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  1. Artikel ; Online: Myasthenia gravis associated with a pelvic follicular lymphoma.

    Willis, Charlotte / Williamson, Max / Constantinou, Nicholas / Johns, Emily

    BMJ case reports

    2022  Band 15, Heft 1

    Abstract: An 81-year-old woman presented with neck weakness, dysarthria, dysphasia and left-sided ptosis. Myasthenia gravis (MG) was strongly suspected. Voltage gated calcium channel (VGCC) antibodies, associated with Lambert-Eaton myasthenic syndrome (LEMS), were ...

    Abstract An 81-year-old woman presented with neck weakness, dysarthria, dysphasia and left-sided ptosis. Myasthenia gravis (MG) was strongly suspected. Voltage gated calcium channel (VGCC) antibodies, associated with Lambert-Eaton myasthenic syndrome (LEMS), were negative. Acetylcholine receptor (AChR) antibody level was 536 nmol/L and diagnosis of MG was confirmed. Imaging revealed a pelvic mass and subsequent biopsy confirmed a pelvic follicular lymphoma. Our searches revealed this to be the first documented case of MG associated with a pelvic follicular lymphoma. She underwent radiotherapy to treat the lymphoma and received both pyridostigmine and immunosuppression to treat the MG. Her AChR antibody level decreased to 38 nmol/L and her MG symptoms resolved aside from head drop which is continuing to improve. Her lymphoma is now in remission. We have presented a case with a successful outcome, which highlights the importance of screening for lymphoma and thymoma in new presentations of MG.
    Mesh-Begriff(e) Aged, 80 and over ; Autoantibodies ; Female ; Humans ; Lambert-Eaton Myasthenic Syndrome ; Lymphoma, Follicular/complications ; Lymphoma, Follicular/diagnosis ; Myasthenia Gravis/complications ; Myasthenia Gravis/diagnosis ; Thymus Neoplasms
    Chemische Substanzen Autoantibodies
    Sprache Englisch
    Erscheinungsdatum 2022-01-13
    Erscheinungsland England
    Dokumenttyp Case Reports ; Journal Article
    ISSN 1757-790X
    ISSN (online) 1757-790X
    DOI 10.1136/bcr-2021-248011
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  2. Artikel ; Online: Measurement of Homologous Recombination at Stalled Mammalian Replication Forks.

    Willis, Nicholas A / Scully, Ralph

    Methods in molecular biology (Clifton, N.J.)

    2020  Band 2153, Seite(n) 329–353

    Abstract: Site-specific replication fork barriers (RFBs) have proven valuable tools for studying mechanisms of repair at sites of replication fork stalling in prokaryotes and yeasts. We adapted the Escherichia coli Tus-Ter RFB for use in mammalian cells and used ... ...

    Abstract Site-specific replication fork barriers (RFBs) have proven valuable tools for studying mechanisms of repair at sites of replication fork stalling in prokaryotes and yeasts. We adapted the Escherichia coli Tus-Ter RFB for use in mammalian cells and used it to trigger site-specific replication fork stalling and homologous recombination (HR) at a defined chromosomal locus in mammalian cells. By comparing HR responses induced at the Tus-Ter RFB with those induced by a site-specific double-strand break (DSB), we have begun to uncover how the mechanisms of mammalian stalled fork repair differ from those underlying the repair of a replication-independent DSB. Here, we outline how to transiently express the Tus protein in mES cells, how to use flow cytometry to score conservative and aberrant repair outcomes, and how to quantify distinct repair outcomes in response to replication fork stalling at the inducible Tus-Ter chromosomal RFB.
    Mesh-Begriff(e) Animals ; Cells, Cultured ; DNA Breaks, Double-Stranded ; DNA Replication ; DNA-Binding Proteins/genetics ; DNA-Binding Proteins/metabolism ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Escherichia coli Proteins/genetics ; Escherichia coli Proteins/metabolism ; Flow Cytometry ; Homologous Recombination ; Mice ; Mouse Embryonic Stem Cells/chemistry ; Mouse Embryonic Stem Cells/cytology ; Transfection
    Chemische Substanzen DNA replication terminus site-binding protein, E coli ; DNA-Binding Proteins ; Escherichia coli Proteins ; tus protein, E coli
    Sprache Englisch
    Erscheinungsdatum 2020-08-24
    Erscheinungsland United States
    Dokumenttyp Comparative Study ; Journal Article
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-0716-0644-5_23
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  3. Artikel ; Online: Exploiting CRISPR/Cas9 to engineer precise segmental deletions in mouse embryonic stem cells.

