+publisher:"Texas A&M University" +contributor:("Ball, Judith M.")

Texas A&M University

1. Schroeder, Megan Elizabeth. Secondary Structural and Functional Studies of Rotavirus NSP4 and Caveolin-1 Peptide-Peptide Interactions.

Degree: PhD, Veterinary Microbiology, 2011, Texas A&M University

URL: http://hdl.handle.net/1969.1/ETD-TAMU-2009-12-7454

► The rotavirus NSP4 protein is the first described viral enterotoxin. Abundant data from our laboratory reveals that NSP4 binds both the N- and C-termini of… (more)

▼ The rotavirus NSP4 protein is the first described viral enterotoxin. Abundant data from our laboratory reveals that NSP4 binds both the N- and C-termini of caveolin- 1 (aa2-31 and 161-178, respectively). Yeast two-hybrid and peptide binding analysis mapped the caveolin-1 binding site to three hydrophobic residues within the amphipathic a-helix, enterotoxic peptide domain (aa114-135). The research studies herein utilized peptides to investigate the interaction between NSP4 and caveolin-1. Peptides were synthesized corresponding to the amphipathic a-helix and caveolin-1 binding domain of NSP4 (aa112-140) and to the N- (aa2-20 and 19-40) and C- (161-178) termini of caveolin-1, and were utilized in structural and functional studies. Fluorescence binding assays revealed that NSP4 (aa112-140) binds to the N-terminus (aa19-40) of caveolin-1 with a stronger affinity than the C-terminus (aa161-178). In addition, this assay further delineated the NSP4 binding domain on caveolin-1 to aa19-40. Secondary structural changes following NSP4-caveolin-1 peptide-peptide interactions were investigated by circular dichroism analysis. Changes in a-helix formation were observed only upon interaction of the NSP4112-140 peptide with the C-terminal caveolin-1 peptide (C-Cav161- 178). The NSP4112-140 peptide contains a potential cholesterol recognition amino acid consensus (CRAC) sequence. Therefore this peptide was examined for cholesterol binding. Results of the binding assay revealed NSP4 binds cholesterol with a Kd of 7.67 +/- 1.49nM and this interaction occurs via aa112-140. Mutation of amino acid residues within the CRAC motif resulted in weaker binding affinities between each of the corresponding mutant peptides and cholesterol. NSP4 peptides containing mutations within the hydrophobic and charged faces of the amphipathic a-helix, enterotoxic peptide and caveolin-1 binding domain of NSP4 were examined for changes in secondary structure as well as diarrhea induction in mouse pups. Circular dichroism analysis revealed that mutation of hydrophobic residues resulted in a decrease in a-helix formation, whereas mutation of acidic and basic charged residues caused little to no change in a-helical content. When tested for diarrhea induction in mouse pups, the peptides containing mutations of either the hydrophobic or basic charged residues did not cause diarrhea. Taken together, the results of this research suggest a complex interplay between NSP4 secondary structure, caveolin-1 and cholesterol binding and diarrheagenic function. Advisors/Committee Members: Ball, Judith M. (advisor), Payne, Susan (committee member), Schroeder, Friedhelm (committee member), Zimmer, Danna (committee member).

Subjects/Keywords: Rotavirus NSP4; caveolin-1; circular dichroism; cholesterol; peptide-peptide interactions

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APA (6^th Edition):

Schroeder, M. E. (2011). Secondary Structural and Functional Studies of Rotavirus NSP4 and Caveolin-1 Peptide-Peptide Interactions. (Doctoral Dissertation). Texas A&M University. Retrieved from http://hdl.handle.net/1969.1/ETD-TAMU-2009-12-7454

Chicago Manual of Style (16^th Edition):

Schroeder, Megan Elizabeth. “Secondary Structural and Functional Studies of Rotavirus NSP4 and Caveolin-1 Peptide-Peptide Interactions.” 2011. Doctoral Dissertation, Texas A&M University. Accessed April 13, 2021. http://hdl.handle.net/1969.1/ETD-TAMU-2009-12-7454.

MLA Handbook (7^th Edition):

Schroeder, Megan Elizabeth. “Secondary Structural and Functional Studies of Rotavirus NSP4 and Caveolin-1 Peptide-Peptide Interactions.” 2011. Web. 13 Apr 2021.

Vancouver:

Schroeder ME. Secondary Structural and Functional Studies of Rotavirus NSP4 and Caveolin-1 Peptide-Peptide Interactions. [Internet] [Doctoral dissertation]. Texas A&M University; 2011. [cited 2021 Apr 13]. Available from: http://hdl.handle.net/1969.1/ETD-TAMU-2009-12-7454.

Council of Science Editors:

Schroeder ME. Secondary Structural and Functional Studies of Rotavirus NSP4 and Caveolin-1 Peptide-Peptide Interactions. [Doctoral Dissertation]. Texas A&M University; 2011. Available from: http://hdl.handle.net/1969.1/ETD-TAMU-2009-12-7454

Texas A&M University

2. Yakshe, Krystle Ann. Functional Analysis of Interactions of Rotavirus NSP4 with Caveolin-1, Cyclophilin A, Cyclophilin 40, Heat Shock Protein 56, and Cholesterol.

