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| 3. |
- Crnalic, S, et al.
(författare)
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Establishment and characterisation of a human clear cell sarcoma model in nude mice
- 2002
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Ingår i: International Journal of Cancer. - : Wiley. - 0020-7136. ; 101:6, s. 505-511
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Tidskriftsartikel (refereegranskat)abstract
- We have established a new experimental model of human clear cell sarcoma, UM-CCSI, using serial subcutaneous transplantation of intact tumour tissue in nude mice. The heterotransplanted nude mouse tumours retained characteristic morphological features of the primary clear cell sarcoma. Immunohistochemical analysis showed the retained expression patterns of S-100 protein, melanoma-associated antigen HMB-45 and vimentin in the xenografts as compared to the primary tumour. DNA index showed low variations both between the xenografts in the same passage and between the serial passages. Cytogenetic analysis of the primary tumour and the xenografts showed the unbalanced translocation der(6)t(6; I 2)(p23;q13). Based on the combined genetic data a reasonable interpretation of our findings is that there was a complex chromosomal rearrangement resulting in a cytogenetically cryptic EWS-ATFI fusion gene. Analysis of cell kinetics using in vivo incorporation of iododeoxyuridine and flow cytometry showed generally short potential doubling time (T-pot) of the xenografts. Volume doubling time showed low variations without correlation with T-pot. The retained phenotypic and genotypic characteristics of the primary tumour and the morphological and structural stability over time makes the model suitable for studies on the tumour biology and treatment of clear cell sarcoma. (C) 2002 Wiley-Liss, Inc.
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- Hamaguchi, I, et al.
(författare)
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Lentivirus vector gene expression during ES cell-derived hematopoietic development in vitro
- 2000
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Ingår i: Journal of Virology. - : American Society for Microbiology. - 0022-538X .- 1098-5514. ; 74:22, s. 84-10778
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Tidskriftsartikel (refereegranskat)abstract
- The murine embryonal stem (ES) cell virus (MESV) can express transgenes from the long terminal repeat (LTR) promoter/enhancer in undifferentiated ES cells, but expression is turned off upon differentiation to embryoid bodies (EBs) and hematopoietic cells in vitro. We examined whether a human immunodeficiency virus type 1-based lentivirus vector pseudotyped with the vesicular stomatitis virus G protein (VSV-G) could transduce ES cells efficiently and express the green fluorescent protein (GFP) transgene from an internal phosphoglycerate kinase (PGK) promoter throughout development to hematopoietic cells in vitro. An oncoretrovirus vector containing the MESV LTR and the GFP gene was used for comparison. Fluorescence-activated cell sorting analysis of transduced CCE ES cells showed 99.8 and 86.7% GPF-expressing ES cells in the VSV-G-pseudotyped lentivirus (multiplicity of infection [MOI] = 59)- and oncoretrovirus (MOI = 590)-transduced cells, respectively. Therefore, VSV-G pseudotyping of lentiviral and oncoretrovirus vectors leads to efficient transduction of ES cells. Lentivirus vector integration was verified in the ES cell colonies by Southern blot analysis. When the transduced ES cells were differentiated in vitro, expression from the oncoretrovirus LTR was severely reduced or extinct in day 6 EBs and ES cell-derived hematopoietic colonies. In contrast, many lentivirus-transduced colonies, expressing the GFP gene in the undifferentiated state, continued to express the transgene throughout in vitro development to EBs at day 6, and many continued to express in cells derived from hematopoietic colonies. This experimental system can be used to analyze lentivirus vector design for optimal expression in hematopoietic cells and for gain-of-function experiments during ES cell development in vitro.
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- Panagopoulos, C.N., et al.
(författare)
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Ni-P-W multilayered alloy coatings produced by pulse plating
- 2000
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Ingår i: Scripta Materialia. - 1359-6462 .- 1872-8456. ; 43:7, s. 677-683
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Tidskriftsartikel (refereegranskat)abstract
- The possibility of producing multilayered alloy coatings consisting of alternate amorphous layers, by the electrodeposition method, using pulsed current was investigated. The alternate metallic amorphous layers were selected to consist of Ni-P-W ternary alloys, in order to study the effect of the incorporation of a third element in the structure of an electrodeposited Ni-P multilayered alloy coating. The well-defined layered structure found in the Ni-P-W multilayered alloy coatings, consisting of X-ray amorphous layers, shows that production of a ternary Ni-P-W multilayered alloy coating with the selected method is feasible and this may expand the potential applications of a Ni-P based multilayered alloy coating.
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- Papachristos, V.D., et al.
