Background: Escherichia coli (E.coli) O86a:K61 belongs to the Enteropathogenic E. coli (EPEC) group of pathogens. Acute gastroenteritis affects 2-4 billion people annually and EPEC is associated with 10-40% of hospitalized diarrhea cases globally. Coagulation Factor (F) V circulates as an inactive procofactor (Mr 330kDa) which upon thrombin activation to the active cofactor, FVa, functions in prothombinase to accelerate prothrombin to thrombin conversion by 300,000-fold. The ability of E.coli O86a:K61 to cause intestinal hemorrhage is of interest because previous research demonstrated that during E.coli O86a:K61 sepsis in baboons, a dose-dependent inactivation of FV was observed as the bacterial dose increased. These results suggested a secreted E.coli protease may have mediated this effect on FV. This research has focused on the purification, identification, and characterization of a secreted E. coli O86a:K61 protease that inactivates FV. The final partially-purified protease inactivated FV to a 250kDa product by immunoblotting, and possessed a 900-fold increase in specific activity versus FV in human plasma compared to the culture supernatant. At least 3 proteins were observed upon SDS-PAGE. Proteolytic inactivation of FV was activated by up to 500-fold with β-mercaptoethanol and 2-fold with 1M urea. The protease was heat stable retaining all of its activity versus FV after 1h at 70°C or 80°C, and partial activity (50%) at 95°C. Proteolysis of FV was blocked by 90% with alpha-1-protease inhibitor; however, the protease was resistant to 1.5 mM PMSF, and unaffected by E64, or iodoacetamide. FV is a major regulator of the coagulation process and its inactivation by the secreted E.coli protease would be expected to result in a net bleeding tendency which may contribute to the mucosal hemorrhage observed in humans with associated hemorrhagic colitis. Proteolytic inactivation of FV is predicted to result in decreased bacterial containment by host fibrin thereby increasing pathogen survival and growth. FV inactivation by the secreted E.coli protease may be part of a novel pathogenic virulence mechanism that deregulates the blood coagulation process to enhance bacterial infectivity and transmission. / UOIT

The success of surface water treatment strongly depends on the effectiveness of coagulant performance. Aluminium sulfate (alum), the most widely used coagulant in water treatment plants in Canada, is well known for its poor performance in cold water. Polyaluminium chloride (PACl), a relatively new polymeric aluminium coagulant increasingly being used in water treatment plants, is found to have many advantages over conventional alum. However, PACl hydrolysis reaction is quite complex and its action is not fully understood. In this research, a series of bench-scale jar tests with alum and PACl was conducted. Alum and PACl coagulation flocs were analyzed for the evaluation of coagulant performances at 19C and 5C for the Split Lake water treatment plant. The results of this research indicated that the settling properties of PACl flocs were superior to those of alum flocs, especially at the lower temperature. The average size of PACl flocs was relatively smaller than that of alum flocs. The density of PACl flocs could be higher than that of alum flocs. And the number of settled PACl flocs could be higher than that of settled alum flocs. The effects of temperature on alum flocs and PACl flocs were different. Alum flocs size decreased at 5C. This is most likely due to the existence of monomeric aluminium species in alum aqueous solution. PACl flocs size did not change significantly at the 5C. This may be due to the existence of polymeric aluminium species in PACl aqueous solution. / February 2006

Surgery precipitates a hypercoagulable state and has been shown to increase the development of cancer metastases in animal models, however mechanism(s) responsible for this are largely unknown. We hypothesize that the prometastatic effect of surgery may be secondary to postoperative hypercoagulable state. Surgical stress was induced in mice by partial hepatectomy or nephrectomy, preceded by intravenous injection of CT26-LacZ or B16F10-LacZ cells to establish pulmonary metastases with or without perioperative anticoagulation and their lung tumor cell emboli (TCE) were quantified. Fibrinogen and platelets were fluorescently labeled prior to surgical stress to evaluate TCE-associated fibrin and platelet clots. Surgery significantly increased metastases while anticoagulation with five different agents attenuated this effect. Fibrin and platelet clots were associated with TCE significantly more frequently in surgically stressed mice. Surgery promotes the formation of fibrin and platelet clots around TCE and this appears to be the mechanism for the increase in metastases seen following surgery.

