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Article: Elusive physiological role of prostatic acid phosphatase (PAP): generation of choline for sperm motility via auto-and paracrine cholinergic signaling.

Hanley, Peter J

2023 Volume 14, Page(s) 1327769

Abstract: Prostatic acid phosphatase (PAP) exists as two splice variants, secreted PAP and transmembrane PAP, the latter of which is implicated in antinociceptive signaling in dorsal root ganglia. However, PAP is predominantly expressed in the prostate gland and ... ...

Abstract	Prostatic acid phosphatase (PAP) exists as two splice variants, secreted PAP and transmembrane PAP, the latter of which is implicated in antinociceptive signaling in dorsal root ganglia. However, PAP is predominantly expressed in the prostate gland and the physiological role of seminal PAP, first identified in 1938, is largely unknown. Here, the author proposes that PAP, following ejaculation, functions to hydrolyze phosphocholine (PC) in seminal fluid and generate choline, which is imported by sperm via a choline transporter and converted to acetylcholine (ACh) by choline acetyltransferase. Auto- and paracrine cholinergic signaling, or choline directly, may subsequently stimulate sperm motility via α7 nicotinic ACh receptors (nAChRs) and contractility of the female reproductive tract through muscarinic ACh receptors (mAChRs). Consistent with a role of PAP in cholinergic signaling, 1) seminal vesicles secrete PC, 2) the prostate gland secretes PAP, 3) PAP specifically catalyzes the hydrolysis of PC into inorganic phosphate and choline, 4) seminal choline levels increase post-ejaculation, 5) pharmacological inhibition of choline acetyltransferase inhibits sperm motility, 6) inhibition or genetic deletion of α7 nAChRs impairs sperm motility, and 7) mAChRs are expressed in the uterus and oviduct (fallopian tube). Notably, PAP does not degrade glycerophosphocholine (GPC), the predominant choline source in the semen of rats and other mammals. Instead, uterine GPC phosphodiesterases may liberate choline from seminal GPC. In summary, the author deduces that PAP in humans, and uterine GPC phosphodiesterases in other mammals, function to generate choline for sperm cholinergic signaling, which promotes sperm motility and possibly contractility of the female reproductive tract.
Language	English
Publishing date	2023-12-22
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2564217-0
ISSN	1664-042X
ISSN	1664-042X
DOI	10.3389/fphys.2023.1327769
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Article: Class IX Myosins: Motorized RhoGAP Signaling Molecules.

Hanley, Peter J / Vollmer, Veith / Bähler, Martin

Advances in experimental medicine and biology

2020 Volume 1239, Page(s) 381–389

Abstract: Class IX myosins are simultaneously motor and signaling molecules. In addition to myosin class-specific functions of the tail region, they feature unique motor properties. Within their motor region they contain a long insertion with a calmodulin- and a F- ...

Abstract	Class IX myosins are simultaneously motor and signaling molecules. In addition to myosin class-specific functions of the tail region, they feature unique motor properties. Within their motor region they contain a long insertion with a calmodulin- and a F-actin-binding site. The rate-limiting step in the ATPase cycle is ATP hydrolysis rather than, typical for other myosins, the release of either product. This means that class IX myosins spend a large fraction of their cycle time in the ATP-bound state, which is typically a low F-actin affinity state. Nevertheless, class IX myosins in the ATP-bound state stochastically switch between a low and a high F-actin affinity state. Single motor domains even show characteristics of processive movement towards the plus end of actin filaments. The insertion thereby acts as an actin tether. The motor domain transports as intramolecular cargo a signaling Rho GTPase-activating protein domain located in the tail region. Rho GTPase-activating proteins catalyze the conversion of active GTP-bound Rho to inactive GDP-bound Rho by stimulating GTP hydrolysis. In cells, Rho activity regulates actin cytoskeleton organization and actomyosin II contractility. Thus, class IX myosins regulate cell morphology, cell migration, cell-cell junctions and membrane trafficking. These cellular functions affect embryonic development, adult organ homeostasis and immune responses. Human diseases associated with mutations in the two class IX myosins, Myo9a and Myo9b, have been identified, including hydrocephalus and congenital myasthenic syndrome in connection with Myo9a and autoimmune diseases in connection with Myo9b.
MeSH term(s)	Actins/metabolism ; GTPase-Activating Proteins/metabolism ; Humans ; Myosins/metabolism ; Protein Binding ; Signal Transduction
Chemical Substances	Actins ; GTPase-Activating Proteins ; MYO9A protein, human ; myosin IXB ; rho GTPase-activating protein ; Myosins (EC 3.6.4.1)
Language	English
Publishing date	2020-05-25
Publishing country	United States
Document type	Journal Article ; Review
ISSN	2214-8019 ; 0065-2598
ISSN (online)	2214-8019
ISSN	0065-2598
DOI	10.1007/978-3-030-38062-5_16
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Article ; Online: Time-lapse Imaging of Mouse Macrophage Chemotaxis.