Degree: PhD, Veterinary Microbiology, 2015, Texas A&M University

URL: http://hdl.handle.net/1969.1/156449

► The rotavirus enterotoxin, NSP4, is responsible for early secretory diarrhea associated with rotavirus infection and is critical for RV replication and morphogenesis. NSP4 interacts directly… (more)

Subjects/Keywords: Rotavirus NSP4; Cyclophilin A; Cyclophilin 40; Heat Shock Protein 56; Caveolin-1; Cholesterol

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APA (6^th Edition):

Yakshe, K. A. (2015). Functional Analysis of Interactions of Rotavirus NSP4 with Caveolin-1, Cyclophilin A, Cyclophilin 40, Heat Shock Protein 56, and Cholesterol. (Doctoral Dissertation). Texas A&M University. Retrieved from http://hdl.handle.net/1969.1/156449

Chicago Manual of Style (16^th Edition):

Yakshe, Krystle Ann. “Functional Analysis of Interactions of Rotavirus NSP4 with Caveolin-1, Cyclophilin A, Cyclophilin 40, Heat Shock Protein 56, and Cholesterol.” 2015. Doctoral Dissertation, Texas A&M University. Accessed April 13, 2021. http://hdl.handle.net/1969.1/156449.

MLA Handbook (7^th Edition):

Yakshe, Krystle Ann. “Functional Analysis of Interactions of Rotavirus NSP4 with Caveolin-1, Cyclophilin A, Cyclophilin 40, Heat Shock Protein 56, and Cholesterol.” 2015. Web. 13 Apr 2021.

Vancouver:

Yakshe KA. Functional Analysis of Interactions of Rotavirus NSP4 with Caveolin-1, Cyclophilin A, Cyclophilin 40, Heat Shock Protein 56, and Cholesterol. [Internet] [Doctoral dissertation]. Texas A&M University; 2015. [cited 2021 Apr 13]. Available from: http://hdl.handle.net/1969.1/156449.

Council of Science Editors:

Yakshe KA. Functional Analysis of Interactions of Rotavirus NSP4 with Caveolin-1, Cyclophilin A, Cyclophilin 40, Heat Shock Protein 56, and Cholesterol. [Doctoral Dissertation]. Texas A&M University; 2015. Available from: http://hdl.handle.net/1969.1/156449

3. Covaleda Salas, Lina M. New Insights Into the Role of Equine Infectious Anemia Virus S2 Protein in Disease Expression.

Degree: PhD, Veterinary Microbiology, 2011, Texas A&M University

URL: http://hdl.handle.net/1969.1/ETD-TAMU-2010-05-7975

► Equine infectious anemia virus (EIAV) is an important animal model to study the contribution of macrophages in viral persistence during lentiviral infections. EIAV is unique… (more)

▼ Equine infectious anemia virus (EIAV) is an important animal model to study the contribution of macrophages in viral persistence during lentiviral infections. EIAV is unique amongst the lentiviruses in that it causes a rapid, rather than the very slow disease progression, characteristic of other lentiviral infections. The accessory gene, S2, unique to EIAV, is an important determinant in viral pathogenesis. A functional S2 gene is required to achieve high-titer viremia and the development of disease in infected horses. Despite its essential role, the mechanisms by which S2 influences EIAV pathogenesis remain elusive. The goal of this research was to gain insight into the role of S2 in pathogenesis. To accomplish this goal we: (i) Examined the effects of EIAV and its S2 protein in the regulation of the cytokine and chemokine responses in macrophages, (ii) Assessed the influence of EIAV infection and the effect of S2 on global gene expression in macrophages and (iii) Identified host cellular proteins that interact with S2 as a starting point for the identification of host factors implicated in S2 function. The results from this study provide evidence for a role of S2 in enhancing a proinflammatory cytokine and chemokine response in infected macrophages. Specifically, S2 enhances the expression of IL-1 alpha, IL-1 beta IL-8, MCP-2, MIP-1 beta, IP-10 and a newly discovered cytokine, IL-34. Involvement of S2 in cytokine and chemokine dysregulation may contribute to disease development by optimizing the host cell environment to promote viral dissemination and replication. Microarray analyses revealed an interesting set of differentially expressed genes upon EIAV infection. Genes affected by EIAV were involved in the immune response, transcription, translation, cell cycle and cell survival. Finally, we used the yeast two-hybrid system to identify S2 host cellular interacting proteins. We identified osteosarcoma amplified 9 (OS-9) and proteasome 26S ATPase subunit 3 (PSMC3) proteins as interacting partners of S2. Additional evidence is needed to demonstrate the physiological relevance of these interactions in vivo. In summary, the results from this study contribute towards our understanding of the role S2 in disease expression and allow the formulation of new hypotheses as to the potential mechanisms of action of S2 during EIAV infection. Advisors/Committee Members: Payne, Susan (advisor), Ball, Judith M. (committee member), Welsh, Jane (committee member), Wilson, Van G. (committee member).

