexpression of glpX+. In contrast, a glpX mutant exhibited a growth-positive phenotype on glycerol, glucose or fructose media. Surprisingly, a double mutant strain glpX pfkA (6-phosphofructokinase I) was more inhibited in growth on glucose and glycerol media than the pfkA parent. Carbohydrate metabolism in the pfkA background may be affected by the glpXmediated change in fructose 6-phosphate/fructose 1,6-bisphosphate levels. FBPase activities of soluble proteins separated by non-denaturing PAGE were visualized, showing a novel (third) FBPase, perhaps encoded by the glpX homolog, yggF. / Master of Science

Partial denitrification/anammox (PdNA) is a biological nitrogen removal technology with significant carbon and aeration savings when compared with conventional nitrification/denitrification. Yet despite these benefits, the use of PdNA in mainstream wastewater treatment remains limited. One of the main barriers to implementation of anammox-based technologies is the slow growth rate of anammox (AMX), which results in a long startup time. To accelerate startup, the typical approach to commissioning AMX-based processes, specifically used for sidestream partial nitritation/AMX, is with biomass augmentation, which is practically unrealistic for full-scale mainstream applications. Thus, this study evaluated startup strategies for mainstream PdNA without AMX inoculation in moving bed biofilm reactors (MBBRs) with two simultaneous experiments. In one experiment, an MBBR was started using IFAS carriers with a preliminary biofilm and no external carbon dosing or AMX biomass inoculation. The feed was controlled to 20°C and included mainstream conditions of nitrite and ammonia controlled to the stoichiometric requirements for AMX growth. After only 84 days of operation, AMX activity was confirmed in the reactor with evidence of activity a few weeks before testing. In the second experiment, four reactors were started with either virgin carriers or integrated fixed-film activated sludge (IFAS) carriers with a preliminary biofilm of heterotrophs and nitrifiers. The reactors were fed mainstream levels of ammonia and nitrate with a temperature control target of 20°C and one reactor of each carrier type was dosed with carbon in the form of either glycerol or methanol. Carbon dosing was based on a feedback proportional-integrative-derivative (PID) control loop with a nitrate residual of 1-1.5 mgNO3-N/L. Of the four reactors, the preliminary biofilm carrier reactor dosed with glycerol achieved AMX activity first after 224 days of operation, but it was determined this was likely limited by synthetic feeding for the first 184 days. These results, along with other recent PdNA work, suggest that growth of AMX on biofilm carriers can be established in mainstream conditions in 50-100 days, depending on media selection and carbon source. Ultimately, this research will help utilities understand methods for starting up mainstream PdNA MBBRs from scratch and make this technology more accessible. / Master of Science / Intensification is the practice by which operational changes and new technologies are employed to reduce economic, resource, energy, and space requirements of wastewater treatment plants. One area of increasing focus involves the use of anaerobic ammonia oxidizing bacteria, or anammox (AMX), to reduce the aeration and carbon dosing needs for treating wastewater. One of the main barriers to implementation of AMX-based technologies is the slow growth rate of AMX, which results in a long startup time. To accelerate startup, the typical approach to commissioning AMX-based processes, specifically used for sidestream partial-nitritation/AMX, is with augmentation of biomass, which is practically unrealistic for full-scale mainstream applications. Thus, this study evaluated startup strategies for mainstream moving bed biofilm reactors (MBBRs) without AMX biomass inoculation in two simultaneous experiments in an AMX MBBR and a partial denitrification/AMX (PdNA) MBBR. In one experiment, idealized stoichiometric conditions for AMX growth were provided to a mainstream MBBR started with carriers from an aerobic integrated fixed-film activated sludge (IFAS) process to determine how fast AMX could potentially grow. In another experiment, different carrier types, virgin or preliminary biofilm carriers from an IFAS process, and different carbon sources, methanol and glycerol, were tested to determine the best methods for encouraging AMX attachment and growth in a PdNA process. These results, along with other recent PdNA work, suggest that growth of AMX on biofilm carriers can be established in mainstream conditions within 50-100 days, depending on media selection and carbon source. Ultimately, this research will help utilities understand methods for starting up mainstream PdNA MBBRs from scratch and make this technology more accessible.

