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Article ; Online: Annulate lamellae and intracellular pathogens.

Eymieux, Sébastien / Blanchard, Emmanuelle / Uzbekov, Rustem / Hourioux, Christophe / Roingeard, Philippe

2021 Volume 23, Issue 8, Page(s) e13328

Abstract: Annulate lamellae (AL) have been observed many times over the years on electron micrographs of rapidly dividing cells, but little is known about these unusual organelles consisting of stacked sheets of endoplasmic reticulum-derived membranes with nuclear ...

Abstract	Annulate lamellae (AL) have been observed many times over the years on electron micrographs of rapidly dividing cells, but little is known about these unusual organelles consisting of stacked sheets of endoplasmic reticulum-derived membranes with nuclear pore complexes (NPCs). Evidence is growing for a role of AL in viral infection. AL have been observed early in the life cycles of the hepatitis C virus (HCV) and, more recently, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), suggesting a specific induction of mechanisms potentially useful to these pathogens. Like other positive-strand RNA viruses, these viruses induce host cells membranes rearrangements. The NPCs of AL could potentially mediate exchanges between these partially sealed compartments and the cytoplasm. AL may also be involved in regulating Ca
MeSH term(s)	Animals ; COVID-19 ; Cytoplasm/virology ; Endoplasmic Reticulum/microbiology ; Endoplasmic Reticulum/parasitology ; Endoplasmic Reticulum/ultrastructure ; Endoplasmic Reticulum/virology ; Host-Pathogen Interactions/physiology ; Humans ; Organelles/microbiology ; Organelles/parasitology ; Organelles/ultrastructure ; Organelles/virology ; SARS-CoV-2/physiology
Language	English
Publishing date	2021-03-25
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	1468320-9
ISSN	1462-5822 ; 1462-5814
ISSN (online)	1462-5822
ISSN	1462-5814
DOI	10.1111/cmi.13328
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Article ; Online: Loose anagen hair syndrome: take a closer look!

Pottier, Cassandre / Eymieux, Sébastien / Blanchard-Laumonnier, Emmanuelle / Robert, Juliette / Maruani, Annabel / Leducq, Sophie

The British journal of dermatology

2022 Volume 187, Issue 4, Page(s) e156

MeSH term(s)	Hair ; Hair Diseases/diagnosis ; Hair Diseases/etiology ; Humans ; Loose Anagen Hair Syndrome
Language	English
Publishing date	2022-05-25
Publishing country	England
Document type	Journal Article
ZDB-ID	80076-4
ISSN	1365-2133 ; 0007-0963
ISSN (online)	1365-2133
ISSN	0007-0963
DOI	10.1111/bjd.21623
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Article ; Online: The double-membrane vesicle (DMV): a virus-induced organelle dedicated to the replication of SARS-CoV-2 and other positive-sense single-stranded RNA viruses.

Roingeard, Philippe / Eymieux, Sébastien / Burlaud-Gaillard, Julien / Hourioux, Christophe / Patient, Romuald / Blanchard, Emmanuelle

Cellular and molecular life sciences : CMLS

2022 Volume 79, Issue 8, Page(s) 425

Abstract: Positive single-strand RNA (+ RNA) viruses can remodel host cell membranes to induce a replication organelle (RO) isolating the replication of their genome from innate immunity mechanisms. Some of these viruses, including severe acute respiratory ... ...