Subjects/Keywords: EIAV; Cytokines; Chemokines; Macrophage; Gene expression; Dysregulation; Real-time PCR; Microarray, Yeast two-hybrid system

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APA (6^th Edition):

Covaleda Salas, L. M. (2011). New Insights Into the Role of Equine Infectious Anemia Virus S2 Protein in Disease Expression. (Doctoral Dissertation). Texas A&M University. Retrieved from http://hdl.handle.net/1969.1/ETD-TAMU-2010-05-7975

Chicago Manual of Style (16^th Edition):

Covaleda Salas, Lina M. “New Insights Into the Role of Equine Infectious Anemia Virus S2 Protein in Disease Expression.” 2011. Doctoral Dissertation, Texas A&M University. Accessed April 13, 2021. http://hdl.handle.net/1969.1/ETD-TAMU-2010-05-7975.

MLA Handbook (7^th Edition):

Covaleda Salas, Lina M. “New Insights Into the Role of Equine Infectious Anemia Virus S2 Protein in Disease Expression.” 2011. Web. 13 Apr 2021.

Vancouver:

Covaleda Salas LM. New Insights Into the Role of Equine Infectious Anemia Virus S2 Protein in Disease Expression. [Internet] [Doctoral dissertation]. Texas A&M University; 2011. [cited 2021 Apr 13]. Available from: http://hdl.handle.net/1969.1/ETD-TAMU-2010-05-7975.

Council of Science Editors:

Covaleda Salas LM. New Insights Into the Role of Equine Infectious Anemia Virus S2 Protein in Disease Expression. [Doctoral Dissertation]. Texas A&M University; 2011. Available from: http://hdl.handle.net/1969.1/ETD-TAMU-2010-05-7975

4. Williams, Cecelia V. Mapping of the rotavirus nonstructural protein-4-caveolin-1 binding site to three hydrophobic residues within the extended, c-terminal amphipathic alpha helix.

Degree: PhD, Veterinary Microbiology, 2009, Texas A&M University

URL: http://hdl.handle.net/1969.1/ETD-TAMU-3258

► Rotavirus NSP4, the first described viral enterotoxin, localizes to the plasma membrane of infected cells, possibly through interaction with caveolin-1. A direct interaction between NSP4… (more)

▼ Rotavirus NSP4, the first described viral enterotoxin, localizes to the plasma membrane of infected cells, possibly through interaction with caveolin-1. A direct interaction between NSP4 and caveolin-1, the structural protein of caveolae, was shown by yeast two-hybrid, peptide binding, and FRET analyses. To dissect the precise NSP4 binding domain to caveolin-1, mutants were prepared by altering either the charged or hydrophobic face of the NSP4 C-terminal amphipathic alpha-helix and examined for binding to caveolin-1. Replacing six charged residues with alanine (FLNSP4Ala) disrupted the charged face, while the hydrophobic face was disrupted by replacing selected hydrophobic residues with charged amino acids (aa) (FLNSP4HydroMut). In yeast two-hybrid and peptide binding assays, FLNSP4Ala retained its binding capacity, whereas FLNSP4HydroMut failed to bind caveolin-1. Mutants were generated with an Nterminal truncated clone (NSP446-175), which removed the hydrophobic domains and aided in yeast-two hybrid assays. These mutants exhibited the same binding pattern as FLNSP4 confirming that the N-terminus of NSP4 lacks the caveolin-1 binding site and NSP446-175 is sufficient for binding. Seven additional mutants were prepared from NSP4HydroMut in which individually charged residues were reverted to the original hydrophobic aa or were replaced with alanine. Analyses of the interaction of these revertants with caveolin-1 localized the NSP4 binding domain to one critical hydrophobic aa (L116) and one or two additional aa (I113, L127, and/or L134) on the hydrophobic face. Those mutants that bound caveolin-1 bound both the N- and C-terminal caveolin-1 peptides, but lacked binding to a centrally located peptide. These data suggest conformational and hydrophobic constraints play a role in the NSP4-caveolin-1 association. The mutant NSP4 molecules also were evaluated for transport to the plasma membrane. Mammalian cells were transfected with FLNSP4, FLNSP41-175Ala, and NSP41-175HydroMut plasmid DNA, surface biotinylated, and examined by IFA or Western blot for NSP4 expression. Epifluorescence revealed FLNSP4 and FLNSP4Ala were exposed on the cell surface in the absence of other viral proteins, whereas NSP4HydroMut remained intracellular. Further, NSP4-transfected cells displayed an intracellular association of with caveolin-1 or the caveolin-1 chaperone complex proteins. These data indicate NSP4 interacts with caveolin-1 in the absence of other viral proteins. Advisors/Committee Members: Ball, Judith M. (advisor), Leibowitz, J (committee member), Parr, Rebecca D. (committee member), Schroeder, Friedhelm (committee member).

