37.3 million Americans (11.2% of the US population) currently have Type 2 diabetes mellitus (T2DM) with over 1.5 million new cases being diagnosed each year. The multifactorial etiology of the patient having neuropathy, overweight/obesity, foot deformities, ischemia, and infection leads to a condition called diabetic foot ulcer (DFU). One in six patients with a DFU will require amputation with infected DFUs have a 155-fold increased risk of amputation. Staphylococcus aureus is the most common bacteria isolated from severe DFU infections that require amputation. Interestingly, diabetics are more heavily colonized with S. aureus compared to non-diabetics suggesting a unique advantageous adaptation to diabetes. The specifics of the underlying molecular mechanisms and triggers by which S. aureus adapts and thrives in the T2DM patient that increase its pathogenicity and colonization compared to non-diabetics with skin ulcer infections are not fully elucidated. Thus, our studies aimed to identify the key virulence components in the pathogenesis of S. aureus infected DFUs and using that information to develop therapeutics aimed at disrupting these components to increase the success rate of conservative treatment and prevent non-traumatic lower extremity amputations in T2DM patients. Our studies found that several different elevated sugars in T2DM patients can trigger virulence factor production in S. aureus. We also found by comparing several different clinical DFU S. aureus isolates that there are clear differences in the ability of each isolate to cause necrotic infections. And lastly, we identified a possible therapeutic, the amino acid L-arginine, that can help prevent/treat S. aureus infections in the Tallyho diabetic mouse model. In conclusion, we have increased the understanding of the pathogenesis of S. aureus infected DFU and have proposed a possible therapeutic to add to the conservative treatment regimen.

No description available.

The Hybrid Norwood (HN) is a relatively new first stage procedure for neonates with Hypoplastic Left Heart Syndrome (HLHS), in which a sustainable univentricular circulation is established in a less invasive manner than with the standard procedure. A computational multiscale model of such HLHS circulation following the HN procedure was used to obtain detailed hemodynamics. Implementation of a reverse-BT shunt (RBTS), a synthetic bypass from the main pulmonary to the innominate artery placed to counteract aortic arch stenosis, and its effects on local and global hemodynamics were studied. A synthetic and a 3D reconstructed, patient derived anatomy after the HN procedure were utilized, with varying degrees of distal arch obstruction, or stenosis, (nominal and 90% reduction in lumen) and varying RBTS diameters (3.0, 3.5, 4.0 mm). A closed lumped parameter model (LPM) for the peripheral or distal circulation coupled to a 3D Computational Fluid Dynamics (CFD) model that allows detailed description of the local hemodynamics was created for each anatomy. The implementation of the RBTS in any of the chosen diameters under severe stenosis resulted in a restoration of arterial perfusion to near-nominal levels. Shunt flow velocity, vorticity, and overall wall shear stress levels are inverse functions of shunt diameter, while shunt perfusion and systemic oxygen delivery correlates positively with diameter. No correlation of shunt diameter with helicity was recorded. In the setting of the hybrid Norwood circulation, our results suggest: (1) the 4.0mm RBTS may be more thrombogenic when implemented in the absence of severe arch stenosis and (2) the 3.0mm and 3.5mm RBTS may be a more suitable alternative, with preference to the latter since it provides similar hemodynamics at lower levels of wall shear stress.

<p>Nonsteroidal anti-inflammatory drugs are among the most commonly used prescription and over-the-counter medications, but they often produce significant gastrointestinal ulceration and bleeding, particularly in elderly patients and patients with certain co-morbidities. Novel anti-inflammatory drugs are seldom tested in animal models that mimic the high-risk human users, leading to an underestimate of the true toxicity of these drugs. In the present study we examined the effects of two novel NSAIDs and two commonly used NSAIDs in models in which mucosal defence was expected to be impaired. Naproxen, celecoxib, ATB-346 (a hydrogen sulfide- and naproxen-releasing compound) and NCX 429 (a nitric oxide- and naproxen-releasing compound) were evaluated in healthy, arthritic, obese, hypertensive rats, and in rats of advanced age (19 months) and rats co-administered low-dose aspirin and/or omeprazole. In all models except hypertension, greater gastric and/or intestinal damage was observed when naproxen was administered in these models than in healthy rats. Celecoxib-induced damage was significantly increased when co-administered with low-dose aspirin and/or omeprazole. In contrast, ATB-346 and NCX 429, when tested at doses that were as effective as naproxen and celecoxib in reducing inflammation and inhibiting cyclooxygenase activity, did not produce significant gastric or intestinal damage in any of the models. These results demonstrate that animal models of human co-morbidities display the same increased susceptibility to NSAID-induced gastrointestinal damage as observed in humans. Moreover, two novel NSAIDs that release mediators of mucosal defence (hydrogen sulfide and nitric oxide) do not induce significant gastrointestinal damage in these models of impaired mucosal defence.</p> / Master of Science (MSc)

