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Book ; Online ; Thesis: Endogen induzierter programmierter Zelltod in Saccharomyces cerevisiae

Wissing, Silke

2005

Author's details	vorgelegt von Silke Wissing
Language	German
Size	Online-Ressource
Document type	Book ; Online ; Thesis
Thesis / German Habilitation thesis	Univ., Diss--Tübingen, 2005
Database	Former special subject collection: coastal and deep sea fishing

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Book ; Online ; Thesis: Endogen induzierter programmierter Zelltod in Saccharomyces cerevisiae

Wissing, Silke [Verfasser]

2005

Author's details	vorgelegt von Silke Wissing
Keywords	Medizin, Gesundheit ; Medicine, Health
Subject code	sg610
Document type	Book ; Online ; Thesis
Database	Digital theses on the web

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Article ; Online: Stable miRNA overexpression in human CAP cells: Engineering alternative production systems for advanced manufacturing of biologics using miR-136 and miR-3074.

Weis, Benjamin L / Guth, Nadine / Fischer, Simon / Wissing, Silke / Fradin, Simon / Holzmann, Karl-Heinz / Handrick, René / Otte, Kerstin

Biotechnology and bioengineering

2018 Volume 115, Issue 8, Page(s) 2027–2038

Abstract: Chinese hamster ovary (CHO) cells still represent the major production host for therapeutic proteins. However, multiple limitations have been acknowledged leading to the search for alternative expression systems. CEVEC's amniocyte production (CAP) cells ... ...

Abstract	Chinese hamster ovary (CHO) cells still represent the major production host for therapeutic proteins. However, multiple limitations have been acknowledged leading to the search for alternative expression systems. CEVEC's amniocyte production (CAP) cells are human production cells demonstrated to enable efficient overexpression of recombinant proteins with human glycosylation pattern. However, CAP cells have not yet undergone any engineering approaches to optimize process parameters for a cheaper and more sustainable production of biopharmaceuticals. Thus, we assessed the possibility to enhance CAP cell production capacity via cell engineering using miRNA technology. Based on a previous high-content miRNA screen in CHO-SEAP cells, selected pro-productive miRNAs including, miR-99b-3p, 30a-5p, 329-3p, 483-3p, 370-3p, 219-1-3p, 3074-5p, 136-3p, 30e-5p, 1a-3p, and 484-5p, were shown to act pro-productive and product independent upon transient transfection in CAP and CHO antibody expressing cell lines. Stable expression of miRNAs established seven CAP cell pools with an overexpression of the pro-productive miRNA strand. Subsequent small-scale screening as well as upscaling batch experiments identified miR-136 and miR-3074 to significantly increase final mAb concentration in CAP-mAb cells. Transcriptomic changes analyzed by microarrays identified several lncRNAs as well as growth and apoptosis-related miRNAs to be differentially regulated in CAP-mAb-miR-136 and -miR-3074. This study presents the first engineering approach to optimize the alternative human expression system of CAP-cells.
MeSH term(s)	Antibodies, Monoclonal/genetics ; Antibodies, Monoclonal/metabolism ; Biological Products/metabolism ; Cell Line ; Humans ; Metabolic Engineering/methods ; MicroRNAs/biosynthesis ; MicroRNAs/genetics ; Recombinant Proteins/genetics ; Recombinant Proteins/metabolism
Chemical Substances	Antibodies, Monoclonal ; Biological Products ; MIRN136 microRNA, human ; MIRN3074 microRNA, human ; MicroRNAs ; Recombinant Proteins
Language	English
Publishing date	2018-05-03
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	280318-5
ISSN	1097-0290 ; 0006-3592
ISSN (online)	1097-0290
ISSN	0006-3592
DOI	10.1002/bit.26715
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: HIV-1 Vif versus the APOBEC3 cytidine deaminases: an intracellular duel between pathogen and host restriction factors.

Wissing, Silke / Galloway, Nicole L K / Greene, Warner C

Molecular aspects of medicine

2010 Volume 31, Issue 5, Page(s) 383–397

Abstract: The Vif protein of HIV is essential for the effective propagation of this pathogenic retrovirus in vivo. Vif acts by preventing virion encapsidation of two potent antiviral factors, the APOBEC3G and APOBEC3F cytidine deaminases. Decreased encapsidation ... ...

