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Article: Editorial: Novel approaches to prevention, diagnosis, and treatment of bacterial and viral infections of clinical relevance.

Leanse, Leon G / Trombetta, Claudia Maria / Carnell, George W

2023 Volume 14, Page(s) 1192435

Language	English
Publishing date	2023-04-13
Publishing country	Switzerland
Document type	Editorial
ZDB-ID	2587354-4
ISSN	1664-302X
ISSN	1664-302X
DOI	10.3389/fmicb.2023.1192435
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Article ; Online: Immune imprinting and next-generation coronavirus vaccines.

Huang, Chloe Qingzhou / Vishwanath, Sneha / Carnell, George William / Chan, Andrew Chun Yue / Heeney, Jonathan Luke

Nature microbiology

2023 Volume 8, Issue 11, Page(s) 1971–1985

Abstract: Vaccines based on historical virus isolates provide limited protection from continuously evolving RNA viruses, such as influenza viruses or coronaviruses, which occasionally spill over between animals and humans. Despite repeated booster immunizations, ... ...

Abstract	Vaccines based on historical virus isolates provide limited protection from continuously evolving RNA viruses, such as influenza viruses or coronaviruses, which occasionally spill over between animals and humans. Despite repeated booster immunizations, population-wide declines in the neutralization of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants have occurred. This has been compared to seasonal influenza vaccinations in humans, where the breadth of immune responses induced by repeat exposures to antigenically distinct influenza viruses is confounded by pre-existing immunity-a mechanism known as imprinting. Since its emergence, SARS-CoV-2 has evolved in a population with partial immunity, acquired by infection, vaccination or both. Here we critically examine the evidence for and against immune imprinting in host humoral responses to SARS-CoV-2 and its implications for coronavirus disease 2019 (COVID-19) booster vaccine programmes.
MeSH term(s)	Animals ; Humans ; COVID-19 Vaccines ; SARS-CoV-2 ; Vaccination ; Orthomyxoviridae
Chemical Substances	COVID-19 Vaccines
Language	English
Publishing date	2023-11-06
Publishing country	England
Document type	Journal Article ; Review
ISSN	2058-5276
ISSN (online)	2058-5276
DOI	10.1038/s41564-023-01505-9
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Article: Correlation of Influenza B Haemagglutination Inhibiton, Single-Radial Haemolysis and Pseudotype-Based Microneutralisation Assays for Immunogenicity Testing of Seasonal Vaccines.

Carnell, George W / Trombetta, Claudia M / Ferrara, Francesca / Montomoli, Emanuele / Temperton, Nigel J

Vaccines

2021 Volume 9, Issue 2

Abstract: Influenza B is responsible for a significant proportion of the global morbidity, mortality and economic loss caused by influenza-related disease. Two antigenically distinct lineages co-circulate worldwide, often resulting in mismatches in vaccine ... ...

Abstract	Influenza B is responsible for a significant proportion of the global morbidity, mortality and economic loss caused by influenza-related disease. Two antigenically distinct lineages co-circulate worldwide, often resulting in mismatches in vaccine coverage when vaccine predictions fail. There are currently operational issues with gold standard serological assays for influenza B, such as lack of sensitivity and requirement for specific antigen treatment. This study encompasses the gold standard assays with the more recent Pseudotype-based Microneutralisation assay in order to study comparative serological outcomes. Haemagglutination Inhibition, Single Radial Haemolysis and Pseudotype-based Microneutralisation correlated strongly for strains in the Yamagata lineage; however, it correlated with neither gold standard assays for the Victoria lineage.
Language	English
Publishing date	2021-01-28
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2703319-3
ISSN	2076-393X
ISSN	2076-393X
DOI	10.3390/vaccines9020100
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Article ; Online: Differential T-cell and antibody responses induced by mRNA versus adenoviral vectored COVID-19 vaccines in patients with immunodeficiencies.

Aguinam, Ernest T / Nadesalingam, Angalee / Chan, Andrew / Smith, Peter / Paloniemi, Minna / Cantoni, Diego / Gronlund, Jessica / Gronlund, Helen / Carnell, George W / Castillo-Olivares, Javier / Temperton, Nigel / Blacklaws, Barbara / Heeney, Jonathan L / Baxendale, Helen

The journal of allergy and clinical immunology. Global

2023 Volume 2, Issue 2, Page(s) 100091

Abstract: Background: Immunodeficient patients (IDPs) are at higher risk of contracting severe coronavirus disease 2019 (COVID-19). Targeted vaccination strategies have been implemented to enhance vaccine-induced protection. In this population, however, clinical ... ...