Abstract	Positive single-strand RNA (+ RNA) viruses can remodel host cell membranes to induce a replication organelle (RO) isolating the replication of their genome from innate immunity mechanisms. Some of these viruses, including severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), induce double-membrane vesicles (DMVs) for this purpose. Viral non-structural proteins are essential for DMV biogenesis, but they cannot form without an original membrane from a host cell organelle and a significant supply of lipids. The endoplasmic reticulum (ER) and the initial mechanisms of autophagic processes have been shown to be essential for the biogenesis of SARS-CoV-2 DMVs. However, by analogy with other DMV-inducing viruses, it seems likely that the Golgi apparatus, mitochondria and lipid droplets are also involved. As for hepatitis C virus (HCV), pores crossing both membranes of SARS-CoV-2-induced DMVs have been identified. These pores presumably allow the supply of metabolites essential for viral replication within the DMV, together with the export of the newly synthesized viral RNA to form the genome of future virions. It remains unknown whether, as for HCV, DMVs with open pores can coexist with the fully sealed DMVs required for the storage of large amounts of viral RNA. Interestingly, recent studies have revealed many similarities in the mechanisms of DMV biogenesis and morphology between these two phylogenetically distant viruses. An understanding of the mechanisms of DMV formation and their role in the infectious cycle of SARS-CoV-2 may be essential for the development of new antiviral approaches against this pathogen or other coronaviruses that may emerge in the future.
MeSH term(s)	COVID-19 ; Endoplasmic Reticulum/metabolism ; Hepacivirus/genetics ; Hepatitis C ; Humans ; RNA, Viral/genetics ; RNA, Viral/metabolism ; SARS-CoV-2 ; Viral Nonstructural Proteins/genetics ; Virus Replication
Chemical Substances	RNA, Viral ; Viral Nonstructural Proteins
Language	English
Publishing date	2022-07-16
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	1358415-7
ISSN	1420-9071 ; 1420-682X
ISSN (online)	1420-9071
ISSN	1420-682X
DOI	10.1007/s00018-022-04469-x
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Article: Crystalline podocytopathy and tubulopathy linked to kappa light chain deposits in a context of smoldering multiple myeloma.

Eymieux, Sébastien / Miquelestorena-Standley, Elodie / Rabot, Nolwenn / Maisons, Valentin / Touchard, Guy / Blanchard, Emmanuelle

Clinical kidney journal

2021 Volume 15, Issue 2, Page(s) 351–353

Abstract: A 42-year-old man with smoldering immunoglobulin G kappa multiple myeloma showed a heavy proteinuria composed of free light chain, prompting performance of a kidney biopsy. Electron microscopy revealed numerous rhomboid-shaped crystals labelled by the ... ...

Abstract	A 42-year-old man with smoldering immunoglobulin G kappa multiple myeloma showed a heavy proteinuria composed of free light chain, prompting performance of a kidney biopsy. Electron microscopy revealed numerous rhomboid-shaped crystals labelled by the anti-kappa in immunogold, notably in the cytoplasm of podocytes, establishing the diagnosis of crystalline podocytopathy. This case illustrates a rare form of monoclonal gammopathy of renal significance, and highlights the key role of electron microscopy and immunogold to better elucidate the location and composition of crystals.
Language	English
Publishing date	2021-10-06
Publishing country	England
Document type	Journal Article
ZDB-ID	2655800-2
ISSN	2048-8513 ; 2048-8505
ISSN (online)	2048-8513
ISSN	2048-8505
DOI	10.1093/ckj/sfab197
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Article ; Online: The double-membrane vesicle (DMV): a virus-induced organelle dedicated to the replication of SARS-CoV-2 and other positive-sense single-stranded RNA viruses

Roingeard, Philippe / Eymieux, Sébastien / Burlaud-Gaillard, Julien / Hourioux, Christophe / Patient, Romuald / Blanchard, Emmanuelle

