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Article ; Online: Maintenance of Human Primordial Germ Cell-Like Cells in a Long-Term Culture System.

Gell, Joanna J / Shioda, Toshi

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

2023 Volume 2677, Page(s) 259–267

Abstract: Primordial germ cells (PGCs) are the earliest form of mammalian germ lineage. In humans, PGCs are present during a very early and limited window in development, limiting the ability to study fundamental developmental steps in human reproductive biology. ... ...

Abstract	Primordial germ cells (PGCs) are the earliest form of mammalian germ lineage. In humans, PGCs are present during a very early and limited window in development, limiting the ability to study fundamental developmental steps in human reproductive biology. However, recent advancements in generating in-vitro models of gametogenesis have allowed the field to generate human primordial germ cell-like cells (hPGCLCs). In this chapter, we will review the generation of hPGCLCs using the incipient mesoderm-like cell (iMeLC) protocol and the subsequent expansion of hPGCLCs in a long-term culture system.
MeSH term(s)	Animals ; Humans ; Cell Differentiation ; Induced Pluripotent Stem Cells ; Germ Cells ; Gametogenesis ; Mammals
Language	English
Publishing date	2023-07-18
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-3259-8_15
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Article: Transgenerational Transcriptomic and DNA Methylome Profiling of Mouse Fetal Testicular Germline and Somatic Cells after Exposure of Pregnant Mothers to Tributyltin, a Potent Obesogen.

Shioda, Keiko / Odajima, Junko / Blumberg, Bruce / Shioda, Toshi

Metabolites

2022 Volume 12, Issue 2

Abstract: Obesogens such as tributyltin (TBT) are xenobiotic compounds that promote obesity, in part by distorting the normal balance of lipid metabolism. The obesogenic effects of TBT can be observed in directly exposed (F1 and F2 generations) and also subsequent ...

Abstract	Obesogens such as tributyltin (TBT) are xenobiotic compounds that promote obesity, in part by distorting the normal balance of lipid metabolism. The obesogenic effects of TBT can be observed in directly exposed (F1 and F2 generations) and also subsequent generations (F3 and beyond) that were never exposed. To address the effects of TBT exposure on germ cells, we exposed pregnant transgenic OG2 mouse dams (F0), which specifically express EGFP in germline cells, to an environmentally relevant dose of TBT or DMSO throughout gestation through drinking water. When fed with a high-fat diet, F3 male offspring of TBT-exposed F0 dams (TBT-F3) accumulated much more body fat than did DMSO-F3 males. TBT-F3 males also lost more body fluid and lean compositions than did DMSO-F3 males. Expression of genes involved in transcriptional regulation or mesenchymal differentiation was up-regulated in somatic cells of TBT-F1 (but not TBT-F3) E18.5 fetal testes, and promoter-associated CpG islands were hyper-methylated in TBT-F1 somatic cells. Global mRNA expression of protein-coding genes in F1 or F3 fetal testicular cells was unaffected by F0 exposure to TBT; however, expression of a subset of endogenous retroviruses was significantly affected in F1 and F3. We infer that TBT may directly target testicular somatic cells in F1 testes to irreversibly affect epigenetic suppression of endogenous retroviruses in both germline and somatic cells.
Language	English
Publishing date	2022-01-20
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2662251-8
ISSN	2218-1989
ISSN	2218-1989
DOI	10.3390/metabo12020095
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Article ; Online: miR-223 Plays a Key Role in Obesogen-Enhanced Adipogenesis in Mesenchymal Stem Cells and in Transgenerational Obesity.

Chang, Richard C / Joloya, Erika M / Li, Zhuorui / Shoucri, Bassem M / Shioda, Toshi / Blumberg, Bruce

Endocrinology

2023 Volume 164, Issue 5

Abstract: Exposure of pregnant F0 mouse dams to the obesogen tributyltin (TBT) predisposes unexposed male descendants to obesity and diverts mesenchymal stem cells (MSCs) toward the adipocytic lineage. TBT promotes adipogenic commitment and differentiation of MSCs ...

