Life Sciences Commons^™

Mucosal Immunization With A Staphylococcus Aureus Isda-Cholera Toxin A2/B Chimera Induces Antigen-Specific Th2-Type Responses In Mice, Britni M. Arlian, Juliette Tinker

Biology Faculty Publications and Presentations

Staphylococcus aureus is a leading cause of opportunistic infection worldwide and a significant public health threat. The iron-regulated surface determinant A (IsdA) adhesin is essential for S. aureus colonization on human nasal epithelial cells and plays an important role in iron acquisition and resistance to human skin defenses. Here we investigated the murine immune response to intranasal administration of a cholera toxin (CT) A₂/B chimera containing IsdA. Plasmids were constructed to express the IsdA-CTA₂/B chimera and control proteins in E. coli. Proper construction of the chimera was verified by SDS-PAGE, western blot, GM1 ELISA, and …

Depletion Of Beclin-1 Due To Proteolytic Cleavage By Caspases In The Alzheimer's Disease Brain, Troy T. Rohn, Ellen Wirawan, Raquel J. Brown, Jordan R. Harris, Eliezer Masliah, Peter Vandenabeele

Biology Faculty Publications and Presentations

The Beclin-1 protein is essential for the initiation of autophagy and recent studies suggest this function may be compromised in Alzheimer’s disease (AD). In addition, in vitro studies have supported a loss of function of Beclin-1 due to proteolytic modification by caspases. In the present study we examined whether caspase-cleavage of Beclin-1 occurs in the AD brain by designing a site-directed caspase-cleavage antibody based upon a known cleavage site within the protein at position D149. We confirmed that Beclin-1 is an excellent substrate for caspase-3 and demonstrate cleavage led to the formation of a 35 kDa C-terminal fragment labeled by …

Immunolocalization Of Influenza A Virus And Markers Of Inflammation In The Human Parkinson’S Disease Brain, Troy T. Rohn, Lindsey W. Catlin

Biology Faculty Publications and Presentations

Although much is known regarding the molecular mechanisms leading to neuronal cell loss in Parkinson’s disease (PD), the initiating event has not been identified. Prevailing theories including a chemical insult or infectious agent have been postulated as possible triggers, leading to neuroinflammation. We present immunohistochemical data indicating the presence of influenza A virus within the substantia nigra pars compacta (SNpc) from postmortem PD brain sections. Influenza A virus labeling was identified within neuromelanin granules as well as on tissue macrophages in the SNpc. Further supporting a role for neuroinflammation in PD was the identification of T-lymphocytes that colocalized with an …

Clinical Significance Of Interleukin (Il)-6 In Cancer Metastasis To Bone: Potential Of Anti-Il-6 Therapies, Ken Tawara, Julia T. Oxford, Cheryl L. Jorcyk

Biology Faculty Publications and Presentations

Metastatic events to the bone occur frequently in numerous cancer types such as breast, prostate, lung, and renal carcinomas, melanoma, neuroblastoma, and multiple myeloma. Accumulating evidence suggests that the inflammatory cytokine interleukin (IL)-6 is frequently upregulated and is implicated in the ability of cancer cells to metastasize to bone. IL-6 is able to activate various cell signaling cascades that include the STAT (signal transducer and activator of transcription) pathway, the PI3K (phosphatidylinositol-3 kinase) pathway, and the MAPK (mitogen-activated protein kinase) pathway. Activation of these pathways may explain the ability of IL-6 to mediate various aspects of normal and pathogenic bone …

Passive Immunization Reduces Behavioral And Neuropathological Deficits In An Alpha-Synuclein Transgenic Model Of Lewy Body Disease, Troy Rohn

Biology Faculty Publications and Presentations

Dementia with Lewy bodies (DLB) and Parkinson's Disease (PD) are common causes of motor and cognitive deficits and are associated with the abnormal accumulation of alpha-synuclein (α-syn). This study investigated whether passive immunization with a novel monoclonal α-syn antibody (9E4) against the C-terminus (CT) of α-syn was able to cross into the CNS and ameliorate the deficits associated with α-syn accumulation. In this study we demonstrate that 9E4 was effective at reducing behavioral deficits in the water maze, moreover, immunization with 9E4 reduced the accumulation of calpain-cleaved α-syn in axons and synapses and the associated neurodegenerative deficits. In vivo studies …

