+publisher:"University of Saskatchewan" +contributor:("Misra, Vikram")

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University of Saskatchewan

1. Balachandran, Yadu. EXPRESSION AND CHARACTERIZATION OF TOLL-LIKE RECEPTOR 10.

Degree: 2016, University of Saskatchewan

URL: http://hdl.handle.net/10388/ETD-2016-03-2457

► Toll-like receptors (TLRs), named after toll proteins identified in Drosophila melanogaster, are the pattern recognition receptors in the innate immune system that detect microbes. TLRs… (more)

▼ Toll-like receptors (TLRs), named after toll proteins identified in Drosophila melanogaster, are the pattern recognition receptors in the innate immune system that detect microbes. TLRs are mono, membrane-spanning, as well as non-catalytic receptors, which are mainly expressed in sentinel cells, such as the dendritic cells, neutrophils and macrophages. While humans have ten TLRs (TLR 1 to 10), the mouse has another three (TLRs 11, 12, 13). TLRs are made up of glycoproteins, which have luminal ligand-binding sites consisting of leucine-rich repeat (LRR) for detection of pathogens leading to activation of immune cells. TLR1, 2, 4, and 6 are responsible for recognition of lipids (such as triacetylated lipopeptide), peptidoglycan, and lipopolysaccharide (LPS). However, the TLR3, 7, 8, and 9 mainly recognize nucleic acids, such as double-stranded RNA (dsRNA) and CpG DNA, while the TLR13 detects ribosomal RNA sequences. So far, there are no data on the localization and immunological functions of TLR10. I studied the expression, localization and role of TLR10 in S. pneumoniae infection. First, I examined the expression of TLR10 in lungs of pig, cattle, dog, rat, and chickens. The light and electron microscopic data show TLR10 expression in vascular endothelium and smooth muscles in lungs of control and inflamed animals. Further, we found altered basal level of expression and localization of TLR10 in bovine neutrophils treated with E. coli lipopolysaccharide. These data show the expression of TLR10 in the lungs of tested animal species, and its alteration by LPS in bovine neutrophils. The next study was designed to investigate the regulation of TLR10 expression and to address its role in neutrophil chemotaxis. E. coli LPS activated human neutrophils showed temporal and spatial change in TLR10 expression. Confocal microscopy showed cytosolic and nuclear distribution of TLR10 in normal and activated neutrophils. TLR10 in E. coli LPS-activated neutrophils colocalized with flotallin-1, a lipid raft marker, and EEA-1, an early endosomal marker, suggested its endocytosis. Live cell imaging of LPS activated neutrophils showed TLR10 translocation to the leading edge. Neutrophils upon TLR10 knockdown were unable for fMLP-induced migration. TLR10 knockdown reduced the number of membrane pseudopods in activated neutrophils without altering the expression of key proteins of actin nucleation process, ARP-3 and Diap1. These data show TLR4-mediated pathway for regulation of TLR10 expression, and that TLR10 may have a role in neutrophil chemotaxis. Next, I examined the role of TLR10 in innate immune response to S. pneumoniae infection in U937 human macrophage cell line. S. pneumoniae are major causative agents of pneumonia, meningitis and bacteremia. A significant increase in TLR10 mRNA expression was found in S. pneumoniae (107 cfu for 6hr) challenged macrophages. TLR10 knockdown significantly reduced production of IL-1β, IL-8, IL-17 and TNF-α and no significant change in IL-10 expression, and also significantly diminished nuclear… Advisors/Committee Members: Singh, Baljit, Misra, Vikram, Mutwiri, George, Simko, Elemir.

Subjects/Keywords: Toll-like receptors; neutrophils; macrophages; chemotaxis; inflammation; Streptococcus pneumoniae

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APA (6^th Edition):

Balachandran, Y. (2016). EXPRESSION AND CHARACTERIZATION OF TOLL-LIKE RECEPTOR 10. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/ETD-2016-03-2457

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Balachandran, Yadu. “EXPRESSION AND CHARACTERIZATION OF TOLL-LIKE RECEPTOR 10.” 2016. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/ETD-2016-03-2457.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Balachandran, Yadu. “EXPRESSION AND CHARACTERIZATION OF TOLL-LIKE RECEPTOR 10.” 2016. Web. 08 Mar 2021.

Vancouver:

Balachandran Y. EXPRESSION AND CHARACTERIZATION OF TOLL-LIKE RECEPTOR 10. [Internet] [Thesis]. University of Saskatchewan; 2016. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/ETD-2016-03-2457.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Balachandran Y. EXPRESSION AND CHARACTERIZATION OF TOLL-LIKE RECEPTOR 10. [Thesis]. University of Saskatchewan; 2016. Available from: http://hdl.handle.net/10388/ETD-2016-03-2457

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Saskatchewan

2. Mohammadzadeh, Nazanin 1990-. ROLE OF APOBEC3 ENZYMES IN HIV-1 EVOLUTION.

Degree: 2018, University of Saskatchewan

URL: http://hdl.handle.net/10388/11676

► There are approximately 36.7 million people living with HIV while 1.8 million become newly infected every year. HIV belongs to the Retroviridae family and is… (more)

▼ There are approximately 36.7 million people living with HIV while 1.8 million become newly infected every year. HIV belongs to the Retroviridae family and is an enveloped, positive sense (+) single stranded RNA virus. HIV has two subtypes, HIV-1 and HIV-2. HIV-1 is responsible for 95% of infections worldwide and is the focus of this thesis. There has not been any report on a conventional cure of this disease and the diagnosed HIV+ individuals need to receive lifelong treatment with antiretroviral drugs. Besides the antiretroviral drugs, there are host restriction factors that fight the infection in addition to the conventional immune barriers and responses. The APOBEC3 (A3) family of enzymes are part of an intrinsic immune system in humans and can act as host restriction factors to restrict the replication of HIV in CD4+ T cells by deaminating cytosine to uracil on the (-) DNA of HIV during reverse transcription. This promutagenic activity, if it occurs frequently enough, can cause viral hypermutation and inactivation. The A3 family contains seven members (A3A to A3H, excluding A3E) some of which developed gene variations that result in protein polymorphisms due to selective pressure over evolutionary time. These polymorphisms acquired enhanced antiretroviral activity to fight off HIV. To combat this, HIV has an accessory protein termed virion infectivity factor (Vif) that interacts with A3 enzymes and components of a Cullin5 E3 ligase complex. This complex causes ubiquitination and degradation of A3 enzymes in HIV infected host cells. HIV also has a high level of genetic variation that allows the virus to escape immunological and pharmacological barriers. Besides the lack of proofreading activity of reverse transcriptase, viral recombination and high rate of replication, the A3 enzymes have a potential role in increasing the genetic diversity of the HIV replicating pool if Vif mediated degradation is not complete. As a result, human A3 enzymes can contribute to this diversity directly by causing sublethal mutations and indirectly when the lethally mutated RNA genomes are “rescued” after virus recombination. While A3 enzymes may have this potential, and some research has found that A3 enzymes cause drug resistance mutations, other research groups have reported that in the presence of Vif the mutation rate of A3s is lower than the mutation rate of reverse transcriptase and their contribution to HIV evolution is not significant. To identify all the contributors to genetic variation of HIV and have a reliable answer for the current controversy in the field we aimed to determine the mutation rate of A3s in wild-type HIV-1 infection and investigate if A3-induced mutations can yield drug resistant HIV-1 variants. Moreover, we aimed to study the restriction abilities of A3F allele variants carrying a single nucleotide polymorphism (SNP). We chose the SNP that causes the A3F 231I/V polymorphism because most people carry the heterozygous alleles. To test if A3 enzymes could contribute to HIV-1 evolution, we established a… Advisors/Committee Members: Chelico, Linda, Bull, Harold, Xiao, Wei, Misra, Vikram.

Subjects/Keywords: APOBEC3F; APOBEC3G; drug resistance; HIV-1; mutagenesis; restriction factors; antiretroviral therapy

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APA (6^th Edition):

Mohammadzadeh, N. 1. (2018). ROLE OF APOBEC3 ENZYMES IN HIV-1 EVOLUTION. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/11676

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Mohammadzadeh, Nazanin 1990-. “ROLE OF APOBEC3 ENZYMES IN HIV-1 EVOLUTION.” 2018. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/11676.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Mohammadzadeh, Nazanin 1990-. “ROLE OF APOBEC3 ENZYMES IN HIV-1 EVOLUTION.” 2018. Web. 08 Mar 2021.

Vancouver:

Mohammadzadeh N1. ROLE OF APOBEC3 ENZYMES IN HIV-1 EVOLUTION. [Internet] [Thesis]. University of Saskatchewan; 2018. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/11676.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Mohammadzadeh N1. ROLE OF APOBEC3 ENZYMES IN HIV-1 EVOLUTION. [Thesis]. University of Saskatchewan; 2018. Available from: http://hdl.handle.net/10388/11676

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Saskatchewan

3. Murcia Rodriguez, Diana 1990-. INFLUENZA A VIRUSES OF SWINE IN WESTERN CANADIAN PIGS AND PEOPLE.

Degree: 2018, University of Saskatchewan

URL: http://hdl.handle.net/10388/9707

► Influenza A viruses (IAV) are well-known for their zoonotic potential, and the health and economic threats they pose to humans and pigs. The complexity of… (more)

▼ Influenza A viruses (IAV) are well-known for their zoonotic potential, and the health and economic threats they pose to humans and pigs. The complexity of influenza virus ecology involving genetically variant viral strains and several natural hosts means that the virus continuously challenges the host-species barrier. Surveillance of IAV is essential as it provides helpful information that can lead to a better understanding of the behavior of the virus at the animal-human interface, the risk factors and the key genetic changes that allow the virus to cross the species barrier. The research aimed to compare the suitability of samples collected for the detection of IAV in swine and to identify the epidemiological and viral factors that might play a fundamental role in the human-swine interface of transmission. The suitability of three types of samples for the detection of IAV in pigs, nasal swabs (NS), oral fluids (OF) and oral swabs (OS), was compared. IAV Matrix gene PCR results showed NS were the most effective method of IAV detection in swine. Compared to NS, OS had a relative sensitivity of 43.6% to 43.8% and relative specificity of 99.3% to 100%. The relative sensitivity and specificity of OF was 57.1% and 95.5%, respectively. Furthermore, the degree of agreement between NS and the other two samples was moderate (k = 0.531-0.583, p < 0.001). Human-swine transmission was evaluated through a pilot project consisting of active surveillance in both swine workers and pigs from 11 farms in Western Canada. Nasal swabs, OS, and surveys assessing flu-like symptoms were collected from 26 swine workers and results were compared with Matrix real-time reverse transcriptase PCR (RT-qPCR) results from swine nasal swabs. There was no statistically significant correlation between the clinical symptoms in humans and the RT-qPCR results from swine samples. However, the IAV Matrix gene PCR results from the NS and OS of the swine workers had a very weak correlation with the results found in swine (r = 0.182-0.200, p = 0.024- 0.040). Transmission among species was not confirmed, but samples with suspect results from human samples coincided with positive swine pool results and the presence of an Alpha H1N2 virus in 4 farms, which is suggestive of a common link between humans and pigs for IAV. Advisors/Committee Members: Detmer, Susan E, Harding, John, Kirychuk, Shelley, Misra, Vikram.

Subjects/Keywords: Influenza A virus; swine; human; surveillance

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APA (6^th Edition):

Murcia Rodriguez, D. 1. (2018). INFLUENZA A VIRUSES OF SWINE IN WESTERN CANADIAN PIGS AND PEOPLE. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/9707

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Murcia Rodriguez, Diana 1990-. “INFLUENZA A VIRUSES OF SWINE IN WESTERN CANADIAN PIGS AND PEOPLE.” 2018. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/9707.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Murcia Rodriguez, Diana 1990-. “INFLUENZA A VIRUSES OF SWINE IN WESTERN CANADIAN PIGS AND PEOPLE.” 2018. Web. 08 Mar 2021.

Vancouver:

Murcia Rodriguez D1. INFLUENZA A VIRUSES OF SWINE IN WESTERN CANADIAN PIGS AND PEOPLE. [Internet] [Thesis]. University of Saskatchewan; 2018. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/9707.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Murcia Rodriguez D1. INFLUENZA A VIRUSES OF SWINE IN WESTERN CANADIAN PIGS AND PEOPLE. [Thesis]. University of Saskatchewan; 2018. Available from: http://hdl.handle.net/10388/9707

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Saskatchewan

4. Subudhi, Sonu 1989-. North American bats and their viruses: The effect of stressors on persistent infections and viral shedding.

Degree: 2019, University of Saskatchewan

URL: http://hdl.handle.net/10388/12098

► There is direct or circumstantial evidence that several viruses that cause no obvious disease in bats have spilled over into humans and other species causing… (more)

▼ There is direct or circumstantial evidence that several viruses that cause no obvious disease in bats have spilled over into humans and other species causing serious and often fatal disease. The reasons for the lack of disease in bats or for the spillover of these viruses from bats are poorly understood. While there is considerable literature on the interactions of these viruses with their secondary hosts or their surrogates, little is known about the interactions of bat viruses in their natural hosts. We used a coronavirus detected in little brown bat (Myotis lucifugus) and a herpesvirus, detected in the big brown bat (Eptesicus fuscus), as models to understand the factors that might alter bat-virus relationships. We demonstrated that a coronavirus (Myotis lucifugus coronavirus – Myl-CoV) detected in the intestines of little brown bats, could persist in them during the 4 months of hibernation. Using this coronavirus-bat model, we showed that the stress of fungal infection by Pseudogymnoascus destructans (Pd), which causes bat white-nose syndrome (WNS), led to a 60-fold increase in viral replication in intestines than bats with virus alone. Increased viral replication correlated with the severity of Pd-related pathology and the intestine of fungus-infected bats showed changes in gene expression suggesting suppressed innate antiviral response and increased apoptotic responses. Our results suggest that the systemic effects of WNS leads to a resurgence of virus replication and increases the potential of virus shedding. Using a bat cell culture model, we showed that viral persistence could be disrupted by artificially suppressing the host cell’s antiviral response and was mediated through similar pathways that were observed during in-vivo experiments. To ascertain whether the effect of stressor could disrupt viral persistence in other bat-virus relationships, I studied the big brown bat herpesvirus. As herpesviruses inherently establish life-long latent infections in their hosts and reactivate periodically in response to stress, we used this model to study the effects of natural stressors on the bat-virus relationship. We characterized the herpesvirus and developed techniques for detecting the virus as well as for monitoring the adaptive antibody response against the virus. We showed that the bat gammaherpesvirus reactivates at the end of hibernation and was accompanied by a lower antibody level, which subsequently increased upon arousal. Our studies on coronavirus and herpesvirus show that bats have a long-term balanced and benign relationships with viruses and a variety of stressors could disrupt this balance allowing an increase in viral replication. Advisors/Committee Members: Misra, Vikram, Zhou, Yan, Falzarano, Darryl, Bollinger, Trent K, Rubin, Joseph.