    Elango, Rajula / Panday, Arvind / Willis, Nicholas A / Scully, Ralph

    STAR protocols

    2022  Band 3, Heft 3, Seite(n) 101551

    Abstract: In this protocol, we use CRISPR/Cas9 to generate large deletions of the entire coding region of a gene of interest, generating a hemizygous cell line. Next, we systematically engineer precise in-frame deletions within the intact wild-type allele, ... ...

    Abstract In this protocol, we use CRISPR/Cas9 to generate large deletions of the entire coding region of a gene of interest, generating a hemizygous cell line. Next, we systematically engineer precise in-frame deletions within the intact wild-type allele, facilitating study of multi-domain proteins. The optimized protocol described here allows us to rapidly screen for effective sgRNA pairs and to engineer either an in-frame deletion or a frameshift mutation in high frequencies in mouse embryonic stem cells. For complete details on the use and execution of this protocol, please refer to Panday et al. (2021).
    Mesh-Begriff(e) Animals ; CRISPR-Cas Systems/genetics ; Mice ; Mouse Embryonic Stem Cells ; Sequence Deletion
    Sprache Englisch
    Erscheinungsdatum 2022-08-19
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
    ISSN 2666-1667
    ISSN (online) 2666-1667
    DOI 10.1016/j.xpro.2022.101551
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  4. Artikel ; Online: A modified CUT&RUN-seq technique for qPCR analysis of chromatin-protein interactions.

    Panday, Arvind / Elango, Rajula / Willis, Nicholas A / Scully, Ralph

    STAR protocols

    2022  Band 3, Heft 3, Seite(n) 101529

    Abstract: Chromatin immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) even with optimization may give low signal-to-background ratio and spatial resolution. Here, we adapted Cleavage Under Targets and Release Using Nuclease (CUT&RUN) (originally ... ...

    Abstract Chromatin immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) even with optimization may give low signal-to-background ratio and spatial resolution. Here, we adapted Cleavage Under Targets and Release Using Nuclease (CUT&RUN) (originally developed by the Henikoff group) to develop CUT&RUN-qPCR. By studying the recruitment of selected proteins (but amenable to other proteins), we find that CUT&RUN-qPCR is more sensitive and gives better spatial resolution than ChIP-qPCR. For complete details on the use and execution of this protocol, please refer to Skene et al. (2018) and Skene and Henikoff (2017).
    Mesh-Begriff(e) Chromatin/genetics ; Chromatin Immunoprecipitation/methods ; Chromosomes/metabolism ; Endonucleases ; Micrococcal Nuclease/metabolism
    Chemische Substanzen Chromatin ; Endonucleases (EC 3.1.-) ; Micrococcal Nuclease (EC 3.1.31.1)
    Sprache Englisch
    Erscheinungsdatum 2022-07-31
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
    ISSN 2666-1667
    ISSN (online) 2666-1667
    DOI 10.1016/j.xpro.2022.101529
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  5. Buch ; Online: Cesium Tiles for High-realism Simulation and Comparing SLAM Results in Corresponding Virtual and Real-world Environments

    Beam, Chris / Zhang, Jincheng / Kakavitsas, Nicholas / Hague, Collin / Wolek, Artur / Willis, Andrew

    2024  

    Abstract: This article discusses the use of a simulated environment to predict algorithm results in the real world. Simulators are crucial in allowing researchers to test algorithms, sensor integration, and navigation systems without deploying expensive hardware. ... ...

