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Book ; Online ; Thesis: Präklinische Untersuchung von Eltrombopag in einem patientenabgeleiteten Xenograftmodell Myelodysplastischer Syndrome

Schmitt, Nanni [Verfasser] / Nowak, Daniel [Akademischer Betreuer]

2023

Author's details	Nanni Schmitt ; Betreuer: Daniel Nowak
Keywords	Medizin, Gesundheit ; Medicine, Health
Subject code	sg610
Language	German
Publisher	Universitätsbibliothek Heidelberg
Publishing place	Heidelberg
Document type	Book ; Online ; Thesis
Database	Digital theses on the web

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Article ; Online: In Silico Pan-Cancer Analysis Reveals Prognostic Role of the Erythroferrone (ERFE) Gene in Human Malignancies.

Xu, Qingyu / Altrock, Eva / Schmitt, Nanni / Streuer, Alexander / Rapp, Felicitas / Nowak, Verena / Obländer, Julia / Weimer, Nadine / Palme, Iris / Göl, Melda / Hofmann, Wolf-Karsten / Nowak, Daniel / Riabov, Vladimir

International journal of molecular sciences

2023 Volume 24, Issue 2

Abstract: The erythroferrone gene ( ...

Abstract	The erythroferrone gene (
MeSH term(s)	Humans ; Prognosis ; Peptide Hormones/genetics ; Hepcidins/metabolism ; Neoplasms/genetics ; Myelodysplastic Syndromes ; Tumor Microenvironment
Chemical Substances	Peptide Hormones ; Hepcidins
Language	English
Publishing date	2023-01-15
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms24021725
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Article ; Online: Treatment with the apoptosis inhibitor Asunercept reduces clone sizes in patients with lower risk Myelodysplastic Neoplasms.

Annals of hematology

2024 Volume 103, Issue 4, Page(s) 1221–1233

Abstract: In low-risk Myelodysplastic Neoplasms (MDS), increased activity of apoptosis-promoting factors such as tumor necrosis factor (TNFα) and pro-apoptotic Fas ligand (CD95L) have been described as possible pathomechanisms leading to impaired erythropoiesis. ... ...

Abstract	In low-risk Myelodysplastic Neoplasms (MDS), increased activity of apoptosis-promoting factors such as tumor necrosis factor (TNFα) and pro-apoptotic Fas ligand (CD95L) have been described as possible pathomechanisms leading to impaired erythropoiesis. Asunercept (APG101) is a novel therapeutic fusion protein blocking CD95, which has previously shown partial efficacy in reducing transfusion requirement in a clinical phase I trial for low-risk MDS patients (NCT01736436; 2012-11-26). In the current study we aimed to evaluate the effect of Asunercept therapy on the clonal bone marrow composition to identify potential biomarkers to predict response. Bone marrow samples of n = 12 low-risk MDS patients from the above referenced clinical trial were analyzed by serial deep whole exome sequencing in a total of n = 58 time points. We could distinguish a mean of 3.5 molecularly defined subclones per patient (range 2-6). We observed a molecular response defined as reductions of dominant clone sizes by a variant allele frequency (VAF) decrease of at least 10% (mean 20%, range: 10.5-39.2%) in dependency of Asunercept treatment in 9 of 12 (75%) patients. Most of this decline in clonal populations was observed after completion of 12 weeks treatment. Particularly early and pronounced reductions of clone sizes were found in subclones driven by mutations in genes involved in regulation of methylation (n = 1 DNMT3A, n = 1 IDH2, n = 1 TET2). Our results suggest that APG101 could be efficacious in reducing clone sizes of mutated hematopoietic cells in the bone marrow of Myelodysplastic Neoplasms, which warrants further investigation.
MeSH term(s)	Humans ; Neoplasms ; Myelodysplastic Syndromes/drug therapy ; Myelodysplastic Syndromes/genetics ; Myelodysplastic Syndromes/pathology ; Clone Cells/pathology ; Bone Marrow/pathology ; Apoptosis ; Mutation
Language	English
Publishing date	2024-02-27
Publishing country	Germany
Document type	Journal Article
ZDB-ID	1064950-5
ISSN	1432-0584 ; 0939-5555 ; 0945-8077
ISSN (online)	1432-0584
ISSN	0939-5555 ; 0945-8077
DOI	10.1007/s00277-024-05664-5
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Article ; Online: Significant improvement of bone marrow-derived MSC expansion from MDS patients by defined xeno-free medium.