Migration Patterns, Use Of Stopover Areas, And Austral Summer Movements Of Swainson's Hawks, Michael N. Kochert, Mark R. Fuller, Linda S. Schueck, Laura Bond, Marc J. Bechard, Brian Woodbridge, Geoff L. Holroyd, Mark S. Martell, Ursula Banasch

Biology Faculty Publications and Presentations

From 1995 to 1998, we tracked movements of adult Swainson’s Hawks (Buteo swainsoni), using satellite telemetry to characterize migration, important stopover areas, and movements in the austral summer. We tagged 46 hawks from July to September on their nesting grounds in seven U.S. states and two Canadian provinces. Swainson’s Hawks followed three basic routes south on a broad front, converged along the east coast of central Mexico, and followed a concentrated corridor to a communal area in central Argentina for the austral summer. North of 20°N, southward and northward tracks differed little for individuals from east of the …

Inhibition Of Proprotein Convertase Ski-1 Blocks Transcription Of Key Extracellular Matrix Genes Regulating Osteoblastic Mineralization, Jeff P. Gorski, Nichole T. Huffman, Sridar Chittur, Ronald J. Midura, Claudine Black, Julia Oxford, Nabil G. Seidah

Biomolecular Research Center Publications and Presentations

Mineralization, a characteristic phenotypic property of osteoblastic lineage cells, was blocked by AEBSF and dec-RRLL-cmk, inhibitors of SKI-1 (site 1; subtilisin kexin like-1) protease. Since SKI-1 is required for activation of SREBP and CREB/ATF family transcription factors, we tested the effect of these inhibitors on gene expression. AEBSF decreased expression of 140 genes by 1.5- to 3.0-fold including Phex, Dmp1, COL1A1, COL11A1 and fibronectin. Direct comparison of AEBSF and dec-RRLL-cmk, a more specific SKI-1 inhibitor, demonstrated that expression of Phex, Dmp1, COL11A1 and fibronectin was reduced by both while COL1A2 and HMGCS1 were reduced …

The Role Of Caspases In Alzheimer's Disease; Potential Novel Therapeutic Opportunities, Troy T. Rohn

Biology Faculty Publications and Presentations

Although apoptosis plays a critical role in molding the CNS into its final appearance and function, inappropriate activation of this pathway in the aging brain may contribute to neurodegeneration. In Alzheimer’s disease (AD), an overwhelming body of evidence supports the activation of apoptosis in general, and caspases specifically as an early event that may not only contribute to neurodegeneration but also promote the underlying pathology associated with this disease. Therefore, caspase inhibitors may provide an effective strategy for treating AD. However, despite the compelling evidence indicating a role for caspases in disease progression, chronic treatment with caspase inhibitors in animal …

The Expression Patterns Of Minor Fibrillar Collagens During Development In Zebrafish, Ming Fang, Jason S. Adams, B. Lane Memahhan, Raquel J. Brown, Julia Oxford

Biomolecular Research Center Publications and Presentations

Minor fibrillar collagens are recognized as the organizers and nucleators during collagen fibrillogenesis but likely serve additional functions. The minor fibrillar collagens include collagens type V and type XI. Mutations of collagen type V and XI can cause Ehlers Danlos, Stickler's, and Marshall's syndromes in human. We have characterized the spatiotemporal expression patterns of Col11a1, Col11a2, Col5a1 as well as Col5a3 in zebrafish embryos by in situ hybridization. Col5a1 is expressed in developing somites, neural crest, the head mesenchyme, developing cranial cartilage, pharyngeal arches and vertebrae. Col5a3 is detected in the notochord, mesenchyme cells in the eyes and lens. Both …

