| 1. |
- Alm, Sofie J., 1988, et al.
(författare)
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Minimal residual disease monitoring in childhood B lymphoblastic leukemia with t(12;21)(p13;q22); ETV6-RUNX1: concordant results using quantitation of fusion transcript and flow cytometry.
- 2017
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Ingår i: International journal of laboratory hematology. - : Wiley. - 1751-553X .- 1751-5521. ; 39:2, s. 121-128
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Tidskriftsartikel (refereegranskat)abstract
- The translocation t(12;21)(p13;q22) resulting in the fusion gene ETV6-RUNX1, is the most frequent gene fusion in childhood B lymphoblastic leukemia. In the Nordic Society of Paediatric Haematology and Oncology ALL-2008 treatment protocol, treatment stratification in B-lineage ALL is based on results of minimal residual disease (MRD) analysis with fluorescence-activated cell sorting (FACS). In this study, we determined whether RT-qPCR of the ETV6-RUNX1 fusion transcript can be a reliable alternative for MRD analysis.Seventy-eight bone marrow samples from 29 children at diagnosis and day 15, 29, and 78 during treatment were analyzed for MRD with FACS and with quantitative reverse transcription polymerase chain reaction (RT-qPCR). Fusion transcript MRD was defined as the ETV6-RUNX1/GUSB ratio at the follow-up time point (day 15/29/78) divided with the ETV6-RUNX1/GUSB ratio at diagnosis (%).MRD analysis with FACS and with RT-qPCR of ETV6-RUNX1 fusion transcript showed strong correlation. All cases showed concordant results at the treatment stratifying time points day 29 and day 78, when comparing the two methods with a cutoff set to 0.1%.RT-qPCR is a valuable addition and could also be an alternative to FACS in cases where FACS is not achievable for MRD analysis.
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| 2. |
- Béné, Marie C., et al.
(författare)
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Unsupervised flow cytometry analysis in hematological malignancies : A new paradigm
- 2021
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Ingår i: International Journal of Laboratory Hematology. - : Wiley. - 1751-5521 .- 1751-553X. ; 43:S1, s. 54-64
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Forskningsöversikt (refereegranskat)abstract
- Ever since hematopoietic cells became “events” enumerated and characterized in suspension by cell counters or flow cytometers, researchers and engineers have strived to refine the acquisition and display of the electronic signals generated. A large array of solutions was then developed to identify at best the numerous cell subsets that can be delineated, notably among hematopoietic cells. As instruments became more and more stable and robust, the focus moved to analytic software. Almost concomitantly, the capacity increased to use large panels (both with mass and classical cytometry) and to apply artificial intelligence/machine learning for their analysis. The combination of these concepts raised new analytical possibilities, opening an unprecedented field of subtle exploration for many conditions, including hematopoiesis and hematological disorders. In this review, the general concepts and progress achieved in the development of new analytical approaches for exploring high-dimensional data sets at the single-cell level will be described as they appeared over the past few years. A larger and more practical part will detail the various steps that need to be mastered, both in data acquisition and in the preanalytical check of data files. Finally, a step-by-step explanation of the solution in development to combine the Bioconductor clustering algorithm FlowSOM and the popular and widely used software Kaluza® (Beckman Coulter) will be presented. The aim of this review was to point out that the day when these progresses will reach routine hematology laboratories does not seem so far away.
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| 3. |
- Hedeland, Ylva, 1974-, et al.
(författare)
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Hemolysis interference in 10 coagulation assays on an instrument with viscosity-based, chromogenic, and turbidimetric clot detection
- 2020
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Ingår i: International Journal of Laboratory Hematology. - : Wiley. - 1751-5521 .- 1751-553X. ; 42:3, s. 341-349
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Tidskriftsartikel (refereegranskat)abstract
- INTRODUCTION: Hemolysate in plasma samples from patients may cause misleading results in coagulation assays. Even though modern coagulation instruments often are equipped with modules that can detect hemolysis, icterus, and lipemia (HIL), studies that report the influence of these interferences are still limited. The present paper focuses on the influence of hemolysis on 10 coagulation assays.METHODS: Artificial hemolysis was created by freezing/thawing, and the hemolysates generated were added to pools of patient plasma. Pathological and normal levels were pooled separately. These spiked samples were analyzed on a STA R Max 2 instrument. The coagulation assays evaluated utilize clot, chromogenic, or immunoturbidimetric detection.RESULTS: Four of the evaluated assays were not influenced by hemolysis: fibrinogen, von Willebrand factor antigen, activated partial thromboplastin time, and factor VIII. Interestingly, normal and slightly elevated prothrombin time (INR < 2.0) was insensitive to hemolysis, whereas samples with a high INR (≥2.0) exhibited falsely high readings. The assays for antithrombin and fibrin D-dimer displayed an intermediate sensitivity to hemolysis. The most sensitive assay turned out to be anti-Xa, followed by protein C and protein S. For the anti-Xa assay, the results are decreased by 10% already at 0.5 g/L hemoglobin.CONCLUSION: The present study shows that hemolysis affects several of commonly used coagulation assays. Since the sensitivity for hemolysis is dependent on the brand of the assay as well as the instrument and principle of measurement, it is necessary to evaluate the influence of each specific combination.