Subjects/Keywords: Rotavirus; NSP4; Caveolin-1; Hydrophobic Face

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

APA (6^th Edition):

Williams, C. V. (2009). Mapping of the rotavirus nonstructural protein-4-caveolin-1 binding site to three hydrophobic residues within the extended, c-terminal amphipathic alpha helix. (Doctoral Dissertation). Texas A&M University. Retrieved from http://hdl.handle.net/1969.1/ETD-TAMU-3258

Chicago Manual of Style (16^th Edition):

Williams, Cecelia V. “Mapping of the rotavirus nonstructural protein-4-caveolin-1 binding site to three hydrophobic residues within the extended, c-terminal amphipathic alpha helix.” 2009. Doctoral Dissertation, Texas A&M University. Accessed April 13, 2021. http://hdl.handle.net/1969.1/ETD-TAMU-3258.

MLA Handbook (7^th Edition):

Williams, Cecelia V. “Mapping of the rotavirus nonstructural protein-4-caveolin-1 binding site to three hydrophobic residues within the extended, c-terminal amphipathic alpha helix.” 2009. Web. 13 Apr 2021.

Vancouver:

Williams CV. Mapping of the rotavirus nonstructural protein-4-caveolin-1 binding site to three hydrophobic residues within the extended, c-terminal amphipathic alpha helix. [Internet] [Doctoral dissertation]. Texas A&M University; 2009. [cited 2021 Apr 13]. Available from: http://hdl.handle.net/1969.1/ETD-TAMU-3258.

Council of Science Editors:

Williams CV. Mapping of the rotavirus nonstructural protein-4-caveolin-1 binding site to three hydrophobic residues within the extended, c-terminal amphipathic alpha helix. [Doctoral Dissertation]. Texas A&M University; 2009. Available from: http://hdl.handle.net/1969.1/ETD-TAMU-3258

5. Storey, Stephen Michael. Intracellular trafficking and plasma membrane microdomain distribution of the NSP4 enterotoxin during rotavirus infection in epithelial cells.

Degree: PhD, Veterinary Microbiology, 2009, Texas A&M University

URL: http://hdl.handle.net/1969.1/ETD-TAMU-1210

► Rotavirus (RV) nonstructural protein 4 (NSP4) is a multifunctional glycoprotein that induces secretory diarrhea in mouse pups in the absence of other viral proteins. The… (more)

▼ Rotavirus (RV) nonstructural protein 4 (NSP4) is a multifunctional glycoprotein that induces secretory diarrhea in mouse pups in the absence of other viral proteins. The intracellular transport route(s) and functional mechanism(s) of NSP4 are poorly understood; however, the recent association of the enterotoxin with cellular caveolin-1 may provide a link between NSP4 transport and function. To determine if NSP4 traffics to a specific subset of lipid rafts at the plasma membrane (PM), we isolated caveolae from a PM-enriched fraction with a new method that yielded endoplasmic reticulum (ER)-free caveolae membranes with a unique membrane structure and composition. Comparison of these caveolae with other detergent- and non-detergent-extracted membranes revealed that each caveolae/raft fraction contained caveolae markers; however, only our PM caveolae fraction mimicked the membrane structure and sterol exchange dynamics of intact PM without ER or non-raft PM contaminants. When these PM caveolae were isolated from RV-infected cells, full-length, high-mannose glycosylated NSP4 was present. Confocal imaging showed association of NSP4 with caveolin-1 moving from perinuclear and cytoplasmic sites toward the PM as the infection progressed. Fluorescent imaging also indicated exposure of the NSP4 Cterminus at the exofacial PM surface without transport of the enterotoxin through the Golgi apparatus. Surface-specific biotinylation was used to confirm NSP4 exposure at the surface of infected MDCK cells and to determine that the exposed protein was fulllength and high-mannose glycosylated. This study presents an ER contaminant-free PM caveolae isolation methodology, identifies the presence of full-length, high-mannose glycosylated NSP4 in both PM caveolae and exposed at the cell surface, and confirms the Golgi-bypassing nature of NSP4 ER to PM transport in RV-infected MDCK cells. Advisors/Committee Members: Ball, Judith M. (advisor), Collisson, Ellen W. (committee member), Leibowitz, Julian (committee member), Schroeder, Friedhelm (committee member).

Subjects/Keywords: caveolae isolation; NSP4; rotavirus; lipid raft

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

APA (6^th Edition):

Storey, S. M. (2009). Intracellular trafficking and plasma membrane microdomain distribution of the NSP4 enterotoxin during rotavirus infection in epithelial cells. (Doctoral Dissertation). Texas A&M University. Retrieved from http://hdl.handle.net/1969.1/ETD-TAMU-1210

Chicago Manual of Style (16^th Edition):

Storey, Stephen Michael. “Intracellular trafficking and plasma membrane microdomain distribution of the NSP4 enterotoxin during rotavirus infection in epithelial cells.” 2009. Doctoral Dissertation, Texas A&M University. Accessed April 13, 2021. http://hdl.handle.net/1969.1/ETD-TAMU-1210.

MLA Handbook (7^th Edition):

Storey, Stephen Michael. “Intracellular trafficking and plasma membrane microdomain distribution of the NSP4 enterotoxin during rotavirus infection in epithelial cells.” 2009. Web. 13 Apr 2021.