Abstract	Background: Immunodeficient patients (IDPs) are at higher risk of contracting severe coronavirus disease 2019 (COVID-19). Targeted vaccination strategies have been implemented to enhance vaccine-induced protection. In this population, however, clinical effectiveness is variable and the duration of protection unknown. Objective: We sought to better understand the cellular and humoral immune responses to mRNA and adenoviral vectored COVID-19 vaccines in patients with immunodeficiency. Methods: Immune responses to severe acute respiratory syndrome coronavirus 2 spike were assessed after 2 doses of homologous ChAdOx1-nCoV-19 or BNT162b2 vaccines in 112 infection-naive IDPs and 131 healthy health care workers as controls. Predictors of vaccine responsiveness were investigated. Results: Immune responses to vaccination were low, and virus neutralization by antibody was not detected despite high titer binding responses in many IDPs. In those exhibiting response, the frequency of specific T-cell responses in IDPs was similar to controls, while antibody responses were lower. Sustained vaccine specific differences were identified: T-cell responses were greater in ChAdOx1-nCoV-19- compared to BNT162b2-immunized IDPs, and antibody binding and neutralization were greater in all cohorts immunized with BNT162b2. The positive correlation between T-cell and antibody responses was weak and increased with subsequent vaccination. Conclusion: Immunodeficient patients have impaired immune responses to mRNA and viral vector COVID-19 vaccines that appear to be influenced by vaccine formulation. Understanding the relative roles of T-cell- and antibody-mediated protection as well as the potential of heterologous prime and boost immunization protocols is needed to optimize the vaccination approach in these high-risk groups.
Language	English
Publishing date	2023-03-15
Publishing country	United States
Document type	Journal Article
ISSN	2772-8293
ISSN (online)	2772-8293
DOI	10.1016/j.jacig.2023.100091
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Article: Development of a Clinical MALDI-ToF Mass Spectrometry Assay for SARS-CoV-2: Rational Design and Multi-Disciplinary Team Work.

Iles, Ray K / Zmuidinaite, Raminta / Iles, Jason K / Carnell, George / Sampson, Alex / Heeney, Jonathan L

Diagnostics (Basel, Switzerland)

2020 Volume 10, Issue 10

Abstract: The COVID-19 pandemic caused by the SARS-CoV-2 coronavirus has stretched national testing capacities to breaking points in almost all countries of the world. The need to rapidly screen vast numbers of a country's population in order to control the spread ...

Abstract	The COVID-19 pandemic caused by the SARS-CoV-2 coronavirus has stretched national testing capacities to breaking points in almost all countries of the world. The need to rapidly screen vast numbers of a country's population in order to control the spread of the infection is paramount. However, the logistical requirement for reagent supply (and associated cost) of RT-PCR based testing (the current front-line test) have been hugely problematic. Mass spectrometry-based methods using swab and gargle samples have been reported with promise, but have not approached the task from a systematic analysis of the entire diagnostic process. Here, the pipeline from sample processing, the biological characteristics of the pathogen in human biofluid, the downstream bio- and physical-chemistry and the all-important data processing with clinical interpretation and reporting, are carefully compiled into a single high-throughput and reproducible rapid process. Utilizing MALDI-ToF mass spectrometric detection to viral envelope glycoproteins in a systems biology-multidisciplinary team approach, we have achieved a multifaceted clinical MALDI ToF MS screening test, primarily (but not limited to) SARS-CoV-2, with direct application to other future epidemics/pandemics that may arise. The clinical information generated not only includes SARS-CoV-2 coronavirus detection-(Spike protein fragments S1, S2b, S2a peaks), but other respiratory viral infections detected as well as an assessment of generalised oral upper respiratory immune response (elevated total Ig light chain peak) and a measure of the viral immune response (elevated intensity of IgA heavy chain peak). The advantages of the method include; (1) ease of sampling, (2) speed of analysis, and much reduced cost of testing. These features reveal the diagnostic utility of MALDI-ToF mass spectrometry as a powerful and economically attractive global solution.
Keywords	covid19
Language	English
Publishing date	2020-09-24
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2662336-5
ISSN	2075-4418
ISSN	2075-4418
DOI	10.3390/diagnostics10100746
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Article ; Online: Immunogenicity of a silica nanoparticle-based SARS-CoV-2 vaccine in mice.

