A Comparison of Two Molecular Methods for Detecting CALR Mutations in Myeloproliferative Neoplasms.

BACKGROUND: Myeloproliferative neoplasms (MPN) are hematopoietic disorders characterized by abnormal proliferation of the myeloid lineage. Three classic subtypes are polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). These disorders are well known for their association with the JAK2 V617F mutation, in addition to mutations in MPL exon 10, and JAK2 exon 12. CALR mutations were detected in approximately 20% to 25% of patients with ET and PMF and not in patients with PV. Most CALR mutations were deletions and insertions in exon 9, which caused frameshift mutations. METHODS: This study included 60 Taiwanese patients with MPN. We identified CALR mutations in patients with MPN using the high-resolution melting (HRM) analysis. Additionally, the HRM analysis was compared with ipsogen CALR RGQ PCR. To confirm the results of HRM and ipsogen CALR RGQ PCR, sequencing analysis was also conducted all the samples. RESULTS: Up to 6.25% of CALR mutations were successfully detected in patients with MPN using HRM analysis. Eight out of 65 patients (12.3%) were positive for the presence of CALR mutation, including p.L367fs*46 and p.K385fs*47. The results proved 100% comparable to those obtained using ipsogen CALR RGQ PCR. CONCLUSIONS: The HRM analysis and ipsogen CALR RGQ PCR are feasible and reliable techniques for the detection of CALR mutation. Furthermore, HRM offers several benefits, for example, it is time-saving, inexpensive, and does not require many personnel.

Effective Three-Stage Demosaicking Method for RGBW CFA Images Using The Iterative Error-Compensation Based Approach.

As the color filter array (CFA)2.0, the RGBW CFA pattern, in which each CFA pixel contains only one R, G, B, or W color value, provides more luminance information than the Bayer CFA pattern. Demosaicking RGBW CFA images I R G B W is necessary in order to provide high-quality RGB full-color images as the target images for human perception. In this letter, we propose a three-stage demosaicking method for I R G B W . In the first-stage, a cross shape-based color difference approach is proposed in order to interpolate the missing W color pixels in the W color plane of I R G B W . In the second stage, an iterative error compensation-based demosaicking process is proposed to improve the quality of the demosaiced RGB full-color image. In the third stage, taking the input image I R G B W as the ground truth RGBW CFA image, an I R G B W -based refinement process is proposed to refine the quality of the demosaiced image obtained by the second stage. Based on the testing RGBW images that were collected from the Kodak and IMAX datasets, the comprehensive experimental results illustrated that the proposed three-stage demosaicking method achieves substantial quality and perceptual effect improvement relative to the previous method by Hamilton and Compton and the two state-of-the-art methods, Kwan et al.'s pansharpening-based method, and Kwan and Chou's deep learning-based method.

Analytical Performance of the VIDAS® High-Sensitivity Troponin I Assay and the Beckman Coulter Unicel® DXI AccuTnI+3 Assay in a Stat Laboratory.

BACKGROUND: The aim of this study was to compare the validity of two different cTnI assay methodologies. METHODS: We collected 82 plasma samples from a stat laboratory. The plasma values of cTnI ranged from 0.012 to 29.715 ng/mL when tested on the Access® platform and from 4.5 to >40,000 ng/L when tested on the VIDAS platform. The patients included 34 females ranging in age from 49 to 100 years of age [76.7 ± 12 years] and 48 males ranging from 29 to 97 years of age [69.7 ± 12 years]. RESULTS: Our results showed that the correlation between the two troponin results was r2 = 0.9836 (p < 0.001). In this study, the kappa statistic (0.89) indicated a high degree of agreement between the VIDAS® High-sensitivity Troponin I assay and the Beckman Coulter Unicel® DXI AccuTnI+3 assay. CONCLUSIONS: In summary, the VIDAS® High-sensitivity Troponin I assay is a reliable and feasible method for determining the levels of cTnI in plasma, but it requires manual operation, hands-on technical expertise, and time.

Transcription Factor Elf3 Modulates Vasopressin-Induced Aquaporin-2 Gene Expression in Kidney Collecting Duct Cells.

Aquaporin-2 (AQP2) is a molecular water channel protein responsible for water reabsorption by the kidney collecting ducts. Many water balance disorders are associated with defects in AQP2 gene expression regulated by the peptide hormone vasopressin. Here, we studied roles of Elf3 (E26 transformation-specific (Ets)-related transcription factor 3) in AQP2 gene expression in the collecting duct cells (mpkCCD). Vasopressin increased AQP2 mRNA and protein levels without affecting AQP2 mRNA degradation, indicative of transcriptional regulation. Elf3 knockdown and overexpression, respectively, reduced and increased AQP2 gene expression under basal and vasopressin-stimulated conditions. However, the vasopressin-to-basal ratios of AQP2 gene expression levels remained constant, indicating that Elf3 does not directly mediate vasopressin response but modulates the level of AQP2 gene expression inducible by vasopressin. The Elf3-modulated AQP2 gene expression was associated with AQP2 promoter activity, in line with Elf3's ability to bind an Ets element in the AQP2 promoter. Mutation in the Ets element reduced both basal and vasopressin-stimulated AQP2 promoter activity, again without affecting vasopressin-to-basal ratios of the AQP2 promoter activity. Lithium chloride reduced both Elf3 and AQP2 mRNA in the mpkCCD cells as well as in mouse kidney inner medulla. We conclude that Elf3 modulates AQP2 promoter activity thereby gauging vasopressin-inducible AQP2 gene expression levels. Our data provide a potential explanation to lithium-induced nephrogenic diabetes insipidus where lithium reduces Elf3 and hence AQP2 abundance.