Subjects/Keywords: bats; virus; stress; persistent infection; viral shedding; coronavirus; gammaherpesvirus; fungal infection; hibernation; eptesicus fuscus; myotis lucifugus; pseudogymnoascus destructans; MAP kinase; interferon

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Subudhi, S. 1. (2019). North American bats and their viruses: The effect of stressors on persistent infections and viral shedding. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/12098

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Subudhi, Sonu 1989-. “North American bats and their viruses: The effect of stressors on persistent infections and viral shedding.” 2019. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/12098.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Subudhi, Sonu 1989-. “North American bats and their viruses: The effect of stressors on persistent infections and viral shedding.” 2019. Web. 08 Mar 2021.

Vancouver:

Subudhi S1. North American bats and their viruses: The effect of stressors on persistent infections and viral shedding. [Internet] [Thesis]. University of Saskatchewan; 2019. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/12098.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Subudhi S1. North American bats and their viruses: The effect of stressors on persistent infections and viral shedding. [Thesis]. University of Saskatchewan; 2019. Available from: http://hdl.handle.net/10388/12098

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Saskatchewan

5. Bodnarchuk, Timothy. Zhangfei suppresses the growth of Medulloblastoma cells and commits them to programmed cell death.

Degree: 2011, University of Saskatchewan

URL: http://hdl.handle.net/10388/etd-06222011-145925

► Medulloblastoma cells do not contain detectable amounts of the bZIP protein Zhangfei. However, previous work has shown that expression of this protein in cells of… (more)

▼ Medulloblastoma cells do not contain detectable amounts of the bZIP protein Zhangfei. However, previous work has shown that expression of this protein in cells of the ONS-76 line, derived from a human medulloblastoma, causes the cells to stop growing and develop processes that resemble neuritis (a characteristic of differentiated neurons). Zhangfei-expressing cells eventually die. My objective was to determine the molecular mechanisms by which Zhangfei influences ONS-76 cells. My strategy was to infect ONS-76 cells with adenovirus vectors expressing either Zhangfei or the control E. coli protein â-galactosidase (LacZ) and then to compare the following parameters in Zhangfei and LacZ-expressing cells: a) markers of apoptosis, autophagy and macropinocytosis (the three main pathways of cell death); b) transcripts for genes involved in neurogenesis and apoptosis; c) phosphorylation of peptide targets of selected cellular protein kinases; and d) active transcription factors. Zhangfei-expressing cells appeared to succumb to apoptosis as determined by the expression of phosphatidylserine on the cell surface and intensity of nuclear staining with the DNA dye Hoechst. Increased staining for autophagic vesicles and upregulated expression of autophagy response genes in these cells indicated that they were undergoing autophagy, possibly associated with apoptosis. My analysis of steady-state transcripts for genes involved in apoptosis and neurogenesis and functional protein kinases in Zhangfei-expressing cells indicated that the mitogen-activated protein kinase (MAPK) pathway was active in these cells. In addition, I found that the transcription factor Brn3a as well as factors implicated in differentiation were also active. These observations led me to hypothesize that Zhangfei enhances the expression of Brn3a, a known inducer of TrkA, the high-affinity receptor for nerve growth factor (NGF). TrkA then binds in an autocrine manner to NGF, triggering the MAPK pathway and leading to differentiation of ONS-76 cells into neuron and glia-like cells, eventually bringing about cell death by apoptosis and autophagy. I tested this hypothesis by showing that Zhangfei could enhance transcription from the isolated Brn3a promoter, that ONS-76 cells produce NGF as detected in a bioassay, and that antibodies against NGF and inhibitors of TrkA and selected components of the MAPK pathway could partially restore the growth of Zhangfei-expressing ONS-76 cells. My work supports previous work highlighting the importance of NGF-TrkA signaling in the outcome of medulloblastomas and shows how Zhangfei is able to trigger this pathway. Advisors/Committee Members: Misra, Vikram, Mousseau, Darrell, Napper, Scott, Bonham, Keith, Hill, Janet.

Subjects/Keywords: Zhangfei; autophagy; apoptosis; Brn3a; medulloblastoma; MapK; TrkA; macropinocytosis

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Bodnarchuk, T. (2011). Zhangfei suppresses the growth of Medulloblastoma cells and commits them to programmed cell death. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/etd-06222011-145925

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Bodnarchuk, Timothy. “Zhangfei suppresses the growth of Medulloblastoma cells and commits them to programmed cell death.” 2011. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/etd-06222011-145925.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Bodnarchuk, Timothy. “Zhangfei suppresses the growth of Medulloblastoma cells and commits them to programmed cell death.” 2011. Web. 08 Mar 2021.

Vancouver:

Bodnarchuk T. Zhangfei suppresses the growth of Medulloblastoma cells and commits them to programmed cell death. [Internet] [Thesis]. University of Saskatchewan; 2011. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/etd-06222011-145925.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Bodnarchuk T. Zhangfei suppresses the growth of Medulloblastoma cells and commits them to programmed cell death. [Thesis]. University of Saskatchewan; 2011. Available from: http://hdl.handle.net/10388/etd-06222011-145925

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Saskatchewan

6. Liu, GuanQun 1988-. THE DYNAMIC INTERPLAY BETWEEN INFLUENZA A VIRUS RNA SIGNATURES AND RETINOIC ACID-INDUCIBLE GENE I.

Degree: 2018, University of Saskatchewan

URL: http://hdl.handle.net/10388/11678

► Influenza A virus (IAV) remains a seemingly undefeatable public health threat owing to its frequent antigenic drift and shift. Seasonal epidemics, occasional pandemics, and avian/zoonotic… (more)

▼ Influenza A virus (IAV) remains a seemingly undefeatable public health threat owing to its frequent antigenic drift and shift. Seasonal epidemics, occasional pandemics, and avian/zoonotic IAV outbreaks cause significant morbidity and mortality among human and other hosts, leading to substantial global economic burdens. Along the path of host-virus coevolution, mammalian hosts have evolved sophisticated cellular networks to combat viral infection, among which the innate immune system constitutes the first line of defense. Its rapid detection of the invading virus initiates an effective antiviral response which in turn primes a specific adaptive immunity. Therefore, an in-depth understanding of the interplay between IAV and host innate immune system not only provides knowledge on IAV pathogenesis but also sheds light on the development of novel approaches to combatting IAV infection. The innate immune recognition of IAV relies on various families of pattern recognition receptors, among which the retinoic acid-inducible gene I (RIG-I), a founding member of the RIG-I-like receptor (RLR) family, is indispensable for the IAV-induced type I interferon (IFN) response. It has been well characterized as a cytoplasmic sensor that recognizes short double-stranded (ds) RNA harboring a 5’-triphosphate moiety. Interestingly, IAV bears a single-stranded RNA genome and does not generate a detectable amount of dsRNA during infection. Over the last decade, the panhandle structure, which is formed from the partial complementarity of the genomic RNA extremities of negative-strand RNA viruses, has been proposed to meet the dsRNA requirement for RIG-I activation. However, it remains unsubstantiated the contribution of panhandle structure in the genomic context of IAV to RIG-I activation. Therefore, in the first part of this thesis, I investigated whether the IAV panhandle structure is directly involved in RIG-I activation and type I IFN induction. Using reconstituted IAV genomic RNA with coding region truncations, it was demonstrated that the viral genomic coding region is dispensable for RIG-I-dependent IFN induction in primary alveolar macrophages. The in vitro-synthesized IAV panhandle RNA directly binds to RIG-I with 1:1 stoichiometry and stimulates RIG-I ATPase activity in vitro and RIG-I-dependent IFNβ promoter activation in DF-1 cells. These lines of evidence demonstrate a direct involvement of the IAV panhandle structure in RIG-I activation. Since the wild-type panhandle structure exhibits an imperfect double-strandedness containing wobble base pairs, mismatch, and bulged nucleotide, it was of particular interest to determine how these elements contribute to RIG-I activation. Elimination of these destabilizing elements from either the 5’ or 3’ arm of the panhandle structure enhances RIG-I binding and ATPase activity in vitro, and promotes RIG-I-dependent IFN induction in DF-1 cells and primary macrophages. Given that the IAV panhandle structure also serves as the viral promoter region, the promoter activity of panhandle-stabilized… Advisors/Committee Members: Zhou, Yan, Tikoo, Suresh, Wilson, Joyce, Liu, Qiang, Misra, Vikram.

Subjects/Keywords: influenza A virus; innate immunity; RIG-I; interferon; panhandle; RNA sensor; nonself; replication; nucleus; Hepatitis B virus; aberrant RNA; RNA polymerase; defective-interfering

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Liu, G. 1. (2018). THE DYNAMIC INTERPLAY BETWEEN INFLUENZA A VIRUS RNA SIGNATURES AND RETINOIC ACID-INDUCIBLE GENE I. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/11678

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Liu, GuanQun 1988-. “THE DYNAMIC INTERPLAY BETWEEN INFLUENZA A VIRUS RNA SIGNATURES AND RETINOIC ACID-INDUCIBLE GENE I.” 2018. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/11678.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Liu, GuanQun 1988-. “THE DYNAMIC INTERPLAY BETWEEN INFLUENZA A VIRUS RNA SIGNATURES AND RETINOIC ACID-INDUCIBLE GENE I.” 2018. Web. 08 Mar 2021.

Vancouver:

Liu G1. THE DYNAMIC INTERPLAY BETWEEN INFLUENZA A VIRUS RNA SIGNATURES AND RETINOIC ACID-INDUCIBLE GENE I. [Internet] [Thesis]. University of Saskatchewan; 2018. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/11678.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Liu G1. THE DYNAMIC INTERPLAY BETWEEN INFLUENZA A VIRUS RNA SIGNATURES AND RETINOIC ACID-INDUCIBLE GENE I. [Thesis]. University of Saskatchewan; 2018. Available from: http://hdl.handle.net/10388/11678

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

7. Costa De Freitas, Aline 1987-. Characterization of the vaginal microbiome in pregnancy.

Degree: 2018, University of Saskatchewan

URL: http://hdl.handle.net/10388/8443

► The vaginal microbiome plays an important role in women's reproductive health. Imbalances in this microbiota are associated with bacterial vaginosis, increased susceptibility to sexually transmitted… (more)

▼ The vaginal microbiome plays an important role in women's reproductive health. Imbalances in this microbiota are associated with bacterial vaginosis, increased susceptibility to sexually transmitted infections, and negative reproductive outcomes. The causes of such variations, however, are poorly understood. A healthy vaginal microbiota is defined as Lactobacillus-dominated and an overgrowth of other species is often associated with unhealthy conditions. An appreciation of "atypical" microbiomes in healthy women, such as Bifidobacterium-dominated, has been gradually increasing. Although bifidobacteria play an important role in gut health, vaginal bifidobacteria have not yet been fully characterized. In this study, a baseline description of the "healthy" vaginal microbiome in pregnancy has been established based on cpn60 gene amplicon sequencing. The vaginal microbiota of pregnant women relative to non-pregnant women had lower richness and diversity, higher Lactobacillus abundance and lower Mollicutes/Ureaplasma prevalence. This gives a better understanding of the vaginal microbiome in healthy pregnancies and provides a control group for a subsequent comparison to women who experienced preterm birth. An association between Mollicutes and preterm was confirmed, and further suggested that a more rich and diverse microbiome is associated with prematurity. To better understand the relationship between reproductive outcomes and microbiota, an improved definition of the healthy microbiome is also needed, which should include evaluation of "atypical" microbiomes, such as Bifidobacterium-dominated. Phenotypic characterization of vaginal bifidobacteria indicated that they have health promoting characteristics similar to beneficial vaginal lactobacilli. Considering the importance of bifidobacteria as one of the primary colonizers of the neonatal gut, the genomes of vaginal and gut isolates of Bifidobacterium breve and Bifidobacterium longum were compared. Results indicated that vaginal and gut microbiomes are colonized by a shared community of Bifidobacterium, which may be transferred from mother to infant. Taken together, the results presented in this thesis provide a better understanding of the vaginal microbiome of pregnant women with low and high risk for preterm birth. It also improves the understanding of a healthy microbiome by phenotypically characterizing vaginal bifidobacteria, and contributes to elucidate aspects of bifidobacteria ecology by comparing the genomes of vaginal and gut bifidobacteria. Advisors/Committee Members: Hill, Janet E, Misra, Vikram, Rubin, Joseph, White, Aaron, Dilon, Jo-Anne.

Subjects/Keywords: Vaginal microbiome; pregnancy; bifidobacteria; genome; preterm birth

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APA (6^th Edition):

Costa De Freitas, A. 1. (2018). Characterization of the vaginal microbiome in pregnancy. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/8443

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Costa De Freitas, Aline 1987-. “Characterization of the vaginal microbiome in pregnancy.” 2018. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/8443.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Costa De Freitas, Aline 1987-. “Characterization of the vaginal microbiome in pregnancy.” 2018. Web. 08 Mar 2021.

Vancouver:

Costa De Freitas A1. Characterization of the vaginal microbiome in pregnancy. [Internet] [Thesis]. University of Saskatchewan; 2018. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/8443.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Costa De Freitas A1. Characterization of the vaginal microbiome in pregnancy. [Thesis]. University of Saskatchewan; 2018. Available from: http://hdl.handle.net/10388/8443

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Saskatchewan

8. Rubin, Joseph Elliot. Prevalence of colonization and antimicrobial resistance among coagulase positive staphylococci in dogs, and the relatedness of canine and human Staphylococcus aureus.

Degree: 2011, University of Saskatchewan

URL: http://hdl.handle.net/10388/etd-06242011-104449

► Coagulase positive staphylococci, Staphylococcus aureus and Staphylococcus pseudintermedius, are important causes of infection in human beings and dogs respectively. The rapid increase in the incidence… (more)

▼ Coagulase positive staphylococci, Staphylococcus aureus and Staphylococcus pseudintermedius, are important causes of infection in human beings and dogs respectively. The rapid increase in the incidence of methicillin resistant S. aureus (MRSA) in people and its emergence in dogs has raised the profile of this organism in the veterinary community. Similarly, human S. pseudintermedius infections have also been recognized as the awareness of bidirectional human-dog transmission increases. Antimicrobial resistance has been complicating the treatment of S. aureus infections since the first penicillin resistance was observed in the 1940s. Methicillin resistance (resistance to the majority of â-lactams), is particularly troublesome as the â-lactams are a safe and effective class of antimicrobials for treating susceptible staphylococcal infections in both human beings and dogs. Additionally, resistance to other antimicrobial classes such as the macrolides, tetracyclines, sulfonamides and chloramphenicol, further complicates the treatment of staphylococcal infections. Particularly in small animal private practice, infections are often treated empirically, requiring knowledge of locally prevalent susceptibility patterns. The emergence of resistance to commonly used drugs necessitates surveillance to monitor the dissemination of resistance, and to guide antimicrobial therapy. In the last decade there have been many studies attempting to address gaps in our knowledge of the ecology of S. aureus and S. pseudintermedius in dogs. In particular, the prevalence of colonization with methicillin resistant staphylococci has been documented in different dog populations. However, failing to sample all relevant sites of colonization, may have decreased the sensitivity of these studies. The sites where coagulase positive staphylococci colonize dogs have not been systematically evaluated. The clinical and infection control implications of S. aureus infections, or colonization in the case of MRSA, requires timely laboratory identification. The tube coagulase test is arguably the most important tool used for identifying of staphylococcal species. Studies dating from the 1970s and 1980s suggested that the use of rabbit plasma, which is the current standard, may not be the ideal media for all situations and that different plasmas may need to be considered in different diagnostic situations. In this thesis, the ecology of coagulase positive staphylococci in dogs was studied from start to finish including sample collection, bacterial identification, antimicrobial susceptibility testing and molecular epidemiological investigations. This thesis will serve as a template to be used for follow up studies or by investigators setting up a surveillance program in their region. We found that multiple sites of colonization (nares, pharynx and rectum), are involved in both S. aureus and S. pseudintermedius carriage in dogs. Single site colonized dogs were identified, suggesting that maximal screening sensitivity requires sampling… Advisors/Committee Members: Sanche, Stephen, Chirino-Trejo, Manuel, Hill, Janet, Wong, Alice, Misra, Vikram, Clark, Chris.