van den Bos, Esther / Walbaum, Stefan / Horsthemke, Markus / Bachg, Anne C / Hanley, Peter J

Journal of visualized experiments : JoVE

2020 , Issue 158

Abstract: Chemotaxis is receptor-mediated guidance of cells along a chemical gradient, whereas chemokinesis is the stimulation of random cell motility by a chemical. Chemokinesis and chemotaxis are fundamental for the mobilization and deployment of immune cells. ... ...

Abstract	Chemotaxis is receptor-mediated guidance of cells along a chemical gradient, whereas chemokinesis is the stimulation of random cell motility by a chemical. Chemokinesis and chemotaxis are fundamental for the mobilization and deployment of immune cells. For example, chemokines (chemotactic cytokines) can rapidly recruit circulating neutrophils and monocytes to extravascular sites of inflammation. Chemoattractant receptors belong to the large family of G protein-coupled receptors. How chemoattractant (i.e., ligand) gradients direct cell migration via G protein-coupled receptor signaling is not yet fully understood. In the field of immunology, neutrophils are popular model cells for studying chemotaxis in vitro. Here we describe a real-time two-dimensional (2D) chemotaxis assay tailored for mouse resident macrophages, which have traditionally been more difficult to study. Macrophages move at a slow pace of ~1 µm/min on a 2D surface and are less well suited for point-source migration assays (e.g., migration towards the tip of a micropipette filled with chemoattractant) than neutrophils or Dictyostelium discoideum, which move an order of magnitude faster. Widely used Transwell assays are useful for studying the chemotactic activity of different substances, but do not provide information on cell morphology, velocity, or chemotactic navigation. Here we describe a time-lapse microscopy-based macrophage chemotaxis assay that allows quantification of cell velocity and chemotactic efficiency and provides a platform to delineate the transducers, signal pathways, and effectors of chemotaxis.
MeSH term(s)	Animals ; Chemotaxis ; Dictyostelium/cytology ; Macrophages/cytology ; Macrophages/metabolism ; Mice ; Monocytes/cytology ; Neutrophils/cytology ; Receptors, G-Protein-Coupled/metabolism ; Signal Transduction ; Time-Lapse Imaging/methods
Chemical Substances	Receptors, G-Protein-Coupled
Language	English
Publishing date	2020-04-02
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Video-Audio Media
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/60750
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Article ; Online: Time-lapse 3D Imaging of Phagocytosis by Mouse Macrophages.

Horsthemke, Markus / Wilden, Janine / Bachg, Anne C / Hanley, Peter J

Journal of visualized experiments : JoVE

2018 , Issue 140

Abstract: Phagocytosis plays a key role in host defense, as well as in tissue development and maintenance, and involves rapid, receptor-mediated rearrangements of the actin cytoskeleton to capture, envelop and engulf large particles. Although phagocytic receptors, ...