Caspase-Cleaved Tar Dna-Binding Protein-43 In Parkinson’S Disease And Dementia With Lewy Bodies, Polina Kokoulina, Troy T. Rohn

Biology Faculty Publications and Presentations

Background: TAR DNA-binding protein-43 (TDP-43) proteinopathies are classified based upon the extent of modified TDP-43 and include a growing number of neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS), frontotemporal lobar degeneration with ubiquitin immunoreactive, tau-negative inclusions (FTLD-U) and FTLD with motor neuron disease (FTLD-MND). Objective: The purpose of the study was to examine whether proteolytic modifications of TDP-43 are a relevant finding in Parkinson’s disease (PD) and dementia with Lewy Bodies (DLB). Methods: A novel site-directed caspase-cleavage antibody, termed TDP caspase-cleavage product antibody (TDPccp), was utilized based upon a known caspase-3 cleavage consensus site within TDP-43 at position 219. …

Differential Expression Of Collagen Type V And Xi Α-1 In Human Ascending Thoracic Aortic Aneurysms, Julia Thom Oxford

Biomolecular Research Center Publications and Presentations

Background—The molecular mechanisms leading to ascending thoracic aortic aneurysms (ATAAs) remain unknown. We hypothesized that alterations in expression levels of specific fibrillar collagens occur during the aneurysmal process.

Methods—Surgical samples from ascending aortas from patients with degenerative ATAAs were subdivided by aneurysm diameter: small, 5 to 6 cm; medium, 6 to 7 cm; and large, greater than 7 cm; and compared with nonaneurysmal aortas (mean diameter, 2.3 cm).

Results—Histology, immunofluorescence, and electron microscopy demonstrated greater disorganization of extracellular matrix constituents in ATAAs as compared with control with an increase in collagen α1(XI) within regions of cystic medial …

Correlation Between Animal And Mathematical Models For Prostate Cancer Progression, Zdzislaw Jackiewiczy, Cheryl L. Jorcyk, Mikhail K. Kolev, Barbara Zubik-Kowal

Biology Faculty Publications and Presentations

This work demonstrates that prostate tumour progression in vivo can be analysed by using solutions of a mathematical model supplemented by initial conditions chosen according to growth rates of cell lines in vitro. The mathematical model is investigated and solved numerically. Its numerical solutions are compared with experimental data from animal models. The numerical results confirm the experimental results with the growth rates in vivo.

Caspase Cleavage Of The Amyloid Precursor Protein Is Prevented After Overexpression Of Bcl-2 In A Triple Transgenic Mouse Model Of Alzheimer’S Disease, Debra K. Kumasaka, Veronica Galvan, Elizabeth Head, Troy T. Rohn

Biology Faculty Publications and Presentations

A recent study demonstrated the lack of beta-amyloid (Aβ) plaque formation and accumulation of the amyloid precursor protein (APP) in a triple transgenic mouse model of Alzheimer’s disease (3xTg-AD) following overexpression of the anti-apoptotic protein, Bcl-2 (Rohn et al., J. Neurosci. 28: 3051-9, 2008). The supposition from that study was the accumulation of APP resulted from a decrease in caspase-mediated processing of APP. To determine a direct role for the caspase-cleavage of APP in 3xTg-AD mice, we designed a site-directed caspasecleavage antibody to APP and demonstrated it is a specific marker for caspase-cleaved APP. Application of this antibody revealed neuronal …

Caspase-Cleaved Tar Dna-Binding Protein-43 In Pick’S Disease, Troy T. Rohn, Polina Kokoulina

Biology Faculty Publications and Presentations

The hyperphosphorylation and proteolytic modification of the TAR DNA binding protein-43 (TDP-43) is a key finding in a number of neurodegenerative diseases including frontotemporal dementia with ubiquitin-positive inclusions (FTLD-U), amyotrophic lateral sclerosis (ALS), and most recently Alzheimer’s disease (AD). To examine whether proteolytic modifications of TDP-43 is a relevant finding in Pick’s disease, we utilized a novel site-directed caspase-cleavage antibody based upon a known caspase-3 cleavage consensus site within TDP-43 at position 219. Application of this antibody, termed TDP caspase-cleavage product (TDPccp) to postmortem Pick’s disease brain sections revealed the presence of caspase-cleaved TDP-43 in Pick and Hirano bodies predominantly …