Subjects/Keywords: zoonoses; methicillin; MRSA; Staphylococcus pseudintermedius; canine; bacteriology; antimicrobial resistance; Staphylococcus aureus

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APA (6^th Edition):

Rubin, J. E. (2011). Prevalence of colonization and antimicrobial resistance among coagulase positive staphylococci in dogs, and the relatedness of canine and human Staphylococcus aureus. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/etd-06242011-104449

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Rubin, Joseph Elliot. “Prevalence of colonization and antimicrobial resistance among coagulase positive staphylococci in dogs, and the relatedness of canine and human Staphylococcus aureus.” 2011. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/etd-06242011-104449.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Vancouver:

Rubin JE. Prevalence of colonization and antimicrobial resistance among coagulase positive staphylococci in dogs, and the relatedness of canine and human Staphylococcus aureus. [Internet] [Thesis]. University of Saskatchewan; 2011. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/etd-06242011-104449.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Rubin JE. Prevalence of colonization and antimicrobial resistance among coagulase positive staphylococci in dogs, and the relatedness of canine and human Staphylococcus aureus. [Thesis]. University of Saskatchewan; 2011. Available from: http://hdl.handle.net/10388/etd-06242011-104449

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Saskatchewan

9. McIntosh, Kathleen. Molecular techniques in the study and control of porcine circovirus type 2.

Degree: 2011, University of Saskatchewan

URL: http://hdl.handle.net/10388/ETD-2011-10-130

► Porcine circovirus type 2 (PCV2) is an emerging virus that may result in devastating disease that affects swine herds worldwide. Only a decade has passed… (more)

▼ Porcine circovirus type 2 (PCV2) is an emerging virus that may result in devastating disease that affects swine herds worldwide. Only a decade has passed since researchers began to study the characteristics of the virus and the resulting diseases, and limited information was available regarding the long term presence or persistence of the virus in healthy swine herds. Our research contributes to the knowledge of the persistence of the virus in serum and semen, and the PCV2 antibody profile in healthy pigs. In addition, we developed a novel quantitative polymerase chain reaction (PCR) assay and quantified the PCV2-shedding in healthy and PCVD (porcine circovirus disease)-affected pigs. Lastly, we determined the efficacy of a novel vaccine in a subset of pigs from a PCVD-affected herd. Porcine serum was assayed by two PCR protocols (nested polymerase chain reaction (nPCR) and non-nested PCR) and a competitive enzyme-linked immunosorbent assay (cELISA) to determine when PCV2 viremia and a rise in the serum level of PCV2-specific antibody (Ab) occurred in pigs raised in a large Canadian farrow-to-finish barn. Eight serial blood samples were collected from each of 40 pigs from 5 to 156 (±1.5) days of age; six pigs were removed from the study for various reasons at various times. Viremia was not detected in the samples collected before 72 days of age but was detected in those collected on or after 72 days: of 33 pigs, seven (21%) had only one serum sample positive for PCV2 deoxyribonucleic acid (DNA) by nPCR after day 72; 11 (33%) were intermittently positive by nPCR, non-nested PCR, or both between 72 and 156 days; and the remaining 15 (45%) were repeatedly positive (in two to four samples). The level of serum Ab against PCV2 declined after weaning and increased between 72 and 107 days of age, only after PCV2 was detected in serum. Our results show that PCV2 viremia persists in the presence of elevated levels of PCV2-specific Ab. In a separate study, we determined the long term presence or persistence of PCV2-shedding in semen from healthy boars and the effects of PCV2 on sperm quality. A nPCR protocol was applied to porcine semen to demonstrate the PCV2-shedding patterns and duration in naturally-infected boars. Sperm morphology analysis was performed on a subset of samples to determine if the presence of PCV2 DNA in semen was associated with reduced semen quality. Semen was collected serially from 43 boars representing six breeds, aged 33.9 to 149.3 weeks. Of the 903 semen samples collected, 30 samples (3.3%) were positive for PCV2 DNA by nPCR from 13 boars. Boars shedding PCV2 DNA in semen ranged between 35.9 and 71.0 weeks of age, and shedding occurred over a period of up to 27.3 weeks. A semen nPCR test was 2.6 times more likely to be positive when collected from pigs that were ≤52 weeks of age and 3.0 times more likely to be positive when collected from pigs that were ≤26 weeks from the time of entry into the stud main unit (Generalized Estimating Equations: P=0.02; 95% confidence interval (CI) of the Odd’s ratio… Advisors/Committee Members: Ellis, John A., Harding, John, Hill, Janet E., Misra, Vikram, Steven, Krakowka.

Subjects/Keywords: polymerase chain reaction (PCR); porcine circovirus type 2 (PCV2); porcine circovirus disease (PCVD); immune stimulating complex (ISCOM); swine.

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APA (6^th Edition):

McIntosh, K. (2011). Molecular techniques in the study and control of porcine circovirus type 2. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/ETD-2011-10-130

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

McIntosh, Kathleen. “Molecular techniques in the study and control of porcine circovirus type 2.” 2011. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/ETD-2011-10-130.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

McIntosh, Kathleen. “Molecular techniques in the study and control of porcine circovirus type 2.” 2011. Web. 08 Mar 2021.

Vancouver:

McIntosh K. Molecular techniques in the study and control of porcine circovirus type 2. [Internet] [Thesis]. University of Saskatchewan; 2011. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/ETD-2011-10-130.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

McIntosh K. Molecular techniques in the study and control of porcine circovirus type 2. [Thesis]. University of Saskatchewan; 2011. Available from: http://hdl.handle.net/10388/ETD-2011-10-130

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Saskatchewan

10. Banerjee, Arinjay 1990-. Dynamics of bat-coronavirus interactions: role of innate antiviral responses.

Degree: 2018, University of Saskatchewan

URL: http://hdl.handle.net/10388/9541

► Bats are speculated to be reservoirs of several emerging viruses including coronaviruses (CoVs) that cause severe acute respiratory syndrome (SARS), Middle-East respiratory syndrome (MERS), porcine… (more)

▼ Bats are speculated to be reservoirs of several emerging viruses including coronaviruses (CoVs) that cause severe acute respiratory syndrome (SARS), Middle-East respiratory syndrome (MERS), porcine epidemic diarrhea and swine acute diarrhea syndrome. These viruses cause significant disease in humans and agricultural animals. MERS-CoV causes serious disease in humans with a thirty-five percent mortality and has evolved proteins that can effectively suppress an innate antiviral response in human cells. Bats that are naturally or experimentally infected with these or similar viruses do not show apparent signs of disease and the molecular mechanisms of protection are not yet known. My doctoral thesis tested the hypothesis that big brown bat cells have unique adaptations in innate antiviral signaling pathways involved in the control of virus replication and coronavirus-induced inflammatory cytokines. To test this hypothesis, we generated the first commercially available North American bat (Eptesicus fuscus; big brown bat) kidney epithelial cell line. Using this cell line, we were able to demonstrate that big brown bat cells have evolved a unique repressor molecule, c-Rel that can effectively suppress double-stranded RNA (poly(I:C)) mediated expression of a key inflammatory cytokine, tumor necrosis factor alpha (TNF). MERS-CoV is thought to have evolved in insectivorous bats before spilling over to camels and eventually to humans. To further our understanding about bat-coronavirus interactions, we demonstrated that big brown bat cells are resistant to MERS-CoV-mediated subversion of antiviral responses. We determined that interferon regulatory factor 3 (IRF3) plays a critical role in controlling MERS-CoV propagation in big brown bat epithelial cells. Indeed, my doctoral thesis has identified two unique adaptations in big brown bat cells that might allow these bats, and probably other species of bats to successfully co-exist with coronaviruses. My thesis supports the hypothesis that bats function as global reservoirs for emerging coronaviruses by providing definitive examples of adaptations that would allow bats to co-exist with these viruses. Future work from my thesis will focus on adapting some of these antiviral strategies in human cells to control coronavirus-mediated disease in humans. Advisors/Committee Members: Misra, Vikram, Griebel, Philip, Ellis, John, Falzarano, Darryl, Bollinger, Trent, Rubin, Joseph.

Subjects/Keywords: Bats; coronavirus; anti-viral defence responses

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APA (6^th Edition):

Banerjee, A. 1. (2018). Dynamics of bat-coronavirus interactions: role of innate antiviral responses. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/9541

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Banerjee, Arinjay 1990-. “Dynamics of bat-coronavirus interactions: role of innate antiviral responses.” 2018. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/9541.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Banerjee, Arinjay 1990-. “Dynamics of bat-coronavirus interactions: role of innate antiviral responses.” 2018. Web. 08 Mar 2021.

Vancouver:

Banerjee A1. Dynamics of bat-coronavirus interactions: role of innate antiviral responses. [Internet] [Thesis]. University of Saskatchewan; 2018. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/9541.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Banerjee A1. Dynamics of bat-coronavirus interactions: role of innate antiviral responses. [Thesis]. University of Saskatchewan; 2018. Available from: http://hdl.handle.net/10388/9541

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Saskatchewan

11. Wisner, Amanda Lynn Stacy. Role of Salmonella enterica subspecies enterica serovar Enteritidis pathogenicity island-2 in chickens.

Degree: 2011, University of Saskatchewan

URL: http://hdl.handle.net/10388/etd-07152011-191604

► Salmonella enterica subspecies enterica serovar Enteritidis (S. Enteritidis) has been identified as a significant cause of salmonellosis in humans. Salmonella pathogenicity islands 1 and 2… (more)

▼ Salmonella enterica subspecies enterica serovar Enteritidis (S. Enteritidis) has been identified as a significant cause of salmonellosis in humans. Salmonella pathogenicity islands 1 and 2 (SPI-1 and SPI-2) each encode a specialized type III secretion system (T3SS) that enables Salmonella to manipulate host cells at various stages of the invasion/infection process. The SPI-2 T3SS has been identified as vital for survival and replication of S. Typhimurium and S. Enteritidis in mouse macrophages, as well as full virulence in mice. In order to test the ability of SE SPI-2 mutants to survive in vivo we used a chicken isolate of SE (Sal18). In one study, we orally co-challenged 35-day-old specific pathogen free (SPF) chickens with two bacterial strains per group. The control group received two versions of the wild-type (WT) strain Sal18: Sal18 attTn7::tet and Sal18 attTn7::cat, while the other two groups received the WT strain (Sal18 attTn7::tet) and one of two mutant strains (Sal18 attTn7::cat ÄspaSÄssaU or Sal18 ÄSPI-1ÄSPI-2::cat). From this study we conclude that S. Enteritidis deficient in the SPI-1 and SPI-2 systems are out-competed by the WT strain. In a second study, groups of SPF chickens were challenged at 1 week of age with four different strains; a WT strain and three other strains missing either one or both of the SPI-1 and SPI-2 regions. On days 1 and 2 post-challenge (PC) we observed a reduced systemic spread of the SPI-2 mutants, but by day 3 the mutants’ systemic distribution levels matched that of the WT strain. Based on these two studies, we conclude that the SPI-2 T3SS facilitates invasion and systemic spread of S. Enteritidis in chickens, but alternative mechanisms for these processes appear to exist. Several structural components of the T3SSs encoded by SPI-1 and SPI-2 are exposed to the host’s immune system prior to/during the infection/invasion process, making them potential vaccine candidates. Several of these candidates genes were cloned, the proteins overproduced, purified, and formulated as vaccines for use in further studies. SPI-2 T3SS proteins used for vaccine studies included the secretin, SsaC, the needle, SsaG, the filament, SseB, and a part of the translocon, SseD, as well as a number of effectors, SseI, SseL, SifA, and SifB. The first vaccine study involved vaccination of SPF chickens with SseB and SseD, followed by challenge with the WT S. Enteritidis strain Sal18. Additional studies evaluated whether hens vaccinated with SPI-2 T3SS structural or effector components could mount a significant humoral immune response (as measured by serum immunoglobulin Y [IgY] titres), whether these antibodies could be transferred to progeny (as measured by egg yolk IgY titres), and whether vaccinates and progeny of vaccinates could be protected against challenge with the WT S. Enteritidis strain Sal8. The results of our studies show that vaccinated chickens do produce high levels of SPI-2 T3SS specific serum IgY that they are able to transfer to their progeny. It was demonstrated that… Advisors/Committee Members: Koester, Wolfgang, Potter, Andrew A, Deneer, Harry, Surette, Michael, Misra, Vikram, Babiuk, Lorne, Griebel, Philip.

Subjects/Keywords: Poultry Vaccine; Salmonella Pathogenicity Island 2; Salmonella; Type III Secretion System

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APA (6^th Edition):

Wisner, A. L. S. (2011). Role of Salmonella enterica subspecies enterica serovar Enteritidis pathogenicity island-2 in chickens. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/etd-07152011-191604

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Wisner, Amanda Lynn Stacy. “Role of Salmonella enterica subspecies enterica serovar Enteritidis pathogenicity island-2 in chickens.” 2011. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/etd-07152011-191604.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Wisner, Amanda Lynn Stacy. “Role of Salmonella enterica subspecies enterica serovar Enteritidis pathogenicity island-2 in chickens.” 2011. Web. 08 Mar 2021.