Abstract	Phagocytosis plays a key role in host defense, as well as in tissue development and maintenance, and involves rapid, receptor-mediated rearrangements of the actin cytoskeleton to capture, envelop and engulf large particles. Although phagocytic receptors, downstream signaling pathways, and effectors, such as Rho GTPases, have been identified, the dynamic cytoskeletal remodeling of specific receptor-mediated phagocytic events remain unclear. Four decades ago, two distinct mechanisms of phagocytosis, exemplified by Fcγ receptor (FcγR)- and complement receptor (CR)-mediated phagocytosis, were identified using scanning electron microscopy. Binding of immunoglobulin G (IgG)-opsonized particles to FcγRs triggers the protrusion of thin membrane extensions, which initially form a so-called phagocytic cup around the particle before it becomes completely enclosed and retracted into the cell. In contrast, complement opsonized particles appear to sink into the phagocyte following binding to complement receptors. These two modes of phagocytosis, phagocytic cup formation and sinking in, have become well established in the literature. However, the distinctions between the two modes have become blurred by reports that complement receptor-mediated phagocytosis may induce various membrane protrusions. With the availability of high resolution imaging techniques, phagocytosis assays are required that allow real-time 3D (three dimensional) visualization of how specific phagocytic receptors mediate the uptake of individual particles. More commonly used approaches for the study of phagocytosis, such as end-point assays, miss the opportunity to understand what is happening at the interface of particles and phagocytes. Here we describe phagocytic assays, using time-lapse spinning disk confocal microscopy, that allow 3D imaging of single phagocytic events. In addition, we describe assays to unambiguously image Fcγ receptor- or complement receptor-mediated phagocytosis.
MeSH term(s)	Animals ; Imaging, Three-Dimensional/methods ; Macrophages/metabolism ; Mice ; Microscopy, Confocal/methods ; Phagocytes/physiology
Language	English
Publishing date	2018-10-19
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Video-Audio Media
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/57566
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Article ; Online: Complement receptor 3 mediates both sinking phagocytosis and phagocytic cup formation via distinct mechanisms.

Walbaum, Stefan / Ambrosy, Benjamin / Schütz, Paula / Bachg, Anne C / Horsthemke, Markus / Leusen, Jeanette H W / Mócsai, Attila / Hanley, Peter J

The Journal of biological chemistry

2021 Volume 296, Page(s) 100256

Abstract: A long-standing hypothesis is that complement receptors (CRs), especially CR3, mediate sinking phagocytosis, but evidence is lacking. Alternatively, CRs have been reported to induce membrane ruffles or phagocytic cups, akin to those induced by Fcγ ... ...

Abstract	A long-standing hypothesis is that complement receptors (CRs), especially CR3, mediate sinking phagocytosis, but evidence is lacking. Alternatively, CRs have been reported to induce membrane ruffles or phagocytic cups, akin to those induced by Fcγ receptors (FcγRs), but the details of these events are unclear. Here we used real-time 3D imaging and KO mouse models to clarify how particles (human red blood cells) are internalized by resident peritoneal F4/80
MeSH term(s)	Animals ; Cell Membrane/metabolism ; Cells, Cultured ; Humans ; Immunoreceptor Tyrosine-Based Activation Motif ; Macrophage-1 Antigen/metabolism ; Macrophages/metabolism ; Mice ; Mice, Knockout ; Phagocytosis ; Pseudopodia/metabolism ; Signal Transduction ; Syk Kinase/metabolism
Chemical Substances	Macrophage-1 Antigen ; SYK protein, human (EC 2.7.10.2) ; Syk Kinase (EC 2.7.10.2)
Language	English
Publishing date	2021-01-08
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1016/j.jbc.2021.100256
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Article ; Online: Complement receptor 3 mediates both sinking phagocytosis and phagocytic cup formation via distinct mechanisms.

Walbaum, Stefan / Ambrosy, Benjamin / Schütz, Paula / Bachg, Anne C / Horsthemke, Markus / Leusen, Jeanette H W / Mócsai, Attila / Hanley, Peter J

The Journal of biological chemistry

2021

Abstract	A long-standing hypothesis is that complement receptors (CRs), especially CR3, mediate sinking phagocytosis, but evidence is lacking. Alternatively, CRs have been reported to induce membrane ruffles or phagocytic cups, akin to those induced by Fcγ receptors (FcγRs), but the details of these events are unclear. Here we used real-time 3D imaging and knockout mouse models to clarify how particles (human red blood cells) are internalized by resident peritoneal F4/80
Language	English
Publishing date	2021-01-04
Publishing country	United States
Document type	Journal Article
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1074/jbc.RA120.015346
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Article: Time-lapse imaging of mouse macrophage chemotaxis

van den Bos, Esther / Walbaum, Stefan / Horsthemke, Markus / Bachg, Anne C / Hanley, Peter J

