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Analysis Of Apoptosis Of Memory T Cells And Dendritic Cells During The Early Stages Of Viral Infection Or Exposure To Toll-Like Receptor Agonists, Kapil Bahl, Anette Hubner, Roger J. Davis, Raymond M. Welsh 2010 Yale University

Analysis Of Apoptosis Of Memory T Cells And Dendritic Cells During The Early Stages Of Viral Infection Or Exposure To Toll-Like Receptor Agonists, Kapil Bahl, Anette Hubner, Roger J. Davis, Raymond M. Welsh

Davis Lab Publications

Profound type I interferon (IFN-I)-dependent attrition of memory CD8 and CD4 T cells occurs early during many infections. It is dramatic at 2 to 4 days following lymphocytic choriomeningitis virus (LCMV) infection of mice and can be elicited by the IFN-inducing Toll receptor agonist poly(I:C). We show that this attrition occurs in many organs, indicating that it is due to T cell loss rather than redistribution. This loss correlated with elevated intracellular staining of T cells ex vivo for activated caspases but with only low levels of ex vivo staining with annexin V, probably due to the ...


Midterm Outcomes Of Autologous Cultivated Limbal Stem Cell Transplantation With Or Without Penetrating Keratoplasty, Hossein Baharvand 2010 university of science & culture

Midterm Outcomes Of Autologous Cultivated Limbal Stem Cell Transplantation With Or Without Penetrating Keratoplasty, Hossein Baharvand

university of science & culture

To report the midterm outcomes of autologous limbal stem cell transplantation cultivated on amniotic membrane (AM) with or without subsequent penetrating keratoplasty (PKP) in patients with total unilateral limbal stem cell deficiency (LSCD). Methods: Eight eyes of 8 consecutive patients with unilateral total LSCD underwent autologous limbal stem cell transplantation cultivated on AM. Four eyes underwent subsequent optical PKP. Main outcome measures were corneal vascularization and transparency. Results: The patients were followed for 34.0 6 13.5 months (6–48 months). Seven cases had a stable corneal epithelium with marked decrease in opacification and vascularization. Progressive sectorial conjunctivalization was ...


Pcdp1 Is A Central Apparatus Protein That Binds Ca2+-Calmodulin And Regulates Ciliary Motility, Christen G. DiPetrillo, Elizabeth F. Smith 2010 Dartmouth College

Pcdp1 Is A Central Apparatus Protein That Binds Ca2+-Calmodulin And Regulates Ciliary Motility, Christen G. Dipetrillo, Elizabeth F. Smith

Open Dartmouth: Faculty Open Access Scholarship

For all motile eukaryotic cilia and flagella, beating is regulated by changes in intraciliary calcium concentration. Although the mechanism for calcium regulation is not understood, numerous studies have shown that calmodulin (CaM) is a key axonemal calcium sensor. Using anti-CaM antibodies and Chlamydomonas reinhardtii axonemal extracts, we precipitated a complex that includes four polypeptides and that specifically interacts with CaM in high [Ca2+]. One of the complex members, FAP221, is an orthologue of mammalian Pcdp1 (primary ciliary dyskinesia protein 1). Both FAP221 and mammalian Pcdp1 specifically bind CaM in high [Ca2+]. Reduced expression of Pcdp1 complex members in ...


Disheveled Hair And Ear (Dhe), A Spontaneous Mouse Lmna Mutation Modeling Human Laminopathies, Paul R. Odgren, Craig H. Pratt, Carole A. MacKay, April Mason-Savas, Michelle Curtain, Lindsay S. Shopland, Tsutomu Ichicki, John P. Sundberg, Leah Rae Donahue 2010 University of Massachusetts Medical School

Disheveled Hair And Ear (Dhe), A Spontaneous Mouse Lmna Mutation Modeling Human Laminopathies, Paul R. Odgren, Craig H. Pratt, Carole A. Mackay, April Mason-Savas, Michelle Curtain, Lindsay S. Shopland, Tsutomu Ichicki, John P. Sundberg, Leah Rae Donahue

Odgren Lab Publications

BACKGROUND: Investigations of naturally-occurring mutations in animal models provide important insights and valuable disease models. Lamins A and C, along with lamin B, are type V intermediate filament proteins which constitute the proteinaceous boundary of the nucleus. LMNA mutations in humans cause a wide range of phenotypes, collectively termed laminopathies. To identify the mutation and investigate the phenotype of a spontaneous, semi-dominant mutation that we have named Disheveled hair and ear (Dhe), which causes a sparse coat and small external ears in heterozygotes and lethality in homozygotes by postnatal day 10.