Cell. Mol. Life Sci.. 2022 Aug., v. 79, no. 8 p.425-425

2022

Abstract	Positive single-strand RNA (+ RNA) viruses can remodel host cell membranes to induce a replication organelle (RO) isolating the replication of their genome from innate immunity mechanisms. Some of these viruses, including severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), induce double-membrane vesicles (DMVs) for this purpose. Viral non-structural proteins are essential for DMV biogenesis, but they cannot form without an original membrane from a host cell organelle and a significant supply of lipids. The endoplasmic reticulum (ER) and the initial mechanisms of autophagic processes have been shown to be essential for the biogenesis of SARS-CoV-2 DMVs. However, by analogy with other DMV-inducing viruses, it seems likely that the Golgi apparatus, mitochondria and lipid droplets are also involved. As for hepatitis C virus (HCV), pores crossing both membranes of SARS-CoV-2-induced DMVs have been identified. These pores presumably allow the supply of metabolites essential for viral replication within the DMV, together with the export of the newly synthesized viral RNA to form the genome of future virions. It remains unknown whether, as for HCV, DMVs with open pores can coexist with the fully sealed DMVs required for the storage of large amounts of viral RNA. Interestingly, recent studies have revealed many similarities in the mechanisms of DMV biogenesis and morphology between these two phylogenetically distant viruses. An understanding of the mechanisms of DMV formation and their role in the infectious cycle of SARS-CoV-2 may be essential for the development of new antiviral approaches against this pathogen or other coronaviruses that may emerge in the future.
Keywords	Golgi apparatus ; Hepatitis C virus ; RNA ; Severe acute respiratory syndrome coronavirus ; Severe acute respiratory syndrome coronavirus 2 ; biogenesis ; endoplasmic reticulum ; genome ; innate immunity ; lipids ; metabolites ; mitochondria ; pathogens ; phylogeny ; virus replication
Language	English
Dates of publication	2022-08
Size	p. 425.
Publishing place	Springer International Publishing
Document type	Article ; Online
Note	Review
ZDB-ID	1358415-7
ISSN	1420-9071 ; 1420-682X
ISSN (online)	1420-9071
ISSN	1420-682X
DOI	10.1007/s00018-022-04469-x
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Article: ELISA-Based Analysis Reveals an Anti-SARS-CoV-2 Protein Immune Response Profile Associated with Disease Severity.

Herrscher, Charline / Eymieux, Sébastien / Gaborit, Christophe / Blasco, Hélène / Marlet, Julien / Stefic, Karl / Roingeard, Philippe / Grammatico-Guillon, Leslie / Hourioux, Christophe

Journal of clinical medicine

2022 Volume 11, Issue 2

Abstract: Since the start of the COVID-19 pandemic, many studies have investigated the humoral response to SARS-CoV-2 during infection. Studies with native viral proteins constitute a first-line approach to assessing the overall immune response, but small peptides ...

Abstract	Since the start of the COVID-19 pandemic, many studies have investigated the humoral response to SARS-CoV-2 during infection. Studies with native viral proteins constitute a first-line approach to assessing the overall immune response, but small peptides are an accurate and valuable tool for the fine characterization of B-cell epitopes, despite the restriction of this approach to the determination of linear epitopes. In this study, we used ELISA and peptides covering a selection of structural and non-structural SARS-CoV-2 proteins to identify key epitopes eliciting a strong immune response that could serve as a biological signature of disease characteristics, such as severity, in particular. We used 213 plasma samples from a cohort of patients well-characterized clinically and biologically and followed for COVID-19 infection. We found that patients developing severe disease had higher titers of antibodies mapping to multiple specific epitopes than patients with mild to moderate disease. These data are potentially important as they could be used for immunological profiling to improve our knowledge of the quantitative and qualitative characteristics of the humoral response in relation to patient outcome.
Language	English
Publishing date	2022-01-14
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2662592-1
ISSN	2077-0383
ISSN	2077-0383
DOI	10.3390/jcm11020405
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Article ; Online: A morphometric analysis of hepatitis B subviral particles shows no correlation of filament proportion and length with clinical stage and genotype.

