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Article ; Online: Selective isoxazolopyrimidine PAT1 (SLC26A6) inhibitors for therapy of intestinal disorders.

Chu, Tifany / Karmakar, Joy / Haggie, Peter M / Tan, Joseph-Anthony / Master, Riya / Ramaswamy, Keerthana / Verkman, Alan S / Anderson, Marc O / Cil, Onur

RSC medicinal chemistry

2023 Volume 14, Issue 11, Page(s) 2342–2347

Abstract: A loss of prosecretory ... ...

Abstract	A loss of prosecretory Cl
Language	English
Publishing date	2023-09-14
Publishing country	England
Document type	Journal Article
ISSN	2632-8682
ISSN (online)	2632-8682
DOI	10.1039/d3md00302g
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Article ; Online: Small molecule inhibitors of intestinal epithelial anion exchanger SLC26A3 (DRA) with a luminal, extracellular site of action.

Cil, Onur / Anderson, Marc O / de Souza Goncalves, Livia / Tan, Joseph-Anthony / Haggie, Peter M / Verkman, Alan S

European journal of medicinal chemistry

2023 Volume 249, Page(s) 115149

Abstract: The anion exchanger protein SLC26A3 (down-regulated in adenoma, DRA) is expressed in the luminal membrane of intestinal epithelial cells in colon, where it facilitates the absorption of ... ...

Abstract	The anion exchanger protein SLC26A3 (down-regulated in adenoma, DRA) is expressed in the luminal membrane of intestinal epithelial cells in colon, where it facilitates the absorption of Cl
MeSH term(s)	Mice ; Animals ; Antiporters/chemistry ; Antiporters/metabolism ; Antiporters/pharmacology ; Molecular Docking Simulation ; Biological Transport ; Constipation ; Anions ; Chlorides/metabolism ; Sulfate Transporters/metabolism
Chemical Substances	Antiporters ; Anions ; Chlorides ; Slc26a3 protein, mouse ; Sulfate Transporters
Language	English
Publishing date	2023-01-27
Publishing country	France
Document type	Journal Article
ZDB-ID	188597-2
ISSN	1768-3254 ; 0009-4374 ; 0223-5234
ISSN (online)	1768-3254
ISSN	0009-4374 ; 0223-5234
DOI	10.1016/j.ejmech.2023.115149
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Article ; Online: Small-molecule inhibitor of intestinal anion exchanger SLC26A3 for treatment of hyperoxaluria and nephrolithiasis.

Cil, Onur / Chu, Tifany / Lee, Sujin / Haggie, Peter M / Verkman, Alan S

JCI insight

2022 Volume 7, Issue 13

Abstract: Nephrolithiasis is a common and recurrent disease affecting 9% of the US population. Hyperoxaluria is major risk factor for calcium oxalate kidney stones, which constitute two-thirds of all kidney stones. SLC26A3 (DRA, downregulated in adenoma) is an ... ...