FINDINGS: Genetic mapping identified a point mutation in the ...


Cellular Immunity In Mouse Models Of Viral Encephalitis, Christina Dawn Steel 2010 Old Dominion University

Cellular Immunity In Mouse Models Of Viral Encephalitis, Christina Dawn Steel

Theses and Dissertations in Biomedical Sciences

Evidence is presented herein that intranasal application of vesicular stomatitis virus (VSV) caused acute infection of the murine central nervous system (CNS) with associated morbidity and significant mortality in mice. However, VSV encephalitis was not invariably fatal, suggesting that the CNS contained a professional antigen-presenting cell (APC) capable of inducing or propagating a protective antiviral immune response. To examine this possibility, we administered VSV via the intranasal route and then characterized the cellular elements that infiltrate the brain as well as the activation status of resident microglia, cells widely believed to represent the major APC population in the CNS. To ...


Legume Phylogeny And The Evolution Of A Unique Contractile Apparatus That Regulates Phloem Transport, Winfried Peters, Claudia Hanakam, Dietmar Haffer, Aart van Bel, Michael Knoblauch 2010 Indiana University - Purdue University Fort Wayne

Legume Phylogeny And The Evolution Of A Unique Contractile Apparatus That Regulates Phloem Transport, Winfried Peters, Claudia Hanakam, Dietmar Haffer, Aart Van Bel, Michael Knoblauch

Winfried S. Peters

Protein bodies called forisomes undergo Ca2+-dependent deformations to occlude sieve tubes reversibly, providing a unique regulatory mechanism of phloem transport. Because forisomes are known exclusively from the Papilionoideae (Leguminosae), the evolution of forisome function may have played a role in the rapid radiation of this huge taxon. The unexpected discovery of a papilionoid species lacking forisomes led us to evaluate a representative set of species covering 33 of the 36 legume tribes traditionally recognized. We found forisomes in Papilionoideae but not in Caesalpinioideae and Mimosoideae. Forisomes were absent from several species of the papilionoid tribe Galegeae. Forisomes with tail-like ...


Reverse Genetic And Cell Biological Approaches To The Study Of Developmental Functions Of Class Xi Myosin In Arabidopsis Thaliana, Eunsook Park 2010 University of Tennessee - Knoxville

Reverse Genetic And Cell Biological Approaches To The Study Of Developmental Functions Of Class Xi Myosin In Arabidopsis Thaliana, Eunsook Park

Doctoral Dissertations

Myosin proteins function as molecular motors that drive the ATP-dependent movement of cellular components along actin filaments. Vascular plants encode two different types of myosin, referred to as class VIII and class XI. Although class XI myosins have been suggested to function in organelle movement and cytoplasmic streaming, little is known about their cellular function in detail.

The Arabidopsis genome encodes 13 class XI myosin genes. The reasons for the relatively large number of myosin XI isoforms present within a single plant species are unknown. To investigate the function of these gene products in the cell, we determined the spatial ...


Yeast Cell Adhesion Molecules Have Functional Amyloid-Forming Sequences, Caleen B. Ramsook, Cho Tan, Melissa C. Garcia, Raymond Fung, Gregory Soybelman, Ryan Henry, Anna Litewka, Shanique O’Meally, Henry N. Otoo, Roy A. Khalaf, Anne M. Dranginis, Nand K. Gaur, Stephen A. Klotz, Jason M. Rauceo, Chong K. Jue, Peter N. Lipke 2010 CUNY Brooklyn College

Yeast Cell Adhesion Molecules Have Functional Amyloid-Forming Sequences, Caleen B. Ramsook, Cho Tan, Melissa C. Garcia, Raymond Fung, Gregory Soybelman, Ryan Henry, Anna Litewka, Shanique O’Meally, Henry N. Otoo, Roy A. Khalaf, Anne M. Dranginis, Nand K. Gaur, Stephen A. Klotz, Jason M. Rauceo, Chong K. Jue, Peter N. Lipke

Publications and Research

The occurrence of highly conserved amyloid-forming sequences in Candida albicans Als proteins (H. N. Otoo et al., Eukaryot. Cell 7:776–782, 2008) led us to search for similar sequences in other adhesins from C. albicans and Saccharomyces cerevisiae. The beta-aggregation predictor TANGO found highly beta-aggregation-prone sequences in almost all yeast adhesins. These sequences had an unusual amino acid composition: 77% of their residues were beta-aggregation aliphatic amino acids Ile, Thr, and Val, which is more than 4-fold greater than their prevalence in the S. cerevisiae proteome. High beta-aggregation potential peptides from S. cerevisiae Flo1p and C. albicans Eap1p rapidly ...