Homeodynamic space (HDS) shrinks as vulnerability increases with aging and repeated damage to the cells. HDS is lost in alcoholic pancreatitis patients due to overconsumption of alcohol, smoking, and high fat diets. Etiologically relevant animal models for study of chronic pancreatitis (CP) are needed. In order to begin filling this gap a central purpose of this dissertation research was to examine relationships between the alcohol and high fat diet (AHF) and pancreatitis with attention to hypersensitivity and anxiety-like behaviors. The AHF diet induced pancreatitis described here etiologically mimics human risk factors of AHF consumption for advancement to alcoholic CP. In this study one group of mice was fed long term with a diet of high fat and alcohol for comparison with a group fed normal chow. Mice consumed a liquid diet containing 6% alcohol and a high fat supplement ad libitum over a period of five months. Each group was evaluated for heat and mechanical hypersensitivity, and histology indicative of CP. The association of pancreatitis pathology with anxiety has been understudied. Anxiety, like pain, is useful as a transient state but when anxiety is prolonged it is termed a disorder. Anxiety is often comorbid with pain and depression. Therefore, it is important to determine anxiety in mice with CP histology. This model was characterized for the interaction of pancreatitis histology, as well as persisting pain-, anxiety-, and fear-like behaviors. The AHF diet mice developed hypersensitivity, demonstrated anxiety-like behaviors, and showed concurrent histology consistent with CP. Nontransgenic mouse models where pancreatitis is induced only by a combination of ad libitum liquid food with added alcohol and lard supplementation do not currently exist, nor has an in-depth study of anxiety-like behaviors been conducted in this mouse model. This dissertation research addresses this knowledge gap.