Abstract	Nephrolithiasis is a common and recurrent disease affecting 9% of the US population. Hyperoxaluria is major risk factor for calcium oxalate kidney stones, which constitute two-thirds of all kidney stones. SLC26A3 (DRA, downregulated in adenoma) is an anion exchanger of chloride, bicarbonate, and oxalate thought to facilitate intestinal oxalate absorption, as evidenced by approximately 70% reduced urine oxalate excretion in knockout mice. We previously identified a small-molecule SLC26A3 inhibitor (DRAinh-A270) that selectively inhibited SLC26A3-mediated chloride/bicarbonate exchange (IC50 ~ 35 nM) and, as found here, oxalate/chloride exchange (IC50 ~ 60 nM). In colonic closed loops in mice, luminal DRAinh-A270 inhibited oxalate absorption by 70%. Following oral sodium oxalate loading in mice, DRAinh-A270 largely prevented the 2.5-fold increase in urine oxalate/creatinine ratio. In a mouse model of oxalate nephropathy produced by a high-oxalate low-calcium diet, vehicle-treated mice developed marked hyperoxaluria with elevated serum creatinine, renal calcium oxalate crystal deposition, and renal injury, which were largely prevented by DRAinh-A270 (10 mg/kg twice daily). DRAinh-A270 administered over 7 days to healthy mice did not show significant toxicity. Our findings support a major role of SLC26A3 in intestinal oxalate absorption and suggest the therapeutic utility of SLC26A3 inhibition for treatment of hyperoxaluria and prevention of calcium oxalate nephrolithiasis.
MeSH term(s)	Animals ; Antiporters ; Bicarbonates ; Calcium Oxalate ; Chlorides/metabolism ; Hyperoxaluria/drug therapy ; Hyperoxaluria/etiology ; Kidney Calculi/complications ; Kidney Calculi/drug therapy ; Kidney Calculi/prevention & control ; Mice ; Oxalates ; Sulfate Transporters
Chemical Substances	Antiporters ; Bicarbonates ; Chlorides ; Oxalates ; Slc26a3 protein, mouse ; Sulfate Transporters ; Calcium Oxalate (2612HC57YE)
Language	English
Publishing date	2022-07-08
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ISSN	2379-3708
ISSN (online)	2379-3708
DOI	10.1172/jci.insight.153359
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Article ; Online: SLC26A6-selective inhibitor identified in a small-molecule screen blocks fluid absorption in small intestine.

Cil, Onur / Haggie, Peter M / Tan, Joseph-Anthony Tapia / Rivera, Amber A / Verkman, Alan S

JCI insight

2021 Volume 6, Issue 11

Abstract: SLC26A6 (also known as putative anion transporter 1 [PAT1]) is a Cl-/HCO3- exchanger expressed at the luminal membrane of enterocytes where it facilitates intestinal Cl- and fluid absorption. Here, high-throughput screening of 50,000 synthetic small ... ...

Abstract	SLC26A6 (also known as putative anion transporter 1 [PAT1]) is a Cl-/HCO3- exchanger expressed at the luminal membrane of enterocytes where it facilitates intestinal Cl- and fluid absorption. Here, high-throughput screening of 50,000 synthetic small molecules in cells expressing PAT1 and a halide-sensing fluorescent protein identified several classes of inhibitors. The most potent compound, the pyrazolo-pyrido-pyrimidinone PAT1inh-B01, fully inhibited PAT1-mediated anion exchange (IC50 ~350 nM), without inhibition of the related intestinal transporter SLC26A3 (also known as DRA). In closed midjejunal loops in mice, PAT1inh-B01 inhibited fluid absorption by 50%, which increased to >90% when coadministered with DRA inhibitor DRAinh-A270. In ileal loops, PAT1inh-B01 blocked fluid absorption by >80%, whereas DRAinh-A270 was without effect. In colonic loops, PAT1inh-B01 was without effect, whereas DRAinh-A270 completely blocked fluid absorption. In a loperamide constipation model, coadministration of PAT1inh-B01 with DRAinh-A270 increased stool output compared with DRAinh-A270 alone. These results provide functional evidence for complementary and region-specific roles of PAT1 and DRA in intestinal fluid absorption, with PAT1 as the predominant anion exchanger in mouse ileum. We believe that PAT1inh-B01 is a novel tool to study intestinal ion and fluid transport and perhaps a drug candidate for small intestinal hyposecretory disorders such as cystic fibrosis-related meconium ileus and distal intestinal obstruction syndrome.
MeSH term(s)	Animals ; Antidiarrheals/pharmacology ; Antiporters/antagonists & inhibitors ; Antiporters/metabolism ; Colon/drug effects ; Colon/metabolism ; Constipation/chemically induced ; Constipation/metabolism ; Dopamine Plasma Membrane Transport Proteins/drug effects ; Drug Evaluation, Preclinical ; HEK293 Cells ; Humans ; Ileum/drug effects ; Ileum/metabolism ; Intestinal Absorption/drug effects ; Intestine, Small/drug effects ; Intestine, Small/metabolism ; Jejunum/drug effects ; Jejunum/metabolism ; Loperamide/pharmacology ; Mice ; Small Molecule Libraries ; Sulfate Transporters/antagonists & inhibitors ; Sulfate Transporters/metabolism
Chemical Substances	Antidiarrheals ; Antiporters ; Dopamine Plasma Membrane Transport Proteins ; Slc26a6 protein, mouse ; Small Molecule Libraries ; Sulfate Transporters ; Loperamide (6X9OC3H4II)
Language	English
Publishing date	2021-06-08
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ISSN	2379-3708
ISSN (online)	2379-3708
DOI	10.1172/jci.insight.147699
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Article ; Online: Small-molecule inhibitor of intestinal anion exchanger SLC26A3 for treatment of hyperoxaluria and nephrolithiasis

