Recirculating aquaculture offers a prospect for successful fish farming, but this form of aquaculture presents a great potential for pathogenic microorganisms to become established in the system through the formation of biofilms. Biofilms are capable of forming on all aquaculture system components, incorporating the various microflora present in the water. Pathogenic microorganisms released from the biofilms are capable of causing recurring exposure to disease in both fish and humans. With the increased consumption of raw and rare fish, the presence of these bacteria in or on the fish could lead to ingestion of pathogens. There is also the possibility of cross-contamination during processing. The objectives of this study were to increase the understanding of pathogen incorporation into biofilms in recirculating aquaculture systems and to determine the effectiveness of various sanitizers in eliminating biofilms. Seven freshwater and two saltwater facilities were sampled, with eight different types of materials tested. Pathogenic bacteria were identified using Bacteriological Analytical Manual methods and rapid commercial test kits. Most of the pathogenic bacteria identified were opportunistic organisms ubiquitous in an aquatic environment. The most significant human pathogens were Bacillus cereus, the Shigella species and the Vibrio species. The major piscine pathogens of concern were Photobacterium damsela, the Vibrio species, and Aeromonas hydrophila. The most significant variation in biofilm pathogens was observed between facilities and not construction material. Buna-N rubber, polyvinyl chloride (PVC), chlorinated PVC, glass, fiberglass and stainless steel disks were suspended in 79.2 liter (20 gallon) aquariums stocked with Nile tilapia (Oreochromus niloticus). The tanks were inoculated with a known amount of green fluorescent protein (GFP) modified Escherichia coli and samples were removed on days 1,3, 7 and 15. The modified E. coli were isolated on Luria Broth Agar and plate counts were performed under ultraviolet light. There was no significant difference in the growth of the surrogate pathogen on the different materials. The GFP E. coli was isolated in the largest numbers 24 hours after inoculation of the tanks, with an approximate 1-log decrease after day 1. Days 3, 7, and 15 showed equivalent growth of the target organism. Two sets of disks were suspended in another six 79.2 liter (20 gallon) aquariums. The tanks were inoculated with a known amount of the surrogate pathogen, GFP E. coli, and after 24 hours one set of disks was removed from each tank. The second set of disks was removed and treated by spraying with water, alkaline cleanser, sodium hypochlorite, quaternary ammonium compound, or peracetic acid. Ozone was bubbled directly into one tank to treat another set of disks. The modified E. coli were isolated and counted. Total aerobic plate counts and Enterobacteriaceae counts were performed. Statistical analysis indicated that the type of material had no significant affect on the effectiveness of the sanitizers. It was determined that sodium hypochlorite (99.4591 overall reduction) and peracetic acid (98.8461 % overall reduction) were the most effective sanitizers overall, and ozone (32.9332 % overall reduction) was the least effective. / Ph. D.

The immune defense is geared to protect against a tremendous array of invaders. The ultimate goal of the immune system is to induce effective and balanced inflammatory responses that enable the efficient elimination of possible threats while avoiding both immunodeficiency and autoimmunity. The skewness towards inflammatory responses causing excessive collateral damage could lead to diverse autoimmune conditions. These conditions could be organ-specific or result from systemic immune dysregulations called systemic autoimmunity. The multifaceted nature and the intricate clinical heterogeneity of systemic autoimmune conditions indicate a strong influence of environmental factors on their immunopathogenesis, where environmental factors could either hinder or contribute to autoimmune development. We focused our research on deciphering the complex effects of environmental factors on the immunopathogenesis of systemic immune dysregulation, taking systemic lupus erythematosus (SLE or Lupus) as a model of systemic autoimmunity. SLE is one of the most mysterious autoimmune disorders with no known cure. In SLE, breaching of tolerance to self-antigens and the subsequent persistent inflammation and collateral tissue damage in multiple organs lead to very diverse clinical manifestations. These manifestations are a result from the interplay between multiple genetic susceptibilities and diverse environmental factors. To date, management plans for SLE are based on non-selective immunosuppressants that could impose significant