131 |
Ross River Virus Infection: Disease Mechanisms and Potential TreatmentRulli, Nestor Ezequiel, na January 2007 (has links)
Ross River virus (RRV) is a mosquito-borne alphavirus and the aetiological agent of
epidemic polyarthritis (EPA). Arthropod borne-Alphaviruses that are related to RRV,
such as Chikungunya virus, Sindbis virus and Barmah Forest virus, are usually
associated with epidemics of infectious arthritides in different parts of the world. In
humans, RRV-induced disease symptoms include fever, rash, myalgia and pain and
stiffness of the joints. Muscle and joint pain are the most debilitating symptoms in RRV
patients and the best treatment available is non-steroidal anti-inflammatory drugs
(NSAID). Previous studies in mice have demonstrated that RRV infection results in
inflammation of skeletal muscle and joints and that macrophages play a primary role in
disease.
The present study was carried out to further elucidate the underlying mechanisms
mediating RRV-induced muscle and joint pathology. Previous studies have reported
that encephalitic alphaviruses trigger apoptosis of brain cells in mice and that blocking
apoptosis reduces mortality rates. In the present study, the ability of RRV to induce
muscle apoptosis was investigated in vitro, using a murine myoblast cell line (C1C12),
and in vivo, using a mouse model of RRV disease. RRV-infected C1C12 myofibres
displayed an array of morphological and biochemical makers of apoptosis. Apoptosis
was also observed in the skeletal muscle of RRV-infected C57BL/6J mice. Blocking
apoptosis by general caspase inhibition resulted in milder disease symptoms, reduced
myofibre damage and decreased inflammation of muscle and joint tissues. The total
number of cell infiltrates as well as the number of macrophages infiltrating muscle was
significantly reduced by the treatment with a caspase inhibitor.
The effects of RRV infection on the skeletal system were also investigated. Primary
human osteoblast cells were infected with RRV and monitored for viral-induced
cytopathic effect. Osteoblasts supported rapid virus growth and, by 48 hours after
infection, succumbed to viral-induced necrosis. In addition, histological examination of
bone tissue from RRV-infected C57BL/6J mice showed clear evidence of bone
resorption. Tibias from infected mice showed an increased number of activated
osteoclasts, a reduction in bone density and thinning of cortical bone.
The expression of host factors involved in inflammatory responses and bone
remodelling was studied in RRV-infected myofibres and osteoblast cell cultures and in
the muscle and joint tissues from infected mice. RRV-infected muscle cells and tissue
showed elevated mRNA levels for the chemokines CCL-2, CCL3, CCL5 and CXCL1,
all of which are known to mediate the migration of monocytic cells. With the exception
of CXCL1, these chemokines were also found to be up-regulated in RRV-infected
osteoblast cultures and in joint tissues from infected mice. Muscle and joint tissue from
infected mice also showed elevated mRNA levels for type I and type II interferons,
TNF- and NOS2. In addition, joint tissues from infected animals contained high levels
of IL-6 and IL-1, two cytokines known to mediate bone remodelling.
Finally, the therapeutic potential of the drug bindarit was investigated using the mouse
model of RRV disease. Bindarit is a known inhibitor of CCL-2 and TNF- and has
been found to prevent protein denaturation. Treatment with bindarit resulted in mice
developing milder disease symptoms, reduced muscle damage and decreased
inflammation of muscle and joint tissues. In particular, bindarit significantly reduced
macrophage infiltration into skeletal muscle tissue.
