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Article ; Online: Micromanipulation in Paramecium: From non-mendelian inheritance to the outlook for versatile micromachines.

Haga, Nobuyuki

2022 Volume 69, Issue 5, Page(s) e12909

Abstract: This review addresses nine areas of knowledge revealed by micromanipulations performed with Paramecium. Microinjection has shown that sexual maturation and senescence of Paramecium caudatum is a programmed process conducted by a specific gene and its ... ...

Abstract	This review addresses nine areas of knowledge revealed by micromanipulations performed with Paramecium. Microinjection has shown that sexual maturation and senescence of Paramecium caudatum is a programmed process conducted by a specific gene and its product protein. In Paramecium tetraurelia, autogamy was revealed to depend on the number of DNA syntheses rather than the number of cell divisions in clonal aging. The cytoplasmic complementarity test established that microinjection of wild-type cytoplasm can correct genetic defects of mutants. The concept of complementarity together with protein chemistry revealed compounds that control membrane excitability. In non-Mendelian inheritance, noncoding small RNAs made from the parental micronucleus regulate the rearrangement of the progeny's macronuclear DNA. The macronucleus has the potential to be used as a factory for genetic engineering. The development and differentiation of progeny's nuclei in mating pairs are controlled by the parental macronucleus. The chemical reaction processes associated with exocytosis have been revealed by microinjection of various enzymes and antibodies. Using the fusion gene of histone H2B and yellow-fluorescence protein, it was revealed that the fusion gene-mRNA is transferred between cells during mating. Experiments with endosymbiotic bacteria and the host shed light on the conditions needed to establish sustainable symbiotic relationships.
MeSH term(s)	Cytoplasm ; Macronucleus/genetics ; Micromanipulation ; Paramecium/physiology ; Paramecium tetraurelia/genetics
Language	English
Publishing date	2022-05-04
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	1147218-2
ISSN	1550-7408 ; 1066-5234
ISSN (online)	1550-7408
ISSN	1066-5234
DOI	10.1111/jeu.12909
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Article: Micromanipulation in Paramecium: From non‐mendelian inheritance to the outlook for versatile micromachines

Haga, Nobuyuki

journal of eukaryotic microbiology. 2022 Sept., v. 69, no. 5

2022

Abstract	This review addresses nine areas of knowledge revealed by micromanipulations performed with Paramecium. Microinjection has shown that sexual maturation and senescence of Paramecium caudatum is a programmed process conducted by a specific gene and its product protein. In Paramecium tetraurelia, autogamy was revealed to depend on the number of DNA syntheses rather than the number of cell divisions in clonal aging. The cytoplasmic complementarity test established that microinjection of wild‐type cytoplasm can correct genetic defects of mutants. The concept of complementarity together with protein chemistry revealed compounds that control membrane excitability. In non‐Mendelian inheritance, noncoding small RNAs made from the parental micronucleus regulate the rearrangement of the progeny's macronuclear DNA. The macronucleus has the potential to be used as a factory for genetic engineering. The development and differentiation of progeny's nuclei in mating pairs are controlled by the parental macronucleus. The chemical reaction processes associated with exocytosis have been revealed by microinjection of various enzymes and antibodies. Using the fusion gene of histone H2B and yellow‐fluorescence protein, it was revealed that the fusion gene‐mRNA is transferred between cells during mating. Experiments with endosymbiotic bacteria and the host shed light on the conditions needed to establish sustainable symbiotic relationships.
Keywords	DNA ; Paramecium caudatum ; Paramecium tetraurelia ; autogamy ; chemical reactions ; cytoplasm ; exocytosis ; genes ; histones ; microbiology ; progeny ; sexual maturity
Language	English
Dates of publication	2022-09
Publishing place	John Wiley & Sons, Ltd
Document type	Article
Note	REVIEW
ZDB-ID	1147218-2
ISSN	1550-7408 ; 1066-5234
ISSN (online)	1550-7408
ISSN	1066-5234
DOI	10.1111/jeu.12909
Database	NAL-Catalogue (AGRICOLA)

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Article: Identification and Characterization of the Gene Responsible for the O

Chiba, Yuta / Takenaka, Yasuhiro / Haga, Nobuyuki

Microorganisms

2024 Volume 12, Issue 3

Abstract: The process of sexual reproduction in eukaryotes starts when gametes from two different sexes encounter each other. ...

