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Following reverse transcription, cDNA was quantified by real-time PCR (ABI PRISM 7500 Real-Time PCR System) at standard conditions using TaqMan Universal PCR Master Mix (Applied Biosystems, Foster City, CA, USA). Expression levels of the ��-subunit of high-affinity IgE receptor (FCER1A) (Hs00175232_m1) were normalized against 18S rRNA endogenous control (Applied Biosystems). All measurements were taken in triplicate for each sample and time point, and the relative expression was analyzed using the ����Ct method. The number of Fc��RI receptors per basophil (CD123?+?/HLA-DR - cells) was analyzed using a FITC-conjugated antibody to Fc��RI (eBioscience, San Diego, CA, USA) and standard curve of Calibration Beads (Dako Cytomation, Glostrup, Denmark) as previously described [20]. For the absolute basophil count (CD123?+?/HLA-DR - cells), we added AccuCount Fluorescent microbeads (Spherotec Inc., Libertyville, [http://www.selleckchem.com/products/bmn-673.html Talazoparib research buy] IL, USA) to fixed samples before analysis. The absolute number of basophils per ��l of whole blood was calculated using the following equation: (number [http://www.selleckchem.com/products/s-gsk1349572.html http://www.selleckchem.com/products/s-gsk1349572.html] of events for basophil region/number of events for microbead region)?��?(number of microbeds used in test/volume of the whole-blood sample initially used). Data distribution was evaluated using the D'Agostino�CPearson normality test. Depending on the distribution of the data, we used either a Wilcoxon's matched pairs test or a paired t-test. Data were expressed as median (IQR) or mean (95% CI). Probability values (P) of?[https://en.wikipedia.org/wiki/CAPNS1 CAPNS1] response following stimulation with submaximal concentrations from 110 to 5.5?ng/ml decreased significantly in both groups of patients. In adults, the decreases before the first MD of 100?��g were the most significant when tested at submaximal concentrations of 55?ng/ml (median 34.1%; IQR 20.1�C59.0), 27.5?ng/ml (3.9%; 2.3�C7.3), and 11?ng/ml (2.6%; 2.1�C3.8), when compared to before ultra-rush VIT (for 55?ng/ml: 49.5%, 17.9�C78.1, P?