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| 5. |
- Karlsson, Lene, et al.
(författare)
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Fusion transcript analysis reveals slower response kinetics than multiparameter flow cytometry in childhood acute myeloid leukaemia
- 2022
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Ingår i: International Journal of Laboratory Hematology. - : Wiley. - 1751-5521 .- 1751-553X. ; 44:6, s. 1094-1101
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Tidskriftsartikel (refereegranskat)abstract
- Introduction Analysis of measurable residual disease (MRD) is increasingly being implemented in the clinical care of children and adults with acute myeloid leukaemia (AML). However, MRD methodologies differ and discordances in results lead to difficulties in interpretation and clinical decision-making. The aim of this study was to compare results from reverse transcription quantitative polymerase chain reaction (RT-qPCR) and multiparameter flow cytometry (MFC) in childhood AML and describe the kinetics of residual leukaemic burden during induction treatment. Methods In 15 children who were treated in the NOPHO-AML 2004 trial and had fusion transcripts quantified by RT-qPCR, we compared MFC with RT-qPCR for analysis of MRD during (day 15) and after induction therapy. Eight children had RUNX1::RUNX1T1, one CBFB::MYH11 and six KMT2A::MLLT3. Results When >= 0.1% was used as cut-off for positivity, 10 of 22 samples were discordant. The majority (9/10) were MRD positive with RT-qPCR but MRD negative with MFC, and several such cases showed the presence of mature myeloid cells. Fusion transcript expression was verified in mature cells as well as in CD34 expressing cells sorted from diagnostic samples. Conclusions Measurement with RT-qPCR suggests slower response kinetics than indicated from MFC, presumably due to the presence of mature cells expressing fusion transcript. The prognostic impact of early measurements with RT-qPCR remains to be determined.
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| 7. |
- Larsson, Sara Marie, et al.
(författare)
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Soluble Transferrin Receptor during infancy and reference intervals for the Roche Cobas platform
- 2021
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Ingår i: International Journal of Laboratory Hematology. - : John Wiley & Sons. - 1751-5521 .- 1751-553X. ; 43:3, s. 378-386
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Tidskriftsartikel (refereegranskat)abstract
- Introduction Infant iron status assessments may be difficult to interpret due to infections. The soluble transferrin receptor (sTfR) has been suggested as a biomarker mainly unaffected by the acute phase response. Reference intervals reflecting dynamics of infant growth first year in life are not well established. Methods The sTfR and CRP concentrations were measured in samples from 451 term infants with the Roche Cobas platform in umbilical cord, at 48-96 hours, 4 and 12 months. Reference values were constructed as the 2.5th and 97.5th percentiles. The relationship between CRP concentrations >1 mg/L and sTfR was tested by Kendall correlation. Results Reference intervals for girls and boys were 2.4-9.5 mg/L at birth, 2.9-8.4 mg/L at 48-96 hours, 2.6-5.7 mg/L at 4 months and 3.0-6.3 mg/L at 12 months. No differences between sexes were observed except for at 4 months. sTfR did not covariate with CRP concentrations >1 mg/L except in 48-96 hours samples. Conclusion This study reports reference intervals for sTfR from birth to 12 months of age in a large group of infants in a low-risk area for iron deficiency. sTfR might add value to infant iron status diagnostics since no covariation with CRP was found at birth, at 4 months or at 12 months.
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| 11. |
- Pettersson, Louise, et al.