    Abstract This article discusses the use of a simulated environment to predict algorithm results in the real world. Simulators are crucial in allowing researchers to test algorithms, sensor integration, and navigation systems without deploying expensive hardware. This article examines how the AirSim simulator, Unreal Engine, and Cesium plugin can be used to generate simulated digital twin models of real-world locations. Several technical challenges in completing the analysis are discussed and the technical solutions are detailed in this article. Work investigates how to assess mapping results for a real-life experiment using Cesium Tiles provided by digital twins of the experimental location. This is accompanied by a description of a process for duplicating real-world flights in simulation. The performance of these methods is evaluated by analyzing real-life and experimental image telemetry with the Direct Sparse Odometry (DSO) mapping algorithm. Results indicate that Cesium Tiles environments can provide highly accurate models of ground truth geometry after careful alignment. Further, results from real-life and simulated telemetry analysis indicate that the virtual simulation results accurately predict real-life results. Findings indicate that the algorithm results in real life and in the simulated duplicate exhibited a high degree of similarity. This indicates that the use of Cesium Tiles environments as a virtual digital twin for real-life experiments will provide representative results for such algorithms. The impact of this can be significant, potentially allowing expansive virtual testing of robotic systems at specific deployment locations to develop solutions that are tailored to the environment and potentially outperforming solutions meant to work in completely generic environments.
    Schlagwörter Computer Science - Robotics ; Computer Science - Computer Vision and Pattern Recognition
    Thema/Rubrik (Code) 629
    Erscheinungsdatum 2024-01-15
    Erscheinungsland us
    Dokumenttyp Buch ; Online
    Datenquelle BASE - Bielefeld Academic Search Engine (Lebenswissenschaftliche Auswahl)

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  6. Artikel ; Online: A modified CUT&RUN-seq technique for qPCR analysis of chromatin-protein interactions

    Arvind Panday / Rajula Elango / Nicholas A. Willis / Ralph Scully

    STAR Protocols, Vol 3, Iss 3, Pp 101529- (2022)

    2022  

    Abstract: Summary: Chromatin immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) even with optimization may give low signal-to-background ratio and spatial resolution. Here, we adapted Cleavage Under Targets and Release Using Nuclease (CUT&RUN) ( ... ...

    Abstract Summary: Chromatin immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) even with optimization may give low signal-to-background ratio and spatial resolution. Here, we adapted Cleavage Under Targets and Release Using Nuclease (CUT&RUN) (originally developed by the Henikoff group) to develop CUT&RUN-qPCR. By studying the recruitment of selected proteins (but amenable to other proteins), we find that CUT&RUN-qPCR is more sensitive and gives better spatial resolution than ChIP-qPCR.For complete details on the use and execution of this protocol, please refer to Skene et al. (2018) and Skene and Henikoff (2017). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
    Schlagwörter Cell Biology ; Cell-based Assays ; Molecular Biology ; Chromatin immunoprecipitation (ChIP) ; Science (General) ; Q1-390
    Sprache Englisch
    Erscheinungsdatum 2022-09-01T00:00:00Z
    Verlag Elsevier
    Dokumenttyp Artikel ; Online
    Datenquelle BASE - Bielefeld Academic Search Engine (Lebenswissenschaftliche Auswahl)

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  7. Artikel ; Online: Recombination and restart at blocked replication forks.

    Scully, Ralph / Elango, Rajula / Panday, Arvind / Willis, Nicholas A

    Current opinion in genetics & development

    2021  Band 71, Seite(n) 154–162

    Abstract: Replication fork stalling occurs when the replisome encounters a barrier to normal fork progression. Replisome stalling events are common during scheduled DNA synthesis, but vary in their severity. At one extreme, a lesion may induce only temporary ... ...