Barbey, Clara / Su, Jinpeng / Billmeier, Martina / Stefan, Nadine / Bester, Romina / Carnell, George / Temperton, Nigel / Heeney, Jonathan / Protzer, Ulrike / Breunig, Miriam / Wagner, Ralf / Peterhoff, David

European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V

2023 Volume 192, Page(s) 41–55

Abstract: Safe and effective vaccines have been regarded early on as critical in combating the COVID-19 pandemic. Among the deployed vaccine platforms, subunit vaccines have a particularly good safety profile but may suffer from a lower immunogenicity compared to ... ...

Abstract	Safe and effective vaccines have been regarded early on as critical in combating the COVID-19 pandemic. Among the deployed vaccine platforms, subunit vaccines have a particularly good safety profile but may suffer from a lower immunogenicity compared to mRNA based or viral vector vaccines. In fact, this phenomenon has also been observed for SARS-CoV-2 subunit vaccines comprising the receptor-binding domain (RBD) of the spike (S) protein. Therefore, RBD-based vaccines have to rely on additional measures to enhance the immune response. It is well accepted that displaying antigens on nanoparticles can improve the quantity and quality of vaccine-mediated both humoral and cell-mediated immune responses. Based on this, we hypothesized that SARS-CoV-2 RBD as immunogen would benefit from being presented to the immune system via silica nanoparticles (SiNPs). Herein we describe the preparation, in vitro characterization, antigenicity and in vivo immunogenicity of SiNPs decorated with properly oriented RBD in mice. We found our RBD-SiNP conjugates show narrow, homogeneous particle distribution with optimal size of about 100 nm for efficient transport to and into the lymph node. The colloidal stability and binding of the antigen was stable for at least 4 months at storage- and in vivo-temperatures. The antigenicity of the RBD was maintained upon binding to the SiNP surface, and the receptor-binding motif was readily accessible due to the spatial orientation of the RBD. The particles were efficiently taken up in vitro by antigen-presenting cells. In a mouse immunization study using an mRNA vaccine and spike protein as benchmarks, we found that the SiNP formulation was able to elicit a stronger RBD-specific humoral response compared to the soluble protein. For the adjuvanted RBD-SiNP we found strong S-specific multifunctional CD4
MeSH term(s)	Animals ; Humans ; Mice ; COVID-19 Vaccines ; COVID-19/prevention & control ; Pandemics ; SARS-CoV-2 ; Viral Vaccines ; Vaccines, Subunit ; Antibodies, Viral ; Antibodies, Neutralizing
Chemical Substances	COVID-19 Vaccines ; Viral Vaccines ; Vaccines, Subunit ; Antibodies, Viral ; Antibodies, Neutralizing
Language	English
Publishing date	2023-09-27
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	1065368-5
ISSN	1873-3441 ; 0939-6411
ISSN (online)	1873-3441
ISSN	0939-6411
DOI	10.1016/j.ejpb.2023.09.015
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Article ; Online: Nanobodies mapped to cross-reactive and divergent epitopes on A(H7N9) influenza hemagglutinin using yeast display.

Gaiotto, Tiziano / Ramage, Walter / Ball, Christina / Risley, Paul / Carnell, George W / Temperton, Nigel / Engelhardt, Othmar G / Hufton, Simon E

Scientific reports

2021 Volume 11, Issue 1, Page(s) 3126

Abstract: Influenza H7N9 virus continues to cause infections in humans and represents a significant pandemic risk. During the most recent 5th epidemic wave in 2016/17 two distinct lineages with increased human infections and wider geographical spread emerged. In ... ...