La présence de cations de métaux de transition en grandes concentrations dans les eaux présente des risques pour la santé humaine et pour l'environnement. Une des méthodes les plus utilisées pour les éliminer est la coagulation-floculation. Il s'agit d'une étape essentielle dans la chaîne de traitement des eaux. Cependant, l'utilisation de cette méthode pour traiter une eau de faible turbidité est difficile. Le recours à l'ajout de montmorillonite est d'un grand intérêt dans ce cas, car cela permet d'intensifier le processus de coagulation-floculation. Afin d'améliorer ce processus, l'ajout d'un floculant, non polluant, non toxique et biodégradable, s'avère nécessaire, d'où l'utilisation du chitosane qui joue deux rôles essentiels: il favorise à la fois la floculation et la complexation des cations des métaux de transition. Le présent travail a pour objet d'étudier l'élimination de trois cations de métaux de transition, à savoir le cobalt, le nickel et le cuivre, par coagulation-floculation du système montmorillonite-chitosane. L'élimination des cations métalliques et de la turbidité de l'eau dépend de plusieurs facteurs, dont le pH et le rapport massique (chitosane/montmorillonite). De même, les concentrations de cations, de chitosane et de montmorillonite peuvent, elles aussi, avoir un effet sur l'élimination de ces cations. Étant donné la multitude des facteurs pouvant influencer l'élimination de ces cations par la montmorillonite et par le chitosane, ainsi que la coagulation-floculation du système montmorillonite-chitosane, l'utilisation d'un plan factoriel s'est avérée nécessaire pour surmonter cette difficulté et déterminer les conditions favorables à une élimination optimale des trois cations étudiés. Les résultats obtenus prouvent qu'il peut exister une synergie entre le chitosane et la montmorillonite dans l'élimination des cations métalliques. La montmorillonite, dont l'efficacité a augmenté après modification chimique, et le chitosane ont montré une affinité différente pour chacun des trois cations métalliques. Cette affinité augmente dans l'ordre suivant: CO²⁺ < Ni²⁺ < Cu²⁺ . Une meilleure coagulation-floculation ne donne pas forcément une meilleure élimination des cations métalliques. Ainsi, l'élimination des cations dans une solution contenant 1 à 3 (g/L) de montmorillonite augmente avec l'augmentation du pH entre 3 et 7, tandis que la turbidité minimale est obtenue à un pH = 5,4 et pour un rapport massique (chitosane/montmorillonite) =0,6%. Ces conditions jugées optimales dans le cadre du présent travail peuvent faire l'objet d'une étude ultérieure plus poussée, visant la mise en oeuvre d'une technologie de démétallisation des eaux. ______________________________________________________________________________ MOTS-CLÉS DE L’AUTEUR : Chitosane, Montmorillonite, Coagulation, Floculation, Cations métalliques, Synergie.