(författare)
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Comparison of RNA- and DNA-based methods for measurable residual disease analysis in NPM1-mutated acute myeloid leukemia
- 2021
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Ingår i: International Journal of Laboratory Hematology. - : Wiley. - 1751-5521 .- 1751-553X. ; 43:4, s. 664-674
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Tidskriftsartikel (refereegranskat)abstract
- Introduction: Reverse transcriptase quantitative PCR (RT-qPCR) is considered the method of choice for measurable residual disease (MRD) assessment in NPM1-mutated acute myeloid leukemia (AML). MRD can also be determined with DNA-based methods offering certain advantages. We here compared the DNA-based methods quantitative PCR (qPCR), droplet digital PCR (ddPCR), and targeted deep sequencing (deep seq) with RT-qPCR. Methods: Of 110 follow-up samples from 30 patients with NPM1-mutated AML were analyzed by qPCR, ddPCR, deep seq, and RT-qPCR. To select DNA MRD cutoffs for bone marrow, we performed receiver operating characteristic analyses for each DNA method using prognostically relevant RT-qPCR cutoffs. Results: The DNA-based methods showed strong intermethod correlation, but were less sensitive than RT-qPCR. A bone marrow cutoff at 0.1% leukemic DNA for qPCR or 0.05% variant allele frequency for ddPCR and deep seq offered optimal sensitivity and specificity with respect to 3 log(10) reduction of NPM1 transcripts and/or 2% mutant NPM1/ABL. With these cutoffs, MRD results agreed in 95% (191/201) of the analyses. Although more sensitive, RT-qPCR failed to detect leukemic signals in 10% of samples with detectable leukemic DNA. Conclusion: DNA-based MRD techniques may complement RT-qPCR for assessment of residual leukemia. DNA-based methods offer high positive and negative predictive values with respect to residual leukemic NPM1 transcripts at levels of importance for response to treatment. However, moving to DNA-based MRD methods will miss a proportion of patients with residual leukemic RNA, but on the other hand some MRD samples with detectable leukemic DNA can be devoid of measurable leukemic RNA.
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| 14. |
- Taxiarchis, Apostolos, et al.
(författare)
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Extracellular vesicles in plasma and cerebrospinal fluid in patients with COVID-19 and neurological symptoms
- 2024
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Ingår i: International Journal of Laboratory Hematology. - : John Wiley & Sons. - 1751-5521 .- 1751-553X. ; 46:1, s. 42-49
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Tidskriftsartikel (refereegranskat)abstract
- Introduction:Increased levels of extracellular vesicles (EVs) are associated with haemostatic disturbances in various clinical settings. However, their role in COVID-19 patients is still not fully clear. In the present study we investigated EVs in plasma from patients with COVID-19 and neurological symptoms in relation to the activation of coagulation.Methods:Nineteen COVID-19 patients with neurological symptoms and twenty-three aged-matched healthy individuals were included. Global coagulation assays were performed and levels of EVs were determined by flow-cytometry in plasma and cerebrospinal fluid (CSF).Results:A procoagulant state characterized by significantly increased overall coagulation- (OCP) and overall haemostatic potential (OHP), diminished overall fibrinolytic potential (OFP) together with a denser fibrin structure was found in patients with COVID-19. Flow cytometry revealed elevated levels of plasma circulating EVs derived from neutrophils (MPO+) and platelets (CD61+), as well as EVs expressing phosphatidylserine (PS+) and complement component C5b-9 (TCC+) in patients with COVID-19 compared with controls. The concentrations of PS+, CD61+ and TCC+ EVs were positively correlated with OCP and OHP in COVID-19 patients. Moreover, we identified CD61+, MPO+ and endothelial cell-derived EVs, as well as EVs exposing PS and TCC in the CSF of patients suffering from neurological symptoms during COVID-19.Conclusion:The unique finding in this study was the presence of EVs in the CSF of COVID-19 patients with neurologic manifestations as well as higher expression of complement protein on circulating plasma EVs. EVs may indicate blood-brain barrier damage during SARS-COV-2 infection.
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| 19. |
- Fleyeh, Hasan, et al.
(författare)
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Eigen-based traffic sign recognition
- 2011
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Ingår i: IET Intelligent Transport Systems. - Stevenage : The Institiution of Engineering and Technology. - 1751-956X .- 1751-9578. ; 5:3, s. 190-196
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Tidskriftsartikel (refereegranskat)abstract
- This paper’s purpose is to introduce Eigen-based traffic sign recognition. This technique is based on invoking the PCA algorithm to choose the most effective components of traffic sign images to classify an unknown traffic sign. A set of weights are computed from the most effective Eigen vectors of the traffic sign. By using the Euclidean distance, unknown traffic sign images are then classified. The approach was tested on two different databases of traffic sign’s borders and speed limit pictograms which were extracted automatically from real-world images. A classification rate of 96.8% and 97.9% was achieved for these two databases. To check the robustness of this approach, non-traffic sign objects and occluded signs were invoked. A performance of 71% was achieved when occluded signs are used. When signs were rotated 10 degrees around their centre, the performance became 89% when traffic signs’ outer shapes were used and for rotated speed limit pictograms the result was 80%.
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| 23. |
- Lee, SH, et al.