First identified over a decade ago, Astrocyte Elevated Gene-1 (AEG-1) has been studied extensively due to early reports of its overexpression in various cancer cell lines. Research groups all over the globe including our own have since identified AEG-1 overexpression in cancers of diverse lineages including cancers of the liver, colon, skin, prostate, breast, lung, esophagus, neurons and neuronal glia as compared to matched normal tissue. A comprehensive and convincing body of data currently points to AEG-1 as an essential component, critical to the progression and perhaps onset of cancer. AEG-1 is a potent activator of multiple pro-tumorigenic signal transduction pathways such as mitogen-activated protein extracellular kinase (MEK)/ extracellular signal-regulated kinase (ERK), phosphotidyl-inositol-3-kinase (PI3K)/Akt/mTOR, NF-κB and Wnt/β-catenin pathway. In addition, studies show that AEG-1 not only alters global gene and protein expression profiles, it also modulates fundamental intracellular processes, such as transcription, translation and RNA interference in cancer cells most likely by functioning as a scaffold protein. The mechanisms by which AEG-1 is overexpressed in cancer have been studied extensively and it is clear that multiple layers of regulation including genomic amplification, transcriptional, posttranscriptional, and posttranslational controls are involved however; the mechanism by which AEG 1 itself induces its oncogenic effects is still poorly understood. Just as questions remain about the exact role of AEG-1 in carcinogenesis, very little is known about the role of AEG-1 in regulating normal physiological functions in the liver. With the help of the Massey Cancer Center Transgenic/Knockout Mouse Core, our lab has successfully created a germline-AEG-1 knockout mouse (AEG-1-/-) as a model to interrogate AEG-1 function in vivo. Here I present the insights gained from efforts to analyze this novel AEG-1-/- mouse model. Aspects of the physiological functions of AEG-1 will be covered in chapter two wherein details of the characterization of the AEG-1-/- mouse are described including the role of AEG-1 in lipid metabolism. Chapter three discusses novel discoveries about the specific role of AEG-1 in mediating hepatocarcinogenesis by modulating NF-κB, a critical inflammatory pathway. First identified over a decade ago, Astrocyte Elevated Gene-1 (AEG-1) has been studied extensively due to early reports of its overexpression in various cancer cell lines. Research groups all over the globe including our own have since identified AEG-1 overexpression in cancers of diverse lineages including cancers of the liver, colon, skin, prostate, breast, lung, esophagus, neurons and neuronal glia as compared to matched normal tissue. A comprehensive and convincing body of data currently points to AEG-1 as an essential component, critical to the progression and perhaps onset of cancer. AEG-1 is a potent activator of multiple pro-tumorigenic signal transduction pathways such as mitogen-activated protein extracellular kinase (MEK)/ extracellular signal-regulated kinase (ERK), phosphotidyl-inositol-3-kinase (PI3K)/Akt/mTOR, NF-κB and Wnt/β-catenin pathway. In addition, studies show that AEG-1 not only alters global gene and protein expression profiles, it also modulates fundamental intracellular processes, such as transcription, translation and RNA interference in cancer cells most likely by functioning as a scaffold protein. The mechanisms by which AEG-1 is overexpressed in cancer have been studied extensively and it is clear that multiple layers of regulation including genomic amplification, transcriptional, posttranscriptional, and posttranslational controls are involved however; the mechanism by which AEG 1 itself induces its oncogenic effects is still poorly understood. Just as questions remain about the exact role of AEG-1 in carcinogenesis, very little is known about the role of AEG-1 in regulating normal physiological functions in the liver. With the help of the Massey Cancer Center Transgenic/Knockout Mouse Core, our lab has successfully created a germline-AEG-1 knockout mouse (AEG-1-/-) as a model to interrogate AEG-1 function in vivo. Here I present the insights gained from efforts to analyze this novel AEG-1-/- mouse model. Aspects of the physiological functions of AEG-1 will be covered in chapter two wherein details of the characterization of the AEG-1-/- mouse are described including the role of AEG-1 in lipid metabolism. Chapter three discusses novel discoveries about the specific role of AEG-1 in mediating hepatocarcinogenesis by modulating NF-κB, a critical inflammatory pathway.