    Abstract Replication fork stalling occurs when the replisome encounters a barrier to normal fork progression. Replisome stalling events are common during scheduled DNA synthesis, but vary in their severity. At one extreme, a lesion may induce only temporary pausing of a DNA polymerase; at the other, it may present a near-absolute barrier to the replicative helicase and effectively block fork progression. Many alternative pathways have evolved to respond to these different types of replication stress. Among these, the homologous recombination (HR) pathway plays an important role, protecting the stalled fork and processing it for repair. Here, we review recent advances in our understanding of how blocked replication forks in vertebrate cells can be processed for recombination and for replication restart.
    Mesh-Begriff(e) Chromosomes ; DNA Helicases/genetics ; DNA Replication/genetics
    Chemische Substanzen DNA Helicases (EC 3.6.4.-)
    Sprache Englisch
    Erscheinungsdatum 2021-08-28
    Erscheinungsland England
    Dokumenttyp Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 1077312-5
    ISSN 1879-0380 ; 0959-437X
    ISSN (online) 1879-0380
    ISSN 0959-437X
    DOI 10.1016/j.gde.2021.08.003
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  8. Artikel: Two-ended recombination at a Flp-nickase-broken replication fork.

    Elango, Rajula / Nilavar, Namrata / Li, Andrew G / Duffey, Erin E / Jiang, Yuning / Nguyen, Daniel / Abakir, Abdulkadir / Willis, Nicholas A / Houseley, Jonathan / Scully, Ralph

    bioRxiv : the preprint server for biology

    2024  

    Abstract: Collision of a replication fork with a DNA nick is thought to generate a one-ended break, fostering genomic instability. Collision of the opposing converging fork with the nick could, in principle, form a second DNA end, enabling conservative repair by ... ...

    Abstract Collision of a replication fork with a DNA nick is thought to generate a one-ended break, fostering genomic instability. Collision of the opposing converging fork with the nick could, in principle, form a second DNA end, enabling conservative repair by homologous recombination (HR). To study mechanisms of nickase-induced HR, we developed the Flp recombinase "step arrest" nickase in mammalian cells. Flp-nickase-induced HR entails two-ended, BRCA2/RAD51-dependent short tract gene conversion (STGC), BRCA2/RAD51-independent long tract gene conversion, and discoordinated two-ended invasions. HR induced by a replication-independent break and by the Flp-nickase differ in their dependence on
    Sprache Englisch
    Erscheinungsdatum 2024-04-10
    Erscheinungsland United States
    Dokumenttyp Preprint
    DOI 10.1101/2024.04.10.588130
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  9. Artikel ; Online: Multi-institutional evaluation of a Pareto navigation guided automated radiotherapy planning solution for prostate cancer.

    Wheeler, Philip A / West, Nicholas S / Powis, Richard / Maggs, Rhydian / Chu, Michael / Pearson, Rachel A / Willis, Nick / Kurec, Bartlomiej / Reed, Katie L / Lewis, David G / Staffurth, John / Spezi, Emiliano / Millin, Anthony E

    Radiation oncology (London, England)

    2024  Band 19, Heft 1, Seite(n) 45

    Abstract: Background: Current automated planning solutions are calibrated using trial and error or machine learning on historical datasets. Neither method allows for the intuitive exploration of differing trade-off options during calibration, which may aid in ... ...