side effects including increased risks of infection and infection-related mortalities. In parallel, environmental factors and the quality of life could significantly impact SLE management strategies. Therefore, delineating the immunomodulatory capacities of environmental factors would likely unravel more effective management strategies for SLE patients. The current research aims to investigate the central hypothesis that dietary and hygienic components modulate the immune dysregulations of SLE in a tissue- and disease stage-specific manner. We have focused on uncovering the complex effects of Vitamin A (VA) as an essential micronutrient with very diverse immunomodulatory capacities, and quaternary ammonium compound (QAC)-based disinfectants as ubiquitously used disinfectants that have been linked to immunotoxicity, on the immunopathogenesis of SLE. Due to the strong female bias of SLE where women especially of childbearing age are more prone to lupus, we have focused our research on delineating how these diverse factors shape the immunopathogenesis of SLE in female mice only. The first project dissected the immunomodulatory effects of VA, a potent immunomodulatory dietary component. Notably, VA exerts its function through a predominant metabolite known as all-trans-retinoic acid (tRA) that, as we have previously shown, has paradoxical and tissue-specific implications on lupus inflammation. Here, we utilized a pristane-induced model of lupus to investigate the disease stage-dependent effects of tRA. Oral supplementation of tRA was given either before pristane induction of lupus from weaning (3 weeks) to 3 months of age or after pristane induction of lupus from 3 to 9 months of age. We found that tRA treatment mediated disease stage-dependent effects and differentially affected the lupus-associated kidney inflammation (lupus nephritis) when given at the initiation vs. continuation phase of lupus. Unlike tRA treatment during active disease, pre-pristane treatment with tRA aggravated glomerulonephritis through potentiating leukocyte activation and trafficking to the kidney and augmenting renal pro-fibrotic signals. Post-pristane tRA treatment, on the other hand, exerted immunosuppressive functions of decreasing circulatory and renal deposition of autoantibodies as well as suppressing the renal expression of proinflammatory cytokines and chemokines. Interestingly, both pre- and post-pristane treatments with tRA reversed the pristane-induced leaky gut and similarly modulated the gut microbiota, suggesting a gut microbiota-independent mechanism by which tRA affects the initiation vs. continuation phase of lupus. As tRA could be protective against lupus nephritis especially during the active disease stage, and previous reports had shown hypovitaminosis A (reduced serum retinol levels) proceeding SLE, we expanded our investigation to decipher whether VA deficiency (VAD) was a contributing factor for severe SLE and to delineate how VAD affected the initiation and/or the progression of lupus nephritis in genetically-prone conditions. For that purpose, we utilized the classical murine lupus-prone model, MRL/lpr, and initiated VAD either during the gestation or after weaning to reveal potential time-dependent effects. VAD exacerbated lupus nephritis by provoking severe neutrophilic tubulointerstitial nephritis, and accelerated renal failure. This was concomitant with significantly higher mortality in all VAD mice. Mechanistically, VAD enhanced early activation of plasma cells and augmented their autoantibodies production. In addition, VAD led to an enhanced expansion of pathogenic T lymphocytes. In parallel, VAD increased renal infiltration of conventional and plasmacytoid dendritic cells. Our findings establish VAD as a driving factor for lupus nephritis progression in genetically predisposed conditions. These findings emphasize the importance of monitoring VA levels in SLE patients and urge for VA supplementations for patients at higher risk for hypovitaminosis A, especially during the maternal-neonatal interface. Additionally, this project warrants further investigations to delineate the molecular targets through which VA modulates cellular functions as well as immunopathogenesis of lupus nephritis. The information obtained from these studies may also benefit women with other autoimmune conditions and will pave the way for VA supplementations to be tested in clinical trials. The second project investigated the effects of ambient exposure to QAC-based disinfectants on the progression of murine SLE in genetically prone mice. We compared the disease progression in MRL/Lpr mice that have been exposed to QACs vs. those kept under a complete QAC-free condition. QAC-based disinfectants CP-64 or Labsan 256 were used under QAC-exposed conditions, while ethanol was used in the