Abstract	The process of sexual reproduction in eukaryotes starts when gametes from two different sexes encounter each other.
Language	English
Publishing date	2024-03-15
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2720891-6
ISSN	2076-2607
ISSN	2076-2607
DOI	10.3390/microorganisms12030588
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Article: Immaturin-Nuclease as a Model System for a Gene-Programmed Sexual Development and Rejuvenescence in

Haga, Nobuyuki / Usui, Toshinori / Takenaka, Yasuhiro / Chiba, Yuta / Abe, Tomoaki

Microorganisms

2022 Volume 11, Issue 1

Abstract: Fertilization-initiated development and adult-onset aging are standard features in the life history of eukaryotes. ... ...

Abstract	Fertilization-initiated development and adult-onset aging are standard features in the life history of eukaryotes. In
Language	English
Publishing date	2022-12-28
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2720891-6
ISSN	2076-2607
ISSN	2076-2607
DOI	10.3390/microorganisms11010082
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Article ; Online: In memoriam: Koichi Hiwatashi (1921-2009).

Haga, Nobuyuki

The Journal of eukaryotic microbiology

2009 Volume 56, Issue 5, Page(s) 492–493

MeSH term(s)	Animals ; Eukaryota/genetics ; Eukaryota/physiology ; History, 20th Century ; History, 21st Century ; Humans ; Male ; Parasitology/history
Language	English
Publishing date	2009-09
Publishing country	United States
Document type	Biography ; Historical Article ; Journal Article
ZDB-ID	1147218-2
ISSN	1550-7408 ; 1066-5234
ISSN (online)	1550-7408
ISSN	1066-5234
DOI	10.1111/j.1550-7408.2009.00426.x
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Article: Studies of Paramecium caudatum by means of scanning electron microscope and projection X-ray microscope.

Yada, Keiji / Abe, Taiki / Haga, Nobuyuki

Bio-medical materials and engineering

2009 Volume 19, Issue 2-3, Page(s) 87–92

Abstract: Samples of Paramecium caudatum are observed by means of a scanning electron microscope (SEM) and a projection X-ray microscope (XRM) with computer tomography (CT) function. The samples are fixed with two kinds of fixatives, glutaraldehyde and osmium- ... ...

Abstract	Samples of Paramecium caudatum are observed by means of a scanning electron microscope (SEM) and a projection X-ray microscope (XRM) with computer tomography (CT) function. The samples are fixed with two kinds of fixatives, glutaraldehyde and osmium-tetra oxide acid. After the fixation and replacement procedure with t-buthyl alcohol, the samples followed by a freeze drying, well retain their structures. Surface structures, cilia and microfibrillar systems including infraciliary lattice structures, are clearly depicted by SEM observation. On the other hand, XRM images give quite different information, namely, in the case of osmium oxide fixation, the structures of internal organelles like the macronucleus placed in the central part of cell body and trichocysts located under the cell membrane of a whole body are visible. In the case of glutaraldehyde fixation, the surface structures and internal structures are both visible but their image contrast is fairly weak. In order to examine toxicological effect, Paramecium caudatum samples treated in the environmental condition containing nano-particles of Ag (17 nm across) and Co-ferrite (300 nm across) are observed with results of certain morphological differences, namely, inner vacuoles increase in number and in volume in Co-ferrite treated cells as compared with Ag treated ones. But then, cilia-less areas increase on the surface of the body of Ag treated cells. In the case of Co-ferrite treated cells, cilia-less areas are not clearly detected. Whether these morphological differences observed in Ag and Co-ferrite treated cells are caused by the differences of materials or particle sizes remain to be examined in future.
MeSH term(s)	Animals ; Image Enhancement/methods ; Microscopy/methods ; Microscopy, Electron, Scanning/methods ; Paramecium/ultrastructure ; Preservation, Biological/methods ; X-Rays
Language	English
Publishing date	2009
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	1066216-9
ISSN	0959-2989
ISSN	0959-2989
DOI	10.3233/BME-2009-0567
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Article ; Online: Identification of two nickel ion-induced genes, NCI16 and PcGST1, in Paramecium caudatum.