(författare)
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Effect of annealing on the structure and hardness of Ni-P-W multilayered alloy coatings produced by pulse plating
- 2000
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Ingår i: Materials Science & Engineering. - 0921-5093 .- 1873-4936. ; 279:1-2, s. 217-230
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Tidskriftsartikel (refereegranskat)abstract
- Ni-P-W multilayered alloy coatings were deposited on copper foils by pulse plating, using a rotating cylinder electrode. They consisted of layers of high (Ni-5 wt.% P-45 wt.% W) and low (Ni-8 wt.% P-15 wt.% W) W content. The wavelengths studied were 8, 40 and 400 nm and the total thickness of the coatings was approximately 25 µm. The multilayered coatings were annealed in the temperature range between 200 and 800°C and the effect of annealing on their structure was studied by using X-ray diffraction, cross-sectional transmission electron microscopy equipped with selected area electron diffraction facility, and differential thermal analysis. The initially amorphous multilayered coatings crystallise in steps as the annealing temperature increases (first the low-W crystallise layers and then the high-W layers), and the layered structure is lost around 600°C. The main crystalline phases formed during annealing are Ni-W solid solution and Ni3P. The wavelength of the coatings seems to affect the onset temperature of the crystallisation processes. The hardness of the coatings initially increases reaching a maximum in the 500-600°C range, and then decreases as the annealing temperature increases. (C) 2000 Elsevier Science S.A. All rights reserved.Ni-P-W multilayered alloy coatings were deposited on copper foils by pulse plating, using a rotating cylinder electrode. They consisted of layers of high (Ni-5 wt.% P-45 wt.% W) and low (Ni-8 wt.% P-15 wt.% W) W content. The wavelengths studied were 8, 40 and 400 nm and the total thickness of the coatings was approximately 25 µm. The multilayered coatings were annealed in the temperature range between 200 and 800°C and the effect of annealing on their structure was studied by using X-ray diffraction, cross-sectional transmission electron microscopy equipped with selected area electron diffraction facility, and differential thermal analysis. The initially amorphous multilayered coatings crystallise in steps as the annealing temperature increases (first the low-W crystallise layers and then the high-W layers), and the layered structure is lost around 600°C. The main crystalline phases formed during annealing are Ni-W solid solution and Ni3P. The wavelength of the coatings seems to affect the onset temperature of the crystallisation processes. The hardness of the coatings initially increases reaching a maximum in the 500-600°C range, and then decreases as the annealing temperature increases.
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- Raddaoui, E, et al.
(författare)
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Fusion of the FUS and ATF1 genes in a large, deep-seated angiomatoid fibrous histiocytoma
- 2002
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Ingår i: Diagnostic Molecular Pathology. - 1052-9551. ; 11:3, s. 157-162
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Tidskriftsartikel (refereegranskat)abstract
- We report a case of a large, deep-seated, diagnostically difficult angiomatoid fibrous histiocytoma. The neoplastic cells were positive for vimentin, calponin, CD99, and, focally, for desmin and contained intertwining cytoplasmic processes joined by desmosomelike junctions. Fusion of codon 175 of the FUS gene to codon 110 of the ATF1 gene was detected by reverse transcription-polymerase chain reaction. Because identical fusion of the FUS and ATF1 genes has been recently reported in another case of angiomatoid fibrous histiocytoma, fusion of these genes may be characteristic for at least a subset of these tumors.
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| 8. |
- Storlazzi, CT, et al.
(författare)
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A novel chromosomal translocation t(3;7)(q26;q21) in myeloid leukemia resulting in overexpression of EVI1
- 2004
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Ingår i: Annals of Hematology. - : Springer Science and Business Media LLC. - 1432-0584 .- 0939-5555. ; 83:2, s. 78-83
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Tidskriftsartikel (refereegranskat)abstract
- The EVI1 proto-oncogene encodes a nuclear zinc finger protein that acts as a transcription repressor factor. In myeloid leukemia it is often activated by chromosomal rearrangements involving band 3q26, where the gene has been mapped. Here we report two leukemia cases [a chronic myeloid leukemia blast crisis (CML-BC) and an acute myeloid leukemia (AML) M4] showing a t(3;7)(q26;q21) translocation in a balanced and unbalanced form, respectively. Fluorescent in situ hybridization (FISH) analysis revealed that both patients showed a breakpoint on chromosome 3 inside the clone RP11-33A1 containing the EVI1 oncogene and, on chromosome 7, inside the clone RP11-322M5, partially containing the CDK6 oncogene which is a D cyclin-dependent kinase gene, observed to be overexpressed and disrupted in many hematological malignancies. Reverse transcriptase polymerase chain reaction (RT-PCR) analysis showed overexpression of EVI1 in both cases, but excluded the presence of any CDK6/EVI1 fusion transcript. CDK6 expression was also detected. Together, these data indicate that EVI1 activation is likely due not to the generation of a novel fusion gene with CDK6 but to a position effect dysregulating its transcriptional pattern.
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