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Special Issue "Frontiers in Imaging"

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A special issue of Viruses (ISSN 1999-4915). This special issue belongs to the section "Animal Viruses".

Deadline for manuscript submissions: closed (31 August 2012)

Special Issue Editors

Guest Editor
Prof. Dr. Thomas Hope

Department of Cell & Molecular Biology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA
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Guest Editor
Dr. Edward M. Campbell

Department of Microbiology and Immunology, Stritch School of Medicine, Loyola University, Chicago, IL, USA
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Published Papers (5 papers)

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Research

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Open AccessArticle High Throughput Method to Quantify Anterior-Posterior Polarity of T-Cells and Epithelial Cells
Viruses 2011, 3(12), 2396-2411; doi:10.3390/v3122396
Received: 23 September 2011 / Revised: 16 November 2011 / Accepted: 23 November 2011 / Published: 28 November 2011
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Abstract
The virologic synapse (VS), which is formed between a virus-infected and uninfected cell, plays a central role in the transmission of certain viruses, such as HIV and HTLV-1. During VS formation, HTLV-1-infected T-cells polarize cellular and viral proteins toward the uninfected T-cell. This
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The virologic synapse (VS), which is formed between a virus-infected and uninfected cell, plays a central role in the transmission of certain viruses, such as HIV and HTLV-1. During VS formation, HTLV-1-infected T-cells polarize cellular and viral proteins toward the uninfected T-cell. This polarization resembles anterior-posterior cell polarity induced by immunological synapse (IS) formation, which is more extensively characterized than VS formation and occurs when a T-cell interacts with an antigen-presenting cell. One measure of cell polarity induced by both IS or VS formation is the repositioning of the microtubule organizing center (MTOC) relative to the contact point with the interacting cell. Here we describe an automated, high throughput system to score repositioning of the MTOC and thereby cell polarity establishment. The method rapidly and accurately calculates the angle between the MTOC and the IS for thousands of cells. We also show that the system can be adapted to score anterior-posterior polarity establishment of epithelial cells. This general approach represents a significant advancement over manual cell polarity scoring, which is subject to experimenter bias and requires more time and effort to evaluate large numbers of cells. Full article
(This article belongs to the Special Issue Frontiers in Imaging)
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Review