Takenaka, Yasuhiro / Haga, Nobuyuki / Inoue, Ikuo / Nakano, Takanari / Ikeda, Masaaki / Katayama, Shigehiro / Awata, Takuya

Eukaryotic cell

2014 Volume 13, Issue 9, Page(s) 1181–1190

Abstract: Here, we describe the isolation of two nickel-induced genes in Paramecium caudatum, NCI16 and PcGST1, by subtractive hybridization. NCI16 encoded a predicted four-transmembrane domain protein (∼16 kDa) of unknown function, and PcGST1 encoded glutathione ... ...

Abstract	Here, we describe the isolation of two nickel-induced genes in Paramecium caudatum, NCI16 and PcGST1, by subtractive hybridization. NCI16 encoded a predicted four-transmembrane domain protein (∼16 kDa) of unknown function, and PcGST1 encoded glutathione S-transferase (GST; ∼25 kDa) with GST and glutathione peroxidase (GPx) activities. Exposing cells to cobalt chloride also caused the moderate upregulation of NCI16 and PcGST1 mRNAs. Both nickel sulfate and cobalt chloride dose dependently induced NCI16 and PcGST1 mRNAs, but with different profiles. Nickel treatment caused a continuous increase in PcGST1 and NCI16 mRNA levels for up to 3 and 6 days, respectively, and a notable increase in H₂O₂ concentrations in P. caudatum. NCI16 expression was significantly enhanced by incubating cells with H₂O₂, implying that NCI16 induction in the presence of nickel ions is caused by reactive oxygen species (ROS). On the other hand, PcGST1 was highly induced by the antioxidant tert-butylhydroquinone (tBHQ) but not by H2O2, suggesting that different mechanisms mediate the induction of NCI16 and PcGST1. We introduced a luciferase reporter vector with an ∼0.42-kb putative PcGST1 promoter into cells and then exposed the transformants to nickel sulfate. This resulted in significant luciferase upregulation, indicating that the putative PcGST1 promoter contains a nickel-responsive element. Our nickel-inducible system also may be applicable to the efficient expression of proteins that are toxic to host cells or require temporal control.
MeSH term(s)	Antioxidants/metabolism ; Glutathione Transferase/biosynthesis ; Glutathione Transferase/genetics ; Glutathione Transferase/isolation & purification ; Hydrogen Peroxide/metabolism ; Ions/metabolism ; Membrane Proteins/genetics ; Nickel/metabolism ; Oxidative Stress/genetics ; Paramecium caudatum/genetics ; Paramecium caudatum/metabolism ; Protozoan Proteins/genetics ; RNA, Messenger/genetics ; Reactive Oxygen Species/metabolism
Chemical Substances	Antioxidants ; Ions ; Membrane Proteins ; Protozoan Proteins ; RNA, Messenger ; Reactive Oxygen Species ; nickel sulfate (4FLT4T3WUN) ; Nickel (7OV03QG267) ; Hydrogen Peroxide (BBX060AN9V) ; Glutathione Transferase (EC 2.5.1.18)
Language	English
Publishing date	2014-07-07
Publishing country	United States
Document type	Journal Article
ZDB-ID	2077635-4
ISSN	1535-9786 ; 1535-9778
ISSN (online)	1535-9786
ISSN	1535-9778
DOI	10.1128/EC.00112-14
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Article: Direct observation of histone H2B-YFP fusion proteins and transport of their mRNA between conjugating Paramecia.