Vancouver:

Storey SM. Intracellular trafficking and plasma membrane microdomain distribution of the NSP4 enterotoxin during rotavirus infection in epithelial cells. [Internet] [Doctoral dissertation]. Texas A&M University; 2009. [cited 2021 Apr 13]. Available from: http://hdl.handle.net/1969.1/ETD-TAMU-1210.

Council of Science Editors:

Storey SM. Intracellular trafficking and plasma membrane microdomain distribution of the NSP4 enterotoxin during rotavirus infection in epithelial cells. [Doctoral Dissertation]. Texas A&M University; 2009. Available from: http://hdl.handle.net/1969.1/ETD-TAMU-1210

6. Baradji, Issa. The Role of Apical Membrane Antigen-1 in Erythrocyte Invasion by the Zoonotic Apicomplexan Babesia microti.

Degree: PhD, Veterinary Microbiology, 2010, Texas A&M University

URL: http://hdl.handle.net/1969.1/ETD-TAMU-2008-08-68

► Babesia microti is a tickborne hemoprotozoan parasite that causes the disease babesiosis in humans. Babesia microti Apical Membrane Antigen-1 (AMA-1) is a micronemal protein suspected… (more)

▼ Babesia microti is a tickborne hemoprotozoan parasite that causes the disease babesiosis in humans. Babesia microti Apical Membrane Antigen-1 (AMA-1) is a micronemal protein suspected to play a role in erythrocyte invasion. To investigate interaction between AMA-1 and the host cell, the ectodomain region of the B. microti ama-1 gene was cloned into an expression vector, expressed as a histidine-tagged fusion protein, and used to probe red blood cell membrane proteins in far Western blot assays. The B. microti ama-1 ectodomain, which excludes the signal peptide and the transmembrane region of the open reading frame, was amplified from a cloned gene sequence. The AMA-1 ectodomain is a membrane bound polypeptide that extends into the extracellular space and is most likely to interact or initiate interaction with the host red blood cell surface receptor(s). The amplicon was ligated into a protein expression vector to produce a 58.1 kDa recombinant His-tagged fusion protein, which was confirmed by Western blot analysis. The recombinant B. microti AMA-1 fusion protein was enriched on nickel affinity columns and then used to probe mouse, human and horse red blood cell membrane proteins in far Western blot assays. Babesia microti AMA-1 consistently reacted strongly with a protein migrating at 49 kDa. A similar reaction occurred between the B. microti AMA-1 and horse red blood cell membrane proteins, suggesting that similar interacting proteins of this size are shared by red blood cells from the three species. The B. microti AMA-1 may bind to red blood cell membrane sialic-acid groups, as shown for other Babesia spp. This may explain the signal at the 49 kDa position observed between B. microti AMA-1 and red blood cell membrane proteins from three different species. Further studies may determine if the binding epitopes of the red blood cell binding partner at this position vary and contribute to the specificity of each parasite AMA-1 for their respective host cells. Advisors/Committee Members: Holman, Patricia J. (advisor), Ball, Judith M. (committee member), Magill, Clint C. (committee member), Womack, James E. (committee member).

Subjects/Keywords: Babesia microti; Apical Membrane Antigen-1; AMA-1; erythrocyte; parasite ligand; red blood cell; receptor; protein-protein interaction

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APA (6^th Edition):

Baradji, I. (2010). The Role of Apical Membrane Antigen-1 in Erythrocyte Invasion by the Zoonotic Apicomplexan Babesia microti. (Doctoral Dissertation). Texas A&M University. Retrieved from http://hdl.handle.net/1969.1/ETD-TAMU-2008-08-68

Chicago Manual of Style (16^th Edition):

Baradji, Issa. “The Role of Apical Membrane Antigen-1 in Erythrocyte Invasion by the Zoonotic Apicomplexan Babesia microti.” 2010. Doctoral Dissertation, Texas A&M University. Accessed April 13, 2021. http://hdl.handle.net/1969.1/ETD-TAMU-2008-08-68.

MLA Handbook (7^th Edition):

Baradji, Issa. “The Role of Apical Membrane Antigen-1 in Erythrocyte Invasion by the Zoonotic Apicomplexan Babesia microti.” 2010. Web. 13 Apr 2021.

Vancouver:

Baradji I. The Role of Apical Membrane Antigen-1 in Erythrocyte Invasion by the Zoonotic Apicomplexan Babesia microti. [Internet] [Doctoral dissertation]. Texas A&M University; 2010. [cited 2021 Apr 13]. Available from: http://hdl.handle.net/1969.1/ETD-TAMU-2008-08-68.

Council of Science Editors:

Baradji I. The Role of Apical Membrane Antigen-1 in Erythrocyte Invasion by the Zoonotic Apicomplexan Babesia microti. [Doctoral Dissertation]. Texas A&M University; 2010. Available from: http://hdl.handle.net/1969.1/ETD-TAMU-2008-08-68

7. Rodriguez Frausto, Heriberto. Development and analytical validation of a gas chromatography-mass spectrometry method for the assessment of gastrointestinal permeability and intestinal absorptive capacity in dogs.

