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  • 1
    Digitale Medien
    Digitale Medien
    Detection of partial cDNA sequences differentially expressed in patients with myelodysplasia (1996)
    Springer
    Annals of hematology 72 (1996), S. 231-236 
    Details
    ISSN: 1432-0584
    Schlagwort(e): Key words Differential display ; Myelodysplastic syndromes ; Chromosomal aberration
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract  The most common chromosomal aberrations in myelodysplastic syndromes (MDS) are complete or partial loss of chromosomes 5 and 7, and trisomy 8. To identify genes important in the pathogenesis of this disease that could be associated with these gross chromosomal defects, we have employed the differential display PCR (DDPCR) procedure developed by Liang and Pardee. This method allows simultaneous comparison of several cDNA sources for the presence of differentially expressed genes. Polymorphonuclear cells (PMNs) from two MDS patients, containing a 5q deletion or a trisomy 8, and three healthy controls were used. Initial screening resulted in the identification of five and three partial cDNA sequences, respectively that were either differentially expressed in both patient samples or in individual patients, as compared with the controls. The authenticity of aberrant expression was verified by reanalyzing the same primer combinations on newly prepared cDNA. Differential expression of the three remaining fragments was subsequently checked on a larger panel of MDS patients, using amplicon-specific primer sets. These were obtained by cloning and sequencing of the fragments. For one partial cDNA (DC3), the original expression pattern, i.e., decreased expression in individual MDS patients, was confirmed. These results demonstrate the utility of the DDPCR procedure to isolate differentially expressed sequences in primary patient samples where the availability of cells is a limiting factor.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/s002770050165
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  • 2
    Digitale Medien
    Digitale Medien
    Biology and treatment of myelodysplastic syndromes — developments in the past decade (1993)
    Springer
    Annals of hematology 66 (1993), S. 107-115 
    Details
    ISSN: 1432-0584
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Conclusions During the past 10 years, MDS has become a topic of considerable interest due to the development of new cytogenetic and molecular techniques and the introduction of the clinical administration of hematopoietic growth factors. The cytological and clinical classifications of the diseases comprising the syndrome are based on two main subtypes, those with a low risk of acquiring acute leukemia and those with a high risk. Which one will prove to be the simplest and most reliable classification system for the individual patient is not yet clear. Studies on clonality have shown that the myeloid cells in most of the patients studied thus far are monoclonal in nature. These methods can be used to evaluate the effect of treatment regimens. The kinetics of the in vitro growth of hematopoietic precursor cells from MDS patients is abnormal, with suboptimal colony formation. Cultures grown in the presence of certain growth factors show preferential growth of normal hematopoietic cells, which may be of clinical relevance in the future. Administration to MDS patients of hematopoietic growth factors such as GM-CSF, G-CSF, and IL-3 improves neutrophil counts in the majority of cases. The effect of erythropoietin is disappointing in most cases. At present, polyclonal hematopoiesis is rarely induced by growth factor treatment alone. More recently identified growth factors, such as MGF, might preferentially affect normal hematopoiesis at the expense of the preleukemic clones, thus changing the natural course of the disease.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF01697618
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  • 3
    Digitale Medien
    Digitale Medien
    In vitro-induced resistance to the deoxycytidine analogues cytarabine (AraC) and 5-aza-2′-deoxycytidine (DAC) in a rat model for acute myeloid leukemia is mediated by mutations in the deoxycytidine kinase (dck) gene (1995)
    Springer
    Annals of hematology 71 (1995), S. 41-47 
    Details
    ISSN: 1432-0584
    Schlagwort(e): AraC and DAC resistance ; dck gene ; Inactivating mutations