Abstract	The Vif protein of HIV is essential for the effective propagation of this pathogenic retrovirus in vivo. Vif acts by preventing virion encapsidation of two potent antiviral factors, the APOBEC3G and APOBEC3F cytidine deaminases. Decreased encapsidation in part involves Vif-mediated recruitment of a ubiquitin E3 ligase complex that promotes polyubiquitylation and proteasome-mediated degradation of APOBEC3G/F. The resultant decline in intracellular levels of these enzymes leads to decreased encapsidation of APOBECG/F into budding virions. This review discusses recent advances in our understanding of the dynamic interplay of Vif with the antiviral APOBEC3 enzymes.
MeSH term(s)	Cytosine Deaminase/metabolism ; HIV-1/growth & development ; HIV-1/metabolism ; Host-Pathogen Interactions ; Humans ; Intracellular Space/metabolism ; Virus Assembly ; vif Gene Products, Human Immunodeficiency Virus/genetics ; vif Gene Products, Human Immunodeficiency Virus/metabolism
Chemical Substances	vif Gene Products, Human Immunodeficiency Virus ; Cytosine Deaminase (EC 3.5.4.1)
Language	English
Publishing date	2010-06-09
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	197640-0
ISSN	1872-9452 ; 0098-2997
ISSN (online)	1872-9452
ISSN	0098-2997
DOI	10.1016/j.mam.2010.06.001
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Article ; Online: Human CAP cells represent a novel source for functional, miRNA-loaded exosome production.

Zeh, Nikolas / Schneider, Helga / Mathias, Sven / Raab, Nadja / Kleemann, Michael / Schmidt-Hertel, Sabine / Weis, Benjamin / Wissing, Silke / Strempel, Nikola / Handrick, René / Otte, Kerstin

PloS one

2019 Volume 14, Issue 8, Page(s) e0221679

Abstract: Exosomes represent a promising delivery tool for nucleic acid-based pharmaceuticals. They are highly suitable for transporting therapeutic miRNAs to tumor cells, due to their natural membrane components. Further, exosomes are capable of effectively ... ...

Abstract	Exosomes represent a promising delivery tool for nucleic acid-based pharmaceuticals. They are highly suitable for transporting therapeutic miRNAs to tumor cells, due to their natural membrane components. Further, exosomes are capable of effectively protecting nucleic acids against ribonucleases and enable the delivery of their content through cell membranes. However, no suitable production host for miRNA containing exosomes of non-tumorigenic origin has yet been identified. In this study we engineered an immortalised human amniocyte cell line (CAP® cells), whose exosomes were enriched and characterised. The cell line modifications not only enabled the production of GFP-labelled but also pro-apoptotic miRNA containing exosomes without negative influence on host cell growth. Furthermore, we demonstrated that pro-apoptotic miRNA containing CAP exosomes are taken up by ovarian cancer cells. Strikingly, delivery of functional exosomal miRNA led to downregulation of several reported target genes in the treated tumor cells. In summary, we revealed CAP cells of non-tumorigenic origin as a novel and efficient exosome production host with the potential to produce functional miRNA-loaded exosomes.
MeSH term(s)	Amnion/cytology ; Apoptosis ; Carcinogenesis/pathology ; Cell Line ; Cell Proliferation ; Cell Survival ; Exosomes/metabolism ; Exosomes/ultrastructure ; Female ; Humans ; MicroRNAs/metabolism ; Ovarian Neoplasms/pathology ; Tetraspanin 30/metabolism
Chemical Substances	MicroRNAs ; Tetraspanin 30
Language	English
Publishing date	2019-08-28
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0221679
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Human CAP cells represent a novel source for functional, miRNA-loaded exosome production.

Nikolas Zeh / Helga Schneider / Sven Mathias / Nadja Raab / Michael Kleemann / Sabine Schmidt-Hertel / Benjamin Weis / Silke Wissing / Nikola Strempel / René Handrick / Kerstin Otte

PLoS ONE, Vol 14, Iss 8, p e

2019 Volume 0221679

Abstract	Exosomes represent a promising delivery tool for nucleic acid-based pharmaceuticals. They are highly suitable for transporting therapeutic miRNAs to tumor cells, due to their natural membrane components. Further, exosomes are capable of effectively protecting nucleic acids against ribonucleases and enable the delivery of their content through cell membranes. However, no suitable production host for miRNA containing exosomes of non-tumorigenic origin has yet been identified. In this study we engineered an immortalised human amniocyte cell line (CAP® cells), whose exosomes were enriched and characterised. The cell line modifications not only enabled the production of GFP-labelled but also pro-apoptotic miRNA containing exosomes without negative influence on host cell growth. Furthermore, we demonstrated that pro-apoptotic miRNA containing CAP exosomes are taken up by ovarian cancer cells. Strikingly, delivery of functional exosomal miRNA led to downregulation of several reported target genes in the treated tumor cells. In summary, we revealed CAP cells of non-tumorigenic origin as a novel and efficient exosome production host with the potential to produce functional miRNA-loaded exosomes.
Keywords	Medicine ; R ; Science ; Q
Subject code	500
Language	English
Publishing date	2019-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Utilization of replication-competent XMRV reporter-viruses reveals severe viral restriction in primary human cells.