QAC-free environment. We found that compared to QAC-free mice, ambient exposure of lupus-prone mice to QACs led to smaller spleens with no change in circulating autoantibodies or the severity of glomerulonephritis. This suggests that QACs may have immunosuppressive effects on lupus. Using a microfluidic device, we showed that ambient exposure to QACs reduced directional migration of bone marrow-derived neutrophils toward an inflammatory chemoattractant ex vivo. Consistent with this, we found decreased infiltration of neutrophils into the spleen. While bone marrow-derived neutrophils appeared to exhibit a pro-inflammatory profile, upregulated expression of PD-L1 was observed on neutrophils that infiltrated the spleen, which in turn interacted with PD-1 on T cells and modulated their fate. Specifically, QAC exposure hindered activation of splenic T cells and increased apoptosis of effector T-cell populations. Collectively, these results suggest that ambient QAC exposure decreases lupus-associated splenomegaly likely through neutrophil-mediated toning of T-cell activation and/or apoptosis. However, our findings also indicate that even ambient exposure could alter immune cell phenotypes, functions, and their fate. Further investigations on how QACs affect immunity under steady-state conditions are warranted. Collectively, the findings of this doctoral research suggest temporal and spatial effects of diet and hygiene on systemic autoimmunity and emphasize the strong influence of environmental factors toning cellular immune responses and subsequently shaping autoimmune outcomes. Our findings could pave the way for more personalized healthcare plans for autoimmune patients that take into consideration tissue involvement, disease stages, and the patient's lifestyle. / Doctor of Philosophy / The immune system is efficiently toned to discriminate between friends and foes. It effectively protects against a wide array of pathogens while at the same time avoiding attacking self-tissues. The inability of immune defenses to achieve this optimal discrimination could lead to the breakdown of tolerance to self in a wide range of autoimmune conditions. Diverse genetic susceptibilities are implicated in the development of autoimmunity. In parallel, during the recent decades, the tremendous increase in the prevalence of autoimmune conditions coincides with evolving dietary and hygiene styles in Westernized societies. This suggests a strong influence of environmental factors such as dietary and hygienic components on the way that the immune system works. Therefore, the current research investigates whether diet and hygiene modulate the immune dysregulations of lupus disease as a model for systemic autoimmunity; and if so, whether such effects are tissue- and/or disease stage-specific. We utilized different mouse models to delineate the mechanisms by which essential nutrients such as vitamin A (VA) and widely used disinfectant compounds known as quaternary ammonium disinfectants (QACs) modulate the systemic autoimmunity in lupus disease. We found that these modulators influence various aspects of the cellular immune responses including (1) leukocyte activation and subsequent expansion of pathogenic (disease contributing) lymphocytes, production of antibodies directed against self-tissue molecules (i.e., autoantibodies), and production of inflammatory mediators (i.e., cytokines and chemokines); (2) cell trafficking and their infiltration into the tissues; (3) signal transduction pathways that modulate cell fate (e.g., PD-1: PD-L1 signaling). Importantly, environmental modulation of autoimmunity during different stages of autoimmune development could significantly impact the disease outcome. VA treatment, for example, differentially modulates the progression of kidney inflammation when given during the initiation vs. progressive disease stages. Similarly, VA deficiency has the most prominent effects on worsening kidney inflammation under genetically prone conditions when the deficiency is initiated early and at the prenatal stage. In parallel, the effects of environmental factors are also tissue-specific. For example, ambient exposure to QAC-based disinfectants exerted immunosuppressive effects on lupus-associated inflammation of lymphoid tissues with no change in circulating autoantibodies or the severity of kidney inflammation. Collectively, the findings of this doctoral research delineated the cellular mechanisms through which environmental factors could shape autoimmune responses. Further studies will dig into the underlying molecular pathways. Ultimately, our research emphasizes the strong influence of exogenous factors on immunity and will pave the way for more effective healthcare management plans and benefit vulnerable populations affected by autoimmune conditions such as lupus.