Eymieux, Sébastien / Hourioux, Christophe / Marlet, Julien / Moreau, Alain / Patient, Romuald / d'Alteroche, Louis / Gaudy-Graffin, Catherine / Blanchard, Emmanuelle / Roingeard, Philippe

Journal of viral hepatitis

2022 Volume 29, Issue 9, Page(s) 719–726

Abstract: It was recently suggested that the composition of circulating hepatitis B subviral particles (SVPs) could be used to differentiate the various stages in chronic hepatitis B virus (HBV) infection, with significantly lower proportions of L and M proteins ... ...

Abstract	It was recently suggested that the composition of circulating hepatitis B subviral particles (SVPs) could be used to differentiate the various stages in chronic hepatitis B virus (HBV) infection, with significantly lower proportions of L and M proteins in inactive carriers than in individuals with chronic hepatitis. L protein is abundant in virions and filamentous SVPs but almost absent from spherical SVPs. We, therefore, performed a morphometric analysis of SVPs in these two groups of patients, by conducting a retrospective analysis on sera from 15 inactive carriers and 11 patients with chronic hepatitis infected with various HBV genotypes. Subviral particles were concentrated by centrifugation on a sucrose cushion, with monitoring by transmission electron microscopy. The percentage of filamentous SVPs and filament length for 100 SVPs was determined with a digital camera. The L protein PreS1 promoter was sequenced from viral genomes by the Sanger method. No marked differences were found between patients, some of whom had only spherical SVPs, whereas others had variable percentages of filamentous SVPs (up to 28%), of highly variable length. High filament percentages were not associated with a particular sequence of the L protein promoter, HBV genotype or even disease stage. High levels of circulating filamentous SVPs are probably more strongly related to individual host factors than to viral strain characteristics or disease stage.
MeSH term(s)	Genotype ; Hepatitis B ; Hepatitis B Surface Antigens ; Hepatitis B virus/genetics ; Hepatitis B, Chronic ; Humans ; Retrospective Studies
Chemical Substances	Hepatitis B Surface Antigens
Language	English
Publishing date	2022-06-07
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1212497-7
ISSN	1365-2893 ; 1352-0504
ISSN (online)	1365-2893
ISSN	1352-0504
DOI	10.1111/jvh.13712
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Article ; Online: ELISA-Based Analysis Reveals an Anti-SARS-CoV-2 Protein Immune Response Profile Associated with Disease Severity

Charline Herrscher / Sébastien Eymieux / Christophe Gaborit / Hélène Blasco / Julien Marlet / Karl Stefic / Philippe Roingeard / Leslie Grammatico-Guillon / Christophe Hourioux

Journal of Clinical Medicine, Vol 11, Iss 405, p

2022 Volume 405

Abstract	Since the start of the COVID-19 pandemic, many studies have investigated the humoral response to SARS-CoV-2 during infection. Studies with native viral proteins constitute a first-line approach to assessing the overall immune response, but small peptides are an accurate and valuable tool for the fine characterization of B-cell epitopes, despite the restriction of this approach to the determination of linear epitopes. In this study, we used ELISA and peptides covering a selection of structural and non-structural SARS-CoV-2 proteins to identify key epitopes eliciting a strong immune response that could serve as a biological signature of disease characteristics, such as severity, in particular. We used 213 plasma samples from a cohort of patients well-characterized clinically and biologically and followed for COVID-19 infection. We found that patients developing severe disease had higher titers of antibodies mapping to multiple specific epitopes than patients with mild to moderate disease. These data are potentially important as they could be used for immunological profiling to improve our knowledge of the quantitative and qualitative characteristics of the humoral response in relation to patient outcome.
Keywords	SARS-CoV-2 antibodies ; SARS-CoV-2 linear epitopes ; COVID-19 ; disease severity ; Medicine ; R
Subject code	610
Language	English
Publishing date	2022-01-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
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Article ; Online: Virus detection by transmission electron microscopy: Still useful for diagnosis and a plus for biosafety.