Stem cell research & therapy

2023 Volume 14, Issue 1, Page(s) 156

Abstract: Background: Robust and reliable in vitro and in vivo models of primary cells are necessary to study the pathomechanisms of Myelodysplastic Neoplasms (MDS) and identify novel therapeutic strategies. MDS-derived hematopoietic stem and progenitor cells ( ... ...

Abstract	Background: Robust and reliable in vitro and in vivo models of primary cells are necessary to study the pathomechanisms of Myelodysplastic Neoplasms (MDS) and identify novel therapeutic strategies. MDS-derived hematopoietic stem and progenitor cells (HSPCs) are reliant on the support of bone marrow (BM) derived mesenchymal stroma cells (MSCs). Therefore, isolation and expansion of MCSs are essential for successfully modeling this disease. For the clinical use of healthy MSCs isolated from human BM, umbilical cord blood or adipose tissue, several studies showed that xeno-free (XF) culture conditions resulted in superior growth kinetics compared to MSCs cultured in the presence of fetal bovine serum (FBS). In this present study, we investigate, whether the replacement of a commercially available MSC expansion medium containing FBS with a XF medium is beneficial for the expansion of MSCs derived from BM of MDS patients which are often difficult to cultivate. Methods: MSCs isolated from BM of MDS patients were cultured and expanded in MSC expansion medium with FBS or XF supplement. Subsequently, the impact of culture media on growth kinetics, morphology, immunophenotype, clonogenic potential, differentiation capacity, gene expression profiles and ability to engraft in immunodeficient mouse models was evaluated. Results: Significant higher cell numbers with an increase in clonogenic potential were observed during culture of MDS MSCs with XF medium compared to medium containing FBS. Differential gene expression showed an increase in transcripts associated with MSC stemness after expansion with XF. Furthermore, immunophenotypes of the MSCs and their ability to differentiate into osteoblasts, adipocytes or chondroblasts remained stable. MSCs expanded with XF media were similarly supportive for creating MDS xenografts in vivo as MSCs expanded with FBS. Conclusion: Our data indicate that with XF media, higher cell numbers of MDS MSCs can be obtained with overall improved characteristics in in vitro and in vivo experimental models.
MeSH term(s)	Animals ; Mice ; Humans ; Culture Media, Serum-Free ; Bone Marrow ; Cell Differentiation ; Mesenchymal Stem Cells/metabolism ; Adipose Tissue ; Cell Proliferation ; Cells, Cultured
Chemical Substances	Culture Media, Serum-Free
Language	English
Publishing date	2023-06-07
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2548671-8
ISSN	1757-6512 ; 1757-6512
ISSN (online)	1757-6512
ISSN	1757-6512
DOI	10.1186/s13287-023-03386-5
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Article ; Online: Bone marrow derived stromal cells from myelodysplastic syndromes are altered but not clonally mutated in vivo.

Jann, Johann-Christoph / Mossner, Maximilian / Riabov, Vladimir / Altrock, Eva / Schmitt, Nanni / Flach, Johanna / Xu, Qingyu / Nowak, Verena / Obländer, Julia / Palme, Iris / Weimer, Nadine / Streuer, Alexander / Jawhar, Ahmed / Darwich, Ali / Jawhar, Mohammad / Metzgeroth, Georgia / Nolte, Florian / Hofmann, Wolf-Karsten / Nowak, Daniel

Nature communications

2021 Volume 12, Issue 1, Page(s) 6170

Abstract: The bone marrow (BM) stroma in myeloid neoplasms is altered and it is hypothesized that this cell compartment may also harbor clonal somatically acquired mutations. By exome sequencing of in vitro expanded mesenchymal stromal cells (MSCs) from n = 98 ... ...

Abstract	The bone marrow (BM) stroma in myeloid neoplasms is altered and it is hypothesized that this cell compartment may also harbor clonal somatically acquired mutations. By exome sequencing of in vitro expanded mesenchymal stromal cells (MSCs) from n = 98 patients with myelodysplastic syndrome (MDS) and n = 28 healthy controls we show that these cells accumulate recurrent mutations in genes such as ZFX (n = 8/98), RANK (n = 5/98), and others. MDS derived MSCs display higher mutational burdens, increased replicative stress, senescence, inflammatory gene expression, and distinct mutational signatures as compared to healthy MSCs. However, validation experiments in serial culture passages, chronological BM aspirations and backtracking of high confidence mutations by re-sequencing primary sorted MDS MSCs indicate that the discovered mutations are secondary to in vitro expansion but not present in primary BM. Thus, we here report that there is no evidence for clonal mutations in the BM stroma of MDS patients.
MeSH term(s)	Adult ; Aged ; Aged, 80 and over ; Bone Marrow/metabolism ; Bone Marrow/pathology ; Cells, Cultured ; Exome/genetics ; Female ; Genotype ; Humans ; Male ; Mesenchymal Stem Cells/metabolism ; Mesenchymal Stem Cells/pathology ; Middle Aged ; Mutation ; Myelodysplastic Syndromes/genetics ; Myelodysplastic Syndromes/pathology ; Phenotype ; Tumor Microenvironment
Language	English
Publishing date	2021-10-25
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-021-26424-3
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Article ; Online: Replication stress signaling is a therapeutic target in myelodysplastic syndromes with splicing factor mutations.