This thesis has contributed to the understanding of RRV pathogenesis. It has identified
novel mechanisms of RRV-induced muscle and bone pathology and provided further
evidence that associate pro-inflammatory host factors to RRV disease. This work has
also demonstrated that bindarit should be considered as a candidate for treating RRV
disease in humans.
|
132 |
Innate immunity to Rhodococcus equi: the response of adult and juvenile equine neutrophilsNerren, Jessica Rachel 15 May 2009 (has links)
Blood was obtained from 5 adult horses and 16 juvenile horses (foals) at the time of birth and subsequently at 2-, 4-, and 8-weeks of age. Neutrophils from adult horses were purified and incubated for 2 h and 4 h with media, avirulent R. equi, virulent R. equi, or recombinant-human granulocyte-macrophage colony stimulating factor (rhGM-CSF). Neutrophils from foals were purified and incubated for 2 h and 4 h with media or virulent R. equi. Total RNA was extracted from both adult and foal neutrophils immediately after purification to measure baseline expression levels (0 h), and immediately after each of the prescribed incubation times. For each sample, 1 µg of total RNA was reverse-transcribed and analyzed for differential gene expression using real-time PCR. After 2 h and 4 h incubation with virulent or avirulent R. equi, neutrophils from adult horses expressed significantly (P< 0.05) greater TNFα, IL-12p40, IL-6, IL-8, and IL-23p19 mRNA relative to expression by unstimulated neutrophils, but not IFNγ or IL-12p35 mRNA. Furthermore, virulent R. equi induced significantly greater IL-23p19 mRNA expression than avirulent R. equi. Stimulation with rhGM-CSF of adult equine neutrophils failed to induce significant changes in cytokine expression. In foal neutrophils, stimulation with virulent R. equi induced significantly greater expression of IFNγ, TNFα, IL-6, IL-8, IL-12p40, and IL-12p35 mRNA relative to expression by unstimulated neutrophils. Furthermore, there were significant effects of age on expression of IL-6, IL-8 and IL-12p40 mRNA. Neutrophil mRNA expression of IL-6 and IL-8 in newborn foals was significantly greater than expression at 2-, 4-, and 8-weeks of age. There was no significant difference between unstimulated and R. equi-stimulated neutrophils from newborn and 2-week-old foals in expression of IL-12p40; however, expression of IL-12p40 by R. equi-stimulated neutrophils from 4- and 8-week-old foals was significantly greater than expression by unstimulated neutrophils. These results demonstrate that R. equi-stimulated neutrophils are a source of many pro-inflammatory cytokines, and that the magnitude of this expression with respect to IL-6, IL-8, and IL-12p40 mRNA expression was influenced by age. Collectively, the data presented indicate a non-phagocytic role for neutrophils that may influence the type of adaptive immune response to R. equi.
|
133 |
Applying Current Methods for Estimating Influenza Burden to an Academic Health Sciences CentreSmith, Tiffany 24 August 2012 (has links)
Public health planning for influenza is based on morbidity and mortality estimates derived from statistical models. Lower than anticipated 2009 H1N1 pandemic death estimates have raised questions about the method. Examining the statistical method is important for future policy and program development. We compared the main methods of estimating influenza burden through a systematic literature review and by comparing statistical estimates of influenza-attributable burden at the Ottawa Hospital (TOH) to clinical estimates validated through chart review. We identified heterogeneity in methods used to estimate influenza-attributable mortality in the literature which resulted in within-season estimate variation by study. We found statistical estimates of influenza burden at TOH to be 4-8 times greater than clinically validated data. We also found no significant association between the outcomes examined and epidemic periods at TOH. The findings of this study suggest discordance between model estimates by model approach and between model estimates and validated findings. Examining reasons for these discordances should be pursued.