Abstract	Exposure of pregnant F0 mouse dams to the obesogen tributyltin (TBT) predisposes unexposed male descendants to obesity and diverts mesenchymal stem cells (MSCs) toward the adipocytic lineage. TBT promotes adipogenic commitment and differentiation of MSCs in vitro. To identify TBT-induced factors predisposing MSCs toward the adipocytic fate, we exposed mouse MSCs to TBT, the peroxisome proliferator activated receptor gamma (PPARγ)-selective agonist rosiglitazone, or the retinoid X receptor (RXR)-selective agonist LG-100268. Then we determined their transcriptomal profiles to determine candidate microRNAs (miR) regulating adipogenic commitment and differentiation. Of the top 10 candidate microRNAs predicted by Ingenuity Pathway Analysis, miR-21, miR-33, and miR-223 were expressed consistent with an ability to differentially regulate target genes during adipogenesis. We found that 24-hour exposure to 50nM TBT caused miR-223 levels in MSCs to increase; expression of its target genes ZEB1, NFIB, and FOXP1 was decreased. Rosiglitazone and TBT increased miR-223 levels. This induction was inhibited by the PPARγ antagonist T0070907 but not by the RXR antagonists HX531 or UVI3003, placing miR-223 downstream of PPARγ. Chromatin immunoprecipitation confirmed TBT-induced binding of PPARγ to regulatory elements in the miR-223 promoter. miR-223 levels were elevated in white adipose tissue of F2 and F3 male descendants of pregnant F0 mouse dams exposed to 50nM TBT throughout gestation. miR-223 levels were potentiated in males fed an increased fat diet. We infer that TBT induced miR-223 expression and increased adipogenesis in MSCs through the PPARγ pathway and that transgenerationally increased expression of miR-223 plays an important role in the development of obesity caused by TBT exposure.
MeSH term(s)	Female ; Pregnancy ; Male ; Animals ; Mice ; Adipogenesis/genetics ; Rosiglitazone/pharmacology ; PPAR gamma/metabolism ; Mesenchymal Stem Cells ; Cell Differentiation/genetics ; Obesity/genetics ; Obesity/metabolism ; MicroRNAs/genetics ; MicroRNAs/metabolism
Chemical Substances	Rosiglitazone (05V02F2KDG) ; PPAR gamma ; tributyltin (4XDX163P3D) ; MicroRNAs ; MIRN223 microRNA, mouse ; Mirn33 microRNA, mouse
Language	English
Publishing date	2023-02-06
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	427856-2
ISSN	1945-7170 ; 0013-7227
ISSN (online)	1945-7170
ISSN	0013-7227
DOI	10.1210/endocr/bqad027
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Article: Evolutionarily diverse mechanisms of germline specification among mammals: what about us?

Mitsunaga, Shino / Shioda, Toshi

Stem cell investigation

2018 Volume 5, Page(s) 12

Language	English
Publishing date	2018-04-24
Publishing country	China
Document type	Editorial ; Comment
ZDB-ID	2884645-X
ISSN	2313-0792 ; 2306-9759
ISSN (online)	2313-0792
ISSN	2306-9759
DOI	10.21037/sci.2018.04.03
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Article: The Crowded Uterine Horn Mouse Model for Examining Postnatal Metabolic Consequences of Intrauterine Growth Restriction vs. Macrosomia in Siblings.

Taylor, Julia A / Coe, Benjamin L / Shioda, Toshi / Vom Saal, Frederick S

Metabolites

2022 Volume 12, Issue 2

Abstract: Differential placental blood flow and nutrient transport can lead to both intrauterine growth restriction (IUGR) and macrosomia. Both conditions can lead to adult obesity and other conditions clustered as metabolic syndrome. We previously showed that ... ...

Abstract	Differential placental blood flow and nutrient transport can lead to both intrauterine growth restriction (IUGR) and macrosomia. Both conditions can lead to adult obesity and other conditions clustered as metabolic syndrome. We previously showed that pregnant hemi-ovariectomized mice have a crowded uterine horn, resulting in siblings whose birth weights differ by over 100% due to differential blood flow based on uterine position. We used this crowded uterus model to compare IUGR and macrosomic male mice and also identified IUGR males with rapid (IUGR-R) and low (IUGR-L) postweaning weight gain. At week 12 IUGR-R males were heavier than IUGR-L males and did not differ from macrosomic males. Rapid growth in IUGR-R males led to glucose intolerance compared to IUGR-L males and down-regulation of adipocyte signaling pathways for fat digestion and absorption and type II diabetes. Macrosomia led to increased fat mass and altered adipocyte size distribution compared to IUGR males, and down-regulation of signaling pathways for carbohydrate and fat digestion and absorption relative to IUGR-R. Clustering analysis of gonadal fat transcriptomes indicated more similarities than differences between IUGR-R and macrosomic males compared to IUGR-L males. Our findings suggest two pathways to adult metabolic disease: macrosomia and IUGR with rapid postweaning growth rate.
Language	English
Publishing date	2022-01-22
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2662251-8
ISSN	2218-1989
ISSN	2218-1989
DOI	10.3390/metabo12020102
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Article: Epigenetic Transgenerational Inheritance of the Effects of Obesogen Exposure.