Organic esters are an important class of industrial and commercial chemicals that can be found in solvents, plasticizer, food flavours, detergents, agrochemicals, and pharmaceuticals. The most common way to synthesis organic esters is with esterification or transesterification. Where esterification was the chosen method in this thesis.  This thesis provides a techno-economic assessment on the production of propyl acetate and glycerol trivalerate through different continuous routes that is than compared with batch production under the same conditions. Simulations was done on trivalerate due to limited literature and data on pentaerythritol tetravalerate. Different continuous technologies that have been assessed in this thesis was plug flow reactor (PFR), Reactive Distillation (RD), and Reactive – Extractive Distillation (RED). The production of mono- and polyol esters with different unit operators was simulated in Aspen Plus V.14. Techno-Economic analysis was conducted with APEA (Aspen Process Economic Analyzer), where cost of raw materials, products, and utilities was inserted to evaluate annual operating cost and product sale. Reaction kinetic for esterification of trivalerate was estimated by obtained values from simulations of a Gibbs reactor in Aspen Plus at four different temperatures. Kinetics was estimated by applying the relation between the chemical equilibrium constant and the Arrhenius equation. Where rate constant and activation energy for forward and revers reaction was obtained by varying min and max values for lsqcurvefit in MATLAB and then validate results with published kinetics.  Results from production of propyl acetate with batch, PFR and RED provides an annual profit of approximately 1 M$ at a capacity of 41.65 kton/year. The three different process provides also approximately an equal capital cost, operating cost, equipment cost, and installed cost according to APEA. RED provided the lowest propyl acetate yield at 93%, batch and PFR provided a propyl acetate yield of 94%.  Results from production of trivalerate with batch, PFR, and RD provides an annual profit of 5.4, 5.78, and 9.2 M$ at a capacity of approximately 5 kton/year. Where RD process provides the lowest capital cost, operating cost, equipment cost, and installed cost compared to batch and PFR processes according to APEA. Obtained results from production of trivalerate can be used to evaluate the economic and technical feasibility of a continuous production plant for pentaerythritol tetravalerate (PETV). Where initial simulations show a good economic and technical viability of a continuous ester production plant.

The aim of this thesis is to investigate molecular mechanism of reactive oxygen species production by flavin dehydrogenases mitochondrial glycerol-3-phosphate dehydrogenase (mGPDH) and succinate dehydrogenase (SDH). Together, they represent important source of reactive oxygen species in mammalian mitochondria, but the mechanism of electron leak is still poorly understood. Because mechanisms of reactive oxygen species production by other complexes of respiratory chain are better characterized, they can serve as case studies to get insight into mechanisms of reactive oxygen species by flavin dehydrogenases. Relevant knowledge is therefore summarized in the first part of the thesis. To study the production of reactive oxygen species by the isolated flavin dehydrogenases, we used brown adipose tissue mitochondria solubilized by digitonin as a model. Enzyme activity measurements, hydrogen peroxide production studies by Amplex UltraRed fluorescence and luminol luminescence revealed flavin as the most likely source of electron leak in SDH under in vivo conditions, while we propose coenzyme Q binding site as the site of reactive oxygen species production in the case of mGPDH. Distinct mechanism of this production by the two dehydrogenases is also apparent from induction of reactive oxygen species...