Jar tests are performed to simulate full-scale pre-treatment and particle removal processes. Operators typically conduct them in an effort to attempt alternative treatment doses and strategies without altering the performance of the full-scale drinking water treatment plant. However, information obtained from these tests must be evaluated judiciously, as they currently focus on reduction of specific water quality parameters (i.e., ultraviolet absorption at 254 nm (UV254) and turbidity), and measuring and understanding the significance of coagulant dose on floc size. Consideration of aggregate structure has been less explored due mainly to a lack of appropriate theories to describe the complex random floc structure. Improving the predictive capacity of bench-scale protocols commonly used for optimizing conventional chemical pre-treatment in full-scale drinking water treatment plants is required. Results from settling tests indicated that the production of larger and more settleable flocs could not be described by floc settling velocities and floc sizes. Settling velocities were not directly related to either UV254 or turbidity reductions. Results of the floc characterization tests indicated that measured values of UV254 and turbidity of the supernatant were generally inversely proportional to aggregate D90; that is, the residual UV254 and/or turbidity decreased as the value of D90 increased, which may have been indicative of flocculent settling. No direct relationship could be discerned between fractal dimension D1 (i.e., floc shape) and the UV254 and turbidity of the supernatant; however, the turbidity after flocculation and a period of settling appeared to be inversely proportional to fractal dimension D2 (i.e., porosity). Overall, the results of the experiments have demonstrated that grain size distributions and fractal dimensions might be used to assess and/or predict pre-treatment and/or particle removal performance. Specifically, the relationship between D90 values calculated from samples of flocculated water prior to settling and UV254 and turbidity values of that water after a period of settling may be a simple tool that can be utilized to describe and potentially better predict flocculent settling performance. At present, this appears to be the first such tool of its kind that has been reported.

Many things might happen in the body when a titanium implant is inserted into bone. Examples are activation of the immune system and imbalance between bone formation and bone resorption, which might lead to damaged bone around the implant and at worse, loosening of the implant. Bisphosphonates, BP’s, is a class of drugs that is able to decrease the osteoclast (bone resorption cell) activity and thereby strengthen the bone. FibMat2.0 is a fibrinogen matrix and consists of a thin protein layer which can be applied on an implant and act as a local drug delivery system. The work in this thesis was divided into two parts where aim of the first part was to study FibMat2.0 with integrated BP’s, and their effect in the presence of blood. The aim for the second part was to determine whether it was possible to incorporate antithrombotic drugs into the fibrinogen matrix. No detection method for the amount of drugs incorporated into the fibrinogen matrix was used but the fact that the drugs gave effect was verifying that it is possible to integrate other drugs than BP’s into FibMat2.0. Methods that have been used in the experiments in presence of blood are imaging of coagulation, fluorescence microscopy and cone-and-plate. For the first part, the results showed that surfaces incubated with fibrinogen and fibrinogen with integrated BP’s act alike in regard to coagulation and platelet adhesion. Compared to titanium, which is known to be a biocompatible material, the surfaces with fibrinogen and fibrinogen with BP’s behave similar in regard to platelet adhesion. When it comes to coagulation, the surfaces coated with fibrinogen with or without an addition of BP’s have shown a longer coagulation time compared to the clean titanium surface. For the second part, some conclusions have been drawn according to the results. Heparin and hirudin have shown anticoagulant effects when integrated in the matrix. The platelet inhibitor cangrelor seemed to have better effect when added in blood and incubated compared to incubation with the platelet inhibitor on the surface before incubation in blood. Finally, when combining heparin and cangrelor, very clear differences in regard to formation of fibrin network could be seen. It seems promising to be able to load different kind of drugs in FibMat2.0.

Introduction: It is recommended to place all the vacuum tubes directly on a sample cradle after vein puncture to prevent analytic error. This recommendation is not always easy to follow because the samples are taken by different professionals under different situations.  The three most common analyses, platelets count, haemoglobin and prothrombin time were tested.  Therefore, it was interesting to compare results from the three most common analyses with or without sample cradle, to evaluate the influence of this step on the result. Methods: Three analyses were preformed, using blood from 50 different persons. Each person gave two vacuum tubes, each contained 4.5mL of venous blood for the study. Tubes containing EDTA were used for platelet counts and measurement of haemoglobin and tubes containing citrate were used for prothrombin time-analysis. One of the tubes was placed, as recommended, directly on the sample cradle while the other tube was placed flat on a bench for 10 minutes before it was placed on the sample cradle. Results: There was a clear difference in platelet counts with and without immediate cradling but only minor difference between the results for haemoglobin and International Normalized Ratio. Conclusion: Some analyses seem to be more sensitive for variation in cradling than others. For platelet count it was important to immediately rock the tubes but for determination of prothrombine time and hemoglobin it had a small impact. The small impact on the results is probably due to the efficiency of the anticoagulant in the vacuum tubes.