Structure And Function Of Glycosylated Tandem Repeats From Candida Albicans Als Adhesins, Aaron T. Frank, Caleen B. Ramsook, Henry N. Otoo, Cho Tan, Gregory Soybelman, Jason M. Rauceo, Nand K. Gaur, Stephen A. Klotz, Peter N. Lipke 2010 University of Michigan-Ann Arbor

Structure And Function Of Glycosylated Tandem Repeats From Candida Albicans Als Adhesins, Aaron T. Frank, Caleen B. Ramsook, Henry N. Otoo, Cho Tan, Gregory Soybelman, Jason M. Rauceo, Nand K. Gaur, Stephen A. Klotz, Peter N. Lipke

Publications and Research

Tandem repeat (TR) regions are common in yeast adhesins, but their structures are unknown, and their activities are poorly understood. TR regions in Candida albicans Als proteins are conserved glycosylated 36-residue sequences with cell-cell aggregation activity (J. M. Rauceo, R. De Armond, H. Otoo, P. C. Kahn, S. A. Klotz, N. K. Gaur, and P. N. Lipke, Eukaryot. Cell 5:1664–1673, 2006). Ab initio modeling with either Rosetta or LINUS generated consistent structures of three-stranded antiparallel beta-sheet domains, whereas randomly shuffled sequences with the same composition generated various structures with consistently higher energies. O- and N-glycosylation patterns showed that ...


C-Jun Nh2-Terminal Kinase Is Required For Lineage-Specific Differentiation But Not Stem Cell Self-Renewal, Ping Xu, Roger J. Davis 2010 University of Massachusetts Medical School

C-Jun Nh2-Terminal Kinase Is Required For Lineage-Specific Differentiation But Not Stem Cell Self-Renewal, Ping Xu, Roger J. Davis

Davis Lab Publications

The c-Jun NH(2)-terminal kinase (JNK) is implicated in proliferation. Mice with a deficiency of either the Jnk1 or the Jnk2 genes are viable, but a compound deficiency of both Jnk1 and Jnk2 causes early embryonic lethality. Studies using conditional gene ablation and chemical genetic approaches demonstrate that the combined loss of JNK1 and JNK2 protein kinase function results in rapid senescence. To test whether this role of JNK was required for stem cell proliferation, we isolated embryonic stem (ES) cells from wild-type and JNK-deficient mice. We found that Jnk1(-/-) Jnk2(-/-) ES cells underwent self-renewal, but these cells proliferated ...


The Saccharomyces Sun Gene, Uth1, Is Involved In Cell Wall Biogenesis, Father Nicanor Austriaco 2010 Providence College

The Saccharomyces Sun Gene, Uth1, Is Involved In Cell Wall Biogenesis, Father Nicanor Austriaco

Rev. Nicanor Austriaco, O.P.

Deletion of the Saccharomyces gene, UTH1, a founding member of the SUN family of fungal genes, has pleiotropic effects. Several phenotypes of Δuth1 cells including their decreased levels of mitochondrial proteins, their impaired autophagic degradation of mitochondria, and their increased viability in the presence of mammalian BAX, a proapoptotic regulator localized to the mitochondria, have prompted others to propose that the Uth1p functions primarily at the mitochondria. In this report, we show that cells lacking UTH1 have more robust cell walls with higher levels of β-d-glucan that allows them to grow in the presence of calcofluor white or sodium dodecyl ...


The Role Of Runx1 N-Terminal Splice Isoforms In Hematopoietic Development, Emmett E. Hedblom 2010 University of Massachusetts Amherst

The Role Of Runx1 N-Terminal Splice Isoforms In Hematopoietic Development, Emmett E. Hedblom

Open Access Dissertations

Runx1/AML1 transcription factor expression in hematopoietic cell lineages is differentially regulated via usage of two distinct promoters. The 5' UTR and a 19 amino acid encoding sequence transcribed from the distal promoter is inserted via alternative splicing into the 5' end of the mRNA transcript, replacing the 5' UTR and a 5 amino acid encoding sequence usually transcribed from the proximal promoter. Expression of proximal Runx1 in 32Dcl.3 cells delays G-CSF induced neutrophil terminal differentiation by increasing viability compared to distal Runx1. We utilized Runx1 Nterminal deletion and point mutants of three evolutionarily conserved residues to describe dual ...