Vancouver:

Wisner ALS. Role of Salmonella enterica subspecies enterica serovar Enteritidis pathogenicity island-2 in chickens. [Internet] [Thesis]. University of Saskatchewan; 2011. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/etd-07152011-191604.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Wisner ALS. Role of Salmonella enterica subspecies enterica serovar Enteritidis pathogenicity island-2 in chickens. [Thesis]. University of Saskatchewan; 2011. Available from: http://hdl.handle.net/10388/etd-07152011-191604

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Saskatchewan

12. Ching, John 1987-. Novel Signaling Function of hCLCA1 in Airway Macrophage Activation.

Degree: 2015, University of Saskatchewan

URL: http://hdl.handle.net/10388/12609

► The CLCA gene family produces both secreted and membrane-associated proteins that modulate ion-channel activity, drive mucus production and have a poorly understood pleiotropic effect on… (more)

Subjects/Keywords: macrophage; CLCA; immune regulation

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APA (6^th Edition):

Ching, J. 1. (2015). Novel Signaling Function of hCLCA1 in Airway Macrophage Activation. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/12609

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Ching, John 1987-. “Novel Signaling Function of hCLCA1 in Airway Macrophage Activation.” 2015. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/12609.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Ching, John 1987-. “Novel Signaling Function of hCLCA1 in Airway Macrophage Activation.” 2015. Web. 08 Mar 2021.

Vancouver:

Ching J1. Novel Signaling Function of hCLCA1 in Airway Macrophage Activation. [Internet] [Thesis]. University of Saskatchewan; 2015. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/12609.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Ching J1. Novel Signaling Function of hCLCA1 in Airway Macrophage Activation. [Thesis]. University of Saskatchewan; 2015. Available from: http://hdl.handle.net/10388/12609

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Saskatchewan

13. Tavares Krause, Ana Rita 1985-. Ovarian Response, Follicular Function and Oocyte Developmental Competence in Gonadotropin Treated Prepubertal Calves.

Degree: 2019, University of Saskatchewan

URL: http://hdl.handle.net/10388/12490

► Sexual immaturity during the prepubertal period in cattle is characterized by low pulse frequency of LH, anovulatory waves of follicle development, absence of corpora lutea… (more)

▼ Sexual immaturity during the prepubertal period in cattle is characterized by low pulse frequency of LH, anovulatory waves of follicle development, absence of corpora lutea (i.e., progesterone) and oocytes of reduced developmental capacity in vitro when compared to oocytes of sexually mature animals. However, calf ovaries are responsive to exogenous gonadotropin treatment early in life, and the use of prepubertal animals as a source of oocytes for in vitro embryo production may have significant potential to decrease generation interval and accelerate the rate of genetic gain. The overall objective of this thesis was to investigate the effect of exogenous FSH treatment on the ovarian response, hormonal profiles, ovulation and oocyte developmental competence in prepubertal calves. In the first study (Chapter 3) the effect of cumulative dose (200 mg vs. 350 mg) and duration (4 vs. 7 days) of FSH treatment on the ovarian response and the number of spontaneous and induced ovulations in 5-month-old calves were compared. Calves (n=24) were selected for gonadotropin treatment from a group of spring-born calves (n=51) based on the antral follicle counts (AFC) at the time of wave emergence. Calves were classified in low, medium and high AFC, and the ones in the medium classification (250.8, range of 20 to 32 follicles) were used in this study. At the end of the FSH treatment and 24 hours after treatment with pLH, the number of follicles  9 mm was greater in the 7-day than in the 4-day treatment group and in calves given a cumulative dose of 350 mg of FSH compared to those given 200 mg. Spontaneous ovulations were observed in 14 calves between Day 4 of FSH treatment and 12-hours post-LH treatment. The number of total and spontaneous ovulations was higher in the 7-day treatment groups than in the 4-day groups, and the number of spontaneous ovulations was higher in calves given a cumulative dose of 200 mg FSH than 350 mg. Numbers of ovulations in response to exogenous LH did not differ among groups. In the second study (Chapter 4), data showed in prepubertal calves (n=46) that the number of follicles at the beginning of a wave was predictive of the number recruited into subsequent waves and that after FSH treatment, the number of medium and large sized follicles available for follicular aspiration was positively associated with the number of follicles  1 mm at the time of wave emergence. In calves with low (n=12) and high (n=10) AFC at wave emergence, 7 days of FSH treatment resulted in a higher number of large than small size follicles than the 4 days of FSH treatment. High AFC at wave emergence resulted in a greater number of follicles  6 mm available for aspiration and a greater number of cumulus oocyte complexes (COC) collected than low AFC. The third study (Chapter 5) was designed to investigate the relationship between the antral follicular counts and plasma concentrations of AMH and FSH at the time of wave emergence in prepubertal calves and to compare the effects of age and duration of gonadotropin treatment on… Advisors/Committee Members: Singh, Jaswant, Adams, Gregg P, Mapletoft, Reuben J, Dias, Fernanda CF, Misra, Vikram, Unniappan, Suraj.

Subjects/Keywords: follicular growth; superstimulation; puberty; oocyte competence; cattle; gonadotropin

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APA (6^th Edition):

Tavares Krause, A. R. 1. (2019). Ovarian Response, Follicular Function and Oocyte Developmental Competence in Gonadotropin Treated Prepubertal Calves. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/12490

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Tavares Krause, Ana Rita 1985-. “Ovarian Response, Follicular Function and Oocyte Developmental Competence in Gonadotropin Treated Prepubertal Calves.” 2019. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/12490.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Tavares Krause, Ana Rita 1985-. “Ovarian Response, Follicular Function and Oocyte Developmental Competence in Gonadotropin Treated Prepubertal Calves.” 2019. Web. 08 Mar 2021.

Vancouver:

Tavares Krause AR1. Ovarian Response, Follicular Function and Oocyte Developmental Competence in Gonadotropin Treated Prepubertal Calves. [Internet] [Thesis]. University of Saskatchewan; 2019. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/12490.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Tavares Krause AR1. Ovarian Response, Follicular Function and Oocyte Developmental Competence in Gonadotropin Treated Prepubertal Calves. [Thesis]. University of Saskatchewan; 2019. Available from: http://hdl.handle.net/10388/12490

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Saskatchewan

14. Arsenault, Ryan. Development and application of genus-specific peptide arrays for kinome analysis.

Degree: 2011, University of Saskatchewan

URL: http://hdl.handle.net/10388/ETD-2011-12-267

► Phosphorylation represents a central mechanism to regulate cell function. Protein phosphorylation is catalyzed by a class of enzymes called kinases, the cellular complement of which… (more)

▼ Phosphorylation represents a central mechanism to regulate cell function. Protein phosphorylation is catalyzed by a class of enzymes called kinases, the cellular complement of which is referred to as its kinome. The study of the kinome has led to the development of peptide arrays as a high throughput tool. Though the approach was effective, it was limited to genera with characterized phosphoproteomes, as well as by the practice of interpreting emerging data through platforms designed for gene expression data. Within this thesis, these pivotal roadblocks are addressed through the presentation of 1) an approach for the development of custom-designed genus-specific arrays based on proteomic information and 2) a data analysis pipeline, Platform for Integrated, Intelligent Kinome Analysis (PIIKA), developed specifically for peptide array kinome data. The utility of these advances is demonstrated through the creation of the customized peptide arrays and subsequent confirmation of PIIKA-enhanced data transformation and mining. These techniques (custom array and PIIKA) were then applied to two complex host-pathogen interactions: prion diseases and Mycobacterium avium subsp. paratuberculosis (MAP) infection. Prion diseases result from the misfolding of the widely expressed and highly conserved cellular prion protein (PrPC) into an infectious and pathological scrapie-like conformation (PrPSc). Little is known about the function of PrPC in either the normal or diseased states, although a role in signal transduction has been suggested. Two PrPC protein-specific ligands, PrP 106-126 prion fragment and the prion-specific monoclonal antibody 6H4, were used to induce signalling in neuronal cells. Kinome analysis revealed distinct signalling pathways and varied signalling responses for each PrPC ligand. This observation is consistent with the emerging concept that PrPC interacts with multiple cellular proteins and plays a multifunctional role in regulating cellular responses. MAP is the causative agent of Johne’s disease in cattle. MAP establishes a persistent infection and has the ability to evade immune responses while replicating inside macrophages. Kinome analysis indicated that MAP is able to inhibit the interferon gamma (IFNγ)-induced JAK-STAT signalling pathway, eliminating the activation of downstream effectors. Further analysis indicated that the JAK-STAT pathway was blocked at the level of the IFNγ receptor, and JAK-STAT suppressor molecules known as SOCS were activated, both novel findings in the field of MAP pathogenesis. Collectively, these investigations highlight the use of custom-designed and genus-specific peptide arrays to address complex biology in distinct genera. Advisors/Committee Members: Napper, Scott K., Lee, Jeremy S., Roesler, William J., Misra, Vikram, Griebel, Philip J., Pato, Mary D..

Subjects/Keywords: kinome; peptide array; genus-specific; kinase; prion; PrPC; PrPSc; Mycobacterium avium subsp. paratuberculosis; MAP; mycobacteria; cell signalling; PIIKA

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Arsenault, R. (2011). Development and application of genus-specific peptide arrays for kinome analysis. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/ETD-2011-12-267

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Arsenault, Ryan. “Development and application of genus-specific peptide arrays for kinome analysis.” 2011. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/ETD-2011-12-267.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Arsenault, Ryan. “Development and application of genus-specific peptide arrays for kinome analysis.” 2011. Web. 08 Mar 2021.

Vancouver:

Arsenault R. Development and application of genus-specific peptide arrays for kinome analysis. [Internet] [Thesis]. University of Saskatchewan; 2011. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/ETD-2011-12-267.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Arsenault R. Development and application of genus-specific peptide arrays for kinome analysis. [Thesis]. University of Saskatchewan; 2011. Available from: http://hdl.handle.net/10388/ETD-2011-12-267

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

15. Yue, Binbin. An analysis of the molecular regulation of tumor necrosis factor-alpha expression in human mast cells.

Degree: 2000, University of Saskatchewan

URL: http://hdl.handle.net/10388/etd-08092012-111047

► The objective of this study was to analyze the molecular regulation of tumour necrosis factor-alpha (TNF-α) gene expression in human cells. Allergic diseases (e.g., asthma)… (more)

▼ The objective of this study was to analyze the molecular regulation of tumour necrosis factor-alpha (TNF-α) gene expression in human cells. Allergic diseases (e.g., asthma) comprise one of the most prominent medical problems in Canada, and their incidence and severity are on the increase here as in most industrialized countries. Allergic responses are initiated by contact of allergens with specific IgE antibodies on mast cells, leading to cellular activation. Such activation induces mast cells to release very large amounts of the pro-inflammatory cytokine TNF-α which, by itself, mediates much of the cellular recruitment associated with the allergic late phase response (LPR). Most of the pathology of allergic diseases is directly attributable to the LPR. The molecular mechanism regulating TNF-α expression in mast cells, as opposed to that in macrophages (which can also produce an abundance of TNF-α) is poorly understood. My hypothesis is that if TNF-α were differentially regulated in mast cells, it might be possible to therapeutically manipulate its highly pathogenic contributions to allergic diseases without affecting the other roles of this cytokine in the body. I examined the kinetics of TNF-α gene transcription and protein expression in mast cell-differentiated KU812 cells which I stimulated with phorbol-12, 13-myristate acetate (PMA) and calcium ionophore A23187 (PMA and A23187 effectively mimic FcεRI stimulation of mast cells). RT-PCR and ELISA methods were used to detect TNF-α mRNA and protein, respectively. My results indicate that low levels of TNF-α is stored in the unstimulated cells (but not in the supernatant fluids), high levels of TNFα were secreted into the culture supernatants within 2 hr after stimulation. The critical regulatory elements in the human TNF-α promoter were studied. A human TNF -a gene promoter fragment (-625 to +19 bp relative to the transcription start site; TSS) was enzymatically digested from the 5' end inward and thereby I generated a series of promoter fragments of varying sizes. Each of these promoter fragments were placed immediately 5' to a cDNA encoding the marker protein luciferase. Then the constructs were transfected into differentiated KU812 cells and I assessed the TNF promoter-driven luciferase expression in the mast cells following PMA/A23187 challenge. A negative (-331 to -205) and several positive (-82 to +19, -131 to -82, -205 to -153) control regions were identified within the TNF-α promoter. Advisors/Committee Members: Gordon, John R., Misra, Vikram, Loh, Lambert, Tabel, Henry.

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Yue, B. (2000). An analysis of the molecular regulation of tumor necrosis factor-alpha expression in human mast cells. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/etd-08092012-111047

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Yue, Binbin. “An analysis of the molecular regulation of tumor necrosis factor-alpha expression in human mast cells.” 2000. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/etd-08092012-111047.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Yue, Binbin. “An analysis of the molecular regulation of tumor necrosis factor-alpha expression in human mast cells.” 2000. Web. 08 Mar 2021.

Vancouver:

Yue B. An analysis of the molecular regulation of tumor necrosis factor-alpha expression in human mast cells. [Internet] [Thesis]. University of Saskatchewan; 2000. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/etd-08092012-111047.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Yue B. An analysis of the molecular regulation of tumor necrosis factor-alpha expression in human mast cells. [Thesis]. University of Saskatchewan; 2000. Available from: http://hdl.handle.net/10388/etd-08092012-111047

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

16. Chapman, Stacy. The effect of histone deacetylase inhibitors on SRC and BCL2L1 gene expression and a potential role for phosphatases in their transcriptional repression.

Degree: 2013, University of Saskatchewan

URL: http://hdl.handle.net/10388/ETD-2013-08-1136

► Histone Deacetylase Inhibitors (HDACi) are a new class of chemotherapeutics which have shown promise in pre-clinical and clinical settings. HDACi have been shown to act… (more)

▼ Histone Deacetylase Inhibitors (HDACi) are a new class of chemotherapeutics which have shown promise in pre-clinical and clinical settings. HDACi have been shown to act by re-programming gene expression, with the transcription of some genes such as p21WAF1 being activated, while others like SRC and BCL2L1 are repressed. The mechanism behind HDACi gene expression changes remains unknown; although it has been shown to involve a direct interaction with gene promoters. Using a quantitative qRT-PCR approach, the effect of various HDACi on the transcription of p21WAF1, SRC and BCL2L1 was examined. TSA and apicidin led to an up regulation of p21WAF1 mRNA levels while c-Src and Bcl-xL mRNA levels were downregulated. Short c-Src mRNA transcripts were unaffected following TSA and apicidin treatments, despite the full length transcripts being repressed. Repression of full length c-Src and Bcl-xL mRNA transcripts was not seen following treatment with MS-275 and MGCD0103, although p21WAF1 mRNA expression was induced. ChIP experiments revealed that following HDACi treatment, histone acetylation levels and RNA Polymerase II occupancy increased in the promoter regions of both the SRC and BCL2L1 genes. RNA Polymerase II occupancy lasted less than 15 minutes in the 3’ regions of the gene following treatment with apicidin and TSA, but was more long-term following MS-275 and MGCD0103 treatment. The protein phosphatase inhibitor Calyculin A completely blocked HDACi mediated repression of c-Src and Bcl-xL mRNA, suggesting a role for protein phosphatases in the mechanism behind HDACi. It is therefore hypothesized that HDACi work through at least two different mechanisms. Whether or not an HDACi leads to gene repression depends on its ability to disrupt an HDAC/protein phosphatase complex and not on their HDAC specificities. The disruption of the complex leads to the release of an active protein phosphatase. The released phosphatase can then presumably act on various factors changing a gene from an active to paused state, possibly through promoter proximal pausing. HDACi unable to disrupt this complex are unable to induce gene repression. Collectively, these studies highlight not only the complexity of HDACi mediated effects within the cell, but also present a new explanation behind HDACi mediated gene repression. Advisors/Committee Members: Bonham, Keith, Moore, Stanley, Wilson, Heather, Misra, Vikram.