Submitted by PPG Zoologia (zoologia-pg@pucrs.br) on 2018-08-01T18:22:00Z No. of bitstreams: 1 Almeida MAB___TESE___VERS?O FINAL.pdf: 2953118 bytes, checksum: 99cbefa9c38c7969abce4bafc4b20d54 (MD5) / Approved for entry into archive by Sheila Dias (sheila.dias@pucrs.br) on 2018-08-02T17:55:11Z (GMT) No. of bitstreams: 1 Almeida MAB___TESE___VERS?O FINAL.pdf: 2953118 bytes, checksum: 99cbefa9c38c7969abce4bafc4b20d54 (MD5) / Made available in DSpace on 2018-08-02T18:39:41Z (GMT). No. of bitstreams: 1 Almeida MAB___TESE___VERS?O FINAL.pdf: 2953118 bytes, checksum: 99cbefa9c38c7969abce4bafc4b20d54 (MD5) Previous issue date: 2018-03-22 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Nonhuman primates (NHP) are susceptible to many arboviruses, including the yellow fever (YF) virus. Although native to Africa, this virus found susceptible NHP and competent mosquito vectors for maintaining its transmission in American forests. A high sensitivity of NHP to YF led health agencies to monitor these animals as a way of monitoring the disease in Brazil. The State of Rio Grande do Sul (RS) began this surveillance in 2002, which has detected the arboviruses Oropouche and Saint Louis (SLEV) and a YF epizootic that killed more than 2,000 NHP (Alouatta caraya and A. guariba clamitans) between 2008 and 2009. The objectives of this PhD thesis research were to generate models of niche suitability for YF based on that epizootic and prospect arboviruses in NHP in northwestern RS. The maximum entropy algorithm - Maxent was used to generate distribution models of Alouatta spp. and the mosquito vector Haemagogus leucocelaenus. Together with climatic, topographic and vegetative variables, these models served as predictor layers to model the occurrence of the disease based on the points of death of NHP of YF. The most influential variables in the YF models were the variation in air humidity, distribution of Alouatta spp. and maximum wind speed followed by mean annual rainfall and maximum temperature. Therefore, support for the influence of the rainfall regime and the ambient temperature on the cycle of jungle YF was found. Wind speed and direction can play an important role in the dispersal of infected mosquitoes and, consequently, the virus. The models based on the occurrence of dead NHP in the first months of the epizootic identified suitable areas to where the disease spread a few months later. In addition, 19 arboviruses were prospected in 40 blood (viral isolation and PCR) and serum (hemagglutination inhibition and neutralization tests [NT]) samples collected from 26 black howler monkeys (A. caraya) belonging to three populations in four field campaigns in the municipality of Santo Ant?nio das Miss?es, RS, between 2014 and 2016. There was no detection of circulating virus, but antibodies to Flavivirus SLEV and Ilh?us and Phlebovirus Icoaraci was found by NT. Evidence of the contact with Ilh?us and Icoaraci are the southernmost records in Brazilian NHP. An increase in antibodies to SLEV detected between two consecutive captures of the same individual is compatible with a recent contact with the virus. An adult male captured in one of the areas presented concomitant infection by the Oropouche, SLEV and YF viruses by NT. Further studies are necessary to understand the role played by NHP and other vertebrates in the circulation of arboviruses in the region, to assess potential risks to NHP and public health, and to identify the driving forces responsible for the dispersal of the YF virus during epizootics in wildlife populations. / Os primatas n?o-humanos (PNH) s?o suscet?veis a diversos arbov?rus, incluindo o v?rus da febre amarela (FA). Embora origin?rio da ?frica, esse v?rus encontrou PNH suscet?veis e mosquitos vetores competentes para sua transmiss?o em matas nas Am?ricas. Uma alta sensibilidade dos PNH ? FA levou ?rg?os de sa?de a monitorar esses animais como forma de vigiar a doen?a no Brasil. O Estado do Rio Grande do Sul (RS) iniciou essa vigil?ncia em 2002, a qual detectou os arbov?rus Oropouche e Saint Louis (SLEV) e uma epizootia de FA que matou mais de 2000 PNH (Alouatta caraya e A. guariba clamitans) entre 2008 e 2009. A presente tese de doutorado teve como objetivos gerar modelos de adequabilidade ambiental para FA com base nessa epizootia e prospectar arbov?rus em PNH no noroeste do RS. Foi utilizado o algoritmo de m?xima entropia ? Maxent para gerar modelos de distribui??o de Alouatta spp. e do mosquito vetor Haemagogus leucocelaenus. Esses modelos serviram como camadas preditoras para, junto a vari?veis clim?ticas, topogr?ficas e vegetacionais, modelar a ocorr?ncia da doen?a baseada nos pontos de morte de PNH por FA. As vari?veis mais influentes nos modelos da FA foram a varia??o na umidade do ar, a distribui??o de Alouatta spp. e a velocidade m?xima dos ventos, seguidas pela precipita??o m?dia anual e a temperatura m?xima. Portanto, foi confirmado suporte para a influ?ncia do regime de chuvas e da temperatura ambiente no ciclo da FA silvestre. A velocidade e a dire??o do vento devem desempenhar um importante papel na dispers?o de mosquitos infectados e, consequentemente, do v?rus. Os modelos baseados na distribui??o espacial de PNH mortos nos primeiros meses da epizootia identificaram ?reas adequadas para onde a doen?a avan?ou poucos meses mais tarde. Tamb?m foram prospectados 19 arbov?rus em 40 amostras de sangue (isolamento viral e PCR) e soro (inibi??o da hemaglutina??o e testes de neutraliza??o [NT]) coletadas em quatro campanhas de campo entre 2014 e 2016 de 26 bugios-pretos (A. caraya) de tr?s popula??es no munic?pio de Santo Ant?nio das Miss?es, RS. N?o houve detec??o de v?rus circulante, mas sim de anticorpos para os Flavivirus SLEV e Ilh?us e o Phlebovirus Icoaraci por NT. As evid?ncias de contato com Ilh?us e Icoaraci s?o as primeiras em PNH no extremo sul do Brasil. Um aumento de anticorpos para SLEV detectado entre duas capturas consecutivas do mesmo indiv?duo ? compat?vel com um contato recente com o v?rus. Um macho adulto capturado em uma das ?reas apresentou infec??o concomitante pelos v?rus Oropouche, SLEV e FA por NT. Mais estudos s?o necess?rios para compreender o papel de PNH e outros vertebrados na circula??o de arbov?rus na regi?o, avaliar poss?veis riscos para PNH e a sa?de humana e identificar as for?as motrizes respons?veis pela dispers?o do v?rus da FA durante epizootias em popula??es selvagens.