Onur Cil / Tifany Chu / Sujin Lee / Peter M. Haggie / Alan S. Verkman

JCI Insight, Vol 7, Iss

2022 Volume 13

Abstract	Nephrolithiasis is a common and recurrent disease affecting 9% of the US population. Hyperoxaluria is major risk factor for calcium oxalate kidney stones, which constitute two-thirds of all kidney stones. SLC26A3 (DRA, downregulated in adenoma) is an anion exchanger of chloride, bicarbonate, and oxalate thought to facilitate intestinal oxalate absorption, as evidenced by approximately 70% reduced urine oxalate excretion in knockout mice. We previously identified a small-molecule SLC26A3 inhibitor (DRAinh-A270) that selectively inhibited SLC26A3-mediated chloride/bicarbonate exchange (IC50 ~ 35 nM) and, as found here, oxalate/chloride exchange (IC50 ~ 60 nM). In colonic closed loops in mice, luminal DRAinh-A270 inhibited oxalate absorption by 70%. Following oral sodium oxalate loading in mice, DRAinh-A270 largely prevented the 2.5-fold increase in urine oxalate/creatinine ratio. In a mouse model of oxalate nephropathy produced by a high-oxalate low-calcium diet, vehicle-treated mice developed marked hyperoxaluria with elevated serum creatinine, renal calcium oxalate crystal deposition, and renal injury, which were largely prevented by DRAinh-A270 (10 mg/kg twice daily). DRAinh-A270 administered over 7 days to healthy mice did not show significant toxicity. Our findings support a major role of SLC26A3 in intestinal oxalate absorption and suggest the therapeutic utility of SLC26A3 inhibition for treatment of hyperoxaluria and prevention of calcium oxalate nephrolithiasis.
Keywords	Nephrology ; Therapeutics ; Medicine ; R
Language	English
Publishing date	2022-07-01T00:00:00Z
Publisher	American Society for Clinical investigation
Document type	Article ; Online
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Article ; Online: 4,8-Dimethylcoumarin Inhibitors of Intestinal Anion Exchanger slc26a3 (Downregulated in Adenoma) for Anti-Absorptive Therapy of Constipation.

Lee, Sujin / Cil, Onur / Haggie, Peter M / Verkman, Alan S

Journal of medicinal chemistry

2019 Volume 62, Issue 17, Page(s) 8330–8337

Abstract: The chloride/bicarbonate exchanger SLC26A3 (downregulated in adenoma) is expressed mainly in colonic epithelium, where it dehydrates the stool by facilitating the final step of chloride and fluid absorption. SLC26A3 inhibition has predicted efficacy in ... ...

Abstract	The chloride/bicarbonate exchanger SLC26A3 (downregulated in adenoma) is expressed mainly in colonic epithelium, where it dehydrates the stool by facilitating the final step of chloride and fluid absorption. SLC26A3 inhibition has predicted efficacy in various types of constipation including that associated with cystic fibrosis. We previously identified, by high-throughput screening, 4,8-dimethylcoumarin inhibitors of murine slc26a3 with IC
MeSH term(s)	Animals ; Antiporters/antagonists & inhibitors ; Antiporters/chemical synthesis ; Antiporters/chemistry ; Antiporters/pharmacology ; Constipation/chemically induced ; Constipation/drug therapy ; Coumarins/chemical synthesis ; Coumarins/chemistry ; Coumarins/pharmacology ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Down-Regulation/drug effects ; Female ; Intestinal Mucosa/drug effects ; Intestinal Mucosa/metabolism ; Loperamide ; Mice ; Molecular Structure ; Structure-Activity Relationship ; Sulfate Transporters/antagonists & inhibitors
Chemical Substances	4,8-dimethylcoumarin ; Antiporters ; Coumarins ; Slc26a3 protein, mouse ; Sulfate Transporters ; Loperamide (6X9OC3H4II)
Language	English
Publishing date	2019-09-03
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	218133-2
ISSN	1520-4804 ; 0022-2623
ISSN (online)	1520-4804
ISSN	0022-2623
DOI	10.1021/acs.jmedchem.9b01192
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Article ; Online: A simple method to generate human airway epithelial organoids with externally orientated apical membranes.