New Insights In The Tssk Family: Studies In The Activity And Function Of The Testis Specific Serine Kinases, Julian Sosnik 2010 University of Massachusetts Amherst

New Insights In The Tssk Family: Studies In The Activity And Function Of The Testis Specific Serine Kinases, Julian Sosnik

Open Access Dissertations

The Testis Specific Serine Kinase (Tssk) family of proteins is a large group of kinases that present high level of conservation within paralogs, as well as within species. In addition, in all reported cases as well as in the analysis of expressed sequence tags available in databases, this family of proteins presents a very strict pattern of either testicular or male-gonadal expression. This high level of conservation prompted the postulate that these kinases ought to be important for either testicular function or fertilization. In this work we attempt a biochemical characterization of one family member (Tssk6) in the mouse. We ...


Beyond Cell Adhesion: Exploring The Role Of Cadherin-11 Extracellular Processing By Adam Metalloproteases In Cranial Neural Crest Migration, Catherine D. McCusker 2010 University of Massachusetts Amherst

Beyond Cell Adhesion: Exploring The Role Of Cadherin-11 Extracellular Processing By Adam Metalloproteases In Cranial Neural Crest Migration, Catherine D. Mccusker

Open Access Dissertations

The migration of the cranial neural crest is an essential part of cranio-facial development in every vertebrate embryo. The cranial neural crest (CNC) is a transient population of cells that forms the lateral border of the anterior neural plate. In the tailbud stage Xenopus embryo, the neural crest cells delaminate from the neural tube, and undergo a large-scale migration from the dorsal to ventral region of the embryo. The CNC travels along distinct pathways, and populates specific regions of the embryos face. Once the CNC ceases migrating, it differentiates into a variety of tissues that are essential for cranio-facial structure ...


Role Of The Hypothalamic-Pituitary-Thyroid Axis In Metabolic Regulation By Jnk1, Guadalupe Sabio, Julie Cavanagh-Kyros, Tamera Barrett, Dae Young Jung, Hwi Jin Ko, Helena Ong, Caroline Morel, Alfonso Mora, Judith Reilly, Jason K. Kim, Roger J. Davis 2010 University of Massachusetts Medical School

Role Of The Hypothalamic-Pituitary-Thyroid Axis In Metabolic Regulation By Jnk1, Guadalupe Sabio, Julie Cavanagh-Kyros, Tamera Barrett, Dae Young Jung, Hwi Jin Ko, Helena Ong, Caroline Morel, Alfonso Mora, Judith Reilly, Jason K. Kim, Roger J. Davis

Davis Lab Publications

The cJun N-terminal kinase 1 (JNK1) is implicated in diet-induced obesity. Indeed, germline ablation of the murine Jnk1 gene prevents diet-induced obesity. Here we demonstrate that selective deficiency of JNK1 in the murine nervous system is sufficient to suppress diet-induced obesity. The failure to increase body mass is mediated, in part, by increased energy expenditure that is associated with activation of the hypothalamic-pituitary-thyroid axis. Disruption of thyroid hormone function prevents the effects of nervous system JNK1 deficiency on body mass. These data demonstrate that the hypothalamic-pituitary-thyroid axis represents an important target of metabolic signaling by JNK1.


Supervillin Slows Cell Spreading By Facilitating Myosin Ii Activation At The Cell Periphery, Norio Takizawa, Reiko Ikebe, Mitsuo Ikebe, Elizabeth J. Luna 2010 University of Massachusetts Medical School

Supervillin Slows Cell Spreading By Facilitating Myosin Ii Activation At The Cell Periphery, Norio Takizawa, Reiko Ikebe, Mitsuo Ikebe, Elizabeth J. Luna

Elizabeth J. Luna

During cell migration, myosin II modulates adhesion, cell protrusion and actin organization at the leading edge. We show that an F-actin- and membrane-associated scaffolding protein, called supervillin (SV, p205), binds directly to the subfragment 2 domains of nonmuscle myosin IIA and myosin IIB and to the N-terminus of the long form of myosin light chain kinase (L-MLCK). SV inhibits cell spreading via an MLCK- and myosin II-dependent mechanism. Overexpression of SV reduces the rate of cell spreading, and RNAi-mediated knockdown of endogenous SV increases it. Endogenous and EGFP-tagged SV colocalize with, and enhance the formation of, cortical bundles of F-actin ...