Quaternary ammonium lipids containing long hydrocarbon-chains can cause deleterious effects when administered to many types of microorganisms. Some compounds have potent activity against spermatozoa and the HIV virus. Consequently, quaternary ammonium lipids are key components in spermicidal and antiseptic formulations. Surface-active quaternary ammonium lipids, containing a hydrophobic and hydrophilic moiety, will aggregate into micelles when dissolved in water above a narrow concentration range, termed the critical micelle concentration (CMC). This project investigates how the CMC relates to spermicidal and microbicidal properties. The CMC’s of the AD-nX and AT-nX compounds [see documents for figures] were determined by conductivity measurements. Relationships among the CMC and factors such as alkyl-chain length, head-group size, counteranion, and biological activity are discussed. [See document for figures of ADn and ATn] / Master of Science

Compostos de amônio quaternário (QACs) têm sido amplamente utilizados como desinfetantes e antissépticos, sendo essenciais na prevenção e controle de infecções bacterianas na medicina humana e veterinária. Embora patógenos prioritários multirresistentes têm sido muito bem caracterizados quanto ao perfil de suscetibilidade e contexto genético da resistência aos antibióticos, dados de resistência aos QACs são limitados. Assim, o objetivo do presente estudo foi avaliar a atividade in vitro dos QACs de uso doméstico e hospitalar [cloreto de benzalcônio (BAC), cloreto de cetilpiridinio (CPC) e brometo de cetiltrimetilamônio (CTAB)], contra patógenos prioritários multirresistentes, identificando os principais genes de resistência associados. Foram estudadas 100 cepas multirresistentes previamente sequenciados usando as plataformas Illumina MiSeq e NextSeq representativas de diferentes hospedeiros (humanos e animais) e fontes (ambientes e alimentos). As cepas foram identificadas como Klebsiella pneumoniae (n= 24), Escherichia coli (n= 30); Pseudomonas aeruginosa (n= 10), Enterobacter spp, (n= 8), Acinetobacter baumannii (n= 11) e Salmonella spp. (n= 17). Genes de resistência aos QACs foram identificados in silico através do alinhamento dos contigs obtidos de cada cepa sequenciada com genes de referência obtidos do GenBank, utilizando o programa Geneious versão 8 (Biomatters Ltd). A identidade de cada gene foi analisada utilizando o programa BLASTx, no qual um critério baseado em ≥90% identidade resultou na identificação dos genes mdfA (77%), qacE (44%), qacEΔ1 (43%), sugE(c) (29%), emrE (21%), qacA (19%), sugE(p) (5%), qacF (7%), qacH (7%) e qacL (7%) em 85 cepas; enquanto que 15 cepas não possuíam nenhum gene de resistência aos QACs. A concentração inibitória mínima (CIM) dos QACs para as 100 cepas foi determinada pelo método de microdiluição em caldo. Os resultados sugeriram que a resistência em patógenos prioritários circulando na interface humano-ambiente-animal não é restrita aos antibióticos, uma vez que a elevada ocorrência de genes qacE, qacEΔ1 e mdfA poderia estar associada com uma redução da suscetibilidade para QACs. Consequentemente, a resistência aos QACs poderia também contribuir para a persistência e adaptação destes patógenos nos seres humanos e outros animais, assim como em ambientes impactados antropogenicamente. / Quaternary ammonium compounds (QACs) have been widely used as disinfectants and antiseptics, being applied as essential compounds in the prevention and control of bacterial infections in human-and veterinary hospital medicine. Although multiresistant priority pathogens have been well characterized with respect to their susceptibility profile and their genetic context of resistance for antibiotics, studies of resistance to QACs are limited. Thus, the objective of the present study was to evaluate the in vitro activity of QACs [(benzalkonium chloride (BAC), cetylpyridinium chloride (CPC) and cetyltrimethylammonium bromide (CTAB)] for household and hospital use against multiresistant priority pathogens, identifying the main resistance genes associated. A hundred multiresistant isolates (previously sequenced using the Illumina MiSeq and NextSeq platforms), representative of different hosts (humans and animals) and sources (environment and food) were studied. Isolates were identified as Klebsiella pneumoniae (n=24), Escherichia coli (n=30), Pseudomonas aeruginosa (n=10), Enterobacter spp. (n=8), Acinetobacter baumannii (n=11) and Salmonella spp. (n=17). In silico analysis for identification of genes conferring resistance to QACs were performed by aligning the contigs obtained from the strains with reference genes deposited in GenBank, using the Geneious version program (Biomatters Ltd). Similarities were analyzed using the BLASTx online program, considering the alignment criteria based on ≥ 90% identity. The result of these analysis revealed the presence of the following QAC genes: mdfA (77%), qacE (44%), qacEΔ1 (43%), sugE(c) (29%), emrE (21%), qacA (19%), sugE (p) (5%), qacF (7%), qacH (7%) e qacL (7%); while 15 strains showed no resistance genes for QACs. Determination of QACs minimum inhibitory concentration (MIC) for the 100 isolates, by the broth microdilution method. These results suggest that resistance to QACs in priority pathogens, circulating at the human-environment-animal interface, is not restricted to antibiotics, since the high occurrence of genes qacE, qacEΔ1 and mdfA were associated with a reduced susceptibility to QACs. Consequently, resistance to QACs could also contribute to the persistence and adaptation of these pathogens in humans and othes animals, as well as in anthropogenically impacted environments.