Introduction: The Aedes aegypti mosquito is responsible for the transmission of Yellow Fever, Dengue, Chikungunya and Zikavirus, making it a deadly vector and global public health threat. Zikavirus and Chikungunya, which were previously restricted to smaller geographic areas, have both appeared in the Western Hemisphere in the past three years and spread to areas where A. aegypti are present. This means that the pathogens have now entered areas in which the population has no previous immunity, which can lead to extensive outbreaks and epidemics. As the effects of global climate change become apparent, the areas of the globe that are suitable for inhabitance by A. aegypti may change. Additionally, this vector prefers human hosts for blood meals and requires standing water to breed, which is often created by water storage containers. This means that increasing urbanization and human population density are likely to put populations at higher risk of exposure to this vector. Methods: To create maps of the future risk of exposure to Aedes aegypti globally, species occurrence data for the vector and the Maxent modeling approach were used. Current and projected climate data were downloaded from WorldClim.org for the four representative concentration pathways (RCPs) used to model future climate change. Human population density, projected to 2050, the same timeframe as the future climate data, were used to model changes in human populations. To identify areas at high risk for future presence of A. aegypti populations, current and future models were compared across areas with at least a 50% probability of increased risk. These results where then used to create maps displaying high risk areas. Results: The AUC, an indicator of model fit, signaled that the models had high predictive power. However, high omission rates indicated that the trade-off of risk mapping may be a need to decrease probability thresholds below 50% to capture the full at-risk population. Future high-risk areas were most often those surrounding current cities, which supports the idea that the combination of urbanization and increasing human population density will work synergistically to increase the disease burden within and around urban centers. Additionally, expansion at the current geographic margins of this species shows that incursion into currently non-endemic areas is possible. Conclusions: Urban and peri-urban populations are likely to be at higher risk of exposure compared to rural areas due to global climate change and changes in population density. Attempts to model expansion of vector habitats should consider how these human population characteristics will change the risk to populations and how to best identify the areas at highest risk. Thresholds for the probability of a population being at risk of exposure to a vector may need to be different from those required to determine whether or not a habitat is suitable for a species. Appropriately determining which areas are high-risk results in maps and models can then be used to identify areas where climate change mitigation and vector control efforts are likely to have the highest impacts.

The Aedes aegypti mosquito is responsible for transmission of four vector-borne diseases that cause considerable global morbidity and mortality. Projections of the future effects of global climate change indicate that expansion of this species due to changing habitats is possible. Furthermore, since A. aegypti is highly dependent on human populations for feeding and egg-laying sites, changing human population characteristics are likely to alter the risk of exposure for humans based on geographic location. This study aims to create future potential risk maps for human exposure to A. aegypti using human population density as a predictor. Using current population density data and future growth trajectories, high-resolution human population density forecasts were created for 2050, then included as variables in ecological niche models developed using Maxent. Species occurrence data and high resolution climate data for current and future conditions (best and worst case scenarios) were included in the model, as well. Model fit indices and variable contributions indicated that the inclusion of human population density improves model accuracy for A. aegypti. Risk maps created by these models showed that areas currently adjacent to large cities within endemic regions, such as central Africa and western Brazil, are likely to see the greatest increase in risk to human populations. This corroborates current projections on increasing urbanization in the future and suggests that these models can be used to target interventions in high risk areas.

No description available.