Le travail de thèse porte sur la synthèse de nouveaux monomères photopolymérisables issus de la biomasse, l’étude de leur photopolymérisation et la caractérisation des matériaux qui en résultent. Les monomères formulés ont été testés dans un procédé d’impression de caractères Braille.Les monomères ont été élaborés en deux étapes en utilisant un chemin réactionnel simple, économique et respectueux de l’environnement. La première étape a consisté à faire réagir des dérivés du glycérol (carbonate de glycérol ou glycidol) avec des acides gras. Dans un deuxième temps, les [alpha]-monoglycérides obtenus ont été fonctionnalisés en vue de les rendre photopolymérisables. Les différents monomères obtenus porteurs de fonctions (méth)acrylate et/ou époxy ont été photopolymérisés en quelques secondes en présence d’un photoamorceur. Les cinétiques de photopolymérisation ont été suivies par spectrométrie IR-TF en mode ATR et différents paramètres tels que la quantité et la nature du photoamorceur, l’intensité d’irradiation et la température ont été optimisés. Les différents matériaux obtenus après photoréticulation ont été caractérisés afin de mettre en évidence leurs principales physico-chimiques. Des relations structure – propriétés ont ainsi été établies.Enfin, une formulation photopolymérisable présentant une viscosité compatible avec le procédé d’impression Braille a été élaborée par ajout de silice nanométrique. Les points Braille obtenus présentent des caractéristiques proches de ceux réalisés par le partenaire industriel. / The work of the PhD deals with the synthesis of polymerizable monomers derived from biomass, the study of their photopolymerization and the characterization of the resulting materials. The formulated monomers were tested in a printing method of Braille characters.The monomers were prepared in a two-steps reaction that is simple, economic and environmentally friendly. The first step comprised a reaction between glycerol derivatives (glycerol carbonate or glycidol) and fatty acids. In the second step, the obtained [alpha]-monoglycerides were functionalized to obtain photopolymerizable monomers. The different obtained monomers that bear (meth)acrylate and/or epoxy groups were photopolymerized in a few seconds in the presence of photoinitiator.The photopolymerization kinetics were followed by FT-IR spectroscopy in ATR mode, and various parameters such as the amount and the nature of the photoinitiator, the irradiation intensity and the temperature were optimized. The different photocured materials were characterized to highlight their main physicochemical properties. Structure-properties relations were then established.Finally, a photopolymerizable formulation having a viscosity compatible with the printing process of Braille characters was developed by adding nanometric silice. The obtained Braille characters exhibited features close those carried out by the industrial partner.

Eastern Africa has experienced strong climatic changes since the last deglaciation (15 000 years ago). The driving mechanisms and teleconnections of these spatially complex climate variations are yet not fully understood. Although previous studies on lake systems have enhanced our knowledge of Holocene precipitation variation in eastern Africa, relatively few studies have reconstructed the terrestrial temperature history of eastern Africa from lake archives. Here, we present (i) a new branched glycerol dialkyl glycerol tetraether (brGDGT) temperature calibration that includes Bale Mountains surface sediments and (ii) a quantitative record of mean annual air temperature (MAT) over the past 12 ka cal BP using brGDGTs in a sediment core collected from Garba Guracha (3950ma.s.l.) in the Bale Mountains. After adding Bale Mountains surface sediment (n = 11) data (Baxter et al., 2019) to the existing East African lake dataset, additional variation in 6-methyl brGDGTs was observed, which necessitated modifying the MBT'5ME calibration (MBT denotes methylation of branched tetraethers) by adding 6-methyl brGDGT IIIa0 (resulting in the MBT Bale Mountains index, r² = 0:93, p < 0:05). Comparing the MBT'5ME and the new MBT Bale Mountains index, our high-altitude Garba Guracha temperature record shows that warming occurred shortly after the Holocene onset when the temperature increased by more than 3.0°C in less than 600 years. The highest temperatures prevailed between 9 and 6 ka cal BP, followed by a temperature decrease until 1.4 ka cal BP. The reconstructed temperature history is linked to supraregional climatic changes associated with insolation forcing and the African Humid Period (AHP), as well as with local anomalies associated with catchment deglaciation and hydrology.

Master of Science / Department of Animal Sciences and Industry / Terry A. Houser / Two experiments were conducted to determine the effects of dried distillers grains with solubles (DDGS) on pork loin and fat quality. In the first experiment, 1,160 barrows (PIC) were used in a 70-d study to determine the influence of DDGS and glycerol on pork loin and fat quality attributes. Barrows were fed a corn-soybean meal based diet with the addition of selected levels of DDGS (0 or 20%) and glycerol (0, 2.5, or 5%) feed stuffs. Loins from the two heaviest pigs in each pen were removed for evaluation of pork loin and fat quality. Experiment two was a pilot study, in which eight non-pregnant sows were fed either 0 or 50% DDGS with a corn soybean meal based diet for 92-d. In the first experiment, there were no DDGS x glycerol interactions for purge loss %, instrumental color (L*a*b*), visual color, marbling score, drip loss %, visual color, pH, Warner-Bratzler shear force (WBSF), cook loss %, myofibrillar tenderness, juiciness, pork flavor intensity, connective tissue amount, and overall tenderness. There was a DDGS x glycerol interaction (P<0.03) for off-flavor intensity. Pigs fed diets with 20% DDGS had higher WBSF values, lower myofibrillar tenderness, lower overall tenderness scores, lower connective tissue scores, and had more off-flavors (P<0.05). Loin fatty acid analysis revealed an increase in palmitoleic, linoleic, and eicosadienoic acids (P<0.05) and iodine value (P<0.03) for pigs fed 20% DDGS. In the second experiment, there were no differences (P>0.64) in BW or backfat change for sows fed either 0 or 50% DDGS. No differences (P>0.23) in lipid oxidation from lean trimmings as measured by 2-thiobarbituric acid reactive substances (TBARS) assay were reported either initially or after 5 d of retail display for sows fed either 0 or 50% DDGS. As expected, lipid oxidation increased (P<0.003) as measured by TBARS assay for both treatments from d 1 to 5. Jowl fatty acid analysis revealed an increase in linoleic acid (P<0.01), total polyunsaturated fatty acids (P<0.01), and the ratio of polyunsaturated fatty acids to saturated fatty acids (P<0.03) for sows fed 50% DDGS.