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract The deoxycytidine kinase (dck) gene encodes the enzyme responsible for the metabolic activation of the antileukemic drugs cytosine arabinoside (AraC) and 5-aza-2′-deoxycytidine (decitabine, DAC). Thedck locus was analyzed at the chromosomal and the molecular level in a model of rat leukemic cell lines, in which AraC and DAC resistance was induced, that was marked bydck deficiency. At the chromosomal level, karyotype analysis of metaphase spreads revealed the presence of an aberrant 2q+ chromosome in the AraC-resistant cell line and a (Xq;11q) translocation in its subclone RA/7. The DAC-resistant lines were identical to the parental RCL/O. Fluorescence in situ hybridization on normal rat fibroblast metaphase spreads localized the ratdck gene to chromosome 14q21-q22, a region that was not involved in any of the observed karyotypic aberrations. Analysis at the molecular level revealed an identical rearrangement of thedck gene in the AraC-resistant cell line RCL/A and its subclone RA/7 that resulted in the absence ofdck expression, as assessed by RT-PCR. No genomic rearrangements were observed in a DAC-resistant cell line RCL/D or in its subclone RD/1. However, detection of a single-stranded conformation polymorphism (SSCP) allowed the identification of a single C to G substitution (His to Gln) in thedck cDNA of the DAC-resistant RD/1 clone. The data demonstrate that exposure to AraC and DAC induces a resistant phenotype marked by functionaldck deficiency that may be the consequence of mutations occurring in thedck gene.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF01696231
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  • 4
    Digitale Medien
    Digitale Medien
    The role of cytokines and hematopoietic growth factors in the autocrine/paracrine regulation of inducible hematopoiesis (1997)
    Springer
    Annals of hematology 75 (1997), S. 27-31 
    Details
    ISSN: 1432-0584
    Schlagwort(e): Key words Cytokines ; Gene expression ; Hematopoiesis ; Bone marrow ; Chemotherapy
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract  To elucidate the role of hematopoietic growth factors (HGFs) and other cytokines in the autocrine or paracrine regulation of inducible hematopoiesis we studied cytokine gene expression in the bone marrow (BM) of patients after myelosuppressive treatment. Furthermore, we studied the cytokine gene expression profile in healthy individuals before and after bone marrow harvesting for the purpose of bone marrow transplantation. We speculated that the bone marrow harvesting procedure might induce changes in cytokine gene expression. No induction of G-CSF, GM-CSF, IL-1α, IL-3, IL-5, IL-8, IL-9, and IL-12 was observed in the BM of patients following intensive chemotherapy. Also, no up-regulation of expression of M-CSF, IL-1β, IL-4, IL-6, TNF-α, TGF-β, IGF-1, EDF, and EPA gene was found, illustrating that the investigated cytokines probably are not relevant in the presumed autocrine/paracrine regulation of the recovery of hematopoiesis following depletion of hematopoietic progenitor cells (HPCs). Concomitantly, elevated G-CSF plasma levels were found in these patients, suggesting that G-CSF has an endocrine regulatory role in inducible hematopoiesis. Induction of GM-CSF and IL-8, but not of G-CSF or IL-3 gene expression and up-regulation of IL-1β and IL-6 gene following BM harvesting was observed. This induction of GM-CSF and IL-8 may be attributed to tissue damage rather than to HPC depletion.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/s002770050308
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  • 5
    Digitale Medien
    Digitale Medien
    Absence of mutations in the RET gene in acute myeloid leukemia (1997)
    Springer
    Annals of hematology 75 (1997), S. 87-90 
    Details
    ISSN: 1432-0584
    Schlagwort(e): Key words Hematopoiesis ; AML ; RET ; Mutation analysis