Engineered organoclays are 2:1 phyllosilicate soils that have been synthesized with a controlled interlayer organic phase to exhibit enhanced strength, lower compressibility, and stronger retention of organic compounds. Engineered organoclays are highly sorptive, and have a variety of potential engineering applications as sorbents or amendments in engineered earthen barrier systems. Previous studies examined the impact of the organic coating on a soil's physical properties; however, the geochemical behaviors of organoclays, especially their interaction with organic compounds at the micro-scale, remained relatively unquantified. This study investigated the engineering behavior of montmorillonite modified with a variety of quaternary ammonium cations (QAC clays) with controlled structure and density of loading. Molecular dynamics simulations were used to model the surfactant arrangement, geochemical processes in the QAC-clay interlayer, including organic compound sorption and mass transport, as well as the surface electrokinetics of suspended QAC-clay particles. All simulations were carried out based on the combined force field of ClayFF and the Consistent-Valence Force Field to ensure the accuracy of the simulation results, and results yielded insight into the prediction of synthesized QAC-clay behaviors as sorptive material for non-polar organic compounds.

Alguns Agentes Farmacêuticos (AF) sólidos podem ser otimizados através de ajustes nas propriedades físicas, permitindo maior controle na solubilidade, estabilidade, biodisponibilidade e farmacocinética. Líquidos iônicos, sais que se apresentam líquidos à temperaturas inferiores a 100º C, representam uma classe de substâncias utilizadas como possível estratégia no planejamento e otimização de fármacos, através da escolha de íons ativos biologicamente. Neste trabalho foram investigadas interações entre pares iônicos formados pelos ânions acetilsalicilato (ASP) e ibuprofenato (IBU), em combinação com os cátions biologicamente ativos, benzil-decil-dimetil-amônio (BDDA) e didecil-dimetil-amônio (DDA) com a membrana biológica formada por palmitoil-oleil-fosfatidil-colina (POPC), pelo método de Dinâmica Molecular. A partir de otimizações geométricas e distribuições de cargas, estabeleceram-se topologias para os íons, definindo-se parâmetros ausentes no campo de força AMBER. Inicialmente simularam-se os pares iônicos solvatados e também sistemas contendo líquidos iônicos puros. Essas simulações indicaram adequada distribuição de cargas para os íons e forneceram informações a respeito da estrutura do líquido, formado por pares iônicos, como densidade e distâncias entre grupos de átomos. As simulações contendo sistemas completos (íons, POPC e água) indicaram que no sistema IBUDDA o par iônico é mantido, enquanto que nos outros sistemas ocorrem maiores variações nas distâncias entre os íons. O sistema ASPDDA apresenta indícios de fuga do ânion através da bicamada. Nos sistemas ASPBDDA e IBUBDDA, apesar da variação das distâncias mínimas, os íons mantêm-se no interior da membrana no tempo simulado. Nos sistemas IBUDDA, IBUBDA e ASPBDDA o grupo carboxilato dos ânions demonstra proximidade e preferência pelo grupo colina da POPC, enquanto que o grupo contendo o nitrogênio do cátion aproxima-se preferencialmente do grupo fosfato da POPC. / Some pharmaceutical agents (AF) solids may be optimized through adjustments in physical properties, allowing greater control on the solubility, stability, bioavailability and pharmacokinetics. Ionic Liquids, salts which are liquids at temperatures lower than 100 ° C, represent a class of substances used as a possible strategy in the design and optimization of drugs through the choice of biologically active ions. In this study were investigated interactions between ion pairs formed by the acetylsalicylate (ASP) and ibuprofenate (IBU) anions, in combination with the biologically active cations benzalkonium (BDDA) and didecyldimethylammonium (DDA) with the biological membrane palmitoyloleylphosphatidylcholine (POPC) by molecular dynamics method. From geometric optimizations and charge distributions, have established topologies for the ions, defining missing parameters in the AMBER force field. The ion pairs were simulated under vacuum and solvated as well as systems containing pure Ionic Liquids. These simulations showed suitable charge distribution for ions and provided information about the structure of the liquid formed by ion pairs, such as density and distances between groups of atoms. The simulations containing entire systems (ions, water and POPC) indicated that in the system IBUDDA the ion pair is maintained while in other systems transients distancing occur between ions. The system containing ASPDDA indicates leakage of the anion through the bilayer. In systems ASPBDDA, IBUDDA and IBUBDDA, despite the variation of minimum distances, the ions remain within the membrane in simulated time. In IBUDDA, IBUBDDA and ASPBDDA the carboxylate group of the anions demonstrate nearness and preference for POPC choline group, whereas the group containing the nitrogen cation preferentially approximates of the POPC phosphate group.