Mohajer, Nicole / Joloya, Erika M / Seo, Jeongbin / Shioda, Toshi / Blumberg, Bruce

Frontiers in endocrinology

2021 Volume 12, Page(s) 787580

Abstract: Obesity and metabolic disorders have become a worldwide pandemic affecting millions of people. Although obesity is a multifaceted disease, there is growing evidence supporting the obesogen hypothesis, which proposes that exposure to a subset of endocrine ...

Abstract	Obesity and metabolic disorders have become a worldwide pandemic affecting millions of people. Although obesity is a multifaceted disease, there is growing evidence supporting the obesogen hypothesis, which proposes that exposure to a subset of endocrine disrupting chemicals (EDCs), known as obesogens, promotes obesity. While these effects can be observed
MeSH term(s)	Animals ; Chromatin/chemistry ; DNA Methylation ; Endocrine Disruptors/adverse effects ; Environmental Exposure/adverse effects ; Epigenesis, Genetic/genetics ; Female ; Genetic Predisposition to Disease ; Histones/metabolism ; Humans ; Inheritance Patterns/genetics ; Male ; Methylation ; Obesity/chemically induced ; Obesity/epidemiology ; Obesity/genetics ; Phenotype
Chemical Substances	Chromatin ; Endocrine Disruptors ; Histones
Language	English
Publishing date	2021-12-16
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Review
ZDB-ID	2592084-4
ISSN	1664-2392
ISSN	1664-2392
DOI	10.3389/fendo.2021.787580
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Article ; Online: Sex Differences in Embryonic Gonad Transcriptomes and Benzo[a]pyrene Metabolite Levels After Transplacental Exposure.

Lim, Jinhwan / Ramesh, Aramandla / Shioda, Toshi / Leon Parada, Kathleen / Luderer, Ulrike

Endocrinology

2021 Volume 163, Issue 1

Abstract: Polycyclic aromatic hydrocarbons like benzo[a]pyrene (BaP) are generated during incomplete combustion of organic materials. Prior research has demonstrated that BaP is a prenatal ovarian toxicant and carcinogen. However, the metabolic pathways active in ... ...

Abstract	Polycyclic aromatic hydrocarbons like benzo[a]pyrene (BaP) are generated during incomplete combustion of organic materials. Prior research has demonstrated that BaP is a prenatal ovarian toxicant and carcinogen. However, the metabolic pathways active in the embryo and its developing gonads and the mechanisms by which prenatal exposure to BaP predisposes to ovarian tumors later in life remain to be fully elucidated. To address these data gaps, we orally dosed pregnant female mice with BaP from embryonic day (E) 6.5 to E11.5 (0, 0.2, or 2 mg/kg/day) for metabolite measurement or E9.5 to E11.5 (0 or 3.33 mg/kg/day) for embryonic gonad RNA sequencing. Embryos were harvested at E13.5 for both experiments. The sum of BaP metabolite concentrations increased significantly with dose in the embryos and placentas, and concentrations were significantly higher in female than male embryos and in embryos than placentas. RNA sequencing revealed that enzymes involved in metabolic activation of BaP are expressed at moderate to high levels in embryonic gonads and that greater transcriptomic changes occurred in the ovaries in response to BaP than in the testes. We identified 490 differentially expressed genes (DEGs) with false discovery rate P-values < 0.05 when comparing BaP-exposed to control ovaries but no statistically significant DEGs between BaP-exposed and control testes. Genes related to monocyte/macrophage recruitment and activity, prolactin family genes, and several keratin genes were among the most upregulated genes in the BaP-exposed ovaries. Results show that developing ovaries are more sensitive than testes to prenatal BaP exposure, which may be related to higher concentrations of BaP metabolites in female embryos.
MeSH term(s)	Animals ; Benzo(a)pyrene/metabolism ; Chromatography, High Pressure Liquid ; Computational Biology ; Female ; Gonads/metabolism ; Inflammation ; Keratins/metabolism ; Macrophages/metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Monocytes/metabolism ; Ovary/metabolism ; Placenta/metabolism ; Pregnancy ; Pregnancy, Animal ; RNA-Seq ; Sex Factors ; Testis/metabolism ; Time Factors ; Transcriptome
Chemical Substances	Benzo(a)pyrene (3417WMA06D) ; Keratins (68238-35-7)
Language	English
Publishing date	2021-11-03
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	427856-2
ISSN	1945-7170 ; 0013-7227
ISSN (online)	1945-7170
ISSN	0013-7227
DOI	10.1210/endocr/bqab228
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Article ; Online: Technical adequacy of bisulfite sequencing and pyrosequencing for detection of mitochondrial DNA methylation: Sources and avoidance of false-positive detection.