Haematologica

2021 Volume 106, Issue 11, Page(s) 2906–2917

Abstract: Somatic mutations in genes coding for splicing factors, e.g. SF3B1, U2AF1, SRSF2, and others are found in approximately 50% of patients with Myelodysplastic Syndromes (MDS). These mutations have been predicted to frequently occur early in the mutational ... ...

Abstract	Somatic mutations in genes coding for splicing factors, e.g. SF3B1, U2AF1, SRSF2, and others are found in approximately 50% of patients with Myelodysplastic Syndromes (MDS). These mutations have been predicted to frequently occur early in the mutational hierarchy of the disease therefore making them particularly attractive potential therapeutic targets. Recent studies in cell lines engineered to carry splicing factor mutations have revealed a strong association with elevated levels of DNA:RNA intermediates (R-loops) and a dependency on proper ATR function. However, data confirming this hypothesis in a representative cohort of primary MDS patient samples have so far been missing. Using CD34+ cells isolated from MDS patients with and without splicing factor mutations as well as healthy controls we show that splicing factor mutation-associated R-loops lead to elevated levels of replication stress and ATR pathway activation. Moreover, splicing factor mutated CD34+ cells are more susceptible to pharmacological inhibition of ATR resulting in elevated levels of DNA damage, cell cycle blockade, and cell death. This can be enhanced by combination treatment with low-dose splicing modulatory compound Pladienolide B. We further confirm the direct association of R-loops and ATR sensitivity with the presence of a splicing factor mutation using lentiviral overexpression of wild-type and mutant SRSF2 P95H in cord blood CD34+ cells. Collectively, our results from n=53 MDS patients identify replication stress and associated ATR signaling to be critical pathophysiological mechanisms in primary MDS CD34+ cells carrying splicing factor mutations, and provide a preclinical rationale for targeting ATR signaling in these patients.
MeSH term(s)	Humans ; Mutation ; Myelodysplastic Syndromes/drug therapy ; Myelodysplastic Syndromes/genetics ; Phosphoproteins/genetics ; RNA Splicing ; RNA Splicing Factors/genetics ; RNA Splicing Factors/metabolism ; Serine-Arginine Splicing Factors/genetics ; Splicing Factor U2AF/genetics
Chemical Substances	Phosphoproteins ; RNA Splicing Factors ; Splicing Factor U2AF ; Serine-Arginine Splicing Factors (170974-22-8)
Language	English
Publishing date	2021-11-01
Publishing country	Italy
Document type	Journal Article
ZDB-ID	2333-4
ISSN	1592-8721 ; 0017-6567 ; 0390-6078
ISSN (online)	1592-8721
ISSN	0017-6567 ; 0390-6078
DOI	10.3324/haematol.2020.254193
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Article ; Online: Inhibition of lysyl oxidases synergizes with 5-azacytidine to restore erythropoiesis in myelodysplastic and myeloid malignancies.

Xu, Qingyu / Streuer, Alexander / Jann, Johann-Christoph / Altrock, Eva / Schmitt, Nanni / Flach, Johanna / Sens-Albert, Carla / Rapp, Felicitas / Wolf, Julia / Nowak, Verena / Weimer, Nadine / Obländer, Julia / Palme, Iris / Kuzina, Mariia / Jawhar, Ahmed / Darwich, Ali / Weis, Cleo-Aron / Marx, Alexander / Wuchter, Patrick /

Costina, Victor / Jäger, Evelyn / Sperk, Elena / Neumaier, Michael / Fabarius, Alice / Metzgeroth, Georgia / Nolte, Florian / Steiner, Laurenz / Levkin, Pavel A / Jawhar, Mohamad / Hofmann, Wolf-Karsten / Riabov, Vladimir / Nowak, Daniel

Nature communications

2023 Volume 14, Issue 1, Page(s) 1497

Abstract: Limited response rates and frequent relapses during standard of care with hypomethylating agents in myelodysplastic neoplasms (MN) require urgent improvement of this treatment indication. Here, by combining 5-azacytidine (5-AZA) with the pan-lysyl ... ...