Journal of visualized experiments. 2020 Apr. 02, , no. 158

2020

Abstract	Chemotaxis is receptor-mediated guidance of cells along a chemical gradient, whereas chemokinesis is the stimulation of random cell motility by a chemical. Chemokinesis and chemotaxis are fundamental for the mobilization and deployment of immune cells. For example, chemokines (chemotactic cytokines) can rapidly recruit circulating neutrophils and monocytes to extravascular sites of inflammation. Chemoattractant receptors belong to the large family of G protein-coupled receptors. How chemoattractant (i.e., ligand) gradients direct cell migration via G protein-coupled receptor signaling is not yet fully understood. In the field of immunology, neutrophils are popular model cells for studying chemotaxis in vitro. Here we describe a real-time two-dimensional (2D) chemotaxis assay tailored for mouse resident macrophages, which have traditionally been more difficult to study. Macrophages move at a slow pace of ~1 µm/min on a 2D surface and are less well suited for point-source migration assays (e.g., migration towards the tip of a micropipette filled with chemoattractant) than neutrophils or Dictyostelium discoideum, which move an order of magnitude faster. Widely used Transwell assays are useful for studying the chemotactic activity of different substances, but do not provide information on cell morphology, velocity, or chemotactic navigation. Here we describe a time-lapse microscopy-based macrophage chemotaxis assay that allows quantification of cell velocity and chemotactic efficiency and provides a platform to delineate the transducers, signal pathways, and effectors of chemotaxis.
Keywords	Dictyostelium discoideum ; G-protein coupled receptors ; cell movement ; cell structures ; chemoattractants ; chemokines ; chemotaxis ; image analysis ; inflammation ; ligands ; macrophages ; mice ; models ; monocytes ; neutrophils ; signal transduction
Language	English
Dates of publication	2020-0402
Size	p. e60750.
Publishing place	Journal of Visualized Experiments
Document type	Article
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/60750
Database	NAL-Catalogue (AGRICOLA)

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Article: Time-lapse 3d imaging of phagocytosis by mouse macrophages

Horsthemke, Markus / Wilden, Janine / Bachg, Anne C / Hanley, Peter J

Journal of visualized experiments. 2018 Oct. 19, , no. 140

2018

Abstract	Phagocytosis plays a key role in host defense, as well as in tissue development and maintenance, and involves rapid, receptor-mediated rearrangements of the actin cytoskeleton to capture, envelop and engulf large particles. Although phagocytic receptors, downstream signaling pathways, and effectors, such as Rho GTPases, have been identified, the dynamic cytoskeletal remodeling of specific receptor-mediated phagocytic events remain unclear. Four decades ago, two distinct mechanisms of phagocytosis, exemplified by Fcγ receptor (FcγR)- and complement receptor (CR)-mediated phagocytosis, were identified using scanning electron microscopy. Binding of immunoglobulin G (IgG)-opsonized particles to FcγRs triggers the protrusion of thin membrane extensions, which initially form a so-called phagocytic cup around the particle before it becomes completely enclosed and retracted into the cell. In contrast, complement opsonized particles appear to sink into the phagocyte following binding to complement receptors. These two modes of phagocytosis, phagocytic cup formation and sinking in, have become well established in the literature. However, the distinctions between the two modes have become blurred by reports that complement receptor-mediated phagocytosis may induce various membrane protrusions. With the availability of high resolution imaging techniques, phagocytosis assays are required that allow real-time 3D (three dimensional) visualization of how specific phagocytic receptors mediate the uptake of individual particles. More commonly used approaches for the study of phagocytosis, such as end-point assays, miss the opportunity to understand what is happening at the interface of particles and phagocytes. Here we describe phagocytic assays, using time-lapse spinning disk confocal microscopy, that allow 3D imaging of single phagocytic events. In addition, we describe assays to unambiguously image Fcγ receptor- or complement receptor-mediated phagocytosis.
Keywords	complement ; guanosinetriphosphatase ; image analysis ; immunoglobulin G ; macrophages ; mice ; microfilaments ; phagocytosis ; receptors ; scanning electron microscopy ; signal transduction ; spinning disk confocal microscopy
Language	English
Dates of publication	2018-1019
Size	p. e57566.
Publishing place	Journal of Visualized Experiments
Document type	Article
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/57566
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Low density lipoprotein interferes with intracellular signaling of monocytes resulting in impaired chemotaxis and enhanced chemokinesis.

Tjaden, Kerstin / Adam, Christina / Godfrey, Rinesh / Hanley, Peter J / Pardali, Evangelia / Waltenberger, Johannes

International journal of cardiology

2018 Volume 255, Page(s) 160–165

Abstract: Background: Hypercholesterolemia (HC) is an important cardiovascular risk factor characterized by elevated low density lipoprotein-cholesterol (LDL-C) plasma levels. HC negatively affects monocyte function by reducing their chemotactic response towards ... ...