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Open AccessReview Application of Live-Cell RNA Imaging Techniques to the Study of Retroviral RNA Trafficking
Viruses 2012, 4(6), 963-979; doi:10.3390/v4060963
Received: 11 May 2012 / Revised: 5 June 2012 / Accepted: 6 June 2012 / Published: 8 June 2012
Cited by 13 | PDF Full-text (488 KB) | HTML Full-text | XML Full-text
Abstract
Retroviruses produce full-length RNA that serves both as a genomic RNA (gRNA), which is encapsidated into virus particles, and as an mRNA, which directs the synthesis of viral structural proteins. However, we are only beginning to understand the cellular and viral factors that
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Retroviruses produce full-length RNA that serves both as a genomic RNA (gRNA), which is encapsidated into virus particles, and as an mRNA, which directs the synthesis of viral structural proteins. However, we are only beginning to understand the cellular and viral factors that influence trafficking of retroviral RNA and the selection of the RNA for encapsidation or translation. Live cell imaging studies of retroviral RNA trafficking have provided important insight into many aspects of the retrovirus life cycle including transcription dynamics, nuclear export of viral RNA, translational regulation, membrane targeting, and condensation of the gRNA during virion assembly. Here, we review cutting-edge techniques to visualize single RNA molecules in live cells and discuss the application of these systems to studying retroviral RNA trafficking. Full article
(This article belongs to the Special Issue Frontiers in Imaging)
Open AccessReview Quantitative Live-Cell Imaging of Human Immunodeficiency Virus (HIV-1) Assembly
Viruses 2012, 4(5), 777-799; doi:10.3390/v4050777
Received: 26 March 2012 / Accepted: 24 April 2012 / Published: 4 May 2012
Cited by 13 | PDF Full-text (2056 KB) | HTML Full-text | XML Full-text
Abstract
Advances in fluorescence methodologies make it possible to investigate biological systems in unprecedented detail. Over the last few years, quantitative live-cell imaging has increasingly been used to study the dynamic interactions of viruses with cells and is expected to become even more indispensable
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Advances in fluorescence methodologies make it possible to investigate biological systems in unprecedented detail. Over the last few years, quantitative live-cell imaging has increasingly been used to study the dynamic interactions of viruses with cells and is expected to become even more indispensable in the future. Here, we describe different fluorescence labeling strategies that have been used to label HIV-1 for live cell imaging and the fluorescence based methods used to visualize individual aspects of virus-cell interactions. This review presents an overview of experimental methods and recent experiments that have employed quantitative microscopy in order to elucidate the dynamics of late stages in the HIV-1 replication cycle. This includes cytosolic interactions of the main structural protein, Gag, with itself and the viral RNA genome, the recruitment of Gag and RNA to the plasma membrane, virion assembly at the membrane and the recruitment of cellular proteins involved in HIV-1 release to the nascent budding site. Full article
(This article belongs to the Special Issue Frontiers in Imaging)
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Open AccessReview The Use of Fluorescence Microscopy to Study the Association Between Herpesviruses and Intrinsic Resistance Factors
Viruses 2011, 3(12), 2412-2424; doi:10.3390/v3122412
Received: 18 October 2011 / Revised: 1 December 2011 / Accepted: 1 December 2011 / Published: 7 December 2011
Cited by 4 | PDF Full-text (2513 KB) | HTML Full-text | XML Full-text
Abstract
Intrinsic antiviral resistance is a branch of antiviral defence that involves constitutively expressed cellular proteins that act within individual infected cells. In recent years it has been discovered that components of cellular nuclear structures known as ND10 or PML nuclear bodies contribute to
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Intrinsic antiviral resistance is a branch of antiviral defence that involves constitutively expressed cellular proteins that act within individual infected cells. In recent years it has been discovered that components of cellular nuclear structures known as ND10 or PML nuclear bodies contribute to intrinsic resistance against a variety of viruses, notably of the herpesvirus family. Several ND10 components are rapidly recruited to sites that are closely associated with herpes simplex virus type 1 (HSV-1) genomes during the earliest stages of infection, and this property correlates with the efficiency of ND10 mediated restriction of HSV-1 replication. Similar but distinct recruitment of certain DNA damage response proteins also occurs during infection. These recruitment events are inhibited in a normal wild type HSV-1 infection by the viral regulatory protein ICP0. HSV‑1 mutants that do not express ICP0 are highly susceptible to repression through intrinsic resistance factors, but they replicate more efficiently in cells depleted of certain ND10 proteins or in which ND10 component recruitment is inefficient. This article presents the background to this recruitment phenomenon and summaries how it is conveniently studied by fluorescence microscopy. Full article
(This article belongs to the Special Issue Frontiers in Imaging)
Open AccessReview Hepatitis C Virus Assembly Imaging
Viruses 2011, 3(11), 2238-2254; doi:10.3390/v3112238
Received: 21 September 2011 / Revised: 3 November 2011 / Accepted: 4 November 2011 / Published: 15 November 2011
Cited by 9 | PDF Full-text (2030 KB)
Abstract
Hepatitis C Virus (HCV) assembly process is the least understood step in the virus life cycle. The functional data revealed by forward and reverse genetics indicated that both structural and non-structural proteins are involved in the assembly process. Using confocal and electron microscopy
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Hepatitis C Virus (HCV) assembly process is the least understood step in the virus life cycle. The functional data revealed by forward and reverse genetics indicated that both structural and non-structural proteins are involved in the assembly process. Using confocal and electron microscopy different groups determined the subcellular localization of different viral proteins and they identified the lipid droplets (LDs) as the potential viral assembly site. Here, we aim to review the mechanisms that govern the viral proteins recruitment to LDs and discuss the current model of HCV assembly process. Based on previous examples, this review will also discuss advanced imaging techniques as potential means to extend our present knowledge of HCV assembly process. Full article
(This article belongs to the Special Issue Frontiers in Imaging)

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