Degree: PhD, Veterinary Microbiology, 2009, Texas A&M University

URL: http://hdl.handle.net/1969.1/ETD-TAMU-3257

► Assessment of gastrointestinal permeability in vivo is considered a suitable method for the evaluation of gastrointestinal mucosal integrity. Probes commonly used include lactulose (L) and… (more)

▼ Assessment of gastrointestinal permeability in vivo is considered a suitable method for the evaluation of gastrointestinal mucosal integrity. Probes commonly used include lactulose (L) and rhamnose (R) for the assessment of intestinal permeability, xylose (X) and 3-O-methylglucose (M) for the evaluation of intestinal absorptive capacity, and sucrose (S) for the assessment of gastric permeability. Traditionally, various methods have been used to quantify these markers in the urine after orogastric administration. However, urine collection is difficult and uncomfortable. A protocol based on the analysis of blood samples would be easier to perform. Thus, the aim of the first part of this project was to develop and validate a new gas chromatography-mass spectrometry (GC-MS) method for the quantification of five sugar probes in canine serum. The method was sensitive, accurate, precise, and reproducible for the simultaneous quantification of 5 sugar probes in serum. The aim of the second part of this project was to assess the kinetic profiles of these 5 sugar probes in serum after orogastric administration in dogs and to determine the optimal time point for sample collection. Dogs received a solution containing L (10 g/L), R (10 g/L), X (10 g/L), M (5 g/L), and S (40 g/L) by orogastric intubation. Baseline blood samples were collected. Subsequent timed blood samples were taken for a 24 hours period. Significant changes in serum concentrations of all 5 sugars were detected after administration of the test dose (p<0.0001 for all 5 probes). Serum concentrations of L and R were significantly different from baseline concentrations from 90 to 240 and from 60 to 300 min post dosing respectively, and those of X, M, and S were significantly different from 30 to 240 min after dosing (p<0.05 for all 5 probes). Variations of the mean sugar concentrations of all dogs at 90, 120, and 180 minutes were analyzed using a Kruskal-Wallis test. Based on the results, only two blood samples, one taken at baseline and a second sample obtained between 90 and 180 after dosing, appear to be sufficient for assessment of intestinal permeability and mucosal absorptive capacity using these sugar probes. Advisors/Committee Members: Steiner, J?M. (advisor), Roussel, Allen J. (committee member), Ball, Judith M. (committee member), Suchodolski, Jan S. (committee member), Williams,David A. (committee member).

Subjects/Keywords: Gas chromatography-Mass Spectrometry; GI Permeabiliy; Intestinal absorption; Sugar probes; Serum

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APA (6^th Edition):

Rodriguez Frausto, H. (2009). Development and analytical validation of a gas chromatography-mass spectrometry method for the assessment of gastrointestinal permeability and intestinal absorptive capacity in dogs. (Doctoral Dissertation). Texas A&M University. Retrieved from http://hdl.handle.net/1969.1/ETD-TAMU-3257

Chicago Manual of Style (16^th Edition):

Rodriguez Frausto, Heriberto. “Development and analytical validation of a gas chromatography-mass spectrometry method for the assessment of gastrointestinal permeability and intestinal absorptive capacity in dogs.” 2009. Doctoral Dissertation, Texas A&M University. Accessed April 13, 2021. http://hdl.handle.net/1969.1/ETD-TAMU-3257.

MLA Handbook (7^th Edition):

Rodriguez Frausto, Heriberto. “Development and analytical validation of a gas chromatography-mass spectrometry method for the assessment of gastrointestinal permeability and intestinal absorptive capacity in dogs.” 2009. Web. 13 Apr 2021.

Vancouver:

Rodriguez Frausto H. Development and analytical validation of a gas chromatography-mass spectrometry method for the assessment of gastrointestinal permeability and intestinal absorptive capacity in dogs. [Internet] [Doctoral dissertation]. Texas A&M University; 2009. [cited 2021 Apr 13]. Available from: http://hdl.handle.net/1969.1/ETD-TAMU-3257.

Council of Science Editors:

Rodriguez Frausto H. Development and analytical validation of a gas chromatography-mass spectrometry method for the assessment of gastrointestinal permeability and intestinal absorptive capacity in dogs. [Doctoral Dissertation]. Texas A&M University; 2009. Available from: http://hdl.handle.net/1969.1/ETD-TAMU-3257

Texas A&M University

8. Mir, Kiran D. The rotavirus nonstructural protein 4 (NSP4) interacts with both the N- and C- termini of caveolin-1.

Degree: MS, Veterinary Microbiology, 2006, Texas A&M University

URL: http://hdl.handle.net/1969.1/3976

► Rotavirus (RV) is an etiologic agent of viral gastroenteritis in children and infants worldwide, accounting for an estimated 500,000 deaths annually. NSP4, the first described… (more)

Subjects/Keywords: rotavirus; NSP4; caveolin-1; yeast two-hybrid

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APA (6^th Edition):

Mir, K. D. (2006). The rotavirus nonstructural protein 4 (NSP4) interacts with both the N- and C- termini of caveolin-1. (Masters Thesis). Texas A&M University. Retrieved from http://hdl.handle.net/1969.1/3976

Chicago Manual of Style (16^th Edition):

Mir, Kiran D. “The rotavirus nonstructural protein 4 (NSP4) interacts with both the N- and C- termini of caveolin-1.” 2006. Masters Thesis, Texas A&M University. Accessed April 13, 2021. http://hdl.handle.net/1969.1/3976.