Boecking, Carolin A / Walentek, Peter / Zlock, Lorna T / Sun, Dingyuan I / Wolters, Paul J / Ishikawa, Hiroaki / Jin, Byung-Ju / Haggie, Peter M / Marshall, Wallace F / Verkman, Alan S / Finkbeiner, Walter E

American journal of physiology. Lung cellular and molecular physiology

2022 Volume 322, Issue 3, Page(s) L420–L437

Abstract: Organoids, which are self-organizing three-dimensional cultures, provide models that replicate specific cellular components of native tissues or facets of organ complexity. We describe a simple method to generate organoid cultures using isolated human ... ...

Abstract	Organoids, which are self-organizing three-dimensional cultures, provide models that replicate specific cellular components of native tissues or facets of organ complexity. We describe a simple method to generate organoid cultures using isolated human tracheobronchial epithelial cells grown in mixed matrix components and supplemented at
MeSH term(s)	Bronchi ; Cell Differentiation ; Cells, Cultured ; Epithelial Cells/metabolism ; Humans ; Organoids/metabolism ; Respiratory Mucosa/metabolism
Language	English
Publishing date	2022-01-26
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1013184-x
ISSN	1522-1504 ; 1040-0605
ISSN (online)	1522-1504
ISSN	1040-0605
DOI	10.1152/ajplung.00536.2020
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Article ; Online: Correction: X-Box Binding Protein 1 (XBP1s) Is a Critical Determinant of Pseudomonas aeruginosa Homoserine Lactone-Mediated Apoptosis.

Valentine, Cathleen D / Anderson, Marc O / Papa, Feroz R / Haggie, Peter M

PLoS pathogens

2016 Volume 12, Issue 5, Page(s) e1005628

Abstract: This corrects the article DOI: 10.1371/journal.ppat.1003576.]. ...

Abstract	[This corrects the article DOI: 10.1371/journal.ppat.1003576.].
Language	English
Publishing date	2016-05
Publishing country	United States
Document type	Published Erratum
ZDB-ID	2205412-1
ISSN	1553-7374 ; 1553-7366
ISSN (online)	1553-7374
ISSN	1553-7366
DOI	10.1371/journal.ppat.1005628
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Article ; Online: Confinement of β(1)- and β(2)-adrenergic receptors in the plasma membrane of cardiomyocyte-like H9c2 cells is mediated by selective interactions with PDZ domain and A-kinase anchoring proteins but not caveolae.

Valentine, Cathleen D / Haggie, Peter M

Molecular biology of the cell

2011 Volume 22, Issue 16, Page(s) 2970–2982

Abstract: The sympathetic nervous system regulates cardiac output by activating adrenergic receptors (ARs) in cardiac myocytes. The predominant cardiac ARs, β(1)- and β(2)AR, are structurally similar but mediate distinct signaling responses. Scaffold protein- ... ...