Systemic acquired resistance (SAR) is a form of inducible defense response triggered upon localized infection that confers broad-spectrum disease resistance against secondary infections. Several factors are known to regulate SAR and these include phenolic phytohormone salicylic acid (SA), phosphorylated sugar glycerol-3-phosphate (G3P), and dicarboxylic acid azelaic acid (AzA). This study evaluated a role for free radicals nitric oxide (NO) and reactive oxygen species (ROS) in SAR. Normal accumulation of both NO and ROS was required for normal SAR and mutations preventing NO/ROS accumulation and/or biosynthesis compromised SAR. A role for NO and ROS was further established using pharmacological approaches. Notably, both NO and ROS conferred SAR in a concentration dependent manner. This was further established using genetic mutants that accumulated high levels of NO. NO/ROS acted upstream of G3P and in parallel to SA. Collectively, these results suggest that NO and ROS are essential components of the SAR pathway.

Background: Trauma is a major cause of death and disability. Adverse events, such as prolonged prehospital time, hypoxia, hypotension and/or hyperventilation have been reported to correlate to poor outcome. Adequate cortisol levels are essential for survival after major trauma. In hypotensive critically ill patients, lack of sufficient amount of cortisol can be suspected, and a concept of critical illness related corticosteroid insufficiency has been proposed. Corticosteroid therapy has many adverse effects in critically ill patients and should only be given if life-saving. Correct measurement of serum cortisol levels is important but difficult in critically ill patients with capillary leakage. Estimation of the free and biologically active cortisol is preferable. In serum less than 10% of cortisol is free and biologically active and not possible to measure with routine laboratory methods. Salivary cortisol can be used as a surrogate for free cortisol, but salivary production is reduced in critically ill patients. Liver resection could reduce cortisol levels due to substrate deficiency. Aims: 1. Evaluate the occurrence of early adverse events in patients with traumatic brain injury and relate them to outcome. 2. Assess cortisol levels over time after trauma and correlate to severity of trauma, sedative/analgesic drugs and cardiovascular function. 3. Evaluate if saliva stimulation could be performed without interfering with salivary cortisol levels. 4. Assess cortisol levels over time after liver resection in comparison to other major surgery. Results: There was no significant correlation between prehospital time ³60 minutes, hypoxia (saturation &lt;95%), hypotension (systolic blood pressure &lt;90 mmHg), or hyperventilation (ETCO2 &lt;4.5 kPa) and a poor outcome (Glasgow Outcome Scale 1-3) in patients with traumatic brain injury. Cortisol levels decreased significantly over time after trauma, but there was no correlation between low (&lt;200 nmol/L) serum cortisol levels and severity of trauma. Infusion of sedative/analgesic drugs was the strongest predictor for a low (&lt;200 nmol/L) serum cortisol. The odds ratio for low serum cortisol levels (&lt;200 nmol/L) was 8.0 for patients receiving continuous infusion of sedative/analgesic drugs. There was no significant difference between unstimulated and stimulated salivary cortisol levels (p=0.06) in healthy volunteers. Liver resection was not associated with significantly lower cortisol levels compared to other major surgery. Conclusion: There was no significant correlation between early adverse events and outcome in patients with traumatic brain injury. Cortisol levels decreased significantly over time in trauma patients. Low cortisol levels (&lt;200 nmol/L) were significantly correlated to continuous infusion of sedative/analgesic drugs. Saliva stimulation could be performed without interfering with salivary cortisol levels. Liver resection was not associated with low cortisol levels compared to other major surgery.