MLA Handbook (7^th Edition):

Mir, Kiran D. “The rotavirus nonstructural protein 4 (NSP4) interacts with both the N- and C- termini of caveolin-1.” 2006. Web. 13 Apr 2021.

Vancouver:

Mir KD. The rotavirus nonstructural protein 4 (NSP4) interacts with both the N- and C- termini of caveolin-1. [Internet] [Masters thesis]. Texas A&M University; 2006. [cited 2021 Apr 13]. Available from: http://hdl.handle.net/1969.1/3976.

Council of Science Editors:

Mir KD. The rotavirus nonstructural protein 4 (NSP4) interacts with both the N- and C- termini of caveolin-1. [Masters Thesis]. Texas A&M University; 2006. Available from: http://hdl.handle.net/1969.1/3976

Texas A&M University

9. Lim, Wah-Seng. Differential cytokine mRNA expression induced by binding of virulent and avirulent molecularly cloned equine infectious anemia viruses to equine macrophages.

Degree: PhD, Veterinary Microbiology, 2004, Texas A&M University

URL: http://hdl.handle.net/1969.1/1225

► Equine infectious anemia virus (EIAV) causes rapid development of acute disease followed by recurring episodes of fever, thrombocytopenia and viremia, termed chronic EIA. Most infected… (more)

▼ Equine infectious anemia virus (EIAV) causes rapid development of acute disease followed by recurring episodes of fever, thrombocytopenia and viremia, termed chronic EIA. Most infected horses control the virus by immune mechanisms and become inapparent carriers. To further our understanding of the equine immune response to EIAV, a multi-probe ribonuclease protection assay (RPA) was developed to quantitate equine-specific cytokine mRNAs. Eleven template plasmids specific to ten equine cytokine genes and the ?-actin gene were generated, from which radiolabeled anti-sense RNA probes were produced. The RPA simultaneously quantitated mRNA levels of interleukin (IL)-1, IL-1, IL-6, IL-8, IL-10, IL-12 p35, IL-12 p40, interferon (IFN)-, transforming growth factor (TGF)-1 and tumor necrosis factor (TNF)- in equine peripheral blood mononuclear cells and equine monocyte-derived macrophages (EMDM). The assay detected as few as 5105 RNA molecules and displayed coefficients of variation of 0.03-0.08 when normalized to -actin expression. Using this RPA, cytokine expression in EMDM infected with 2 molecularly cloned viruses (EIAV17 and EIAV19) was determined. EIAV17 varies from EIAV19 only in env, rev and LTR and causes fatal disease in Shetland ponies. When added to EMDM cultures, virulent EIAV17 stimulated expression of IL-1, IL-1, IL-6, IL-10 and TNF-. These cytokine mRNAs were significantly elevated by 0.5 to 1 hr post infection (hpi) and returned to basal levels by 12 to 24 hpi, indicating modulation by early event(s), such as receptor binding. In contrast to EIAV17, EIAV19 is avirulent in vivo and failed to induce any of the tested cytokines in EMDM. These data show a direct correlation between the virulence of the EIAV clone and the induction of cytokines. The cytokines stimulated by EIAV17 may contribute to EIA-associated symptoms, enhance viral replication in the host, and regulate the host immune response. To determine whether cytokine induction requires EIAV17 replication, EMDM cultures were exposed to UV-inactivated EIAV17 and cytokine induction was monitored. UV-inactivation did not block cytokine induction by EIAV17, suggesting dispensability of viral replication. Given that EIAV17 induces cytokines in a rapid and replication-independent manner, the activation of cytokine expression is likely mediated by binding of EIAV17 to equine macrophage receptor(s). Advisors/Committee Members: Ball, Judith M. (advisor), Welsh, C. Jane (committee member), Collisson, Ellen W. (committee member), Smith, Roger, III (committee member).

Subjects/Keywords: equine infectious anemia viruse; macrophage; cytokine; virulence

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

APA (6^th Edition):

Lim, W. (2004). Differential cytokine mRNA expression induced by binding of virulent and avirulent molecularly cloned equine infectious anemia viruses to equine macrophages. (Doctoral Dissertation). Texas A&M University. Retrieved from http://hdl.handle.net/1969.1/1225

Chicago Manual of Style (16^th Edition):

Lim, Wah-Seng. “Differential cytokine mRNA expression induced by binding of virulent and avirulent molecularly cloned equine infectious anemia viruses to equine macrophages.” 2004. Doctoral Dissertation, Texas A&M University. Accessed April 13, 2021. http://hdl.handle.net/1969.1/1225.