Abstract	Limited response rates and frequent relapses during standard of care with hypomethylating agents in myelodysplastic neoplasms (MN) require urgent improvement of this treatment indication. Here, by combining 5-azacytidine (5-AZA) with the pan-lysyl oxidase inhibitor PXS-5505, we demonstrate superior restoration of erythroid differentiation in hematopoietic stem and progenitor cells (HSPCs) of MN patients in 20/31 cases (65%) versus 9/31 cases (29%) treated with 5-AZA alone. This effect requires direct contact of HSPCs with bone marrow stroma components and is dependent on integrin signaling. We further confirm these results in vivo using a bone marrow niche-dependent MN xenograft model in female NSG mice, in which we additionally demonstrate an enforced reduction of dominant clones as well as significant attenuation of disease expansion and normalization of spleen sizes. Overall, these results lay out a strong pre-clinical rationale for efficacy of combination treatment of 5-AZA with PXS-5505 especially for anemic MN.
MeSH term(s)	Humans ; Female ; Mice ; Animals ; Azacitidine/pharmacology ; Azacitidine/therapeutic use ; Erythropoiesis ; Protein-Lysine 6-Oxidase ; Hematopoietic Stem Cells ; Myelodysplastic Syndromes/drug therapy ; Myelodysplastic Syndromes/pathology ; Myeloproliferative Disorders/pathology ; Neoplasms/pathology
Chemical Substances	Azacitidine (M801H13NRU) ; Protein-Lysine 6-Oxidase (EC 1.4.3.13)
Language	English
Publishing date	2023-03-17
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-023-37175-8
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Article ; Online: ASXL1

Riabov, Vladimir / Xu, Qingyu / Schmitt, Nanni / Streuer, Alexander / Ge, Guo / Bolanos, Lyndsey / Wunderlich, Mark / Jann, Johann-Christoph / Wein, Alina / Altrock, Eva / Demmerle, Marie / Mukherjee, Sanjay / Ali, Abdullah Mahmood / Rapp, Felicitas / Nowak, Verena / Weimer, Nadine / Obländer, Julia / Palme, Iris / Göl, Melda /

Jawhar, Ahmed / Darwich, Ali / Wuchter, Patrick / Weiss, Christel / Raza, Azra / Foulks, Jason M / Starczynowski, Daniel T / Yang, Feng-Chun / Metzgeroth, Georgia / Steiner, Laurenz / Jawhar, Mohamad / Hofmann, Wolf-Karsten / Nowak, Daniel

Haematologica

2023

Abstract: Inhibitors of anti-apoptotic BCL-2 family proteins in combination with chemotherapy and hypomethylating agents (HMAs) are promising therapeutic approaches in acute myeloid leukemia (AML) and high-risk myelodysplastic syndromes (MDS). Alvocidib, a cyclin- ... ...

Abstract	Inhibitors of anti-apoptotic BCL-2 family proteins in combination with chemotherapy and hypomethylating agents (HMAs) are promising therapeutic approaches in acute myeloid leukemia (AML) and high-risk myelodysplastic syndromes (MDS). Alvocidib, a cyclin-dependent kinase 9 (CDK9) inhibitor and indirect transcriptional repressor of the anti-apoptotic factor MCL-1, has previously shown clinical activity in AML. Availability of biomarkers for response to the alvocidib + 5- AZA could also extend the rationale of this treatment concept to high-risk MDS. In this study, we performed a comprehensive in vitro assessment of alvocidib and 5-AZA effects in n=45 high-risk MDS patients. Our data revealed additive cytotoxic effects of the combination treatment. Mutational profiling of MDS samples identified ASXL1 mutations as predictors of response. Further, increased response rates were associated with higher gene-expression of the pro-apoptotic factor NOXA in ASXL1 mutated samples. The higher sensitivity of ASXL1 mutant cells to the combination treatment was confirmed in vivo in ASXL1Y588X transgenic mice. Overall, our study demonstrated augmented activity for the alvocidib + 5-AZA combination in higher-risk MDS and identified ASXL1 mutations as a biomarker of response for potential stratification studies.
Language	English
Publishing date	2023-11-02
Publishing country	Italy
Document type	Journal Article
ZDB-ID	2333-4
ISSN	1592-8721 ; 0017-6567 ; 0390-6078
ISSN (online)	1592-8721
ISSN	0017-6567 ; 0390-6078
DOI	10.3324/haematol.2023.282921
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Article ; Online: Comparative analysis of clonal hematopoiesis of multipotent stem cells in healthy elderly in blood and bone marrow.