Subjects/Keywords: Histone Deacetylase Inhibitors; SRC; BCL2L1

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Chapman, S. (2013). The effect of histone deacetylase inhibitors on SRC and BCL2L1 gene expression and a potential role for phosphatases in their transcriptional repression. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/ETD-2013-08-1136

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Chapman, Stacy. “The effect of histone deacetylase inhibitors on SRC and BCL2L1 gene expression and a potential role for phosphatases in their transcriptional repression.” 2013. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/ETD-2013-08-1136.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Chapman, Stacy. “The effect of histone deacetylase inhibitors on SRC and BCL2L1 gene expression and a potential role for phosphatases in their transcriptional repression.” 2013. Web. 08 Mar 2021.

Vancouver:

Chapman S. The effect of histone deacetylase inhibitors on SRC and BCL2L1 gene expression and a potential role for phosphatases in their transcriptional repression. [Internet] [Thesis]. University of Saskatchewan; 2013. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/ETD-2013-08-1136.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Chapman S. The effect of histone deacetylase inhibitors on SRC and BCL2L1 gene expression and a potential role for phosphatases in their transcriptional repression. [Thesis]. University of Saskatchewan; 2013. Available from: http://hdl.handle.net/10388/ETD-2013-08-1136

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

17. Yousefi, Iran. Selective activation of unfolded protein response (UPR) by herpes simplex virus type 1 (HSV-1) in permissive and non permissive cells.

Degree: 2011, University of Saskatchewan

URL: http://hdl.handle.net/10388/etd-08182011-142259

► The unfolded protein response (UPR) is induced by a variety of external and internal stimuli, including accumulation of misfolded proteins in the endoplasmic reticulum (ER).… (more)

▼ The unfolded protein response (UPR) is induced by a variety of external and internal stimuli, including accumulation of misfolded proteins in the endoplasmic reticulum (ER). Viruses such as Herpes Simplex Virus type 1 (HSV-1) induce host cells to produce viral proteins many of which undergo glycosylation and other modifications in the ER, causing stress to the ER and consequently UPR activation. I have tested the hypothesis that HSV-1 has evolved strategies to regulate the UPR in order to suppress aspects of the UPR that might interfere with viral replication and to promote pathways that aid its own survival and replication. The purpose of this study was to test the hypothesis that HSV-1 selectively modulates the three pathways (PERK, ATF6, and IRE-1) of the UPR in epithelial and neuronal cells and to examine the similarities and the differences between these two types of cells in their responses to ER stress. Vero and ONS-76 cells were used as models of epithelial and neuronal cells respectively and qRT PCR technique was used for analyzing RNA levels of transcripts of spliced Xbp1, HERP, CHOP and BIP, selected target genes for three pathways of the UPR. HSV-1 DNA synthesis and infectious virus production in infected cells showed that compared to the permissive Vero cells, ONS-76 cells seemed to be semi-permissive to HSV-1 infection with limited viral DNA synthesis and infectious virus production. The kinetics of transcript and protein synthesis for genes representing immediate early, early and late classes of viral genes was also monitored. Expression of the immediate early gene, ICP0, was similar in both cell types but the expression of the early gene, TK and late genes VP16 and VP 5 was different. My work reveals that HSV-1 infection in cells of epithelial and neuronal origins results in activation of the UPR, but through cell type selective regulation of the three signal transduction pathways of the UPR (PERK, ATF6, and IRE-1). While HSV-1 infection resulted in upregulation of Spliced Xbp1 and its target gene HERP (IRE1 pathway) and downregulation of BIP (ATF6 pathway) in both cell types, CHOP (PERK pathway) was upregulated only in ONS cells. My results suggest that some aspects of the UPR are regulated differently in cells representing the sites for HSV-1 lytic and latent infections. This may indicate the need for increasing the capacity for protein folding and degradation (Xbp1 and ATF6-induced) in both cells but a requirement for suppressing apoptosis (PERK-induced) only in epithelial cells. As well, I show that HSV-1 infection not only selectively activates the UPR pathways in different cell types, but also inactivates some components of the UPR pathways activated by the drug thapsigargin. Advisors/Committee Members: Misra, Vikram, Verge, Valerie, Tikoo, Suresh, Wilson, Joyce, Hill, Janet.

Subjects/Keywords: ONS-76 cells; Verp cells; UPR; HSV-1

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APA (6^th Edition):

Yousefi, I. (2011). Selective activation of unfolded protein response (UPR) by herpes simplex virus type 1 (HSV-1) in permissive and non permissive cells. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/etd-08182011-142259

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Yousefi, Iran. “Selective activation of unfolded protein response (UPR) by herpes simplex virus type 1 (HSV-1) in permissive and non permissive cells.” 2011. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/etd-08182011-142259.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Yousefi, Iran. “Selective activation of unfolded protein response (UPR) by herpes simplex virus type 1 (HSV-1) in permissive and non permissive cells.” 2011. Web. 08 Mar 2021.

Vancouver:

Yousefi I. Selective activation of unfolded protein response (UPR) by herpes simplex virus type 1 (HSV-1) in permissive and non permissive cells. [Internet] [Thesis]. University of Saskatchewan; 2011. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/etd-08182011-142259.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Yousefi I. Selective activation of unfolded protein response (UPR) by herpes simplex virus type 1 (HSV-1) in permissive and non permissive cells. [Thesis]. University of Saskatchewan; 2011. Available from: http://hdl.handle.net/10388/etd-08182011-142259

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

18. Just, Natasha. Characterization of Bioaerosols from Cage-housed and Floor-housed Poultry Operations.

Degree: 2012, University of Saskatchewan

URL: http://hdl.handle.net/10388/ETD-2012-12-855

► Background. Dust and endotoxin levels are higher in bioaerosols from floor-housed (FH) poultry operations than cage-housed (CH) poultry facilities. Workers from CH operations have reported… (more)

▼ Background. Dust and endotoxin levels are higher in bioaerosols from floor-housed (FH) poultry operations than cage-housed (CH) poultry facilities. Workers from CH operations have reported a greater prevalence of respiratory symptoms than FH workers. The negative respiratory symptoms observed in workers are typically attributed to endotoxin. However, other components of poultry bioaerosols and their effects on human health, such as bacteria, antibiotics and archaea, are poorly understood. Bacteria have been detected in intestinal, fecal, litter, and air samples from poultry operations. Chicken fecal bacteria differ depending on bird age and antibiotic use, which differ between CH and FH facilities. Antibiotics are used in CH and FH poultry operations to lower the likeliness of disease transmission. In FH facilities, antibiotics may also be used at sub-therapeutic levels for growth promotion. Low levels of antibiotic create a selective pressure towards antimicrobial resistance (AMR) in chicken fecal bacteria. Archaea have been detected in ceca, fecal, litter and house fly samples from poultry facilities but have not been investigated in bioaerosols. However, archaea have been detected in swine and dairy bioaerosols and can induce airway inflammation. Further understanding of poultry bioaerosols, with a comparison of those from CH and FH operations, will aid in the development of management practices to reduce worker exposure and response. Objective. The objective of these studies was to compare bioaerosols from CH and FH poultry facilities. Specifically, levels of dust, endotoxin, total bacteria, bacterial species, antimicrobial resistance genes and methanogenic archaea were examined. Methods. Bioaerosols were collected from fifteen CH and fifteen FH poultry operations using stationary area samplers as well as personal sampling devices. Dust was measured by gravimetric analyses. Limulus Amoebocyte Lysate (LAL) assays were used to quantify endotoxin. Bacteria and archaea concentrations were measured by quantitative PCR. Bacterial and archaeal diversity was investigated using PCR followed by denaturing gradient gel electrophoresis (DGGE) and sequencing. AMR genes were detected using end-point PCR. Results. Dust (p<0.001), endotoxin (p<0.05), total bacteria (p<0.05), Enterococcus (p<0.001), E. coli (p<0.001) and Staphylococcus (p<0.001) were more concentrated in bioaerosols from FH poultry operations than CH bioaerosols. Methanogenic archaea (p<0.001) and C. perfringens (p<0.05) were significantly higher in bioaerosols from CH facilities than FH bioaerosols. Zinc bacitracin resistance gene (bcrR), erythromycin resistance gene (ermA), and tetracycline resistance gene (tetA/C), were more prevalent in bioaerosols from FH facilities than CH bioaerosols (p<0.01, p<0.01 and p<0.05, respectively). Conclusions. Bioaerosols from CH and FH poultry operations are significantly different, suggesting that CH and FH workers are exposed to significantly different environments. Bacterial diversity,… Advisors/Committee Members: Singh, Baljit, Duchaine, Caroline, Misra, Vikram, Dosman, James, Hill, Janet.

Subjects/Keywords: bioaerosols; air sampling; poutlry; bacteria; endotoxin; dust; antimicrobial resistance; archaea

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APA (6^th Edition):

Just, N. (2012). Characterization of Bioaerosols from Cage-housed and Floor-housed Poultry Operations. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/ETD-2012-12-855

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Just, Natasha. “Characterization of Bioaerosols from Cage-housed and Floor-housed Poultry Operations.” 2012. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/ETD-2012-12-855.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Just, Natasha. “Characterization of Bioaerosols from Cage-housed and Floor-housed Poultry Operations.” 2012. Web. 08 Mar 2021.

Vancouver:

Just N. Characterization of Bioaerosols from Cage-housed and Floor-housed Poultry Operations. [Internet] [Thesis]. University of Saskatchewan; 2012. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/ETD-2012-12-855.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Just N. Characterization of Bioaerosols from Cage-housed and Floor-housed Poultry Operations. [Thesis]. University of Saskatchewan; 2012. Available from: http://hdl.handle.net/10388/ETD-2012-12-855

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

19. Dias, Fernanda. Effects of follicular aging and duration of superstimulation on oocyte competence and granulosa cell gene expression in cattle.

Degree: 2013, University of Saskatchewan

URL: http://hdl.handle.net/10388/ETD-2013-06-1110

► A prolonged growth phase of the ovulatory follicle results in follicular aging. Whether follicular aging is detrimental or beneficial to oocyte competence is not fully… (more)

▼ A prolonged growth phase of the ovulatory follicle results in follicular aging. Whether follicular aging is detrimental or beneficial to oocyte competence is not fully known. The objective of this thesis is to investigate the effects of follicular aging on oocyte competence and granulosa cell gene expression in cattle. Four sets of experiments were designed to address the objective. The following hypotheses were tested during the course of these studies: 1) oocyte competence will improve by the longer growing phase but will be adversely affected by FSH starvation, 2) follicles that undergo superstimulation will have different gene expression than dominant follicles from a natural cycle, 3) extending the superstimulation protocol by 3 days will allow follicles to mature better and 4) markers of maturity, cellular health and survival will be turned off by FSH starvation. The objective of the first study (Chapter 3) was to determine the effects of extending the length of superstimulation and follicular aging on oocyte competence by in vitro embryo production. Multiple follicles were allowed to grow for 4 (Short FSH) or 7 days (Long FSH) under the treatment of 8 or 14 injections of FSH (at 12-hour intervals), respectively. Multiple follicles in the FSH starvation group were allowed to grow for 7 days but FSH was provided for only the first 4 days of superstimulation. Extending the duration of follicular growth by superstimulation resulted in a greater number of ≥9 mm follicles and in 2.5 more transferable embryos per animal (morulae+blastocysts) at Day 9 of in vitro embryo culture. The FSH starvation resulted in a greater proportion of poor quality oocytes lower cleavage rate and lower embryonic development. Microarray analysis was used to assess the effect of superstimulation (Chapter 4), follicular aging (Chapter 5) and FSH starvation (Chapter 6) on the gene expression profile of superstimulated granulosa cells. Gene expression of granulosa cells from the post-LH preovulatory dominant follicle was compared (Chapter 4) with those from follicles of the same status after a standard 4-day superstimulation (same protocol as Short FSH group from Chapter 3). A total of 190 genes were down-regulated and 280 genes were upregulated in the superstimulated group when compared with the reference (non-superstimulated control). Data analysis showed that superstimulated follicles are still in a growing phase compared to untreated dominant follicles (most of the upregulated genes are related to matrix remodeling due to tissue proliferation) and did not respond to LH properly (down regulation of LH gene markers). Four-day superstimulation also disturbed genes related to angiogenesis and activated oxidative stress response genes. Extending the superstimulation protocol (7 days; same protocol as Long FSH from Chapter 3) allowed more time for follicles to leave the growing stage and properly respond to LH surge (most of the upregulated genes in the Long FSH group are markers of post LH surge) when compared to the standard 4 day… Advisors/Committee Members: Singh, Jaswant, Adams, Gregg P., Misra, Vikram, Sirard, Marc-Andre.

Subjects/Keywords: cattle; FSH; follicle dynamics; follicular aging; gene expression; granulosa cells; oocyte competence; superstimulation

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APA (6^th Edition):

Dias, F. (2013). Effects of follicular aging and duration of superstimulation on oocyte competence and granulosa cell gene expression in cattle. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/ETD-2013-06-1110

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Dias, Fernanda. “Effects of follicular aging and duration of superstimulation on oocyte competence and granulosa cell gene expression in cattle.” 2013. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/ETD-2013-06-1110.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Dias, Fernanda. “Effects of follicular aging and duration of superstimulation on oocyte competence and granulosa cell gene expression in cattle.” 2013. Web. 08 Mar 2021.

Vancouver:

Dias F. Effects of follicular aging and duration of superstimulation on oocyte competence and granulosa cell gene expression in cattle. [Internet] [Thesis]. University of Saskatchewan; 2013. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/ETD-2013-06-1110.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Dias F. Effects of follicular aging and duration of superstimulation on oocyte competence and granulosa cell gene expression in cattle. [Thesis]. University of Saskatchewan; 2013. Available from: http://hdl.handle.net/10388/ETD-2013-06-1110

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

20. Di Marzo, Andrea. Is Latent Equine Herpesvirus Type 1 (EHV-1) Reactivated by Triggering Activation of the Unfolded Protein Response in Equine Peripheral Blood Leukocytes?.