Roingeard, Philippe / Raynal, Pierre-Ivan / Eymieux, Sébastien / Blanchard, Emmanuelle

Reviews in medical virology

2018 Volume 29, Issue 1, Page(s) e2019

Abstract: Transmission electron microscopy (TEM) is the only imaging technique allowing the direct visualization of viruses, due to its nanometer-scale resolution. Between the 1960s and 1990s, TEM contributed to the discovery of many types of viruses and served as ...

Abstract	Transmission electron microscopy (TEM) is the only imaging technique allowing the direct visualization of viruses, due to its nanometer-scale resolution. Between the 1960s and 1990s, TEM contributed to the discovery of many types of viruses and served as a diagnostic tool for identifying viruses directly in biological samples, either in suspension or in sections of tissues or mammalian cells grown in vitro in contact with clinical samples. The diagnosis of viral infections improved considerably during the 1990s, with the advent of highly sensitive techniques, such as enzyme-linked immunosorbent assay (ELISA) and PCR, rendering TEM obsolete for this purpose. However, the last 20 years have demonstrated the utility of this technique in particular situations, due to its "catch-all" nature, making diagnosis possible through visualization of the virus, without the need of prior assumptions about the infectious agent sought. Thus, in several major outbreaks in which molecular techniques failed to identify the infectious agent, TEM provided the answer. TEM is also still occasionally used in routine diagnosis to characterize infections not diagnosed by molecular assays. It is also used to check the microbiological safety of biological products. Many biopharmaceuticals are produced in animal cells that might contain little-known, difficult-to-detect viruses. In this context, the "catch-all" properties of TEM make it possible to document the presence of viruses or virus-like particles in these products.
MeSH term(s)	Animals ; Containment of Biohazards/methods ; Diagnostic Tests, Routine/methods ; Disease Transmission, Infectious/prevention & control ; Humans ; Microscopy, Electron, Transmission/methods ; Technology, Pharmaceutical/methods ; Virion/ultrastructure ; Virus Diseases/diagnosis ; Viruses/isolation & purification ; Viruses/ultrastructure
Keywords	covid19
Language	English
Publishing date	2018-11-09
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	1086043-5
ISSN	1099-1654 ; 1052-9276
ISSN (online)	1099-1654
ISSN	1052-9276
DOI	10.1002/rmv.2019
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Zs.A 3308: Show issues

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Article ; Online: Secretory Vesicles Are the Principal Means of SARS-CoV-2 Egress.

Eymieux, Sébastien / Uzbekov, Rustem / Rouillé, Yves / Blanchard, Emmanuelle / Hourioux, Christophe / Dubuisson, Jean / Belouzard, Sandrine / Roingeard, Philippe

Cells

2021 Volume 10, Issue 8

Abstract: The mechanisms of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) egress, similar to those of other coronaviruses, remain poorly understood. The virus buds in intracellular compartments and is therefore thought to be released by the ... ...

Abstract	The mechanisms of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) egress, similar to those of other coronaviruses, remain poorly understood. The virus buds in intracellular compartments and is therefore thought to be released by the biosynthetic secretory pathway. However, several studies have recently challenged this hypothesis. It has been suggested that coronaviruses, including SARS-CoV-2, use lysosomes for egress. In addition, a focused ion-beam scanning electron microscope (FIB/SEM) study suggested the existence of exit tunnels linking cellular compartments rich in viral particles to the extracellular space resembling those observed for the human immunodeficiency (HIV) in macrophages. Here, we analysed serial sections of Vero cells infected with SARS-CoV-2 by transmission electron microscopy (TEM). We found that SARS-CoV-2 was more likely to exit the cell in small secretory vesicles. Virus trafficking within the cells involves small vesicles, with each generally containing a single virus particle. These vesicles then fuse with the plasma membrane to release the virus into the extracellular space. This work sheds new light on the late stages of the SARS-CoV-2 infectious cycle of potential value for guiding the development of new antiviral strategies.
Language	English
Publishing date	2021-08-10
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells10082047
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