Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-04-11T19:27:27Z No. of bitstreams: 1 daianaaparecidarodrigues.pdf: 8225936 bytes, checksum: 96ec323f343f92c319f4e261145f9c6a (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-04-24T03:34:16Z (GMT) No. of bitstreams: 1 daianaaparecidarodrigues.pdf: 8225936 bytes, checksum: 96ec323f343f92c319f4e261145f9c6a (MD5) / Made available in DSpace on 2016-04-24T03:34:16Z (GMT). No. of bitstreams: 1 daianaaparecidarodrigues.pdf: 8225936 bytes, checksum: 96ec323f343f92c319f4e261145f9c6a (MD5) Previous issue date: 2013-08-29 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Fenômenos biológicos são todo e qualquer evento que possa ser observado nos seres vivos. O estudo desses fenômenos permite propor explicações para o seu mecanismo, a m de entender as causas e efeitos. Pode-se citar como exemplos de fenômenos biológicos o comportamento das células como respiração, reprodução, metabolismo e morte celular. Equações de reação-difusão são frequentemente utilizadas para modelar fenômenos bioló- gicos. Sistemas de reação-difusão podem produzir padrões espaciais estáveis a partir de uma distribuição inicial uniforme esse fenômeno é conhecido como instabilidade de Turing. Este trabalho apresenta a análise da instabilidade de Turing bem como resultados numéricos para a solução de três modelos biológicos, modelo de Schnakenberg, modelo de glicólise e modelo da coagulação sanguínea. O modelo de Schnakenberg é utilizado para descrever uma reação química autocatalítica e o modelo de glicólise é relativo ao processo de degradação metabólica da molécula de glicose para proporcionar energia para o metabolismo celular, esses dois modelos são frequentemente relatados na literatura. O terceiro modelo é mais recente e descreve o fenômeno da coagulação sanguínea. Nas soluções numéricas se utiliza o método das linhas onde a discretização espacial é feita através de um esquema de diferenças nitas. O sistema de equações diferencias ordinárias resultante é resolvido por um esquema de integração adaptativo, com a utilização de pacote para computação cientí ca da linguagem Python, Scipy. / Biological phenomena are all and any event that can be observed in living beings. The study of these phenomena enables us to propose explanations for its mechanisms in order to understand causes and e ects. One can cite as examples of biological phenomena the behavior of cells as respiration, reproduction, metabolism and cell death. Reactiondi usion equations are often used to model biological phenomena. Reaction-di usion systems can produce stable spatial patterns from a uniform initial distribution, this phenomenon is known as Turing instability. This dissertation presents an analysis of the Turing instability as well as numerical results for the solution of three biological models, model Schnakenberg, model of glycolysis and model of blood coagulation. The Schnakenberg model is used to describe an autocatalytic chemical reaction and glycolysis model refers to the process of metabolic breakdown of the glucose molecule to provide energy for cellular metabolism, these two models are frequently reported in the literature. The third model is newer and describes the phenomenon of blood coagulation. The method of lines is used in the numerical solutions, where the spatial discretization is done through a nite di erence scheme. The resulting system of ordinary di erential equations is then solved by an adaptive integration scheme with the use of the package for scienti c computing of Python language, Scipy.

A proteína miostatina, é um regulador negativo do crescimento muscular e a modulação de sua expressão pode consistir em tratamento para distrofias musculares. Nós estudamos expressão endógena da miostatina no músculos gastrocnêmio e diafragma de 4 modelos murinos de degeneração muscular: os camundongos Dmdmdx, SJL/J, Largemyd e Lama2dy-2J/J. Observamos que a miostatina é menos expressa no músculo gastrocnêmio do que diafragma normal, refletindo um músculo mais sujeito a lesão. Nas quatro linhagens distróficas a miostatina é menos expressa do que em camundongos normais, tanto no músculo gastrocnêmio como diafragma, sem diferença entre os dois. A analise comparativa da degeneração e regeneração muscular mostrou maior correlação da inibição da miostatina com o padrão de degeneração. Nossos resultados sugerem que o processo de degeneração, quando iniciado, e independentemente de seu grau, causa molecular primária, ou músculo afetado, parece atuar de forma similar na inibição da expressão da miostatina, possivelmente como estimulo a regeneração do dano. / Myostatin is a negative regulator of muscle growth, and its inhibition has been considered a therapeutic strategy for muscular dystrophies. We evaluated the endogenous expression of myostatin in the gastrocnemius and diaphragm muscles from 4 mouse dystrophic models including Dmdmdx, SJL/J>, Largemyd and Lama2dy2J/J. In normal mice, we observed that myostatin is less expressed in the gastrocnemius than in the diaphragm, reflecting a muscle most prone to lesions. In the 4 dystrophic models, myostatin expression was reduced, in both gastrocnemius and diaphragm muscles. The comparative analysis of the histopathology of the muscles with the expression of myostatin showed a stronger correlation with the pattern of degeneration then regeneration. Our results suggest that, when started, the process of degeneration of the muscle, independently of the primary molecular defect, or degree, seems to act in a similar pathway leading to the inhibition of the expression of myostatin in the affected muscles, possibly as a stimulus to regeneration of damage.