Abstract	Influenza H7N9 virus continues to cause infections in humans and represents a significant pandemic risk. During the most recent 5th epidemic wave in 2016/17 two distinct lineages with increased human infections and wider geographical spread emerged. In preparation for any future adaptations, broadly reactive antibodies against H7N9 are required for surveillance, therapy and prophylaxis. In this study we have isolated a panel of nanobodies (Nbs) with broad reactivity across H7 influenza strains, including H7N9 strains between 2013 and 2017. We also describe Nbs capable of distinguishing between the most recent high and low pathogenicity Yangtze River Delta lineage H7N9 strains. Nanobodies were classified into 5 distinct groups based on their epitope footprint determined using yeast display and mutational scanning. The epitope footprint of Nbs capable of distinguishing high pathogenic (HP) A/Guangdong/17SF003/2016 from low pathogenic (LP) A/Hong Kong/125/2017 (H7N9) were correlated to natural sequence divergence in the head domain at lysine 164. Several Nbs binding to the head domain were capable of viral neutralisation. The potency of one nanobody NB7-14 could be increased over 1000-fold to 113 pM by linking two Nbs together. Nbs specific for distinct epitopes on H7N9 may be useful for surveillance or therapy in human or veterinary settings.
MeSH term(s)	Amino Acid Sequence ; Animals ; Antibodies, Neutralizing/biosynthesis ; Antibodies, Neutralizing/isolation & purification ; Antibodies, Viral/biosynthesis ; Antibodies, Viral/isolation & purification ; Binding Sites ; Birds/virology ; Epitopes/chemistry ; Epitopes/genetics ; Epitopes/immunology ; Hemagglutinin Glycoproteins, Influenza Virus/chemistry ; Hemagglutinin Glycoproteins, Influenza Virus/genetics ; Hemagglutinin Glycoproteins, Influenza Virus/immunology ; Humans ; Influenza A Virus, H7N9 Subtype/drug effects ; Influenza A Virus, H7N9 Subtype/genetics ; Influenza A Virus, H7N9 Subtype/immunology ; Influenza in Birds/immunology ; Influenza in Birds/prevention & control ; Influenza in Birds/transmission ; Influenza in Birds/virology ; Influenza, Human/immunology ; Influenza, Human/prevention & control ; Influenza, Human/transmission ; Influenza, Human/virology ; Models, Molecular ; Peptide Library ; Protein Binding ; Protein Conformation ; Protein Interaction Domains and Motifs ; Saccharomyces cerevisiae/genetics ; Saccharomyces cerevisiae/metabolism ; Single-Domain Antibodies/biosynthesis ; Single-Domain Antibodies/isolation & purification
Chemical Substances	Antibodies, Neutralizing ; Antibodies, Viral ; Epitopes ; Hemagglutinin Glycoproteins, Influenza Virus ; Peptide Library ; Single-Domain Antibodies ; hemagglutinin, avian influenza A virus
Language	English
Publishing date	2021-02-04
Publishing country	England
Document type	Journal Article ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-021-82356-4
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Article ; Online: An Optimized Method for the Production Using PEI, Titration and Neutralizationof SARS-CoV Spike Luciferase Pseudotypes.

Carnell, George / Grehan, Keith / Ferrara, Francesca / Molesti, Eleonora / Temperton, Nigel

Bio-protocol

2017 Volume 7, Issue 16, Page(s) e2514

Abstract: The protocol outlined represents a cost-effective, rapid and reliable method for the generation of high-titre viral pseudotype particles with the wild-type SARS-CoV spike protein on a lentiviral vector core using the widely available transfection reagent ...

Abstract	The protocol outlined represents a cost-effective, rapid and reliable method for the generation of high-titre viral pseudotype particles with the wild-type SARS-CoV spike protein on a lentiviral vector core using the widely available transfection reagent PEI. This protocol is optimized for transfection in 6-well plates; however it can be readily scaled to different production volumes according to application. This protocol has multiple benefits including the use of readily available reagents, consistent, high pseudotype virus production Relative Luminescence Units (RLU) titres and rapid generation of novel coronavirus pseudotypes for research into strain variation, tropism and immunogenicity/sero-prevalence.
Language	English
Publishing date	2017-08-20
Publishing country	United States
Document type	Journal Article
ZDB-ID	2833269-6
ISSN	2331-8325 ; 2331-8325
ISSN (online)	2331-8325
ISSN	2331-8325
DOI	10.21769/BioProtoc.2514
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Article ; Online: Inhibition of the Lectin Pathway of Complement Activation Reduces Acute Respiratory Distress Syndrome Severity in a Mouse Model of SARS-CoV-2 Infection.

Ali, Youssif M / Carnell, George W / Fumagalli, Stefano / Mercurio, Domenico / Seminara, Serena / Lynch, Nicholas J / Khatri, Priyanka / Arachchilage, Chanuka H / Mascheroni, Luca / Kaminski, Clemens / George, Charlotte L / Stewart, Hazel / Yabuki, Munehisa / Demopulos, Gregory / Heeney, Jonathan L / Schwaeble, Wilhelm

The Journal of infectious diseases

2023 Volume 229, Issue 3, Page(s) 680–690

Abstract: Most patients with COVID-19 in the intensive care unit develop an acute respiratory distress syndrome characterized by severe hypoxemia, decreased lung compliance, and high vascular permeability. Activation of the complement system is a hallmark of ... ...