Degree: 2013, University of Saskatchewan

URL: http://hdl.handle.net/10388/ETD-2013-06-1138

► Equine Herpesvirus type 1 (EHV-1) is a worldwide threat to the health of horses. It can cause mild respiratory disease, abortions and deaths of newborn… (more)

▼ Equine Herpesvirus type 1 (EHV-1) is a worldwide threat to the health of horses. It can cause mild respiratory disease, abortions and deaths of newborn foals as well as a potentially fatal neurologic disorder known as Equine Herpesvirus Myeloencephalopathy (EHM). The virus is maintained in populations by stress-induced periodic reactivation of virus in long-term latently infected horses and transmission of the reactivated virus to susceptible individuals. In horses, peripheral blood leukocytes (PBLs) are thought to be an important site for EHV-1 latent genomes. Since the Unfolded Protein Response (UPR) is a cellular response to a variety of stressors that has been linked to reactivation of herpes simplex virus in humans, a virus closely related to EHV-1, I tested the hypothesis that latent EHV-1 relies on the UPR as a pluripotent stress sensor and uses it to reactivate lytic gene expression. Since little work has been done in defining the UPR in horses, I first successfully developed a quantitative real-time polymerase chain reaction (RT-qPCR) assay to detect and quantitate transcripts for selected UPR genes in equine dermal (E.Derm) cells and PBLs. Activation of the UPR was achieved in both cell types using thapsigargin and a difference in gene expression after activation of the UPR in two equine cell types was found. A nested PCR assay to detect and distinguish latent EHV-1 and EHV-4 was evaluated and the sensitivity of the technique to detect EHV-1 was determined. I discovered that the nested PCR technique was not sensitive enough to detect the estimated one latent viral genome in 50,000 PBLs. Lytic EHV-1 infection was characterized by single step growth curve in E.Derm cells and consistent detection of temporal EHV-1 gene expression by RT-qPCR was achieved. The relationship between EHV-1 gene expression and UPR gene expression during lytic infection was investigated. While EHV-1 infection had no effect on UPR gene expression, activation of the UPR appeared to decrease the expression of EHV-1 genes temporarily and reversibly during the first 4 h after infection. Finally, detection of EHV-1 in PBLs from horses presumed to be latently infected by co-cultivation with E. Derm cells permissive to EHV-1 infection was attempted. To detect viral DNA, PBLs were stimulated with thapsigargin or interleukin 2 (IL-2) which was previously reported to induce reactivation of latent EHV-1. I was not able to reproduce previously published experiments of reactivation in vitro of latent EHV-1 by stimulation with IL-2, and virus reactivation did not occur after stimulation of PBLs with thapsigargin. In summary, a RT-qPCR assay to measure the expression of equine UPR genes was developed and activation of the UPR by treatment of E.Derm cells and PBLs with thapsigargin was successfully achieved. A difference in gene expression after activation of the UPR in two equine cell types was found. In contrast to what has been reported for other alphaherpesviruses, there appears to be no, or only little, interaction between the UPR and EHV-1… Advisors/Committee Members: Lohmann, Katharina, Misra, Vikram, Haines, Deborah, Stookey, Joseph.

Subjects/Keywords: Equine herpesvirus type 1; EHV-1; Latency; Unfolded Protein response; UPR

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APA (6^th Edition):

Di Marzo, A. (2013). Is Latent Equine Herpesvirus Type 1 (EHV-1) Reactivated by Triggering Activation of the Unfolded Protein Response in Equine Peripheral Blood Leukocytes?. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/ETD-2013-06-1138

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Di Marzo, Andrea. “Is Latent Equine Herpesvirus Type 1 (EHV-1) Reactivated by Triggering Activation of the Unfolded Protein Response in Equine Peripheral Blood Leukocytes?.” 2013. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/ETD-2013-06-1138.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Vancouver:

Di Marzo A. Is Latent Equine Herpesvirus Type 1 (EHV-1) Reactivated by Triggering Activation of the Unfolded Protein Response in Equine Peripheral Blood Leukocytes?. [Internet] [Thesis]. University of Saskatchewan; 2013. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/ETD-2013-06-1138.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Di Marzo A. Is Latent Equine Herpesvirus Type 1 (EHV-1) Reactivated by Triggering Activation of the Unfolded Protein Response in Equine Peripheral Blood Leukocytes?. [Thesis]. University of Saskatchewan; 2013. Available from: http://hdl.handle.net/10388/ETD-2013-06-1138

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

21. Johnson, Lisa Anne 1991-. Detection and characterization of pathogenic and non-pathogenic Brachyspira.

Degree: 2017, University of Saskatchewan

URL: http://hdl.handle.net/10388/8212

► Brachyspira is associated with mucohaemorrhagic diarrhea in pigs, though the pathogenesis and virulence factors behind the disease are not currently understood. This thesis aimed to… (more)

▼ Brachyspira is associated with mucohaemorrhagic diarrhea in pigs, though the pathogenesis and virulence factors behind the disease are not currently understood. This thesis aimed to identify putative virulence factors of Brachyspira through a comparative genomics approach, and to determine the frequency of the occurrence of mixed Brachyspira species infections within clinical case material. “B. hampsonii” strain 30446 was originally isolated from colon contents of a pig with mucohaemorrhagic diarrhea, and through inoculation trials has been demonstrated to be pathogenic in swine. “B. hampsonii” strain KL-180 was isolated from a Lesser Snow Goose, and is remarkably genetically similar to strain 30446. However, inoculation trials indicate “B. hampsonii” strain KL-180 is not pathogenic in swine. Their differing pathogenicity, despite their genetic similarity, allows comparison to identify gene content differences, which may correspond to putative virulence factors. Genome sequences for each strain were available, and were annotated. The basic genome features of strain 30446 and strain KL-180 were similar, with an average nucleotide identity (ANI) of 97%. Comparison of the annotated genomes revealed genes unique to each strain. Any gene differences could be important to disease outcome, including hypothetical proteins. “B. hampsonii” strain 30446, contained a set of genes absent in strain KL-180 which correspond to the Streptolysin S (sag) operon. The presence of this operon was confirmed with PCR and primers designed for each gene. The presence of this operon in pathogenic Brachyspira species, and absence in the non-pathogenic “B. hampsonii” strain KL-180 supports the possibility of the operon being a virulence factor and contributing to disease outcome. Current diagnostic methods implemented on clinical case material often indicate the presence of more than one Brachyspira species. It is not currently known if co-infection with multiple species is common, or if the presence of multiple species contributes to Brachyspira pathogenesis. To investigate the frequency of the occurrence of multiple species in Brachyspira clinical cases, deep sequencing of the nox gene target was used on clinical case material from Western Canada, Mexico and Brazil. Synthetically created communities were also sequenced, to provide insight as to what levels of Brachyspira can be detected through this method. Pigs with clinical disease were very frequently colonized by multiple species of Brachyspira. “B. hampsonii” was detected in pigs from both Mexico and Brazil, though often at very low levels. Canadian clinical cases were more diverse than those from Mexico and Brazil, and were dominated by a range of Brachyspira species. Sequencing of the synthetic communities revealed a negative primer bias towards “B. hampsonii” strain 30599 (Clade I). Strain 30599 may therefore be present at higher levels and be more widely distributed than demonstrated by current diagnostic methods based on PCR of the nox gene. The results of this thesis demonstrate… Advisors/Committee Members: Hill, Janet E, White, Aaron, Luby, Chris, Misra, Vikram.

Subjects/Keywords: Swine dysentery; comparative genomics; virulence; diagnostics; sequencing; polymerase chain reaction; NADH-oxidase; Brachyspira

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APA (6^th Edition):

Johnson, L. A. 1. (2017). Detection and characterization of pathogenic and non-pathogenic Brachyspira. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/8212

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Johnson, Lisa Anne 1991-. “Detection and characterization of pathogenic and non-pathogenic Brachyspira.” 2017. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/8212.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Johnson, Lisa Anne 1991-. “Detection and characterization of pathogenic and non-pathogenic Brachyspira.” 2017. Web. 08 Mar 2021.

Vancouver:

Johnson LA1. Detection and characterization of pathogenic and non-pathogenic Brachyspira. [Internet] [Thesis]. University of Saskatchewan; 2017. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/8212.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Johnson LA1. Detection and characterization of pathogenic and non-pathogenic Brachyspira. [Thesis]. University of Saskatchewan; 2017. Available from: http://hdl.handle.net/10388/8212

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

22. Alam, Md Kausar. Galactofuranose biosynthesis is important for maintaining normal growth and cell wall properties in Aspergillus nidulans.

Degree: 2014, University of Saskatchewan

URL: http://hdl.handle.net/10388/ETD-2014-02-1434

► The cell wall is essential for fungal survival in natural environments. Galactofuranose (Galf) decorates certain carbohydrates and lipids of Aspergillus cell wall, is absent in… (more)

▼ The cell wall is essential for fungal survival in natural environments. Galactofuranose (Galf) decorates certain carbohydrates and lipids of Aspergillus cell wall, is absent in humans and appears to play a role in fungal cell wall maturation. Previous studies in our lab showed that deletion of any of three sequential-acting genes (ugeA, ugmA, and ugtA) of Galf pathway caused substantially reduced growth and spore production. Two genes upstream of the Galf pathway, galD and galE are essential for galactose metabolism in many systems including the budding yeast, Saccharomyces cerevisiae. Interestingly, characterization of galD and galE in A. nidulans using cell and molecular techniques showed that unlike yeast, neither of these genes was essential for growth at physiological pH 7.5. Nevertheless for each case, their expressions were up-regulated by growth on galactose, revealing the relative complexity of galactose metabolism in A. nidulans. Our study also showed that repression of the three sequentially acting Galf pathway genes by conditional promoters phenocopied previously characterized deletion morphology. Using anti-Galf (L10) we also showed that deletion and repression of these genes caused no Galf in the hyphal wall. Gene deletion or repression also increased sensitivity to the wall-targeting drug, caspofungin. Related results from qPCR showed that deletion or repression of ugmA increased gene expression of α-glucan synthase agsB and decreased that of β-glucan synthase fksA. Therefore, Galf is non-essential but important for many aspects of Aspergillus growth, sporulation, and wall maturation. Aspergillosis, the most common airborne systemic fungal disease, is typically caused by Aspergillus fumigatus. Several A. fumigatus UgmA (AfUgmA) mutants with altered enzyme activity due to single amino acid changes were used to assess their effect on growth and wall composition in A. nidulans. Wild type AfugmA complemented the phenotypic defects in an A. nidulans ugmAΔ strain, consistent with these two genes being homologous. The AfUgmA crystal structure has been solved, and the in vitro enzymatic effects of specific mutations in the enzyme active site have been published. AfUgmA mutated strains with reduced activity in vitro impaired A. nidulans growth in a manner substantially similar to gene deletion and gene down-regulation. Site directed mutagenesis showed that AfUgmA residues R182 and R327 were critical for Galf generation both in vivo and in vitro. This supports previous results showing that UgmA is essential for Galf biosynthesis. Using fluorescent latex beads, we showed that reduction of wall Galf increased hyphal surface adhesion. Consistent with qPCR studies, immunofluorescence and ELISA results showed that loss or absence of Galf increased wall α-glucan but reduced wall β -glucan. Galf is important for wall surface integrity and for maintaining dynamic co-ordination with other pathways. To begin to assess this dynamic co-ordination, Tandem Affinity Purification (TAP) tagging combined with LC-MS/MS was used… Advisors/Committee Members: Kaminskyj, Susan, Wei, Yangdou, Todd, Christopher, Misra, Vikram, Moore, Margo.

Subjects/Keywords: Aspergillus nidulans; Cell wall; galactose; Galactofuranose; drug sensitivity

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APA (6^th Edition):

Alam, M. K. (2014). Galactofuranose biosynthesis is important for maintaining normal growth and cell wall properties in Aspergillus nidulans. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/ETD-2014-02-1434

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Alam, Md Kausar. “Galactofuranose biosynthesis is important for maintaining normal growth and cell wall properties in Aspergillus nidulans.” 2014. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/ETD-2014-02-1434.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Alam, Md Kausar. “Galactofuranose biosynthesis is important for maintaining normal growth and cell wall properties in Aspergillus nidulans.” 2014. Web. 08 Mar 2021.

Vancouver:

Alam MK. Galactofuranose biosynthesis is important for maintaining normal growth and cell wall properties in Aspergillus nidulans. [Internet] [Thesis]. University of Saskatchewan; 2014. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/ETD-2014-02-1434.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Alam MK. Galactofuranose biosynthesis is important for maintaining normal growth and cell wall properties in Aspergillus nidulans. [Thesis]. University of Saskatchewan; 2014. Available from: http://hdl.handle.net/10388/ETD-2014-02-1434

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

23. Waugh, Taryn. Regulation of Ion Channel Physiology in Airway Epithelial cells in response to Influenza A Virus Infection.

Degree: 2013, University of Saskatchewan

URL: http://hdl.handle.net/10388/ETD-2013-08-1133

► Epithelial cells lining the upper airways are characterized by low sodium absorption and elevated chloride secretion. Together, the movement of these ions creates the osmotic… (more)

▼ Epithelial cells lining the upper airways are characterized by low sodium absorption and elevated chloride secretion. Together, the movement of these ions creates the osmotic drive to hydrate the airways. Recent studies indicate that influenza is capable of directly modulating the vectorial transport of sodium and chloride ions. However, the direct impact of influenza has not been studied with respect to potassium channels. This is significant because potassium conductance creates the driving force for chloride secretion. Disruptions to this process leads to edema formation in the lungs and can subsequently cause Acute Respiratory Distress Syndrome. Additionally, it has been demonstrated that the induction of pro-inflammatory cytokines in infected cells may contribute to altered ion channel function, further exacerbating edema formation. The purpose of this study was to assess the direct and indirect effects of influenza virus infection on potassium and chloride ion channel function in a secretory epithelial cell model. In order to assess the direct effects we exposed polarized epithelial cell monolayers to varying doses of H1N1 virus. Potassium and chloride channel function was measured by means of short-circuit current in an Ussing chamber. The immune response to viral infection was determined by RT-qPCR and Bioplex suspension array. Virus conditioned media (CM), and IL-8 were used to characterize the indirect effects on non-infected cells. We observed an increase in chloride secretion, consistent with edema formation, when 60% of the epithelium was infected, and after CM treatment. This observation correlated with increased potassium channel conductance through the calcium-activated (KCNN4) and cAMP-activated potassium channels (KCNQ1), which was ameliorated upon specific inhibition of these channels. The data suggest that the mixture of pro-inflammatory cytokines induced by viral infection directly up-regulate these potassium channels. However, treatment with IL-8 also appears to increase chloride secretion, although the underlying mechanism remains to be determined, as it is not mediated through KCNN4 and KCNQ1. We conclude that the strong induction of cytokines in infected cells act in a paracrine manner on non-infected cells to increase potassium channel conductance. This up-regulation of potassium channels subsequently drives an increase in chloride secretion, leading to fluid build-up in the lungs and edema formation. Advisors/Committee Members: Loewen, Matthew, Misra, Vikram, Blakley, Barry, Wobeser, Bruce.