(författare)
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Definition of ringed sideroblasts
- 2010
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Ingår i: INTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY. - 1751-5521. ; 32:1, s. E185-E185
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Tidskriftsartikel (övrigt vetenskapligt/konstnärligt)
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| 24. |
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| 26. |
- Dahlbäck, Björn
(författare)
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Pro- and anticoagulant properties of factor V in pathogenesis of thrombosis and bleeding disorders
- 2016
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Ingår i: International Journal of Laboratory Hematology. - : Wiley. - 1751-5521. ; 38, s. 4-11
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Tidskriftsartikel (refereegranskat)abstract
- Factor V (FV) serves an important role in the regulation of blood coagulation, having both pro- and anticoagulant properties. The circulating high molecular weight single-chain FV molecule undergoes a series of proteolytic cleavages during both activation of coagulation and during anticoagulant regulation of coagulation by activated protein C (APC). It is noteworthy that mutations in the factor V gene (F5) either cause thrombosis or bleeding. New insights into the importance and complexity of FV functions have been generated from elucidation of the pathogenic mechanisms of two familial mutations in the F5 gene. The first mutation was identified as a result of the discovery of APC resistance as the most common risk factor for venous thrombosis. The mutation (FV Leiden) predicts the Arg506Gln replacement, which impairs the normal regulation of FVa by APC, as the Arg506 site is an important APC cleavage site. In addition, elucidation of APC resistance resulted in the discovery of the anticoagulant APC cofactor activity of FV. The second FV mutation (FVA2440G), identified in a family with an autosomal dominant bleeding disorder, has led to the discovery of an alternative splicing generating a previously unidentified FV isoform (FV-Short), which inhibits coagulation via an unexpected and intriguing mechanism involving the coagulation inhibitor TFPI-α. These are naturally occurring mutations in the F5 gene that have generated new knowledge on the role of FV in regulation of coagulation and the importance of genetic risk factors for thrombosis and bleeding.
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| 30. |
- Rajab, Amr, et al.
(författare)
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Ten-color 15-antibody flow cytometry panel for immunophenotyping of lymphocyte population
- 2017
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Ingår i: International Journal of Laboratory Hematology. - : Wiley. - 1751-5521. ; 39, s. 76-85
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Forskningsöversikt (refereegranskat)abstract
- We have developed a lymphoproliferative disorder screening tube (LPD-ST) with the aim to provide comprehensive immunophenotyping of lymphocyte subsets with minimal need for additional testing. The LPD-ST consists of CD4/kappa FITC, CD8/lambda PE, CD3/CD14ECD, CD38PC5.5, CD20/CD56PC7, CD10APC, CD19APC-A700, CD5APC-A750, CD57/CD23PB and CD45KO. The LPD-ST was validated against previously used lymphocyte subset panels in Canada (n=60) and in Sweden (n=43) and against the OneFlow™ LST (n=60). The LPD-ST panel was then implemented in clinical practice using dried monoclonal antibody reagents (Duraclone®) on 649 patient samples in Sweden. In 204 of 649 samples (31%), a monotypic B-cell population was found. Of these cases, a final diagnosis could be rendered in 106 cases (52%), and in the remainder, additional B-cell immunophenotyping was performed. In 20 (3%) samples, an aberrant T-cell population was confirmed by additional testing. Of 425 samples diagnosed as normal/reactive lymphoid tissue, 50 (12%) required additional immunophenotyping, mostly due to an abnormal CD4/CD8 ratio. The LPD-ST tube significantly minimizes the need for additional testing, improves the turn-around time, and reduces the cost of LPD immunophenotyping. It is also suitable for investigating paucicellular samples such as cerebrospinal fluid or fine needle aspirates.
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| 31. |
- Tesfa, D
(författare)
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ACQUIRED NEUTROPENIAS
- 2014
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Ingår i: INTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY. - 1751-5521. ; 36, s. 14-14
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)
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| 32. |
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| 33. |
- Pham, Tuan A., et al.
(författare)
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Analytical model for the design of piled embankments considering cohesive soils
- 2022
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Ingår i: Geosynthetics International. - : Emerald. - 1072-6349 .- 1751-7613. ; 29:4, s. 369-388
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Tidskriftsartikel (refereegranskat)abstract
- Geosynthetic-reinforced and pile-supported (GRPS) systems have already proven their good performance in supporting embankments constructed over soft soil. The load transfer mechanism in GRPS embankments depends on the complex interaction between the soil in place, the structural elements and the embankment's soil type (cohesive or cohesionless). However, the cohesion influence of the embankment soil has not been well investigated as it is often not considered in the design of such systems. The main aim of this study is to present an analytical model for GRPS embankments that combines several phenomena such as the concentric arches model in cohesive fill soils, the hyperbolic model for the isochrone geogrid curve, and subsoil's consolidation. Three-dimensional numerical analyses are also conducted to evaluate the embankment soil cohesion influence on the soil arching. Both the numerical and analytical results agree that the cohesive embankment fills strengthen the soil arching effect and increase the efficacy if compared with cohesionless embankment fills. A comparison of the analytical model with measured data and other design methods for full-scale field tests proved the proposed model efficiency. The proposed analytical model therefore can be applicable for GRPS embankments with cohesive and non-cohesive fill soils.
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