Owa, Chie / Poulin, Matthew / Yan, Liying / Shioda, Toshi

PloS one

2018 Volume 13, Issue 2, Page(s) e0192722

Abstract: The existence of cytosine methylation in mammalian mitochondrial DNA (mtDNA) is a controversial subject. Because detection of DNA methylation depends on resistance of 5'-modified cytosines to bisulfite-catalyzed conversion to uracil, examined parameters ... ...

Abstract	The existence of cytosine methylation in mammalian mitochondrial DNA (mtDNA) is a controversial subject. Because detection of DNA methylation depends on resistance of 5'-modified cytosines to bisulfite-catalyzed conversion to uracil, examined parameters that affect technical adequacy of mtDNA methylation analysis. Negative control amplicons (NCAs) devoid of cytosine methylation were amplified to cover the entire human or mouse mtDNA by long-range PCR. When the pyrosequencing template amplicons were gel-purified after bisulfite conversion, bisulfite pyrosequencing of NCAs did not detect significant levels of bisulfite-resistant cytosines (brCs) at ND1 (7 CpG sites) or CYTB (8 CpG sites) genes (CI95 = 0%-0.94%); without gel-purification, significant false-positive brCs were detected from NCAs (CI95 = 4.2%-6.8%). Bisulfite pyrosequencing of highly purified, linearized mtDNA isolated from human iPS cells or mouse liver detected significant brCs (~30%) in human ND1 gene when the sequencing primer was not selective in bisulfite-converted and unconverted templates. However, repeated experiments using a sequencing primer selective in bisulfite-converted templates almost completely (< 0.8%) suppressed brC detection, supporting the false-positive nature of brCs detected using the non-selective primer. Bisulfite-seq deep sequencing of linearized, gel-purified human mtDNA detected 9.4%-14.8% brCs for 9 CpG sites in ND1 gene. However, because all these brCs were associated with adjacent non-CpG brCs showing the same degrees of bisulfite resistance, DNA methylation in this mtDNA-encoded gene was not confirmed. Without linearization, data generated by bisulfite pyrosequencing or deep sequencing of purified mtDNA templates did not pass the quality control criteria. Shotgun bisulfite sequencing of human mtDNA detected extremely low levels of CpG methylation (<0.65%) over non-CpG methylation (<0.55%). Taken together, our study demonstrates that adequacy of mtDNA methylation analysis using methods dependent on bisulfite conversion needs to be established for each experiment, taking effects of incomplete bisulfite conversion and template impurity or topology into consideration.
MeSH term(s)	Animals ; CpG Islands ; Cytosine/metabolism ; DNA Methylation ; DNA, Mitochondrial/genetics ; DNA, Mitochondrial/metabolism ; Humans ; Mice ; Sequence Analysis, DNA ; Sulfites/chemistry
Chemical Substances	DNA, Mitochondrial ; Sulfites ; Cytosine (8J337D1HZY) ; hydrogen sulfite (OJ9787WBLU)
Language	English
Publishing date	2018-02-08
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0192722
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Article ; Online: RARγ is required for mesodermal gene expression prior to gastrulation in

Janesick, Amanda / Tang, Weiyi / Shioda, Toshi / Blumberg, Bruce

Development (Cambridge, England)

2018 Volume 145, Issue 18

Abstract: The developing vertebrate embryo is exquisitely sensitive to retinoic acid (RA) concentration, particularly during anteroposterior patterning. In contrast to Nodal and Wnt signaling, RA was not previously considered to be an instructive signal in ... ...

