| 1. |
- Andersson, Charlotta, et al.
(författare)
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Reduction in WT1 Gene Expression During Early Treatment Predicts the Outcome in Patients With Acute Myeloid Leukemia
- 2012
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Ingår i: Diagnostic molecular pathology (Print). - : Lippincott Williams & Wilkins. - 1052-9551 .- 1533-4066. ; 21:4, s. 225-233
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Tidskriftsartikel (refereegranskat)abstract
- Wilms tumor gene 1 (WT1) expression has been suggested as an applicable minimal residual disease marker in acute myeloid leukemia (AML). We evaluated the use of this marker in 43 adult AML patients. Quantitative assessment of WT1 gene transcripts was performed using real-time quantitative-polymerase chain reaction assay. Samples from both the peripheral blood and the bone marrow were analyzed at diagnosis and during follow-up. A strong correlation was observed between WT1 normalized with 2 different control genes (beta-actin and ABL1, P < 0.001). WT1 mRNA level at diagnosis was of no prognostic relevance (P > 0.05). A >= 1-log reduction in WT1 expression in bone marrow samples taken < 1 month after diagnosis significantly correlated with an improved overall survival (P = 0.004) and freedom from relapse (P = 0.010) when beta-actin was used as control gene. Furthermore, a reduction in WT1 expression by >= 2 logs in peripheral blood samples taken at a later time point significantly correlated with a better outcome for overall survival (P = 0.004) and freedom from relapse (P = 0.012). This result was achieved when normalizing against both b-actin and ABL1. These results therefore suggest that WT1 gene expression can provide useful information for minimal residual disease detection in adult AML patients and that combined use of control genes can give more informative results.
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| 2. |
- Botling, Johan, et al.
(författare)
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Impact of thawing on RNA integrity and gene expression analysis in fresh frozen tissue
- 2009
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Ingår i: Diagnostic molecular pathology (Print). - 1052-9551 .- 1533-4066. ; 18:1, s. 44-52
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Tidskriftsartikel (refereegranskat)abstract
- Biobanks of fresh, unfixed human tissue represent a valuable source for gene expression analysis in translational research and molecular pathology. The aim of this study was to evaluate the impact of thawing on RNA integrity and gene expression in fresh frozen tissue specimens. Portions of snap frozen tonsil tissue, unfixed or immersed in RNAlater, were thawed at room temperature for 0 minute, 5 minutes, 30 minutes, 45 minutes, 1 hour, 3 hours, 6 hours, and 16 hours before RNA extraction. Additionally, tonsil tissue underwent repetitive freezing and thawing cycles. RNA integrity was analyzed by microchip gel electrophoresis and gene expression by quantitative real-time polymerase chain reaction for selected genes (FOS, TGFB1, HIF1A, BCL2, and PCNA). Minimal RNA degradation was detected after 30 minutes of thawing in unfixed samples. This degradation was accompanied by relevant changes in gene expression for FOS and BCL2 at 45 minutes. Modified primer design or the use of different housekeeping genes could not rectify the changes for FOS. Repetitive thawing cycles had similar effects on RNA integrity. The incubation of the tissue in RNAlater efficiently prevented RNA degradation. In conclusion, degradation of RNA in frozen tissue occurs first after several minutes of thawing. Already minimal decrease in RNA quality may result in significant changes in gene expression patterns in clinical tissue samples.
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| 3. |
- Karlsson, Mats G., 1960-, et al.
(författare)
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Nasal messenger RNA expression of interleukins 2, 4, and 5 in patients with allergic rhinitis
- 1995
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Ingår i: Diagnostic molecular pathology (Print). - Philadelphia, USA : Lippincott Williams & Wilkins. - 1052-9551 .- 1533-4066. ; 4:2, s. 85-92
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Tidskriftsartikel (refereegranskat)abstract
- In nasal biopsies from 17 adult patients with seasonal allergic rhinitis and from 10 healthy controls, cytokines were analyzed by reverse-transcriptase polymerase chain reaction (RT-PCR). The time-course study during winter included repeated local allergen provocation with subsequent nasal biopsies as well as biopsies taken during pollen season. The RT-PCR for CD44 yielded positive bands in 65 of 71 cases, in which cases mRNA for interleukins 2, 4, and 5 (IL-2, IL-4, and IL-5) were thus investigated by means of seminested PCR. IL-4 mRNA was found almost exclusively in the allergic patients. During provocation a significant increase in IL-4 was noticed compared with controls (p = 0.043). Equally, during the natural pollen season, IL-4 mRNA expression was significantly higher in patients not using nasal corticosteroids compared with those who did (p = 0.011). No differences in IL-2 or IL-5 were observed between the groups. These findings also indicate, together with earlier observations of T-cell activation, a phenotype switch toward T-helper 2 (Th2) cells, and the accumulation (homing) of these T cells in the nasal mucosa, that T cells constitute the main source for IL-4 in the nasal mucosa. Therefore, allergic patients have an increased synthesis of IL-4 when provoked with the allergen, and during natural pollen season this synthesis can be downregulated by corticosteroids. Furthermore, this study exemplifies the versatility of molecular biology in surgical pathology and that even low-copy-number cytokine mRNA can be examined in routinely snap-frozen surgical specimens.
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| 4. |
- Yajloo, Mohsen Mohammadian, et al.
(författare)
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Rapid alpha-1-antitrypsin M-variant genotyping by primer-induced restriction analysis.
- 2007
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Ingår i: Diagnostic molecular pathology : the American journal of surgical pathology, part B. - : Ovid Technologies (Wolters Kluwer Health). - 1052-9551 .- 1533-4066. ; 16:1, s. 54-6
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Tidskriftsartikel (refereegranskat)abstract
- The 4 normal alleles of M1, M2, M3, and M4 are the most common gene products of the human alpha-1-antitrypsin (hAAT). Two single substitutions in M1 are responsible for M3 and M4, whereas 2 substitutions in M1 produce M2. Polymerase chain reaction-restriction fragment length polymorphism analysis of the Arg(101)/His(101) sequence variation can separate M1 and M3 from M2 and M4 alleles. To complete the genotyping procedure of hAAT M variants, the exon-V Glu(376)/Asp(376) sequence variation was directly analyzed using a designer primer with a single-base substitution in its sequence. This substitution induced an artificial site for the same restriction enzyme in the polymerase chain reaction product. The new restriction site was present in M1 and M4 but absent in M2 and M3, which can be applied as a rapid reliable means for the M-variant genotyping of hAAT.
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| 5. |
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| 6. |
- 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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