Myeloid-derived suppressor cells (MDSCs) numbers increase significantly in sepsis and are associated with high mortality rates. These myeloid cell precursors promote immunosuppression, especially in the late (post sepsis) stage. However, the mechanisms that underlie MDSC expansion and programming are not completely understood. To investigate these mechanisms, we used a cecal-ligation and puncture (CLP) mouse model of polymicrobial sepsis that progresses from an early/acute proinflammatory phase to a late/chronic immunosuppressive phase. Previous studies in our laboratory showed that microRNA (miR)-21 and miR-181b elevate levels of the transcription factor nuclear factor 1 (NFI-A) that promotes MDSC expansion. We report here that miR-21 and miR-181b regulate NFI-A expression via a post-transcriptional regulatory mechanism by recruiting RNA-binding proteins HuR and Ago1 to stabilize NFI-A mRNA, thus increasing its protein levels. Studies in our laboratory also showed that inflammatory mediator S100A9 accumulates in the nucleus in Gr1+CD11b+ myeloid precursors in the later phases of sepsis and is necessary for their expansion and programming into immunosuppressive MDSCs. We demonstrate here that nuclear S100A9 associates with specific transcription factors that activate miR-21 and miR-181b expressions. In our final manuscript, we uncover another layer of the mechanisms of MDSC expansion and programming. We found that long non-coding RNA (lncRNA) Hotairm1 binds to and recruits S100A9 to the nucleus to program Gr1+CD11b+ myeloid precursors into MDSCs in the later phases of sepsis. Together, our results reveal three regulatory layers involving NFI-A, S100A9 and Hotairm1 in the pathway leading to MDSCs development in sepsis and suggest that therapeutically targeting these molecular switches might improve sepsis survival.

<p>The need to understand and accurately assess the health risks of low dose ionizing radiation is more important now than ever before. The global applications of ionizing radiation in medicine, mining, manufacturing, and the nuclear industry have increased exponentially in recent years. Parallel to this increase are the health concerns regarding occupational and medical exposures to radiation. The research presented here investigates the biological and health effects of ionizing radiation, specifically from medical diagnostic exposures.</p> <p>Medical diagnostic procedures such as x-rays and computed tomography (CT) scans account for a notable portion of the public's exposure to ionizing radiation. The health risk to humans associated with these low dose exposures is unknown. Often times they are correlated with risk estimates derived from much higher radiation doses. There is no doubt that very high dose ionizing radiation can be harmful; however, the same notion does not exist regarding exposures to low dose ionizing radiation such as that from medical diagnostic CT exposures.</p> <p>The objective of this research is to address the effects and risks associated with diagnostic CT scans. This research focuses on the biological outcome of cancer which remains a primary concern in health care and the development of radiation risk policies. The investigation utilized various mouse models that have differing sensitivities to radiation and susceptibilities to developing radiation-induced cancer.</p> <p>Results from this research found that low-dose diagnostic CT scans do not increase risk and can, in fact, induce protective effects. The hypothesis that harmful effects increase linearly with radiation dose is not supported by this research. With low doses of CT scans, protective biological effects such as reduced chromosomal aberrations, decreased radiation-induced oxidative DNA damage, and enhanced clearance of damaged cells have been observed. In cancer-prone mice, CT scans can increase longevity and reduce cancer risk by delaying the latency of specific cancers.</p> <p>This research advances the understanding of the biological effects and health risk associated with low-dose medical diagnostic procedures. This research is timely and important to allow medical practitioners, policy makers, and regulators to make informed decisions about using ionizing radiation in the clinic. Such knowledge is valuable as better, more complex, and perhaps more damaging modalities are being used to image and manage disease.</p> / Doctor of Philosophy (PhD)