Takenaka, Yasuhiro / Yanagi, Akira / Masuda, Hiromi / Mitsui, Youji / Mizuno, Hiroshi / Haga, Nobuyuki

Gene

2007 Volume 395, Issue 1-2, Page(s) 108–115

Abstract: Cytoplasmic exchange between conjugating cells of Paramecium caudatum has been implicated by mating experiments using wild-type and behavioral mutant cells. To observe macromolecular transport between mating cells, we cloned and expressed the P. caudatum ...

Abstract	Cytoplasmic exchange between conjugating cells of Paramecium caudatum has been implicated by mating experiments using wild-type and behavioral mutant cells. To observe macromolecular transport between mating cells, we cloned and expressed the P. caudatum histone H2B gene as a fusion protein attached to an enhanced yellow fluorescent protein (YFP) named PcVenus. Significant fluorescent signals derived from histone H2B-PcVenus were detected throughout the macro- and micronuclei of transformant cells after microinjection of the expression vector. The normal growth and high mating reactivity of the transformants indicated that H2B-PcVenus functioned normally. Seven hours after a transformant cell expressing histone H2B-PcVenus was mated with an untransformed complementary mating-type cell, fluorescence derived from histone H2B-PcVenus was emitted from the macronuclei of the untransformed cell. About 48 h later, the fluorescent signal was detected not only in the macro- and micronuclei of untransformed cells but also in the macronuclear anlagen of both mating cells. This suggests that conjugant cells share parental histones during meiosis and subsequent DNA rearrangement. Single-cell RT-PCR analysis demonstrated the presence of H2B-PcVenus mRNA in untransformed cells 15 and 24 h after conjugation. We concluded that at least the mRNA of histone H2B-PcVenus was transferred from the transformed, to the untransformed cell during conjugation.
MeSH term(s)	Amino Acid Sequence ; Animals ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Base Sequence ; Biological Transport, Active ; Cloning, Molecular ; Conjugation, Genetic ; DNA, Protozoan/genetics ; Genes, Protozoan ; Genetic Vectors ; Histones/genetics ; Histones/metabolism ; Luminescent Proteins/genetics ; Luminescent Proteins/metabolism ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Paramecium caudatum/genetics ; Paramecium caudatum/metabolism ; Protozoan Proteins/genetics ; Protozoan Proteins/metabolism ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; RNA, Protozoan/genetics ; RNA, Protozoan/metabolism ; Recombinant Fusion Proteins/genetics ; Recombinant Fusion Proteins/metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Homology, Amino Acid
Chemical Substances	Bacterial Proteins ; DNA, Protozoan ; Histones ; Luminescent Proteins ; Protozoan Proteins ; RNA, Messenger ; RNA, Protozoan ; Recombinant Fusion Proteins ; yellow fluorescent protein, Bacteria
Language	English
Publishing date	2007-06-15
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	391792-7
ISSN	1879-0038 ; 0378-1119
ISSN (online)	1879-0038
ISSN	0378-1119
DOI	10.1016/j.gene.2007.02.013
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Article: Transformation of Paramecium caudatum with a novel expression vector harboring codon-optimized GFP gene.

Takenaka, Yasuhiro / Haga, Nobuyuki / Harumoto, Terue / Matsuura, Tadashi / Mitsui, Youji

Gene

2002 Volume 284, Issue 1-2, Page(s) 233–240

Abstract: We have developed a novel expression vector, pTub-tel3, for transformation in Paramecium caudatum. The vector was constructed by cloning P. caudatum alpha-tubulin 5' and 3' non-coding regions. To examine transformation with the pTub-tel3 construct, we ... ...