Trichomycetes From Lentic And Lotic Aquatic Habitats In Ontario, Canada, D. B. Strongman, Merlin M. White

Biology Faculty Publications and Presentations

Fungi and protists make up an ecological group, trichomycetes, that inhabit the guts of invertebrates, mostly aquatic insects. Trichomycetes are reported herein from arthropods collected in lotic habitats (fast flowing streams) and lentic environments (ponds, ditches, seeps, and lakes) from 11 sites in Algonquin Park and 6 other sites in Ontario, Canada. Thirty-two trichomycete species were recovered, including 7 new species: Legeriomyces algonquinensis, Legeriosimilis leptocerci, Legeriosimilis whitneyi and Paramoebidium umbonatum are described from mayfly nymphs (Ephemeroptera); Pennella digitata and Glotzia incilis from black fly and midge larvae (Diptera), respectively; and Arundinula opeongoensis from a crayfish (Crustacea). Legeriomyces rarus Lichtw. & …

Ambient Temperature Influences Diet Selection And Physiology Of An Herbivorous Mammal, Neotoma Albigula, M. D. Dearing, Jennifer S. Forbey, J. D. Mclister, L. Santos

Biology Faculty Publications and Presentations

The whitethroat woodrat (Neotoma albigula) eats juniper (Juniperus monosperma), but the amount of juniper in its diet varies seasonally. We tested whether changes in juniper consumption are due to changes in ambient temperature and what the physiological consequences of consuming plant secondary compounds (PSCs) at different ambient temperatures might be. Woodrats were acclimated to either 20ºC or 28ºC. Later, they were given two diets to choose from (50% juniper and a nontoxic control) for 7 d. Food intake, resting metabolic rate (RMR), and body temperature (T_b) were measured over the last 2 d. Woodrats at 28ºC …

Caspase-Cleaved Tar Dna Binding Protein-43 Is A Major Pathological Finding In Alzheimer’S Disease, Troy T. Rohn

Biology Faculty Publications and Presentations

The TAR DNA binding protein-43 (TDP-43) has been identified as a major constituent of inclusions found in frontotemporal dementia with ubiquitin-positive inclusions (FTLD-U) and amyotrophic lateral sclerosis (ALS). To determine a possible role for TDP-43 in Alzheimer’s disease (AD), a site-directed caspase-cleavage antibody to TDP-43 based upon a known caspase-3 cleavage consensus site within TDP-43 at position D219 was designed. In vitro, this antibody labeled the predicted 25 kDa caspase-cleavage fragment of TDP-43 without labeling full-length TDP-43 following digestion of recombinant TDP-43 with caspase-3 or treatment of Hela cells with staurosporine. Application of this antibody in postmortem brain sections …

Lack Of Pathology In A Triple Transgenic Mouse Model Of Alzheimer’S Disease After Overexpression Of The Anti-Apoptotic Protein Bcl-2, Troy T. Rohn, Veera Vyas, Tatiana Hernandez-Estrada, Kathryn E. Nichol, Lori-Ann Christie, Elizabeth Head

Biology Faculty Publications and Presentations

Alzheimer’s disease (AD) is characterized by the accumulation of plaques containing ß-amyloid (Aß) and neurofibrillary tangles (NFTs) consisting of modified tau. Although Aß deposition is thought to precede the formation of NFTs in AD, the molecular steps connecting these two pathologies is not known. Previous studies have suggested that caspase activation plays an important role in promoting the pathology associated with AD. To further understand the contribution of caspases in disease progression, a triple transgenic Alzheimer’s mouse model overexpressing the anti-apoptotic protein Bcl-2 was generated. Here we show that overexpression of Bcl-2 limited caspase-9 activation and reduced the caspase cleavage …