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract  Expression of the tyrosine kinase receptor RET has previously been detected in normal hematopoietic cells, and especially in cells of the myeloid lineage. Furthermore, RET was shown to be differentially expressed in acute myeloid leukemia (AML), a disease characterized by excessive cell growth and aberrant maturation of cells, with the highest levels of expression in leukemias with monocytic differentiation. RET is known to be expressed in cells from the excretory system and from the developing central and peripheral nervous system. Both activating and inactivating aberrations in the RET gene have been detected in disorders derived from these tissues. To investigate whether the differential expression is a primary defect in AML, the presence of RET alterations was scanned by Southern blot analysis on DNA of blasts obtained from 17 AML patients. However, no RET gene aberrations were found. Subsequently, denaturing gradient gel electrophoresis (DGGE) analysis was performed on the DNA of blasts from ten selected cases. All five variants detected turned out to represent neutral DNA polymorphisms, including a novel polymorphism in exon 14. Since we were unable to detect mutations of RET in AML, it is unlikely that it plays an important role in leukemogenesis.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/s002770050319
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  • 6
    Digitale Medien
    Digitale Medien
    Extramedullary hematopoietic growth factor and cytokine gene expression indicate endocrine regulation of hematopoiesis in patients with acute appendicitis (1997)
    Springer
    Annals of hematology 75 (1997), S. 201-205 
    Details
    ISSN: 1432-0584
    Schlagwort(e): Key words Hematopoiesis ; Hematopoietic growth factors ; Cytokines ; Appendicitis ; Bone marrow
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract  To analyze the role of hematopoietic growth factors (HGFs) and other cytokines in the regulation of hematopoiesis in vivo, we investigated HGFs and cytokine gene expression in appendices obtained from patients who underwent surgery for suspected appendicitis. Concomitantly, HGF gene expression was studied in bone marrow (BM) biopsy specimens and plasma HGF levels were measured. G-CSF gene expression was detected in inflamed but not in normal appendices. With one exception, GM-CSF was detectable in all appendices whether inflamed or not, whereas IL-3, except for one case, was not expressed in appendices. None of the investigated HGFs appeared to be expressed in BM biopsy specimens concurrently obtained with the appendices. Plasma G-CSF levels were significantly elevated in patients with appendicitis compared with patients without inflamed appendices. Circulating levels of GM-CSF and IL-3 were not increased. Significant up-regulation of IL-8 and IL-6 gene expression was observed in response to inflammation, in contrast to IL-1α and IL-1β expression, which appeared not to be influenced by the inflammatory state. These data indicate that G-CSF, and not GM-CSF or IL-3, is essential for the regulation of inducible granulopoiesis in acute inflammatory conditions, and that G-CSF acts in an endocrine fashion.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/s002770050343
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  • 7
    Digitale Medien
    Digitale Medien
    The human macrophage colony-stimulating factor gene is localized at chromosome 1 band p21 and not at 5q33.1 (1992)
    Springer
    Annals of hematology 64 (1992), S. 110-111 
    Details
    ISSN: 1432-0584
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF01715356
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  • 8
    Digitale Medien
    Digitale Medien
    In vitro-induced resistance to the deoxycytidine analogues cytarabine (AraC) and 5-aza-2′-deoxycytidine (DAC) in a rat model for acute myeloid leukemia is mediated by mutations in the deoxycytidine kinase (dck) gene (1995)
    Springer
    Annals of hematology 71 (1995), S. 41-47 
    Details
    ISSN: 1432-0584
    Schlagwort(e): Key words AraC and DAC resistance ; dck gene ; Inactivating mutations
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract  The deoxycytidine kinase (dck) gene encodes the enzyme responsible for the metabolic activation of the antileukemic drugs cytosine arabinoside (AraC) and 5-aza-2′-deoxycytidine (decitabine, DAC). The dck locus was analyzed at the chromosomal and the molecular level in a model of rat leukemic cell lines, in which AraC and DAC resistance was induced, that was marked by dck deficiency. At the chromosomal level, karyotype analysis of metaphase spreads revealed the presence of an aberrant 2q+ chromosome in the AraC-resistant cell line and a (Xq;11q) translocation in its subclone RA/7. The DAC-resistant lines were identical to the parental RCL/O. Fluorescence in situ hybridization on normal rat fibroblast metaphase spreads localized the rat dck gene to chromosome 14q21–q22, a region that was not involved in any of the observed