Abstract	Background: Hypercholesterolemia (HC) is an important cardiovascular risk factor characterized by elevated low density lipoprotein-cholesterol (LDL-C) plasma levels. HC negatively affects monocyte function by reducing their chemotactic response towards different growth factors. We aimed to elucidate the molecular mechanisms by which LDL induces monocyte dysfunction. Methods and results: Human monocytes exposed to either native (nLDL) or oxidized LDL (oxLDL) in vitro showed reduced chemotactic responses towards vascular endothelial growth factor A (VEGFA) and monocyte chemotactic protein-1 (MCP-1), but displayed enhanced random migration (chemokinesis). Mechanistically, the exposure to LDL resulted in the activation of p38 mitogen-activated protein kinase (MAPK) and modulated MCP-1 and VEGFA-induced signaling in human monocytes. Furthermore, the aberrant p38 activation induced by oxLDL is due to the functional impairment of Dual Specificity Phosphatase-1 (DUSP-1). In the absence of LDL, the pharmacological inhibition of DUSP-1 alone was sufficient to recapitulate the accelerated chemokinetic and blunted chemotactic phenotype of monocytes. Finally, p38 MAPK inhibition in monocytes isolated from hyperlipidemic mice prevented the aberrant chemokinetic phenotype. Conclusions: Our data demonstrate that LDL induces monocyte chemokinesis of human monocytes by inducing mononuclear cell activation through the aberrant modulation of DUSP-1-p38/MAPK signaling axis. Moreover, our findings suggest that MCP-1/VEGFA-induced chemotaxis is reduced by LDL secondary to the impairment of ligand-induced signaling. These findings provide novel insight into hypercholesterolemia-associated vascular dysfunction and its potential involvement in the pathogenesis of atherosclerosis.
MeSH term(s)	Animals ; Cells, Cultured ; Chemokines/metabolism ; Chemotaxis/drug effects ; Chemotaxis/physiology ; Humans ; Intracellular Fluid/drug effects ; Intracellular Fluid/metabolism ; Lipoproteins, LDL/toxicity ; Mice ; Mice, Knockout ; Monocytes/drug effects ; Monocytes/metabolism ; Signal Transduction/drug effects ; Signal Transduction/physiology
Chemical Substances	Chemokines ; Lipoproteins, LDL ; oxidized low density lipoprotein
Language	English
Publishing date	2018-02-09
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	779519-1
ISSN	1874-1754 ; 0167-5273
ISSN (online)	1874-1754
ISSN	0167-5273
DOI	10.1016/j.ijcard.2017.11.109
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Article: Treatment of tinnitus with a customized, dynamic acoustic neural stimulus: underlying principles and clinical efficacy.

Hanley, Peter J / Davis, Paul B

Trends in amplification

2008 Volume 12, Issue 3, Page(s) 210–222

Abstract: Tinnitus has been challenging to treat with consistently positive results. The Neuromonics Tinnitus Treatment is a newly available approach to the treatment of clinically significant, problematic tinnitus (and reduced sound tolerance) that was developed ... ...

Abstract	Tinnitus has been challenging to treat with consistently positive results. The Neuromonics Tinnitus Treatment is a newly available approach to the treatment of clinically significant, problematic tinnitus (and reduced sound tolerance) that was developed with the intention of simultaneously addressing the auditory, attentional, and emotional processes underlying the condition. It uses a prescribed acoustic stimulus, customized for each patient's individual audiometric profile, which provides a broad frequency stimulus to address the effects of auditory deprivation, promotes relief and relaxation with the intention of reducing engagement of the limbic system/amygdala and autonomic nervous system, and applies the principles of systematic desensitization to address the attentional processes. This article describes the underlying principles behind this approach. It also summarizes evidence for clinical efficacy from controlled clinical studies and from a private practice clinical setting, where it has been shown to provide consistently positive outcomes for patients meeting suitability criteria.
MeSH term(s)	Acoustic Stimulation/methods ; Cochlear Nerve/physiology ; Humans ; Tinnitus/therapy ; Treatment Outcome
Language	English
Publishing date	2008-07-09
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2213216-8
ISSN	1940-5588 ; 1084-7138
ISSN (online)	1940-5588
ISSN	1084-7138
DOI	10.1177/1084713808319942
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