Jann, Johann-Christoph / Nolte, Florian / Mossner, Maximilian / Flach, Johanna / Altrock, Eva / Schmitt, Nanni / Röhl, Henning / Jawhar, Ahmed / Neumann, Uwe / Nowak, Verena / Danner, Justine / Obländer, Julia / Palme, Iris / Hofmann, Wolf-Karsten / Nowak, Daniel

Leukemia research

2019 Volume 82, Page(s) 15–18

MeSH term(s)	Aged ; Aged, 80 and over ; Antigens, CD/genetics ; Antigens, CD/immunology ; Arthroplasty, Replacement, Hip ; Biomarkers/metabolism ; Bone Marrow Cells/cytology ; Bone Marrow Cells/immunology ; Cell Differentiation ; Clone Cells ; Female ; Gene Expression ; Hematopoiesis/genetics ; Hematopoiesis/immunology ; Humans ; Immunophenotyping ; Male ; Middle Aged ; Multipotent Stem Cells/cytology ; Multipotent Stem Cells/immunology ; Primary Cell Culture
Chemical Substances	Antigens, CD ; Biomarkers
Language	English
Publishing date	2019-05-16
Publishing country	England
Document type	Comparative Study ; Letter ; Research Support, Non-U.S. Gov't
ZDB-ID	752396-8
ISSN	1873-5835 ; 0145-2126
ISSN (online)	1873-5835
ISSN	0145-2126
DOI	10.1016/j.leukres.2019.05.005
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Article ; Online: Preclinical evaluation of eltrombopag in a PDX model of myelodysplastic syndromes.

Schmitt, Nanni / Jann, Johann-Christoph / Altrock, Eva / Flach, Johanna / Danner, Justine / Uhlig, Stefanie / Streuer, Alexander / Knaflic, Antje / Riabov, Vladimir / Xu, Qingyu / Mehralivand, Arwin / Palme, Iris / Nowak, Verena / Obländer, Julia / Weimer, Nadine / Haselmann, Verena / Jawhar, Ahmed / Darwich, Ali / Weis, Cleo-Aron /

Marx, Alexander / Steiner, Laurenz / Jawhar, Mohamad / Metzgeroth, Georgia / Boch, Tobias / Nolte, Florian / Hofmann, Wolf-Karsten / Nowak, Daniel

Leukemia

2021 Volume 36, Issue 1, Page(s) 236–247

Abstract: Preclinical research of myelodysplastic syndromes (MDSs) is hampered by a lack of feasible disease models. Previously, we have established a robust patient-derived xenograft (PDX) model for MDS. Here we demonstrate for the first time that this model is ... ...

Abstract	Preclinical research of myelodysplastic syndromes (MDSs) is hampered by a lack of feasible disease models. Previously, we have established a robust patient-derived xenograft (PDX) model for MDS. Here we demonstrate for the first time that this model is applicable as a preclinical platform to address pending clinical questions by interrogating the efficacy and safety of the thrombopoietin receptor agonist eltrombopag. Our preclinical study included n = 49 xenografts generated from n = 9 MDS patient samples. Substance efficacy was evidenced by FACS-based human platelet quantification and clonal bone marrow evolution was reconstructed by serial whole-exome sequencing of the PDX samples. In contrast to clinical trials in humans, this experimental setup allowed vehicle- and replicate-controlled analyses on a patient-individual level deciphering substance-specific effects from natural disease progression. We found that eltrombopag effectively stimulated thrombopoiesis in MDS PDX without adversely affecting the patients' clonal composition. In conclusion, our MDS PDX model is a useful tool for testing new therapeutic concepts in MDS preceding clinical trials.
MeSH term(s)	Aged ; Aged, 80 and over ; Animals ; Apoptosis ; Benzoates/therapeutic use ; Cell Proliferation ; Female ; Humans ; Hydrazines/therapeutic use ; Male ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Middle Aged ; Myelodysplastic Syndromes/drug therapy ; Myelodysplastic Syndromes/pathology ; Prognosis ; Pyrazoles/therapeutic use ; Tumor Cells, Cultured ; Xenograft Model Antitumor Assays
Chemical Substances	Benzoates ; Hydrazines ; Pyrazoles ; eltrombopag (S56D65XJ9G)
Language	English
Publishing date	2021-06-25
Publishing country	England
Document type	Evaluation Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	807030-1
ISSN	1476-5551 ; 0887-6924
ISSN (online)	1476-5551
ISSN	0887-6924
DOI	10.1038/s41375-021-01327-w
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