karyotypic aberrations. Analysis at the molecular level revealed an identical rearrangement of the dck gene in the AraC-resistant cell line RCL/A and its subclone RA/7 that resulted in the absence of dck expression, as assessed by RT-PCR. No genomic rearrangements were observed in a DAC-resistant cell line RCL/D or in its subclone RD/1. However, detection of a single-stranded conformation polymorphism (SSCP) allowed the identification of a single C to G substitution (His to Gln) in the dck cDNA of the DAC-resistant RD/1 clone. The data demonstrate that exposure to AraC and DAC induces a resistant phenotype marked by functional dck deficiency that may be the consequence of mutations occurring in the dck gene.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF01696231
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  • 9
    Digitale Medien
    Digitale Medien
    IL-4 down-regulates IL-2-, IL-3-, and GM-CSF-induced cytokine gene expression in peripheral blood monocytes (1994)
    Springer
    Annals of hematology 68 (1994), S. 293-298 
    Details
    ISSN: 1432-0584
    Schlagwort(e): Cytokines ; Monocytes ; GM-CSF ; IL-2 ; IL-3 ; IL-4
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary IL-4, a product of the T-helper 0 (Th0) and 2 (Th2) subset, was originally described as a B-cell stimulatory factor and has subsequently been found to suppress IL-1α, IL-1Β, IL-6, IL-8, and TNF-α gene expression in monocytes stimulated with LPS, and to upregulate IL-1 receptor antagonist (IL1-RA) gene expression. In this study we investigated the effect of IL-4 on the expression of cytokine genes in monocytes evoked by other T-helper cell cytokines: IL-2, IL-3, and GM-CSF. IL-4 down-regulated mRNA accumulation of the proinflammatory cytokines IL-1Β, IL-8, and TNF-α in monocytes stimulated with IL-2, IL-3, and GM-CSF. IL-4 also suppressed the IL-2-induced IL-6 mRNA expression. Temporal analysis of the IL-4 down-regulatory effect on the IL-2-, IL-3-, or GM-CSF — induced proinflammatory cytokine gene expression in monocytes provided evidence that IL-4 acts predominantly oh the post-transcriptional level. This was supported by the observation that the down-regulatory capacity of IL-4 appeared to be dependent on de novo protein synthesis. IL-4 did not exert significant influence on the induction of expression of IL-1-RA or various CSFs by IL-2, IL-3, and GM-CSF. We hypothesize that owing to its being a down-regulator of the expression of proinflammatory cytokines induced by IL-2, IL-3, and GM-CSF — products of activated ThO or Th1 cells —together with its capacity as a B-cell stimulatory factor, IL-4 has an important role in directing the immune response.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF01695035
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  • 10
    Digitale Medien
    Digitale Medien
    Order of human hematopoietic growth factor and receptor genes on the long arm of chromosome 5, as determined by fluorescence in situ hybridization (1993)
    Springer
    Annals of hematology 66 (1993), S. 15-20 
    Details
    ISSN: 1432-0584
    Schlagwort(e): Growth factors/receptors ; Gene order ; FISH ; Chromosome 5q31
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary A large number of human hematopoietic growth factor and growth factor receptor genes are localized at the long arm of chromosome 5. In this study we have determined the order of the human interleukin-3 (IL3), IL4, IL5, IL9, granulocyte macrophage-colony stimulating factor (GMCSF), and the MCSF receptor (MCSFR) genes by fluorescence in situ hybridization. Genomic λ-clones were isolated using polymerase chain reaction (PCR)-generated probes and labeled with biotin and/or digoxigenin. These clones were first individually mapped: IL3, IL4, IL5, IL9, and GMCSF to 5q31 and MCSFR to 5q33. For ordering purposes multiple probe combinations were hybridized to metaphase chromosomes and interphase nuclei. The interphase hybridizations were evaluated by image analysis, which also allowed the measurement of the physical distance between the hybridization spots. These mapping results suggest the gene order 5cen-IL3/GMCSF-IL5-IL4-IL9-MCSFR-qter. The known genomic distance between the IL4 and IL5 genes allowed the estimation of the physical distances between the 5q31-specific genes, demonstrating that they are all within about 1.5 Mb of DNA.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF01737684
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