|
134 |
Host Inflammatory Pathways in Malaria Infection: Potential Therapeutic Targets and Biomarkers of Disease SeverityErdman, Laura Kelly 06 January 2012 (has links)
Severe malaria infections cause almost 1 million deaths annually, mostly among non-immune African children. The pathogenesis of severe malaria is poorly understood. It is increasingly appreciated that while host innate immune responses such as inflammation and phagocytosis are critical for control of parasite replication, they can become dysregulated and contribute to severe disease. The goals of this work were: (1) to characterize inflammatory responses to malaria by defining their relationship to phagocytosis and identifying novel molecular mediators, and (2) to evaluate the utility of biomarkers of inflammation and other host responses for predicting outcome in severe malaria infection. Using an in vitro model of the malaria-macrophage interaction, inflammatory and phagocytic responses to Plasmodium falciparum were found to be partially coupled. Activation of Toll-like receptors (TLRs) by purified parasite components increased internalization of parasitized erythrocytes, but uptake of parasitized erythrocytes did not require TLRs, nor did it trigger cytokine production via TLRs or other receptors. Two candidate molecules – Triggering receptor expressed on myeloid cells-1 (TREM-1) and Chitinase-3 like-1 (CHI3L1) – did not appear to critically modulate inflammation to malaria in vitro or in murine models. However, exogenous TREM-1 activation enhanced the pro-
inflammatory nature of the response to P. falciparum, with potential implications for malarial-bacterial co-infection. CHI3L1-deficient mice showed a trend towards earlier death in experimental cerebral malaria, suggesting that CHI3L1 may protect against severe malaria; however, further investigation in more informative models is required. Admission levels of plasma TREM-1, CHI3L1, and other biomarkers of inflammation and endothelial activation were increased in Ugandan children with severe malaria. Simple combinations of these biomarkers predicted mortality among severe malaria patients with high accuracy, warranting larger validation studies. Taken together, these findings identify host responses as putative targets for adjunctive therapies, and suggest the utility of host biomarker combinations as prognostic tests for severe malaria.
|
135 |
Host Inflammatory Pathways in Malaria Infection: Potential Therapeutic Targets and Biomarkers of Disease SeverityErdman, Laura Kelly 06 January 2012 (has links)
Severe malaria infections cause almost 1 million deaths annually, mostly among non-immune African children. The pathogenesis of severe malaria is poorly understood. It is increasingly appreciated that while host innate immune responses such as inflammation and phagocytosis are critical for control of parasite replication, they can become dysregulated and contribute to severe disease. The goals of this work were: (1) to characterize inflammatory responses to malaria by defining their relationship to phagocytosis and identifying novel molecular mediators, and (2) to evaluate the utility of biomarkers of inflammation and other host responses for predicting outcome in severe malaria infection. Using an in vitro model of the malaria-macrophage interaction, inflammatory and phagocytic responses to Plasmodium falciparum were found to be partially coupled. Activation of Toll-like receptors (TLRs) by purified parasite components increased internalization of parasitized erythrocytes, but uptake of parasitized erythrocytes did not require TLRs, nor did it trigger cytokine production via TLRs or other receptors. Two candidate molecules – Triggering receptor expressed on myeloid cells-1 (TREM-1) and Chitinase-3 like-1 (CHI3L1) – did not appear to critically modulate inflammation to malaria in vitro or in murine models. However, exogenous TREM-1 activation enhanced the pro-
inflammatory nature of the response to P. falciparum, with potential implications for malarial-bacterial co-infection. CHI3L1-deficient mice showed a trend towards earlier death in experimental cerebral malaria, suggesting that CHI3L1 may protect against severe malaria; however, further investigation in more informative models is required. Admission levels of plasma TREM-1, CHI3L1, and other biomarkers of inflammation and endothelial activation were increased in Ugandan children with severe malaria. Simple combinations of these biomarkers predicted mortality among severe malaria patients with high accuracy, warranting larger validation studies. Taken together, these findings identify host responses as putative targets for adjunctive therapies, and suggest the utility of host biomarker combinations as prognostic tests for severe malaria.