Subjects/Keywords: Airway epithelium; ion channels; CFTR; KCNN4; KCNQ1; influenza virus; cytokines

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Waugh, T. (2013). Regulation of Ion Channel Physiology in Airway Epithelial cells in response to Influenza A Virus Infection. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/ETD-2013-08-1133

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Waugh, Taryn. “Regulation of Ion Channel Physiology in Airway Epithelial cells in response to Influenza A Virus Infection.” 2013. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/ETD-2013-08-1133.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Waugh, Taryn. “Regulation of Ion Channel Physiology in Airway Epithelial cells in response to Influenza A Virus Infection.” 2013. Web. 08 Mar 2021.

Vancouver:

Waugh T. Regulation of Ion Channel Physiology in Airway Epithelial cells in response to Influenza A Virus Infection. [Internet] [Thesis]. University of Saskatchewan; 2013. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/ETD-2013-08-1133.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Waugh T. Regulation of Ion Channel Physiology in Airway Epithelial cells in response to Influenza A Virus Infection. [Thesis]. University of Saskatchewan; 2013. Available from: http://hdl.handle.net/10388/ETD-2013-08-1133

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

24. Schroeder, Kristen. Characterizing the use of differentiated medulloblastoma cells to examine Herpes Simplex Virus latency and reactivation.

Degree: 2013, University of Saskatchewan

URL: http://hdl.handle.net/10388/ETD-2013-06-1089

► In human infection, herpes simplex virus (HSV) navigates two distinct life cycles; lytic and latent. The latent cycle takes place in sensory neurons, and is… (more)

▼ In human infection, herpes simplex virus (HSV) navigates two distinct life cycles; lytic and latent. The latent cycle takes place in sensory neurons, and is characterized as a dormant period punctuated by stress-induced episodes of viral reactivation. Understanding the mechanisms by which HSV latency and reactivation occur has been hindered by the lack of a model that faithfully recapitulates the environment of a human sensory neuron. Systems ranging from rat neurons to human fibroblasts have been developed to host HSV latency, however few available models have been able to investigate the role of human neuron-specific factors. To address this need, human medulloblastoma tumour cell lines, which derive from neuronal precursor cells, were differentiated and examined for their ability to host the HSV latency-reactivation cycle—in a manner similar to the differentiated PC-12 cell model. ONS-76 and UW228 medulloblastoma cell lines were screened for differentiation capacity. The differentiated cells were demonstrated to possess neuronal character as several neuron-specific proteins were found to be expressed. Differentiated ONS-76 cells were not compatible with hosting HSV latency, however, infection with a viral mutant impaired for lytic cycle initiation exhibited a deviant pattern of gene expression that resembles what has been observed in reactivation. Differentiated UW228 cells were found to host a low frequency, stable infection with the HSV mutant, characterized by the absence of infectious virus and viral lytic gene expression in the presence of persisting viral DNA. This DNA could further be induced to re-enter the lytic cycle through heat shock treatment and removal of differentiating agents from cell cultures. These results depict differentiated medulloblastoma cells as a novel tool in the study of HSV latency and reactivation, as these cells derive from the central nervous system and provide a new cellular perspective through which HSV biology can be viewed. Advisors/Committee Members: Misra, Vikram, Wilson, Joyce, Hill, Janet, Wobeser, Bruce.

Subjects/Keywords: HSV; latency; reactivation; medulloblastoma; VP16

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APA (6^th Edition):

Schroeder, K. (2013). Characterizing the use of differentiated medulloblastoma cells to examine Herpes Simplex Virus latency and reactivation. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/ETD-2013-06-1089

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Schroeder, Kristen. “Characterizing the use of differentiated medulloblastoma cells to examine Herpes Simplex Virus latency and reactivation.” 2013. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/ETD-2013-06-1089.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Schroeder, Kristen. “Characterizing the use of differentiated medulloblastoma cells to examine Herpes Simplex Virus latency and reactivation.” 2013. Web. 08 Mar 2021.

Vancouver:

Schroeder K. Characterizing the use of differentiated medulloblastoma cells to examine Herpes Simplex Virus latency and reactivation. [Internet] [Thesis]. University of Saskatchewan; 2013. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/ETD-2013-06-1089.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Schroeder K. Characterizing the use of differentiated medulloblastoma cells to examine Herpes Simplex Virus latency and reactivation. [Thesis]. University of Saskatchewan; 2013. Available from: http://hdl.handle.net/10388/ETD-2013-06-1089

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

25. Hoffman, Brett. Investigating the modulation of viral translation by the Hepatitis C virus nonstructural protein 5A.

Degree: 2015, University of Saskatchewan

URL: http://hdl.handle.net/10388/ETD-2015-04-2030

► Hepatitis C virus NS5A is a multi-functional viral protein essential for viral replication and assembly, although the exact role the protein plays in the viral… (more)

▼ Hepatitis C virus NS5A is a multi-functional viral protein essential for viral replication and assembly, although the exact role the protein plays in the viral lifecycle remains unclear. A vast array of functions have been attributed to NS5A in recent years, despite the lack of enzymatic activity. NS5A has been found to interact with over 130 host proteins including many which are central to cellular signaling pathways. NS5A is composed of three domains separated by regions of low complexity. All three domains perform important functions in the viral lifecycle. Domains I and II are essential for viral replication whereas domain III is required for viral assembly. However, the role that NS5A and its individual domains may play in modulating viral translation remains controversial. Previous studies have utilized translation reporter systems that do not accurately reflect the role of the viral 3´-UTR in modulating viral translation. We and others have shown that NS5A binds to the poly-U/UC region of the 3´-UTR. In addition to serving as the initiation site for negative strand synthesis the 3´-UTR functions to significantly enhance viral translation. The mechanism of translation enhancement remains unclear but may involve long range RNA-RNA interaction with the IRES, the binding of cellular proteins which stimulate translation and/or the recycling of ribosomes. Therefore, the protein-RNA interaction between NS5A and the poly-U/UC region has the potential to modulate viral translation. Therefore we set out to determine the role of NS5A and its individual domains in modulating viral translation and the role of the NS5A-poly-U/UC region interaction in this modulation. Utilizing monocistronic RNA reporters which contain the viral 5´- and 3´-UTRs and an internal Renilla luciferase reporter gene, we determined that NS5A specifically down-regulates viral translation in a dose-dependent manner through a mechanism dependent upon the presence of the poly-U/UC region in the viral 3´-UTR. Furthermore, we have re-tested the effect using full-length HCV genomic RNA reporters. These results suggest that NS5A is able to interfere with the stimulation of viral translation exerted by the 3´-UTR. This down-regulatory function of NS5A may function in mediating a switch from translation to replication, a step required in the lifecycle of a positive sensed RNA virus. Having established a role for NS5A in modulating viral translation, we then aimed to determine which region of NS5A was responsible for this effect. We found that each of NS5A domains was capable of this modulatory effect on viral translation independently. Although surprising, this finding is not entirely unexpected as each domain has been shown to retain the ability to bind to the poly-U/UC region. Within NS5A domain I we identified a 61 aa. region sufficient for translation down-regulation. Furthermore, we have identified a number of positively charged residues within this region involved in the modulation of viral translation, in particular arginine 112 (R112). This… Advisors/Committee Members: Liu, Qiang, Anderson, Deborah, Napper, Scott, Misra, Vikram.

Subjects/Keywords: Hepatitis C virus; protein translation; NS5A

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APA (6^th Edition):

Hoffman, B. (2015). Investigating the modulation of viral translation by the Hepatitis C virus nonstructural protein 5A. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/ETD-2015-04-2030

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Hoffman, Brett. “Investigating the modulation of viral translation by the Hepatitis C virus nonstructural protein 5A.” 2015. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/ETD-2015-04-2030.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Hoffman, Brett. “Investigating the modulation of viral translation by the Hepatitis C virus nonstructural protein 5A.” 2015. Web. 08 Mar 2021.

Vancouver:

Hoffman B. Investigating the modulation of viral translation by the Hepatitis C virus nonstructural protein 5A. [Internet] [Thesis]. University of Saskatchewan; 2015. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/ETD-2015-04-2030.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Hoffman B. Investigating the modulation of viral translation by the Hepatitis C virus nonstructural protein 5A. [Thesis]. University of Saskatchewan; 2015. Available from: http://hdl.handle.net/10388/ETD-2015-04-2030

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

26. Khan, Muhammad. Effect of Maternal Age on Transcriptome of Granulosa Cells from Bovine Dominant Follicles.

Degree: 2014, University of Saskatchewan

URL: http://hdl.handle.net/10388/ETD-2014-01-1364

► Advanced maternal age has been shown to influence follicular and luteal dynamics in bovine ovary resulting in reduced fertility. The overall objective of the four… (more)

▼ Advanced maternal age has been shown to influence follicular and luteal dynamics in bovine ovary resulting in reduced fertility. The overall objective of the four studies presented in this thesis is to identify the maternal age-associated transcriptional changes in granulosa cells of the dominant follicles during follicle development. In the first study, mRNA expression levels of housekeeping genes were measured by real–time quantitative PCR (RT-qPCR) in granulosa cells of dominant follicles and FSH-stimulated follicles to select and validate suitable reference genes for relative gene expression analyses during maternal and follicular aging. Stability of six reference genes (GAPDH, ACTB, EIF2B2, UBE2D2, SF3A1 and RNF20) was analyzed using GeNorm, DeltaCT and NormFinder programs and comprehensive ranking order was determined based on these programs. Geometric mean of multiple genes (UBE2D2, EIF2B2, GAPDH and SF3A1) was more appropriate reference control than individual genes for the comparison of relative gene expression among dominant and FSH-stimulated follicles during maternal and/or follicular aging studies. In the second study, maternal age-associated changes in the transcriptome of granulosa cells recovered at the time of selection of the dominant follicle from aged (n=3) and young cows (n=3) were determined by EmbryoGENE bovine oligo-microarrays (EMBV3, Agilent Technology). The mRNA expression of five transcripts (CYP19A1, PCNA, GJA1, TPM2, and VNN1) was confirmed in a different set of granulosa cell samples by RT-qPCR to validate microarray data. A total of 169 genes/isoforms were differentially expressed (≥ 2-fold-change; P ≤ 0.05) in aged cows vs. young cows. These transcripts revealed inefficient 1) control of gonadotropins, and gonadotropin-induced changes in the cytoskeleton and extracellular matrix, 2) lipid metabolism and steroidogenesis 3) cell proliferation, cell cycle control and intercellular communication, and 4) higher oxidative stress responses in aged cows vs. young cows. In the third study, changes in the transcriptome of granulosa cells of the preovulatory follicle 24 h after LH treatment from aged (n= 3) and young (n=3) were determined. A total of 1340 genes were expressed differentially (≥ 2-fold change; P ≤ 0.05) in aged cows vs. young cows. The mRNA expression of five transcripts (RGS2, PTGS2, TNFAIP6, VNN1, NR5A2 and GADD45B) was confirmed in a different set of granulosa cell samples to validate microarray data. These transcripts were related to delayed 1) response to LH treatment 2) cellular differentiation and luteinization and 3) progesterone synthesis. Intra-follicle levels of progesterone were lower (P < 0.05) in aged cows compared to young and mid-aged cows. The fourth study compared the aged-associated changes in the transcriptome of granulosa cells during follicle development from the time of dominant follicle selection to preovulatory stage (24 h after LH). In comparison to young cows, aged cows expressed fewer differentially expressed genes/isoforms (1206 vs. 2260, respectively)… Advisors/Committee Members: Singh, Jaswant, Adams, Gregg P., Sirard, Marc A., Misra, Vikram, Anzar, Muhammad.

Subjects/Keywords: Maternal age; Granulosa cells; Dominant follicle at selection; Preovulatory follicle; Reference genes; Microarrays; RT-qPCR; Bovine; Luteinizing hormone; Ovariectomy; Follicular aspiration; Transcriptome analysis; Ingenuity Pathway Analysis; Upstream regulators; Estradiol; Progesterone; Follicular fluid; Ultrasonography; Ovulation

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APA (6^th Edition):

Khan, M. (2014). Effect of Maternal Age on Transcriptome of Granulosa Cells from Bovine Dominant Follicles. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/ETD-2014-01-1364

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Khan, Muhammad. “Effect of Maternal Age on Transcriptome of Granulosa Cells from Bovine Dominant Follicles.” 2014. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/ETD-2014-01-1364.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Khan, Muhammad. “Effect of Maternal Age on Transcriptome of Granulosa Cells from Bovine Dominant Follicles.” 2014. Web. 08 Mar 2021.

Vancouver:

Khan M. Effect of Maternal Age on Transcriptome of Granulosa Cells from Bovine Dominant Follicles. [Internet] [Thesis]. University of Saskatchewan; 2014. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/ETD-2014-01-1364.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Khan M. Effect of Maternal Age on Transcriptome of Granulosa Cells from Bovine Dominant Follicles. [Thesis]. University of Saskatchewan; 2014. Available from: http://hdl.handle.net/10388/ETD-2014-01-1364

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

27. Huys, Adam. miR-122 binding of Hepatitis C virus 5'untranslated region augments the HCV life cycle independent from the p-body protein DDX6, and represents a novel target for siRNA targeted therapy.

Degree: 2014, University of Saskatchewan

URL: http://hdl.handle.net/10388/ETD-2014-08-1657

► Generally Hepatitis C Virus tropism is limited to hepatocytes. This limited tropism is a result of the receptors HCV requires for cellular entry and other… (more)

▼ Generally Hepatitis C Virus tropism is limited to hepatocytes. This limited tropism is a result of the receptors HCV requires for cellular entry and other host cellular factors including, uniquely, a liver specific miRNA, miR-122. The relationship between HCV and miR-122 is interesting, as commonly, miRNA are associated with suppression of function, but in the case of HCV, miR-122 actively promotes HCV proliferation. In-depth studies have demonstrated that miR-122 along with the RNA induced silencing complex (RISC) protein Argonaute 2 (Ago2) binds directly to two seed sequences separated by 8-9 nucleotides on HCV 5’UTR. Binding to the 5’UTR results in an increase in viral replication and translation. The method by which miR-122 promotes HCV translation and replication is not fully understood but evidence suggests that part of the function of miR-122 is to stabilize the HCV genome and protect it from exonuclease degradation by Xrn1, but other mechanisms remain to be identified. The reliance of HCV on miR-122 is best exemplified by the fact that removal of miR-122 by a miR-122 antagonist drastically impedes HCV viral titers in Chimpanzees and humans with no indication of escape mutants. The observation that HCV augmentation of the HCV life cycle by miR-122 requires Ago2 suggests that other components downstream in the miRNA suppression pathway may also be part of the mechanism of action. Our studies focused specifically on the processing body (p-body) associated DEAD-box helicase DDX6. DDX6 is essential for p-body assembly, required for robust miRNA suppression activity and elevated in HCV associated hepatocellular carcinomas. As such we hypothesized that DDX6 and p-bodies were directly or in-directly associated with the mechanism of action of miR-122. Knocking down DDX6 with siRNA indicated that DDX6 augments both HCV replication and translation. To examine whether DDX6 augmentation of HCV replication was related to the effects of miR-122 on the HCV life cycle, HCV replication and translation were assessed in the presence or absence of miR-122 when DDX6 was knocked down. Our data indicated that HCV replication and translation were augmented equally by miR-122 whether DDX6 was present or not. Our data also demonstrated that HCV replication and translation that was occurring independent of miR-122 was also still affected by DDX6 knockdown. Taken together our observations strongly suggest that the role DDX6 has on HCV is independent of HCV and miR-122’s relationship. In order to better understand miR-122’s relationship with HCV, we hypothesized that targeting the miR-122 binding region with siRNA would inhibit HCV replication initially, but that over the course of several rounds of treatment with the same siRNA, HCV would mutate to escape the siRNA, producing escape mutants that replicate without a dependency on miR-122. These escape mutants could be evaluated on how they replicate without using miR-122, shedding light on miR-122 and HCV’s relationship. Conversely if no escape mutants arose the siRNA could be… Advisors/Committee Members: Wilson, Joyce A., Bretscher, Peter A., van den Hurk, Sylvia, Tikoo, Suresh K., Misra, Vikram.