Alguns Agentes Farmacêuticos (AF) sólidos podem ser otimizados através de ajustes nas propriedades físicas, permitindo maior controle na solubilidade, estabilidade, biodisponibilidade e farmacocinética. Líquidos iônicos, sais que se apresentam líquidos à temperaturas inferiores a 100º C, representam uma classe de substâncias utilizadas como possível estratégia no planejamento e otimização de fármacos, através da escolha de íons ativos biologicamente. Neste trabalho foram investigadas interações entre pares iônicos formados pelos ânions acetilsalicilato (ASP) e ibuprofenato (IBU), em combinação com os cátions biologicamente ativos, benzil-decil-dimetil-amônio (BDDA) e didecil-dimetil-amônio (DDA) com a membrana biológica formada por palmitoil-oleil-fosfatidil-colina (POPC), pelo método de Dinâmica Molecular. A partir de otimizações geométricas e distribuições de cargas, estabeleceram-se topologias para os íons, definindo-se parâmetros ausentes no campo de força AMBER. Inicialmente simularam-se os pares iônicos solvatados e também sistemas contendo líquidos iônicos puros. Essas simulações indicaram adequada distribuição de cargas para os íons e forneceram informações a respeito da estrutura do líquido, formado por pares iônicos, como densidade e distâncias entre grupos de átomos. As simulações contendo sistemas completos (íons, POPC e água) indicaram que no sistema IBUDDA o par iônico é mantido, enquanto que nos outros sistemas ocorrem maiores variações nas distâncias entre os íons. O sistema ASPDDA apresenta indícios de fuga do ânion através da bicamada. Nos sistemas ASPBDDA e IBUBDDA, apesar da variação das distâncias mínimas, os íons mantêm-se no interior da membrana no tempo simulado. Nos sistemas IBUDDA, IBUBDA e ASPBDDA o grupo carboxilato dos ânions demonstra proximidade e preferência pelo grupo colina da POPC, enquanto que o grupo contendo o nitrogênio do cátion aproxima-se preferencialmente do grupo fosfato da POPC. / Some pharmaceutical agents (AF) solids may be optimized through adjustments in physical properties, allowing greater control on the solubility, stability, bioavailability and pharmacokinetics. Ionic Liquids, salts which are liquids at temperatures lower than 100 ° C, represent a class of substances used as a possible strategy in the design and optimization of drugs through the choice of biologically active ions. In this study were investigated interactions between ion pairs formed by the acetylsalicylate (ASP) and ibuprofenate (IBU) anions, in combination with the biologically active cations benzalkonium (BDDA) and didecyldimethylammonium (DDA) with the biological membrane palmitoyloleylphosphatidylcholine (POPC) by molecular dynamics method. From geometric optimizations and charge distributions, have established topologies for the ions, defining missing parameters in the AMBER force field. The ion pairs were simulated under vacuum and solvated as well as systems containing pure Ionic Liquids. These simulations showed suitable charge distribution for ions and provided information about the structure of the liquid formed by ion pairs, such as density and distances between groups of atoms. The simulations containing entire systems (ions, water and POPC) indicated that in the system IBUDDA the ion pair is maintained while in other systems transients distancing occur between ions. The system containing ASPDDA indicates leakage of the anion through the bilayer. In systems ASPBDDA, IBUDDA and IBUBDDA, despite the variation of minimum distances, the ions remain within the membrane in simulated time. In IBUDDA, IBUBDDA and ASPBDDA the carboxylate group of the anions demonstrate nearness and preference for POPC choline group, whereas the group containing the nitrogen cation preferentially approximates of the POPC phosphate group.