Supervillin Reorganizes The Actin Cytoskeleton And Increases Invadopodial Efficiency, Jessica Lynn Crowley, Tara C. Smith, Zhiyou Fang, Norio Takizawa, Elizabeth J. Luna 2010 University of Massachusetts Medical School

Supervillin Reorganizes The Actin Cytoskeleton And Increases Invadopodial Efficiency, Jessica Lynn Crowley, Tara C. Smith, Zhiyou Fang, Norio Takizawa, Elizabeth J. Luna

Elizabeth J. Luna

Tumor cells use actin-rich protrusions called invadopodia to degrade extracellular matrix (ECM) and invade tissues; related structures, termed podosomes, are sites of dynamic ECM interaction. We show here that supervillin (SV), a peripheral membrane protein that binds F-actin and myosin II, reorganizes the actin cytoskeleton and potentiates invadopodial function. Overexpressed SV induces redistribution of lamellipodial cortactin and lamellipodin/RAPH1/PREL1 away from the cell periphery to internal sites and concomitantly increases the numbers of F-actin punctae. Most punctae are highly dynamic and colocalize with the podosome/invadopodial proteins, cortactin, Tks5, and cdc42. Cortactin binds SV sequences in vitro and contributes ...


Chapter 27 – Studying Plus-End Tracking At Single Molecule Resolution Using Tirf Microscopy, Ram Dixit, Jennifer L. Ross 2010 Washington University in St Louis

Chapter 27 – Studying Plus-End Tracking At Single Molecule Resolution Using Tirf Microscopy, Ram Dixit, Jennifer L. Ross

Biology Faculty Publications & Presentations

The highly dynamic microtubule plus-ends are key sites of regulation that impact the organization and function of the microtubule cytoskeleton. Much of this regulation is performed by the microtubule plus-end tracking (+TIP) family of proteins. +TIPs are a structurally diverse group of proteins that bind to and track with growing microtubule plus-ends in cells. +TIPs regulate microtubule dynamics as well as mediate interactions between microtubule tips and other cellular structures. Most +TIPs can directly bind to microtubules in vitro; however, the mechanisms for their plus-end specificity are not fully understood. Cellular studies of +TIP activity are complicated by the fact ...


Chapter 26 – Multiple Color Single Molecule Tirf Imaging And Tracking Of Maps And Motors, Jennifer L. Ross, Ram Dixit 2010 Washington University in St Louis

Chapter 26 – Multiple Color Single Molecule Tirf Imaging And Tracking Of Maps And Motors, Jennifer L. Ross, Ram Dixit

Biology Faculty Publications & Presentations

Microtubules are part of a complex mechano-chemical network inside cells. In order to understand how the components of these systems work together, careful in vitro experiments must be performed with added complexity. These experiments can ideally image all the interacting species. In order to image these molecules, multiple-color fluorescence imaging can be performed. In this chapter, we describe some methods for performing multiple-color single molecule fluorescence imaging using total internal reflection fluorescence microscopy. We give several specific examples of species of microtubule-associate proteins and motors that can be examined with detailed protocols for labeling, purification, and imaging.


Real-Time Pcr Method For The Quantification Of Burkholderia Cepacia Complex Attached To Lung Epithelial Cells And Inhibitionn Of That Attachment, Ciara Wight, Gillian Herbert, Ruth Pilkington, Máire Callaghan, Siobhan McClean 2010 Institute of Technology Tallaght

Real-Time Pcr Method For The Quantification Of Burkholderia Cepacia Complex Attached To Lung Epithelial Cells And Inhibitionn Of That Attachment, Ciara Wight, Gillian Herbert, Ruth Pilkington, Máire Callaghan, Siobhan Mcclean

Articles

To develop a rapid method to quantify the attachment of the cystic fibrosis pathogen, Burkholderia multivorans, to lung epithelial cells (16HBE14o(-)) using real-time PCR with a view to monitoring potential inhibition of lung cell attachment. Mammalian and bacterial DNA were purified from bacteria attached to lung epithelial cells. The relative amount of bacteria attached was determined by amplification of the recA gene relative to the human GAPDH gene, in the presence of SYBR Green. The method was thoroughly validated and shown to correlate well with traditional plating techniques. Inhibition of bacterial attachment with simple sugars was then evaluated by real-time ...


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