Subjects/Keywords: Hepatitis C virus; DDX6; miR-122; siRNA; Treatment; P-bodies; miRNA; Virus

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APA (6^th Edition):

Huys, A. (2014). miR-122 binding of Hepatitis C virus 5'untranslated region augments the HCV life cycle independent from the p-body protein DDX6, and represents a novel target for siRNA targeted therapy. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/ETD-2014-08-1657

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Huys, Adam. “miR-122 binding of Hepatitis C virus 5'untranslated region augments the HCV life cycle independent from the p-body protein DDX6, and represents a novel target for siRNA targeted therapy.” 2014. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/ETD-2014-08-1657.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Vancouver:

Huys A. miR-122 binding of Hepatitis C virus 5'untranslated region augments the HCV life cycle independent from the p-body protein DDX6, and represents a novel target for siRNA targeted therapy. [Internet] [Thesis]. University of Saskatchewan; 2014. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/ETD-2014-08-1657.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Huys A. miR-122 binding of Hepatitis C virus 5'untranslated region augments the HCV life cycle independent from the p-body protein DDX6, and represents a novel target for siRNA targeted therapy. [Thesis]. University of Saskatchewan; 2014. Available from: http://hdl.handle.net/10388/ETD-2014-08-1657

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

28. Munasinghe, Lilani Indika. Complement receptor 2 (CR2/CD21) in experimental African trypanosomiasis.

Degree: 2009, University of Saskatchewan

URL: http://hdl.handle.net/10388/etd-04232009-123902

► African trypanosomes are protozoan blood parasites that infect both humans and livestock. BALB/c mice are highly susceptible to experimental infections by Trypanosoma congolense while C57BL/6… (more)

▼ African trypanosomes are protozoan blood parasites that infect both humans and livestock. BALB/c mice are highly susceptible to experimental infections by Trypanosoma congolense while C57BL/6 mice are relatively resistant, as measured by degree and pattern of parasitemia and survival time. Rapid death observed in highly susceptible BALB/c mice is due to a systemic inflammatory response syndrome (SIRS). A small subset of pathogenic, MHC class II-restricted CD4+ T cells, activated during the course of T. congolense infections, mediates early mortality in infected highly susceptible BALB/c mice via excessive synthesis of the cytokine IFN-gamma. Since these pathogenic T cells are matrix–adherent, they could be distinguished from conventional Th1 cells. There is a possibility that this subpopulation of T cells has unique surface markers. The complement system is highly activated in African trypanosomiasis, leading to persistent hypocomplementemia. Amplification of the alternative pathway of complement is faster in BALB/c mice than in C57BL/6 mice and the degradation of complement component C3b to complement component C3d, during the amplification of the alternative pathway of complement, proceeds faster in BALB/c than in C57BL/6 mice (Ogunremi et al., 1993). T. congolense-infected BALB/c mice have more immune complexes containing trypanosomal variant surface glycoprotein (VSG) than C57BL/6 mice in their plasma (Pan & Tabel, unpublished). T. congolense-infected BALB/c mice might have more VSG-C3d immune complexes than infected C57BL/6 mice. The receptor for complement component C3d is the cell surface molecule CR2, also referred to as CD21. It is known that CR2 is widely expressed on B lymphocytes and follicular dendritic cells. There is also some evidence that CR2 is expressed on a subpopulation of activated T cells. Binding of VSG-C3d immune complexes to the complement receptor CR2 might costimulate the CR2+ T cells to produce IFN-ã. I hypothesize that IFN-ã-producing T cells in T. congolense-infected BALB/c mice are CR2+ and that the CR2+ T cells increase in numbers in experimental murine T. congolense infections. Kinetic studies were carried out by staining spleen cells of T. congolense-infected BALB/c mice for the presence of CR2 on T cells (CD3+ cells). Total numbers of spleen cells showed a 5-fold increase with progressive T. congolense infections. The total numbers of T cells in the spleen showed a 7-fold increase at day 8 post infection. The total numbers of CR2+ T cells in the spleen showed a 3 to 7-fold increase with progressive infection. Parallel studies on B lymphocytes (CD19+ cells) showed that absolute numbers of B cells in the spleen had a 5 to 6-fold increase with progressive infection. Absolute numbers of CR2+ B cells in the spleen showed a 4-fold increase at day 7 post infection. The total numbers of CR2+ cells in the spleen showed an increase while the mean numbers of CR2 molecules per cell showed a reduction with progressive infection. These results show that CR2+ T cells in the spleen… Advisors/Committee Members: Tabel, Henry, Havele, Calliopi, Gordon, John R., Bretscher, Peter A., Misra, Vikram.

Subjects/Keywords: CR2; Trypanosomiasis; kinetic studies; spleen cells

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APA (6^th Edition):

Munasinghe, L. I. (2009). Complement receptor 2 (CR2/CD21) in experimental African trypanosomiasis. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/etd-04232009-123902

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Munasinghe, Lilani Indika. “Complement receptor 2 (CR2/CD21) in experimental African trypanosomiasis.” 2009. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/etd-04232009-123902.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Munasinghe, Lilani Indika. “Complement receptor 2 (CR2/CD21) in experimental African trypanosomiasis.” 2009. Web. 08 Mar 2021.

Vancouver:

Munasinghe LI. Complement receptor 2 (CR2/CD21) in experimental African trypanosomiasis. [Internet] [Thesis]. University of Saskatchewan; 2009. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/etd-04232009-123902.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Munasinghe LI. Complement receptor 2 (CR2/CD21) in experimental African trypanosomiasis. [Thesis]. University of Saskatchewan; 2009. Available from: http://hdl.handle.net/10388/etd-04232009-123902

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

29. Zhang, Rui. The Effect of Transcription Factor Zhangfei/CREBZF on Osteosarcoma Cells and the Mechanisms Responsible.

Degree: 2014, University of Saskatchewan

URL: http://hdl.handle.net/10388/ETD-2014-06-1557

► Osteosarcoma (OS) is the most common primary malignant bone tumour in humans and dogs. Although medicine has made dramatic progress in treating osteosarcoma by surgery,… (more)

▼ Osteosarcoma (OS) is the most common primary malignant bone tumour in humans and dogs. Although medicine has made dramatic progress in treating osteosarcoma by surgery, with chemotherapy given before and after surgery, drug resistance and highly metastatic spread are often responsible for the failure of current therapies. Thus, more effective therapeutic approaches for treating osteosarcoma are needed. Previous results from our laboratory and others had shown that the basic-leucine zipper (bLZip) containing transcription factor, Zhangfei/CREBZF is a potent inhibitor of a variety of other transcription factors and has a dramatic effect on the growth of several cancer cell lines, including dog OS and human medulloblastoma cells. The objective of the studies described in this thesis was to determine the molecular mechanisms by which Zhangfei exerts its effect on dog and human OS cells. Several stressors in the microenvironment of cancer cells directly or indirectly perturb the endoplasmic reticulum (ER), which then activates the Unfolded Protein Response (UPR). The UPR modulates the effects of stress and allows tumours to survive, develop, metastasize and escape therapy. The UPR is regulated by three bLZip transcription factors—ATF6, ATF4 and Xbp1s. Since Zhangfei inhibits Luman/CREB3, a bLZip structurally similar to and closely related to ATF6 and ATF4, I initially focused my efforts on this pathway. I hypothesized that Zhangfei interacts with UPR-related bLZip transcription factors and inhibits their ability to activate the UPR signaling pathways, thereby suppressing the growth of cancer cells and increasing their susceptibility to ER stressors. To test this hypothesis, we monitored cell growth as well as levels of UPR gene transcripts and proteins in several dog and human osteosarcoma cell lines infected with adenovirus vectors expressing Zhangfei, and studied the interactions between Zhangfei and the UPR-mediator, Xbp1s. The results showed that the ectopic expression of Zhangfei in cell lines derived from dog osteosarcomas potently suppressed cell growth and inhibited their ability to activate the UPR. Further studies demonstrated that Zhangfei inhibited the UPR, at least partially, by binding to Xbp1s and suppressing its ability to activate transcription from a promoter containing unfolded protein response elements (UPRE). The leucine zipper of Zhangfei was required for this interaction, which led to the subsequent proteasomal degradation of Xbp1s. However, we also found that the effects of Zhangfei were not universal. While Zhangfei had a profound effect on the growth and UPR in some OS cell lines, it either had only a partial effect, or no effect on others. This suggested that susceptibility (or resistance) to Zhangfei may be an inherent property of OS cell lines. Since the suppressive effects of Zhangfei were not universal, and it had no obvious effects on untransformed cells and some cancer cell lines, I proposed that Zhangfei mediates its effect on cell growth and the UPR through an intermediary that is… Advisors/Committee Members: Misra, Vikram, Wobeser, Bruce, Haines, Deborah, Xiang, Jim, MacDonald Dickinson, Valerie, Hill, Janet.

Subjects/Keywords: Zhangfei/CREBZF; osteosarcoma; cell growth; UPR; p53

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APA (6^th Edition):

Zhang, R. (2014). The Effect of Transcription Factor Zhangfei/CREBZF on Osteosarcoma Cells and the Mechanisms Responsible. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/ETD-2014-06-1557

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Zhang, Rui. “The Effect of Transcription Factor Zhangfei/CREBZF on Osteosarcoma Cells and the Mechanisms Responsible.” 2014. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/ETD-2014-06-1557.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Zhang, Rui. “The Effect of Transcription Factor Zhangfei/CREBZF on Osteosarcoma Cells and the Mechanisms Responsible.” 2014. Web. 08 Mar 2021.

Vancouver:

Zhang R. The Effect of Transcription Factor Zhangfei/CREBZF on Osteosarcoma Cells and the Mechanisms Responsible. [Internet] [Thesis]. University of Saskatchewan; 2014. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/ETD-2014-06-1557.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Zhang R. The Effect of Transcription Factor Zhangfei/CREBZF on Osteosarcoma Cells and the Mechanisms Responsible. [Thesis]. University of Saskatchewan; 2014. Available from: http://hdl.handle.net/10388/ETD-2014-06-1557

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

30. Links, Matthew. Microbial profiling using metagenomic assembly.

Degree: 2013, University of Saskatchewan

URL: http://hdl.handle.net/10388/ETD-2013-09-1206

► The application of second generation sequencing technology to the characterization of complex microbial communities has profoundly affected our appreciation of microbial diversity. The explosive growth… (more)

▼ The application of second generation sequencing technology to the characterization of complex microbial communities has profoundly affected our appreciation of microbial diversity. The explosive growth of microbial sequence data has also necessitated advances in bioinformatic methods for profiling microbial communities. Data aggregation strategies should allow the relation of metagenomic sequence data to our understanding of microbial taxonomy, while also facilitating the discovery of novel taxa. For eukaryotes, a method has been established that links DNA sequences to the identification of organisms: DNA Barcoding. A similar approach has been developed for prokaryotes using target genic regions as markers for species identification and to profile communities. A key difference in these efforts is that within DNA barcoding there is a formalized framework for the evaluation of barcoding targets, whereas for prokaryotes the 16S rRNA gene target has become the de facto barcode without formal evaluation. Using the framework developed for evaluating DNA barcodes in eukaryotes, a study was undertaken to formally evaluate 16S rRNA and cpn60 as DNA barcodes for Bacteria. Both 16S rRNA and cpn60 were found to meet the criteria for DNA barcodes, with cpn60 a preferred barcode based on its superior resolution of closely related taxa. The high resolution of cpn60 enabled a method of sequence data aggregation through sequence assembly: microbial profiling using metagenomic assembly (mPUMA). The scoring of metagenomic assemblies in terms of sensitivity and specificity of the operational taxonomic units formed was used to evaluate and optimize the assembly of cpn60 barcodes. Using optimized parameters, mPUMA was demonstrated to faithfully reconstruct a synthetic community in terms of richness and abundance. To facilitate the use of mPUMA, a software package was developed and released under an open source license. The utility of mPUMA was further examined through the characterization of the epiphytic seed microbiomes of Triticum and Brassica species. A microbiome shared across both crop genera including fungi and bacteria was detected: a particularly important observation as it implies that seeds may serve as a vector for microbes that could include both pathogenic and beneficial organisms. The relative abundances of taxa identified by mPUMA were confirmed by qPCR for multiple cases of both fungal and bacterial taxa. By culturing isolates of both bacteria and fungi from the seed surfaces it was demonstrated that mPUMA faithfully assembled consensus sequences for OTUs that were 100% identical to isolated fungi and bacteria. Patterns observed in the relative abundances of the shared microbiome OTUs were used to generate the hypothesis that an Pantoea-like bacterium and an Alternaria-like fungus had an antagonistic relationship, since sequences corresponding to these organisms showed reciprocal abundance patterns on Triticum and Brassica seeds. Studies of the interactions of cultured isolates revealed fungistatic interactions that… Advisors/Committee Members: Hill, Janet E., Hemmingsen, Sean M., Parkin, Isobel A., Wilkinson, Mark, Misra, Vikram.

Subjects/Keywords: mPUMA; microbial profiling using metagenomic assembly; microbial profiling; metagenomics; microbiome; bioinformatics; computational biology; genomics; microbiota

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APA (6^th Edition):

Links, M. (2013). Microbial profiling using metagenomic assembly. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/ETD-2013-09-1206

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Links, Matthew. “Microbial profiling using metagenomic assembly.” 2013. Thesis, University of Saskatchewan. Accessed March 08, 2021. http://hdl.handle.net/10388/ETD-2013-09-1206.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Links, Matthew. “Microbial profiling using metagenomic assembly.” 2013. Web. 08 Mar 2021.

Vancouver:

Links M. Microbial profiling using metagenomic assembly. [Internet] [Thesis]. University of Saskatchewan; 2013. [cited 2021 Mar 08]. Available from: http://hdl.handle.net/10388/ETD-2013-09-1206.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Links M. Microbial profiling using metagenomic assembly. [Thesis]. University of Saskatchewan; 2013. Available from: http://hdl.handle.net/10388/ETD-2013-09-1206

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

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