|
136 |
Innate immunity to Rhodococcus equi: the response of adult and juvenile equine neutrophilsNerren, Jessica Rachel 15 May 2009 (has links)
Blood was obtained from 5 adult horses and 16 juvenile horses (foals) at the time of birth and subsequently at 2-, 4-, and 8-weeks of age. Neutrophils from adult horses were purified and incubated for 2 h and 4 h with media, avirulent R. equi, virulent R. equi, or recombinant-human granulocyte-macrophage colony stimulating factor (rhGM-CSF). Neutrophils from foals were purified and incubated for 2 h and 4 h with media or virulent R. equi. Total RNA was extracted from both adult and foal neutrophils immediately after purification to measure baseline expression levels (0 h), and immediately after each of the prescribed incubation times. For each sample, 1 µg of total RNA was reverse-transcribed and analyzed for differential gene expression using real-time PCR. After 2 h and 4 h incubation with virulent or avirulent R. equi, neutrophils from adult horses expressed significantly (P< 0.05) greater TNFα, IL-12p40, IL-6, IL-8, and IL-23p19 mRNA relative to expression by unstimulated neutrophils, but not IFNγ or IL-12p35 mRNA. Furthermore, virulent R. equi induced significantly greater IL-23p19 mRNA expression than avirulent R. equi. Stimulation with rhGM-CSF of adult equine neutrophils failed to induce significant changes in cytokine expression. In foal neutrophils, stimulation with virulent R. equi induced significantly greater expression of IFNγ, TNFα, IL-6, IL-8, IL-12p40, and IL-12p35 mRNA relative to expression by unstimulated neutrophils. Furthermore, there were significant effects of age on expression of IL-6, IL-8 and IL-12p40 mRNA. Neutrophil mRNA expression of IL-6 and IL-8 in newborn foals was significantly greater than expression at 2-, 4-, and 8-weeks of age. There was no significant difference between unstimulated and R. equi-stimulated neutrophils from newborn and 2-week-old foals in expression of IL-12p40; however, expression of IL-12p40 by R. equi-stimulated neutrophils from 4- and 8-week-old foals was significantly greater than expression by unstimulated neutrophils. These results demonstrate that R. equi-stimulated neutrophils are a source of many pro-inflammatory cytokines, and that the magnitude of this expression with respect to IL-6, IL-8, and IL-12p40 mRNA expression was influenced by age. Collectively, the data presented indicate a non-phagocytic role for neutrophils that may influence the type of adaptive immune response to R. equi.
|
137 |
Estimating Network Features and Associated Measures of Uncertainty and Their Incorporation in Network Generation and AnalysisGoyal, Ravi 19 November 2012 (has links)
The efficacy of interventions to control HIV spread depends upon many features of the communities where they are implemented, including not only prevalence, incidence, and per contact risk of transmission, but also properties of the sexual or transmission network. For this reason, HIV epidemic models have to take into account network properties including degree distribution and mixing patterns. The use of sampled data to estimate properties of a network is a common practice; however, current network generation methods do not account for the uncertainty in the estimates due to sampling. In chapter 1, we present a framework for constructing collections of networks using sampled data collected from ego-centric surveys. The constructed networks not only target estimates for density, degree distributions and mixing frequencies, but also incorporate the uncertainty due to sampling. Our method is applied to the National Longitudinal Study of Adolescent Health and considers two sampling procedures. We demonstrate how a collection of constructed networks using the proposed methods are useful in investigating variation in unobserved network topology, and therefore also insightful for studying processes that operate on networks. In chapter 2, we focus on the degree to which impact of concurrency on HIV incidence in a community may be overshadowed by differences in unobserved, but local, network properties. Our results demonstrate that even after controlling for cumulative ego-centric properties, i.e. degree distribution and concurrency, other network properties, which include degree mixing and clustering, can be very influential on the size of the potential epidemic. In chapter 3, we demonstrate the need to incorporate information about degree mixing patterns in such modeling. We present a procedure to construct collections of bipartite networks, given point estimates for degree distribution, that either makes use of information on the degree mixing matrix or assumes that no such information is available. These methods permit a demonstration of the differences between these two network collections, even when degree sequence is fixed. Methods are also developed to estimate degree mixing patterns, given a point estimate for the degree distribution.