Abstract	Most patients with COVID-19 in the intensive care unit develop an acute respiratory distress syndrome characterized by severe hypoxemia, decreased lung compliance, and high vascular permeability. Activation of the complement system is a hallmark of moderate and severe COVID-19, with abundant deposition of complement proteins in inflamed tissue and on the endothelium during COVID-19. Using a transgenic mouse model of SARS-CoV-2 infection, we assessed the therapeutic utility of an inhibitory antibody (HG4) targeting MASP-2, a key enzyme in the lectin pathway. Treatment of infected mice with HG4 reduced the disease severity score and improved survival vs mice that received an isotype control antibody. Administration of HG4 significantly reduced the lung injury score, including alveolar inflammatory cell infiltration, alveolar edema, and alveolar hemorrhage. The ameliorating effect of MASP-2 inhibition on the severity of COVID-19 pathology is reflected by a significant reduction in the proinflammatory activation of brain microglia in HG4-treated mice.
MeSH term(s)	Humans ; Animals ; Mice ; COVID-19 ; Mannose-Binding Protein-Associated Serine Proteases/metabolism ; SARS-CoV-2/metabolism ; Complement Activation ; Respiratory Distress Syndrome ; Disease Models, Animal ; Complement System Proteins
Chemical Substances	Mannose-Binding Protein-Associated Serine Proteases (EC 3.4.21.-) ; Complement System Proteins (9007-36-7)
Language	English
Publishing date	2023-10-05
Publishing country	United States
Document type	Journal Article
ZDB-ID	3019-3
ISSN	1537-6613 ; 0022-1899
ISSN (online)	1537-6613
ISSN	0022-1899
DOI	10.1093/infdis/jiad462
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Article ; Online: Tetherin antagonism by SARS-CoV-2 ORF3a and spike protein enhances virus release.

Stewart, Hazel / Palmulli, Roberta / Johansen, Kristoffer H / McGovern, Naomi / Shehata, Ola M / Carnell, George W / Jackson, Hannah K / Lee, Jin S / Brown, Jonathan C / Burgoyne, Thomas / Heeney, Jonathan L / Okkenhaug, Klaus / Firth, Andrew E / Peden, Andrew A / Edgar, James R

EMBO reports

2023 Volume 24, Issue 12, Page(s) e57224

Abstract: The antiviral restriction factor, tetherin, blocks the release of several different families of enveloped viruses, including the Coronaviridae. Tetherin is an interferon-induced protein that forms parallel homodimers between the host cell and viral ... ...

Abstract	The antiviral restriction factor, tetherin, blocks the release of several different families of enveloped viruses, including the Coronaviridae. Tetherin is an interferon-induced protein that forms parallel homodimers between the host cell and viral particles, linking viruses to the surface of infected cells and inhibiting their release. We demonstrate that SARS-CoV-2 infection causes tetherin downregulation and that tetherin depletion from cells enhances SARS-CoV-2 viral titres. We investigate the potential viral proteins involved in abrogating tetherin function and find that SARS-CoV-2 ORF3a reduces tetherin localisation within biosynthetic organelles where Coronaviruses bud, and increases tetherin localisation to late endocytic organelles via reduced retrograde recycling. We also find that expression of Spike protein causes a reduction in cellular tetherin levels. Our results confirm that tetherin acts as a host restriction factor for SARS-CoV-2 and highlight the multiple distinct mechanisms by which SARS-CoV-2 subverts tetherin function.
MeSH term(s)	Humans ; Bone Marrow Stromal Antigen 2/antagonists & inhibitors ; Bone Marrow Stromal Antigen 2/metabolism ; COVID-19/virology ; GPI-Linked Proteins/genetics ; GPI-Linked Proteins/metabolism ; SARS-CoV-2/physiology ; Spike Glycoprotein, Coronavirus/genetics ; Virus Release
Chemical Substances	Bone Marrow Stromal Antigen 2 ; GPI-Linked Proteins ; Spike Glycoprotein, Coronavirus ; spike protein, SARS-CoV-2 ; ORF3a protein, SARS-CoV-2
Language	English
Publishing date	2023-10-11
Publishing country	England
Document type	Journal Article
ZDB-ID	2020896-0
ISSN	1469-3178 ; 1469-221X
ISSN (online)	1469-3178
ISSN	1469-221X
DOI	10.15252/embr.202357224
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