    Abstract Background: Current automated planning solutions are calibrated using trial and error or machine learning on historical datasets. Neither method allows for the intuitive exploration of differing trade-off options during calibration, which may aid in ensuring automated solutions align with clinical preference. Pareto navigation provides this functionality and offers a potential calibration alternative. The purpose of this study was to validate an automated radiotherapy planning solution with a novel multi-dimensional Pareto navigation calibration interface across two external institutions for prostate cancer.
    Methods: The implemented 'Pareto Guided Automated Planning' (PGAP) methodology was developed in RayStation using scripting and consisted of a Pareto navigation calibration interface built upon a 'Protocol Based Automatic Iterative Optimisation' planning framework. 30 previous patients were randomly selected by each institution (I
    Results: PGAP led to marked improvements across the majority of rectal dose metrics, with D
    Conclusions: PGAP enabled intuitive adaptation of automated protocols to an institution's planning aims and yielded plans more congruent with the institution's clinical preference than the locally produced manual clinical plans.
    Mesh-Begriff(e) Male ; Humans ; Radiotherapy, Intensity-Modulated/methods ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted/methods ; Urinary Bladder ; Prostatic Neoplasms/radiotherapy ; Organs at Risk
    Sprache Englisch
    Erscheinungsdatum 2024-04-08
    Erscheinungsland England
    Dokumenttyp Multicenter Study ; Journal Article
    ZDB-ID 2224965-5
    ISSN 1748-717X ; 1748-717X
    ISSN (online) 1748-717X
    ISSN 1748-717X
    DOI 10.1186/s13014-024-02404-x
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  10. Artikel ; Online: Humanization with CD34-positive hematopoietic stem cells in NOG-EXL mice results in improved long-term survival and less severe myeloid cell hyperactivation phenotype relative to NSG-SGM3 mice.

    Willis, Elinor / Verrelle, Jillian / Banerjee, Esha / Assenmacher, Charles-Antoine / Tarrant, James C / Skuli, Nicholas / Jacobson, Moriah L / O'Rouke, Donald M / Binder, Zev A / Radaelli, Enrico

    Veterinary pathology

    2024  , Seite(n) 3009858231222216

    Abstract: NSG-SGM3 and NOG-EXL mice combine severe immunodeficiency with transgenic expression of human myeloid stimulatory cytokines, resulting in marked expansion of myeloid populations upon humanization with CD34+ hematopoietic stem cells (HSCs). Humanized NSG- ... ...

    Abstract NSG-SGM3 and NOG-EXL mice combine severe immunodeficiency with transgenic expression of human myeloid stimulatory cytokines, resulting in marked expansion of myeloid populations upon humanization with CD34+ hematopoietic stem cells (HSCs). Humanized NSG-SGM3 mice typically develop a lethal macrophage activation syndrome and mast cell hyperplasia that limit their use in long-term studies (e.g., humanization followed by tumor xenotransplantation). It is currently unclear to what extent humanized NOG-EXL mice suffer from the same condition observed in humanized NSG-SGM3 mice. We compared the effects of human CD34+ HSC engraftment in these two strains in an orthotopic patient-derived glioblastoma model. NSG-SGM3 mice humanized in-house were compared to NOG-EXL mice humanized in-house and commercially available humanized NOG-EXL mice. Mice were euthanized at humane or study endpoints, and complete pathological assessments were performed. A semiquantitative multiparametric clinicopathological scoring system was developed to characterize chimeric myeloid cell hyperactivation (MCH) syndrome. NSG-SGM3 mice were euthanized at 16 weeks after humanization because of severe deterioration of clinical conditions. Humanized NOG-EXL mice survived to the study endpoint at 22 weeks after humanization and showed less-severe MCH phenotypes than NSG-SGM3 mice. Major differences included the lack of mast cell expansion and limited tissue/organ involvement in NOG-EXL mice compared to NSG-SGM3 mice. Engraftment of human lymphocytes, assessed by immunohistochemistry, was similar in the two strains. The longer survival and decreased MCH phenotype severity in NOG-EXL mice enabled their use in a tumor xenotransplantation study. The NOG-EXL model is better suited than the NSG-SGM3 model for immuno-oncology studies requiring long-term survival after humanization.
    Sprache Englisch
    Erscheinungsdatum 2024-01-10
    Erscheinungsland United States
    Dokumenttyp Journal Article
    ZDB-ID 188012-3
    ISSN 1544-2217 ; 0300-9858
    ISSN (online) 1544-2217
    ISSN 0300-9858
    DOI 10.1177/03009858231222216
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