MLA Handbook (7^th Edition):

Lim, Wah-Seng. “Differential cytokine mRNA expression induced by binding of virulent and avirulent molecularly cloned equine infectious anemia viruses to equine macrophages.” 2004. Web. 13 Apr 2021.

Vancouver:

Lim W. Differential cytokine mRNA expression induced by binding of virulent and avirulent molecularly cloned equine infectious anemia viruses to equine macrophages. [Internet] [Doctoral dissertation]. Texas A&M University; 2004. [cited 2021 Apr 13]. Available from: http://hdl.handle.net/1969.1/1225.

Council of Science Editors:

Lim W. Differential cytokine mRNA expression induced by binding of virulent and avirulent molecularly cloned equine infectious anemia viruses to equine macrophages. [Doctoral Dissertation]. Texas A&M University; 2004. Available from: http://hdl.handle.net/1969.1/1225

Texas A&M University

10. Styles, Darrel Keith. Psittacid herpesvirus associated with internal papillomatous disease and other tumors in psittacine birds.

Degree: PhD, Veterinary Microbiology, 2005, Texas A&M University

URL: http://hdl.handle.net/1969.1/2646

► Internal papillomatous disease (IPD) is characterized by mucosal papillomas occurring primarily in the oral cavity and cloaca of Neotropical parrots. These lesions can cause considerable… (more)

▼ Internal papillomatous disease (IPD) is characterized by mucosal papillomas occurring primarily in the oral cavity and cloaca of Neotropical parrots. These lesions can cause considerable morbidity, and in some cases result in mortality. Efforts to demonstrate papillomavirus DNA or proteins in the lesions have been largely unsuccessful. However, increasing evidence suggests that mucosal papillomas may contain psittacid herpesviruses (PsHVs). In this study, PsHV 1 genotype 1, 2, and 3 DNA was found in 100% of mucosal papillomas from 30 Neotropical parrots by PCR using PsHV specific primers. However, Psittacus erithacus papillomavirus and finch papillomavirus DNA were not detected. Additionally, a novel PsHV sequence related to, but phylogenetically distinct from PsHV 1, was identified in 4 African grey parrots (Psittacus erithacus), two of which exhibited papillomas. These findings suggest that mucosal papillomas may develop in parrots latently infected with PsHV. Tumors of the bile and pancreatic ducts have also been observed in parrots with IPD. Other mucosal tumors including carcinomas of the proventriculus and ventriculus may be coincident with bile duct tumors, but cloacal carcinomas usually develop as solitary lesions. To test whether PsHV was associated with these tumors, the fresh tissues from 11 parrots and the formalin-fixed paraffin-embedded (FFPE) tissues of 5 parrots exhibiting mucosal tumors were examined by PCR. All tumors were found to contain PsHV 1 genotype 3 DNA except one bird with a cloacal carcinoma that contained genotype 4. Histologically normal tissues available from six parrots did not contain PsHV DNA. Experiments were performed using the FFPE tissues of 5 parrots with IPD related tumors known to contain PsHV by PCR, to show that the virus was in significantly higher concentration in the neoplastic tissue compared to adjacent histologically normal tissue. Neoplastic and adjacent unaffected cells were dissected from the tissues using laser capture microdissection and the DNA was examined by PCR. In situ hybridization using PsHV specific probes and direct in situ PCR were also performed on the tissues. A strong association was shown between infection by PsHV 1 genotype 3 and birds manifesting IPD related tumors and other neoplasms of the digestive tract. Advisors/Committee Members: Phalen, David N. (advisor), Ball, Judith M. (committee member), Tizard, Ian R. (committee member), Wilson, Van G. (committee member).

Subjects/Keywords: Parrots; Psittacid Herpesviruses; Papillomas; Tumors

Record Details Similar Records Cite « Share

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

APA (6^th Edition):

Styles, D. K. (2005). Psittacid herpesvirus associated with internal papillomatous disease and other tumors in psittacine birds. (Doctoral Dissertation). Texas A&M University. Retrieved from http://hdl.handle.net/1969.1/2646

Chicago Manual of Style (16^th Edition):

Styles, Darrel Keith. “Psittacid herpesvirus associated with internal papillomatous disease and other tumors in psittacine birds.” 2005. Doctoral Dissertation, Texas A&M University. Accessed April 13, 2021. http://hdl.handle.net/1969.1/2646.

MLA Handbook (7^th Edition):

Styles, Darrel Keith. “Psittacid herpesvirus associated with internal papillomatous disease and other tumors in psittacine birds.” 2005. Web. 13 Apr 2021.

Vancouver:

Styles DK. Psittacid herpesvirus associated with internal papillomatous disease and other tumors in psittacine birds. [Internet] [Doctoral dissertation]. Texas A&M University; 2005. [cited 2021 Apr 13]. Available from: http://hdl.handle.net/1969.1/2646.

Council of Science Editors:

Styles DK. Psittacid herpesvirus associated with internal papillomatous disease and other tumors in psittacine birds. [Doctoral Dissertation]. Texas A&M University; 2005. Available from: http://hdl.handle.net/1969.1/2646

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