Abstract	The developing vertebrate embryo is exquisitely sensitive to retinoic acid (RA) concentration, particularly during anteroposterior patterning. In contrast to Nodal and Wnt signaling, RA was not previously considered to be an instructive signal in mesoderm formation during gastrulation. Here, we show in
MeSH term(s)	Aldehyde Dehydrogenase 1 Family ; Aldehyde Oxidase/biosynthesis ; Aldehyde Oxidase/genetics ; Animals ; Body Patterning/genetics ; CD56 Antigen/metabolism ; Cell Adhesion/genetics ; Gastrulation/genetics ; Mesoderm/embryology ; Muscle, Skeletal/embryology ; MyoD Protein/metabolism ; Receptors, Retinoic Acid/genetics ; Receptors, Retinoic Acid/metabolism ; Retinal Dehydrogenase ; Retinoic Acid 4-Hydroxylase/biosynthesis ; Retinoic Acid 4-Hydroxylase/genetics ; Signal Transduction/genetics ; Transcriptional Activation/genetics ; Tretinoin/metabolism ; Xenopus Proteins/biosynthesis ; Xenopus Proteins/genetics ; Xenopus laevis/embryology ; Xenopus laevis/genetics ; Retinoic Acid Receptor gamma
Chemical Substances	CD56 Antigen ; MyoD Protein ; Receptors, Retinoic Acid ; Xenopus Proteins ; Tretinoin (5688UTC01R) ; Retinoic Acid 4-Hydroxylase (EC 1.14.14.1) ; Aldehyde Dehydrogenase 1 Family (EC 1.2.1) ; Retinal Dehydrogenase (EC 1.2.1.36) ; aldh1a2 protein, Xenopus (EC 1.2.1.36) ; Aldehyde Oxidase (EC 1.2.3.1)
Language	English
Publishing date	2018-09-17
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	90607-4
ISSN	1477-9129 ; 0950-1991
ISSN (online)	1477-9129
ISSN	0950-1991
DOI	10.1242/dev.147769
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Article ; Online: Transgenerational Self-Reconstruction of Disrupted Chromatin Organization After Exposure To An Environmental Stressor in Mice.

Diaz-Castillo, Carlos / Chamorro-Garcia, Raquel / Shioda, Toshi / Blumberg, Bruce

Scientific reports

2019 Volume 9, Issue 1, Page(s) 13057

Abstract: Exposure to environmental stressors is known to increase disease susceptibility in unexposed descendants in the absence of detectable genetic mutations. The mechanisms mediating environmentally-induced transgenerational disease susceptibility are poorly ... ...

Abstract	Exposure to environmental stressors is known to increase disease susceptibility in unexposed descendants in the absence of detectable genetic mutations. The mechanisms mediating environmentally-induced transgenerational disease susceptibility are poorly understood. We showed that great-great-grandsons of female mice exposed to tributyltin (TBT) throughout pregnancy and lactation were predisposed to obesity due to altered chromatin organization that subsequently biased DNA methylation and gene expression. Here we analyzed DNA methylomes and transcriptomes from tissues of animals ancestrally exposed to TBT spanning generations, sexes, ontogeny, and cell differentiation state. We found that TBT elicited concerted alterations in the expression of "chromatin organization" genes and inferred that TBT-disrupted chromatin organization might be able to self-reconstruct transgenerationally. We also found that the location of "chromatin organization" and "metabolic" genes is biased similarly in mouse and human genomes, suggesting that exposure to environmental stressors in different species could elicit similar phenotypic effects via self-reconstruction of disrupted chromatin organization.
MeSH term(s)	Animals ; Chromatin/genetics ; DNA Methylation ; Environmental Exposure ; Epigenesis, Genetic ; Gene Expression Profiling ; Gene-Environment Interaction ; Humans ; Mice ; Miosis/genetics ; Mitosis/genetics ; Obesity/genetics ; Stress, Physiological ; Transcriptome
Chemical Substances	Chromatin
Language	English
Publishing date	2019-09-10
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-019-49440-2
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