Jar tests are performed to simulate full-scale pre-treatment and particle removal processes. Operators typically conduct them in an effort to attempt alternative treatment doses and strategies without altering the performance of the full-scale drinking water treatment plant. However, information obtained from these tests must be evaluated judiciously, as they currently focus on reduction of specific water quality parameters (i.e., ultraviolet absorption at 254 nm (UV254) and turbidity), and measuring and understanding the significance of coagulant dose on floc size. Consideration of aggregate structure has been less explored due mainly to a lack of appropriate theories to describe the complex random floc structure. Improving the predictive capacity of bench-scale protocols commonly used for optimizing conventional chemical pre-treatment in full-scale drinking water treatment plants is required. Results from settling tests indicated that the production of larger and more settleable flocs could not be described by floc settling velocities and floc sizes. Settling velocities were not directly related to either UV254 or turbidity reductions. Results of the floc characterization tests indicated that measured values of UV254 and turbidity of the supernatant were generally inversely proportional to aggregate D90; that is, the residual UV254 and/or turbidity decreased as the value of D90 increased, which may have been indicative of flocculent settling. No direct relationship could be discerned between fractal dimension D1 (i.e., floc shape) and the UV254 and turbidity of the supernatant; however, the turbidity after flocculation and a period of settling appeared to be inversely proportional to fractal dimension D2 (i.e., porosity). Overall, the results of the experiments have demonstrated that grain size distributions and fractal dimensions might be used to assess and/or predict pre-treatment and/or particle removal performance. Specifically, the relationship between D90 values calculated from samples of flocculated water prior to settling and UV254 and turbidity values of that water after a period of settling may be a simple tool that can be utilized to describe and potentially better predict flocculent settling performance. At present, this appears to be the first such tool of its kind that has been reported.

To achieve predictive capability for complex environmental systems with coagulation and arsenic sorption, a unified improved coagulation model coupled with arsenic sorption was developed. A unified coagulation model coupled with arsenic sorption was achieved by the following steps: (1) an improved discretized population balance equation (PBE) was developed to obtain the exact solution of conventional coagulation, (2) the improved PBE was extended to an adjustable geometric size interval having higher numerical stability, accuracy, and computational efficiency than existing models for fractal aggregate coagulation that includes agglomeration and fragmentation, (3) a surface complexation equilibrium model and a sorption kinetic model was introduced to predict arsenic sorption behavior onto hydrous metal oxide surfaces, and (4) an improved discretized PBE was coupled with arsenic sorption kinetics and equilibrium models by aid of collision efficiency ?? depending on surface charge (potential) on the hydrous metal oxide particles, colliding particle size ratio, and fluid strain-rate in applied flow system. The collision efficiency ?? into the improved (r,r)ij(r,r)ijdiscretized coagulation model for fractal aggregate yielded a unified improved coagulation model coupled with arsenic sorption kinetics and the equilibrium model. Thus, an improved unified coagulation model could provide high statistical accuracy, numerical stability, and computational efficiency to enhance predictive capability for behavior of arsenic sorption and fractal colloid particle aggregation and break-up, simultaneously. From the investigation, it is anticipated that the unified coagulation model coupled with arsenic sorption kinetics and equilibrium will provide a more complete understanding of the arsenic removal mechanism and its application to water/wastewater treatment. Further, this coupled model can be applied to other water and wastewater treatment systems combined with sorption and filtration processes. These combined processes can be optimized by the coupled model that was developed in this study. By simulating the arsenic sorption and particle size distribution as a pretreatment before filtration (sand filtration or membrane filtration), the overall arsenic removal efficiency and operation cost can be estimated.

Thesis (M.S.)--Oregon State University, 2010. / Printout. Includes bibliographical references (leaves 42-44). Also available on the World Wide Web.