|
138 |
Turning the tide against TB: Remaking ineffective host defenses into mechanisms for tuberculosis controlZhang, Yanjia Jason 07 June 2014 (has links)
Most antibiotics, including the drugs currently used for treating tuberculosis (TB), were first discovered as molecules that inhibit bacterial growth in laboratory culture conditions and later translated to infection models and clinical use. Mycobacterium tuberculosis (Mtb) has evolved specifically to survive in its human host, and it is in this infectious context that new drugs need to work. The host environment is characterized by a multitude of antimicrobial defenses induced by the immune system, and we can leverage these defenses to kill Mtb in vivo. Mtb employs a diverse set of responses to survive host defenses. By blocking these responses, we can make Mtb more susceptible to host immunity, turning these previously impotent defenses into effective strategies of immune control.
|
139 |
Computational Gains Via a Discretization of the Parameter Space in Individual Level Models of Infectious DiseaseFANG, XUAN 13 January 2012 (has links)
The Bayesian Markov Chain Monte Carlo(MCMC) approach to inference is commonly used to estimate the parameters in spatial infectious disease models. However, such MCMC analyses can pose a hefty computational burden. Here we present new method to reduce the computing time cost in such MCMC analyses and study its usefulness. This method is based a round the discretization of the spatial parameters in the infectious disease model. A normal approximation of the posterior density of the output from the original model will be compared to that of the modified model, using the Kullback-Leibler(KL) divergence measure.
|
140 |
Is Porcine Periweaning Failure-to-Thrive Syndrome an Infectious Diseases?2013 December 1900 (has links)
Porcine Periweaning Failure-to-Thrive syndrome (PFTS) is a clinical syndrome of newly weaned pigs with unknown etiology and characterized by anorexia, lethargy and progressive debilitation. The hypothesis of this thesis is that PFTS is an infectious disease. Investigation in an index farm affected by PFTS from Saskatchewan Canada ruled out most common swine pathogens as the etiology and identified several lesions that were consistent across many cases. A larger study including multiple farms in North America was then undertaken. A total of 8 farms were investigated, within which 5 met the clinical definition of PFTS. Gross and histological examinations were performed on 8 case and 4 control pigs on each farm. Detection of relevant porcine pathogens, complete blood count, serum chemistry, and serum cytokine analysis were performed on each pig. Thymic atrophy, superficial gastritis and small intestinal villous atrophy were significantly more prevalent in case pigs compared to control pigs. All case pigs had at least two of these three lesions. All case and control pigs were negative for porcine reproductive and respiratory syndrome virus, swine influenza virus and were free of porcine circovirus associated diseases. Although several pathogens, such as porcine cytomegalovirus, haemagglutinating encephalomyelitis virus, porcine enteric calicivirus, group A rotavirus, enteroviruses and Cystoisospora suis were detected in some of the case and control pigs, none were associated with clinical status. Clinical pathology findings of case pigs was consistent with anorexia and dehydration, such as increases in haematocrit, blood urea, serum bilirubin, albumin, beta-hydroxybutyrate and decreases in blood glucose, calcium and phosphorous. Case pigs had similar levels to IL1-β than control pigs, which suggested that PFTS was not a result of excessive cytokines. In subsequent experiments, a snatched-farrowed porcine-colostrum-deprived (SF-pCD) pig model was developed and tissue homogenates were used to inoculate SF-pCD pigs in an attempt to reproduce the clinical signs of PFTS. The SF-pCD pigs were immunologically characterized and shown to be suitable for inoculation studies. However, inoculation of tissue homogenate from PFTS pigs failed to reproduce the clinical signs of PFTS in SF-pCD pigs. All together, PFTS is a clinical syndrome with consistent pathological and serum analytical changes among affected pigs. Despite the efforts of this research to establish an infectious etiology, there is a lack of evidence that PFTS is an infectious disease.
|
Page generated in 0.0534 seconds