Abstract	The sympathetic nervous system regulates cardiac output by activating adrenergic receptors (ARs) in cardiac myocytes. The predominant cardiac ARs, β(1)- and β(2)AR, are structurally similar but mediate distinct signaling responses. Scaffold protein-mediated compartmentalization of ARs into discrete, multiprotein complexes has been proposed to dictate differential signaling responses. To test the hypothesis that βARs integrate into complexes in live cells, we measured receptor diffusion and interactions by single-particle tracking. Unstimulated β(1)- and β(2)AR were highly confined in the membrane of H9c2 cardiomyocyte-like cells, indicating that receptors are tethered and presumably integrated into protein complexes. Selective disruption of interactions with postsynaptic density protein 95/disks large/zonula occludens-1 (PDZ)-domain proteins and A-kinase anchoring proteins (AKAPs) increased receptor diffusion, indicating that these scaffold proteins participate in receptor confinement. In contrast, modulation of interactions between the putative scaffold caveolae and β(2)AR did not alter receptor dynamics, suggesting that these membrane domains are not involved in β(2)AR confinement. For both β(1)- and β(2)AR, the receptor carboxy-terminus was uniquely responsible for scaffold interactions. Our data formally demonstrate that distinct and stable protein complexes containing β(1)- or β(2)AR are formed in the plasma membrane of cardiomyocyte-like cells and that selective PDZ and AKAP interactions are responsible for the integration of receptors into complexes.
MeSH term(s)	A Kinase Anchor Proteins/metabolism ; Actin Cytoskeleton/metabolism ; Actins/metabolism ; Animals ; Caveolae/metabolism ; Cell Line ; Chlorocebus aethiops ; Fluorescence Recovery After Photobleaching ; Humans ; Microscopy, Fluorescence ; Myocytes, Cardiac/metabolism ; PDZ Domains ; Peptide Fragments/metabolism ; Protein Binding ; Rats ; Receptors, Adrenergic, beta-1/chemistry ; Receptors, Adrenergic, beta-1/genetics ; Receptors, Adrenergic, beta-1/metabolism ; Receptors, Adrenergic, beta-2/chemistry ; Receptors, Adrenergic, beta-2/genetics ; Receptors, Adrenergic, beta-2/metabolism ; Single-Cell Analysis ; Time-Lapse Imaging
Chemical Substances	A Kinase Anchor Proteins ; Actins ; Peptide Fragments ; Receptors, Adrenergic, beta-1 ; Receptors, Adrenergic, beta-2
Language	English
Publishing date	2011-06-16
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1098979-1
ISSN	1939-4586 ; 1059-1524
ISSN (online)	1939-4586
ISSN	1059-1524
DOI	10.1091/mbc.E11-01-0034
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Article ; Online: Nanomolar-potency 'co-potentiator' therapy for cystic fibrosis caused by a defined subset of minimal function CFTR mutants.

Phuan, Puay-Wah / Tan, Joseph-Anthony / Rivera, Amber A / Zlock, Lorna / Nielson, Dennis W / Finkbeiner, Walter E / Haggie, Peter M / Verkman, Alan S

Scientific reports

2019 Volume 9, Issue 1, Page(s) 17640

Abstract: Available CFTR modulators provide no therapeutic benefit for cystic fibrosis (CF) caused by many loss-of-function mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel, including N1303K. We previously introduced the ...

Abstract	Available CFTR modulators provide no therapeutic benefit for cystic fibrosis (CF) caused by many loss-of-function mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel, including N1303K. We previously introduced the concept of 'co-potentiators' (combination-potentiators) to rescue CFTR function in some minimal function CFTR mutants. Herein, a screen of ~120,000 drug-like synthetic small molecules identified active co-potentiators of pyrazoloquinoline, piperidine-pyridoindole, tetrahydroquinoline and phenylazepine classes, with EC
MeSH term(s)	Cystic Fibrosis/drug therapy ; Cystic Fibrosis/genetics ; Cystic Fibrosis/metabolism ; Cystic Fibrosis Transmembrane Conductance Regulator/genetics ; Cystic Fibrosis Transmembrane Conductance Regulator/metabolism ; Drug Discovery ; Drug Synergism ; High-Throughput Screening Assays ; Humans ; Mutation ; Piperidines/therapeutic use ; Pyrazoles/therapeutic use ; Structure-Activity Relationship
Chemical Substances	CFTR protein, human ; Piperidines ; Pyrazoles ; Cystic Fibrosis Transmembrane Conductance Regulator (126880-72-6)
Language	English
Publishing date	2019-11-27
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-019-54158-2
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