<p>NB: I had another committee member, Dr. Mark Larché; and would like to have his name included in the document.</p> <p>Thank you.</p> / <p><strong>Introduction: </strong>Increasing incidence of chronic immune diseases are mirrored by changing disease risk factors, which include maternal stress during pregnancy. To date, no studies have investigated the impact of prenatal stress challenge (PNS) on the fetal immune system. Fetal liver and bone marrow represent major sources of hematopoietic stem cells (HSC) at mid gestation, which differentiate and mature in the thymus. Disturbance of immune development may cause immune impairment in later life. Further, progesterone is recognized as a critical part of feto-maternal interaction. This study aimed to determine if PNS interferes with normal fetal immune development in mice and the impact of progesterone supplementation on stress effects. <strong>Methods: </strong>DBA/2J-mated BALB/c dams were sorted into three groups: control, PNS (gestation days (GDs) 12.5 and 14.5) and PNS plus progesterone supplementation (DHD). Fetal tissue was collected on GDs 16.5 and 18.5. Flow cytometric analysis examined frequency and phenotype of fetal immune cell populations: HSC in fetal liver and bone marrow, and different stages of T cell maturation and regulatory T (Treg) cells in the thymus. Fetal tails were collected to determine fetal sex by PCR analysis. <strong>Results: </strong>PNS induced a decrease in organ size on GD16.5, which was not seen on GD18.5 and was reversed by DHD treatment. PNS altered the percentage and absolute number of HSC within the liver and bone marrow populations, on GD16.5 and 18.5. There was a significant lag in T cell maturation as demonstrated by the altered expression of CD3 and skewed CD3-:CD3+ ratio. There was a significant decrease in Treg cells within CD3+ thymic cells in response to PNS. PNS effects in the thymus were ameliorated by DHD treatment. There was no PNS-induced sex bias. <strong>Conclusions: </strong>These results indicate that PNS compromises the developing fetal immune system, which could account for impaired immune responses in adults with chronic immune disease, and provide evidence for a therapeutic role of progesterone supplementation.</p> / Master of Science (MSc)

As the prevalence of cardiovascular diseases continues to exponentially grow in populations across the globe, the necessity of determining underlying factors, effective methods of diagnoses, and universally available preventive measures also grows. Early detection of endothelial dysfunction, a proven precursor of cardiovascular diseases, can be extremely impactful in encouraging preventative measures and early intervention before medical conditions become chronic. In recent years, ice plunging, a form of cryotherapy involving full body immersion in cold water, has gained popularity within circles of fitness and health practitioners, gaining the interest of people of all backgrounds. Certain parallels observed between the human physiological response to cold exposure and endothelial function encourage further study of the effects of ice plunging on cardiovascular health. Calibrated cuff plethysmography is a promising method of reflecting on endothelial function by measuring arterial compliance of select blood vessels. In this study, a calibrated cuff plethysmography device was built and tested for efficiency as it was used to measure compliance and cross-sectional area of the brachial artery of 14 participants 30 minutes before, immediately after, and 30 minutes after a 5-minute cold plunge in a temperature of 10°C - 15°C. Results found some significant differences between baseline measurements recorded immediately after the ice plunge and measurements recorded during reactive hyperemia conditions at normal body temperature but did not conclude that 5-minute cold-water immersion intervention had a significant impact on arterial compliance or area overall since this was a short term experiment with only acute intervention methods. The device used was concluded to effectively measure arterial compliance and area.

Non-small cell lung cancers (NSCLC) constitute 80-85% of lung cancers and are the leading cause of cancer-related deaths globally. The most common cause is prolonged smoking. Current treatment options for NSCLC include surgery, radiation, chemotherapy, targeted drug therapy, and immunotherapy. Although these medications are effective in the short term, patients often face issues of drug resistance and debilitating side effects with prolonged use. Currently, the use of Zika virus (ZIKV) is being researched as a possible alternative treatment for cancer, which minimizes side effects and the risk of drug resistance. TIM/TAM proteins are identified as the putative ZIKV receptors on the surface of human cells that mediate viral entry through apoptotic mimicry. Once engulfed, the virus can hijack the host cell’s machinery to replicate and propagate the infection. Previous research has shown the potential of using Zika virus as an oncolytic agent in glioblastoma and neuroblastoma cell lines. The success of Zika-induced oncolysis in these cancers opens doors for expanding into other cancers, including NSCLC. Infection of six diverse NSCLC cell lines with ZIKV revealed that three cell lines were sensitive to ZIKV-induced oncolysis while the remaining were resistant. Transcriptome data analysis of TIM/TAM and CD24 mRNA expression levels were compared between ZIKV-sensitive and resistant cell lines, revealing AXL and TIM-1 as potential players in increasing or decreasing ZIKV infection. High AXL (TAM) expression correlated with increased sensitivity to ZIKV, while high TIM-1 (TIM) expression correlated with increased resistance. Experiments with AXL silencing in ZIKV-sensitive cell lines provided evidence of the role of AXL in increasing ZIKV sensitivity. Although further studies with TIM-1 must be done to determine its role in conferring resistance, AXL and TIM-1 have the potential to be biomarkers in predicting tumor sensitivity to ZIKV-induced oncolytic therapy.