Stürzel, Christina Martina / Palesch, David / Khalid, Mohammad / Wissing, Silke / Fischer, Nicole / Münch, Jan

PloS one

2013 Volume 8, Issue 9, Page(s) e74427

Abstract: The gammaretrovirus termed xenotropic murine leukemia virus-related virus (XMRV) was described to be isolated from prostate cancer tissue biopsies and from blood of patients suffering from chronic fatigue syndrome. However, many studies failed to detect ... ...

Abstract	The gammaretrovirus termed xenotropic murine leukemia virus-related virus (XMRV) was described to be isolated from prostate cancer tissue biopsies and from blood of patients suffering from chronic fatigue syndrome. However, many studies failed to detect XMRV and to verify these disease associations. Data suggesting the contamination of specimens in particular by PCR-based methods and recent reports demonstrating XMRV generation via recombination of two murine leukemia virus precursors raised serious doubts about XMRV being a genuine human pathogen. To elucidate cell tropism of XMRV, we generated replication competent XMRV reporter viruses encoding a green fluorescent protein or a secretable luciferase as tools to analyze virus infection of human cell lines or primary human cells. Transfection of proviral DNAs into LNCaP prostate cancer cells resulted in readily detectably reporter gene expression and production of progeny virus. Inoculation of known XMRV susceptible target cells revealed that these virions were infectious and expressed the reporter gene, allowing for a fast and highly sensitive quantification of XMRV infection. Both reporter viruses were capable of establishing a spreading infection in LNCaP and Raji B cells and could be easily passaged. However, after inoculation of primary human blood cells such as CD4 T cells, macrophages or dendritic cells, infection rates were very low, and a spreading infection was never established. In line with these results we found that supernatants derived from these XMRV infected primary cell types did not contain infectious virus. Thus, although XMRV efficiently replicated in some human cell lines, all tested primary cells were largely refractory to XMRV infection and did not support viral spread. Our results provide further evidence that XMRV is not a human pathogen.
MeSH term(s)	Animals ; Cells, Cultured ; Gene Expression ; Genes, Reporter ; Humans ; Mice ; Proviruses/physiology ; Retroviridae Infections/pathology ; Retroviridae Infections/virology ; Virion/metabolism ; Virus Replication/physiology ; Xenotropic murine leukemia virus-related virus/physiology
Language	English
Publishing date	2013-09-13
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0074427
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Rapid intracellular competition between hepatitis C viral genomes as a result of mitosis.

Webster, Brian / Wissing, Silke / Herker, Eva / Ott, Melanie / Greene, Warner C

Journal of virology

2012 Volume 87, Issue 1, Page(s) 581–596

Abstract: Cells infected with hepatitis C virus (HCV) become refractory to further infection by HCV (T. Schaller et al., J. Virol. 81:4591-4603, 2007; D. M. Tscherne et al., J. Virol. 81:3693-3703, 2007). This process, termed superinfection exclusion, does not ... ...

Abstract	Cells infected with hepatitis C virus (HCV) become refractory to further infection by HCV (T. Schaller et al., J. Virol. 81:4591-4603, 2007; D. M. Tscherne et al., J. Virol. 81:3693-3703, 2007). This process, termed superinfection exclusion, does not involve downregulation of surface viral receptors but instead occurs inside the cell at the level of RNA replication. The originally infecting virus may occupy replication niches or sequester host factors necessary for viral growth, preventing effective growth of viruses that enter the cell later. However, there appears to be an additional level of intracellular competition between viral genomes that occurs at or shortly following mitosis. In the setting of cellular division, when two viral replicons of equivalent fitness are present within a cell, each has an equal opportunity to exclude the other. In a population of dividing cells, the competition between viral genomes proceeds apace, randomly clearing one or the other genome from cells in the span of 9 to 12 days. These findings demonstrate a new mechanism of intracellular competition between HCV strains, which may act to further limit HCV's genetic diversity and ability to recombine in vivo.
MeSH term(s)	Cell Line ; Hepacivirus/growth & development ; Hepacivirus/physiology ; Hepatocytes/virology ; Host-Pathogen Interactions ; Humans ; Mitosis ; Viral Interference
Language	English
Publishing date	2012-10-24
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	80174-4
ISSN	1098-5514 ; 0022-538X
ISSN (online)	1098-5514
ISSN	0022-538X
DOI	10.1128/JVI.01047-12
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Article ; Online: Utilization of replication-competent XMRV reporter-viruses reveals severe viral restriction in primary human cells.