Alguns Agentes Farmacêuticos (AF) sólidos podem ser otimizados através de ajustes nas propriedades físicas, permitindo maior controle na solubilidade, estabilidade, biodisponibilidade e farmacocinética. Líquidos iônicos, sais que se apresentam líquidos à temperaturas inferiores a 100º C, representam uma classe de substâncias utilizadas como possível estratégia no planejamento e otimização de fármacos, através da escolha de íons ativos biologicamente. Neste trabalho foram investigadas interações entre pares iônicos formados pelos ânions acetilsalicilato (ASP) e ibuprofenato (IBU), em combinação com os cátions biologicamente ativos, benzil-decil-dimetil-amônio (BDDA) e didecil-dimetil-amônio (DDA) com a membrana biológica formada por palmitoil-oleil-fosfatidil-colina (POPC), pelo método de Dinâmica Molecular. A partir de otimizações geométricas e distribuições de cargas, estabeleceram-se topologias para os íons, definindo-se parâmetros ausentes no campo de força AMBER. Inicialmente simularam-se os pares iônicos solvatados e também sistemas contendo líquidos iônicos puros. Essas simulações indicaram adequada distribuição de cargas para os íons e forneceram informações a respeito da estrutura do líquido, formado por pares iônicos, como densidade e distâncias entre grupos de átomos. As simulações contendo sistemas completos (íons, POPC e água) indicaram que no sistema IBUDDA o par iônico é mantido, enquanto que nos outros sistemas ocorrem maiores variações nas distâncias entre os íons. O sistema ASPDDA apresenta indícios de fuga do ânion através da bicamada. Nos sistemas ASPBDDA e IBUBDDA, apesar da variação das distâncias mínimas, os íons mantêm-se no interior da membrana no tempo simulado. Nos sistemas IBUDDA, IBUBDA e ASPBDDA o grupo carboxilato dos ânions demonstra proximidade e preferência pelo grupo colina da POPC, enquanto que o grupo contendo o nitrogênio do cátion aproxima-se preferencialmente do grupo fosfato da POPC. / Some pharmaceutical agents (AF) solids may be optimized through adjustments in physical properties, allowing greater control on the solubility, stability, bioavailability and pharmacokinetics. Ionic Liquids, salts which are liquids at temperatures lower than 100 ° C, represent a class of substances used as a possible strategy in the design and optimization of drugs through the choice of biologically active ions. In this study were investigated interactions between ion pairs formed by the acetylsalicylate (ASP) and ibuprofenate (IBU) anions, in combination with the biologically active cations benzalkonium (BDDA) and didecyldimethylammonium (DDA) with the biological membrane palmitoyloleylphosphatidylcholine (POPC) by molecular dynamics method. From geometric optimizations and charge distributions, have established topologies for the ions, defining missing parameters in the AMBER force field. The ion pairs were simulated under vacuum and solvated as well as systems containing pure Ionic Liquids. These simulations showed suitable charge distribution for ions and provided information about the structure of the liquid formed by ion pairs, such as density and distances between groups of atoms. The simulations containing entire systems (ions, water and POPC) indicated that in the system IBUDDA the ion pair is maintained while in other systems transients distancing occur between ions. The system containing ASPDDA indicates leakage of the anion through the bilayer. In systems ASPBDDA, IBUDDA and IBUBDDA, despite the variation of minimum distances, the ions remain within the membrane in simulated time. In IBUDDA, IBUBDDA and ASPBDDA the carboxylate group of the anions demonstrate nearness and preference for POPC choline group, whereas the group containing the nitrogen cation preferentially approximates of the POPC phosphate group.