Abstract	We have developed a novel expression vector, pTub-tel3, for transformation in Paramecium caudatum. The vector was constructed by cloning P. caudatum alpha-tubulin 5' and 3' non-coding regions. To examine transformation with the pTub-tel3 construct, we chose the green fluorescent protein (GFP) as a selection marker. When a linearized pTub-tel3 vector containing a GFP open reading frame was injected into the macronucleus, the GFP transcript was expressed in many clones whereas protein expression was detected only after extensive optimization of original GFP codons. GFP-derived fluorescence was distributed throughout the nuclei and cytoplasm except for contractile and food vacuoles. Upon continuous cell division, notable heterogeneity of GFP fluorescence among descendants from the same transformant has emerged. This expression vector can be applied to the analysis of protein trafficking and localization in addition to exogenous gene expression in P. caudatum.
MeSH term(s)	Animals ; Base Sequence ; Cloning, Molecular ; Gene Expression ; Genetic Vectors/genetics ; Green Fluorescent Proteins ; Luminescent Proteins/genetics ; Luminescent Proteins/metabolism ; Microscopy, Fluorescence ; Molecular Sequence Data ; Paramecium/genetics ; Sequence Homology, Nucleic Acid ; Transformation, Genetic ; Tubulin/genetics
Chemical Substances	Luminescent Proteins ; Tubulin ; Green Fluorescent Proteins (147336-22-9)
Language	English
Publishing date	2002-02-06
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	391792-7
ISSN	1879-0038 ; 0378-1119
ISSN (online)	1879-0038
ISSN	0378-1119
DOI	10.1016/s0378-1119(01)00886-1
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Article: FBN2, FBN1, TGFBR1, and TGFBR2 analyses in congenital contractural arachnodactyly.

Nishimura, Akira / Sakai, Haruya / Ikegawa, Shiro / Kitoh, Hiroshi / Haga, Nobuyuki / Ishikiriyama, Satoshi / Nagai, Toshiro / Takada, Fumio / Ohata, Takako / Tanaka, Fumihiko / Kamasaki, Hotaka / Saitsu, Hirotomo / Mizuguchi, Takeshi / Matsumoto, Naomichi

American journal of medical genetics. Part A

2007 Volume 143A, Issue 7, Page(s) 694–698

Abstract: FBN2, FBN1, TGFBR1, and TGFBR2 were analyzed by direct sequencing in 15 probands with suspected congenital contractural arachnodactyly (CCA). A total of four novel FBN2 mutations were found in four probands (27%, 4/15), but remaining the 11 did not show ... ...

Abstract	FBN2, FBN1, TGFBR1, and TGFBR2 were analyzed by direct sequencing in 15 probands with suspected congenital contractural arachnodactyly (CCA). A total of four novel FBN2 mutations were found in four probands (27%, 4/15), but remaining the 11 did not show any abnormality in either of the genes. This study indicated that FBN2 mutations were major abnormality in CCA, and TGFBR and FBN1 defects may not be responsible for the disorder. FBN2 mutations were only found at introns 30, 31, and 35 in this study. Thus analysis of a mutational hotspot from exons 22 to 36 (a middle part) of FBN2 should be prioritized in CCA as previously suggested.
MeSH term(s)	Adolescent ; Child ; Child, Preschool ; Female ; Fibrillin-1 ; Fibrillin-2 ; Fibrillins ; Humans ; Infant ; Infant, Newborn ; Male ; Marfan Syndrome/genetics ; Microfilament Proteins/genetics ; Protein-Serine-Threonine Kinases/genetics ; Receptor, Transforming Growth Factor-beta Type I ; Receptor, Transforming Growth Factor-beta Type II ; Receptors, Transforming Growth Factor beta/genetics
Chemical Substances	FBN1 protein, human ; FBN2 protein, human ; Fibrillin-1 ; Fibrillin-2 ; Fibrillins ; Microfilament Proteins ; Receptors, Transforming Growth Factor beta ; Protein-Serine-Threonine Kinases (EC 2.7.11.1) ; Receptor, Transforming Growth Factor-beta Type I (EC 2.7.11.30) ; Receptor, Transforming Growth Factor-beta Type II (EC 2.7.11.30) ; TGFBR1 protein, human (EC 2.7.11.30)
Language	English
Publishing date	2007-03-07
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2108614-X
ISSN	1552-4825
ISSN	1552-4825
DOI	10.1002/ajmg.a.31639
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