Christina Martina Stürzel / David Palesch / Mohammad Khalid / Silke Wissing / Nicole Fischer / Jan Münch

PLoS ONE, Vol 8, Iss 9, p e

2013 Volume 74427

Abstract	The gammaretrovirus termed xenotropic murine leukemia virus-related virus (XMRV) was described to be isolated from prostate cancer tissue biopsies and from blood of patients suffering from chronic fatigue syndrome. However, many studies failed to detect XMRV and to verify these disease associations. Data suggesting the contamination of specimens in particular by PCR-based methods and recent reports demonstrating XMRV generation via recombination of two murine leukemia virus precursors raised serious doubts about XMRV being a genuine human pathogen. To elucidate cell tropism of XMRV, we generated replication competent XMRV reporter viruses encoding a green fluorescent protein or a secretable luciferase as tools to analyze virus infection of human cell lines or primary human cells. Transfection of proviral DNAs into LNCaP prostate cancer cells resulted in readily detectably reporter gene expression and production of progeny virus. Inoculation of known XMRV susceptible target cells revealed that these virions were infectious and expressed the reporter gene, allowing for a fast and highly sensitive quantification of XMRV infection. Both reporter viruses were capable of establishing a spreading infection in LNCaP and Raji B cells and could be easily passaged. However, after inoculation of primary human blood cells such as CD4 T cells, macrophages or dendritic cells, infection rates were very low, and a spreading infection was never established. In line with these results we found that supernatants derived from these XMRV infected primary cell types did not contain infectious virus. Thus, although XMRV efficiently replicated in some human cell lines, all tested primary cells were largely refractory to XMRV infection and did not support viral spread. Our results provide further evidence that XMRV is not a human pathogen.
Keywords	Medicine ; R ; Science ; Q
Subject code	570
Language	English
Publishing date	2013-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: The inhibitor-of-apoptosis protein Bir1p protects against apoptosis in S. cerevisiae and is a substrate for the yeast homologue of Omi/HtrA2.

Walter, David / Wissing, Silke / Madeo, Frank / Fahrenkrog, Birthe

Journal of cell science

2006 Volume 119, Issue Pt 9, Page(s) 1843–1851

Abstract: Inhibitor-of-apoptosis proteins (IAPs) play a crucial role in the regulation of metazoan apoptosis. IAPs are typically characterized by the presence of one to three baculovirus IAP repeat (BIR) domains that are essential for their anti-apoptotic activity. ...

Abstract	Inhibitor-of-apoptosis proteins (IAPs) play a crucial role in the regulation of metazoan apoptosis. IAPs are typically characterized by the presence of one to three baculovirus IAP repeat (BIR) domains that are essential for their anti-apoptotic activity. Bir1p is the sole BIR-protein in yeast and has been shown to participate in chromosome segregation events. Here, we show that Bir1p is a substrate for Nma111p, which is the homologue of the human pro-apoptotic serine protease Omi/HtrA2 and which is known to mediate apoptosis in yeast. Bir1p is a cytoplasmic and nuclear protein, and yeast cells lacking bir1 are more sensitive to apoptosis induced by oxidative stress. Consistently, overexpression of Bir1p reduces apoptosis-like cell death, whereas this protective effect can be antagonized in vivo by simultaneous overexpression of Nma111p. Moreover, chronologically aged cells that constitutively overexpress Bir1p show a delayed onset of cell death. Therefore, Bir1p, like its closest metazoan homologues deterin and survivin, has dual functions: it participates in chromosome segregation events and cytokinesis and exhibits anti-apoptotic activity.
MeSH term(s)	Aging/physiology ; Animals ; Apoptosis/physiology ; Fungal Proteins/genetics ; Fungal Proteins/metabolism ; High-Temperature Requirement A Serine Peptidase 2 ; Humans ; Mitochondrial Proteins/metabolism ; Recombinant Fusion Proteins/genetics ; Recombinant Fusion Proteins/metabolism ; Saccharomyces cerevisiae/physiology ; Saccharomyces cerevisiae/ultrastructure ; Saccharomyces cerevisiae Proteins/genetics ; Saccharomyces cerevisiae Proteins/metabolism ; Serine Endopeptidases/genetics ; Serine Endopeptidases/metabolism
Chemical Substances	Fungal Proteins ; Mitochondrial Proteins ; Recombinant Fusion Proteins ; Saccharomyces cerevisiae Proteins ; NMA111 protein, S cerevisiae (EC 3.4.21.-) ; Serine Endopeptidases (EC 3.4.21.-) ; HTRA2 protein, human (EC 3.4.21.108) ; High-Temperature Requirement A Serine Peptidase 2 (EC 3.4.21.108)
Language	English
Publishing date	2006-05-01
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2993-2
ISSN	1477-9137 ; 0021-9533
ISSN (online)	1477-9137
ISSN	0021-9533
DOI	10.1242/jcs.02902
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