Calreticulin Mutations in Myeloproliferative Neoplasms: Comparison of Three Diagnostic Methods.

Calreticulin (CALR) mutations have recently been reported in 70-84% of JAK2V617F-negative myeloproliferative neoplasms (MPN), and this detection has become necessary to improve the diagnosis of MPN. In a large single-centre cohort of 298 patients suffering from Essential Thrombocythemia (ET), the JAK2V617F, CALR and MPL mutations were noted in 179 (60%), 56 (18.5%) and 13 (4.5%) respectively. For the detection of the CALR mutations, three methods were compared in parallel: high-resolution melting-curve analysis (HRM), product-sizing analysis and Sanger sequencing. The sensitivity for the HRM, product-sizing analysis and Sanger sequencing was 96.4%, 98.2% and 89.3% respectively, whereas the specificity was 96.3%, 100% and 100%. In our cohort, the product-sizing analysis was the most sensitive method and was the easiest to interpret, while the HRM was sometimes difficult to interpret. In contrast, when large series of samples were tested, HRM provided results more quickly than did the other methods, which required more time. Finally, the sequencing method, which is the reference method, had the lowest sensitivity but can be used to describe the type of mutation precisely. Altogether, our results suggest that in routine laboratory practice, product-sizing analysis is globally similar to HRM for the detection of CALR mutations, and that both may be used as first-line screening tests. If the results are positive, Sanger sequencing can be used to confirm the mutation and to determine its type. Product-sizing analysis provides sensitive and specific results, moreover, with the quantitative measurement of CALR, which might be useful to monitor specific treatments.

Feasibility study of date (Phoenix dactylifera L.) fruit syrup-based natural jelly using central composite design.

A feasibility study of natural fruit jelly from three Algerian raw materials, namely date (Phoenix dactylifera l.) fruit syrup and suspension of orange albedo powder (OAP) in lemon juice (LJ) was performed by response surface methodology (RSM) based on central composite design (CCD). The textural properties of the final jelly were investigated through two dependent variables: hardness and stickiness. The cooking temperature (X1), corresponding to that of thermo stated oil bath, and cooking time (X 2), taken for heating the initial fruit mixture in the oil bath (from ambient temperature without fixing however, the final temperature), were found to be the most influent factors, compared to °Brix of date syrup (X3) and temperature (X4) of the cooling stage following the cooking process. Results have also shown that the second-degree polynomial models correctly fit experimental data (R(2), adjusted R(2) (R(2) adj) and cross-validation (Q(2)) ≈ 1). Considering textural properties of commercial jellies as a reference, it was found that the cooking temperature of 155 °C for 10 min gave a jelly with suitable textural properties. On the other hand, FT-IR spectra revealed that the structure of such jelly was partially close to that of pectin molecules. Finally, the color analysis in the CIELab system of the fruit mixture over the cooking process showed that both lightness (L(*)) and a*/b* ratio were not affected by the experienced temperature range (80-155 °C).