Submitted by Ubirajara Cruz (ubirajara.cruz@gmail.com) on 2016-09-23T14:52:44Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_francine_souza.pdf: 890441 bytes, checksum: 8e77c3780d75f3009137594794500fc2 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2016-09-23T18:43:00Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_francine_souza.pdf: 890441 bytes, checksum: 8e77c3780d75f3009137594794500fc2 (MD5) / Made available in DSpace on 2016-09-23T18:43:00Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_francine_souza.pdf: 890441 bytes, checksum: 8e77c3780d75f3009137594794500fc2 (MD5) Previous issue date: 2014-02-21 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / O uso de produtos naturais com fins medicinais, para tratamento, cura e prevenção de doenças é uma das formas mais antigas de prática medicinal da humanidade. A própolis é uma substância resinosa natural, produzida por abelhas melíferas a partir de exsudatos coletados em diferentes partes das plantas, que vem sendo utilizada desde a antiguidade na medicina popular, tanto humana quanto veterinária, devido as suas propriedades terapêuticas. A atividade antimicrobiana é relatada em diversos estudos que utilizam como modelo experimental diferentes gêneros de vírus, bactérias, fungos e parasitas, porém são raros os estudos utilizando-a como desinfetante. Avaliou-se um extrato etanólico de própolis verde (EEPV), em comparação com um desinfetante comercial à base de amônia quaternária, quanto à capacidade desinfetante in situ em bebedouros utilizados para frangos de corte até 28 dias de idade das aves. O EEPV também foi avaliado, in vitro, contra as bactérias isoladas a partir de suabes dos bebedouros utilizados no experimento. Aos 28 dias de idade dos frangos, quando o desafio microbiano foi maior, a ação do EEPV como desinfetante foi semelhante à ação do desinfetante comercial a base de amônia quaternária. Nesta mesma idade das aves, o EEPV, assim como o desinfetante comercial, inibiu por completo o crescimento fúngico nos bebedouros. O EEPV apresentou ação in vitro contra as bactérias gram positivas Staphylococcus sp. coagulase negativa, S. aureus, Corynebacterium sp., isoladas a partir de suabe coletado dos bebedouros, além da bactéria gram negativa Escherichia coli. / The use of natural products for medicinal purposes for treatment, cure and prevention of diseases is one of the most antique forms of medical practice of humanity. Propolis is a natural resinous substance produced by honeybees from exudates collected of different parts of the plant and has been used as a therapeutic compound since antiquity in the popular medicine both human and veterinary due to its therapeutics properties. The antimicrobial activity is mentioned in various studies applying different genus of viruses, fungus, bacterias and parasites as a experimental model. An ethanolic extract from green propolis was evaluated as to the sanitizer capacity in situ over drinking fountains used for broilers in comparison with a commercial disinfectant based on quaternary ammonia. The extract was also evaluated in vitro against bacterias isolated from the drinking fountains used in the experiment. At 28 days of broiler age when the microbial challenge was greater the action of the extract as a disinfectant was similar to the action of the commercial disinfectant based on quaternary ammonium. In the same age of the birds both the propolis extract as well as the commercial disinfectant the fungal growth was completely inhibited in the drinking fountains. The extract presented action in vitro against gram positive bacteria Staphylococcus sp. negative coagulase, S. aureus, Corynebacterium sp. isolated from swabs collected from the drinking fountains and also the gram negative bacteria Escherichia coli.

Leather wastewater is one of the most polluting industrial emissions. An in-situ, green, and innovative strategy that limits dye emissions is required to replace subsequent waste management. A novel cationic protein with a high quaternary ammonium degree was designed and synthesized. The results show that at concentrations ranging from 3 to 15 wt%, this cationic protein rapidly and completely adsorbs Direct Purple N and Acid Black 24 within 5 min. A remarkable efficiency in removing Acid Red 73, Acid Golden G, Acid Lake Blue A, Acid Green, and Acid Orange II, with >96% removal rates, was achieved. The cationic protein was most accurately represented by the pseudo-second-order kinetic model. Acid Orange II (2000 mg L-1) and 15 wt% cationic protein were used in an actual tanning process. The residual concentration of Acid Orange II in the wastewater was 23.1 mg L-1. These results reflect that the emission reduction targets have been effectively achieved.