nonparametric statistics were used to compare the levels of repeated measurements and post hoc non-parametric analysis was applied for between time-point comparisons

nonparametric statistics were used to compare the levels of repeated measurements and post hoc non-parametric analysis was applied for between time-point comparisons. A constant relationship was found between the levels of Ig A in blood and saliva (L-Asparaginase (5000?U/m2/d) (Medac/KYOWA, Germany) during the next twenty-six days, regardless of the stratification to risk group. post hoc non-parametric analysis was applied for between time-point comparisons. A constant relationship was found between the levels of Ig A in blood and saliva (L-Asparaginase (5000?U/m2/d) (Medac/KYOWA, Germany) during the next twenty-six days, regardless of the stratification to risk group. The daunorubicin and vincristine were delivered intravenously on days 8 and 15. The vincristine dose was repeated again on days 22 and 29. On day 11, L-asparaginase was introduced intravenously; the doses were repeated 7 occasions, every 3 days (Fig. ?(Fig.11). Open in a separate window Physique 1 Remission induction treatment regimen (the first 33 days of therapy) for child years acute lymphoblastic leukaemia according to the ALL-IC BFM 2009 protocol with denoted time-points when saliva was collected. 2.3. Collecting samples and laboratory screening Saliva and blood samples were taken at 3 time points: around the first, eighth and 22nd day of treatment (Fig. ?(Fig.1).1). The saliva was collected in the morning, before breakfast and before chemotherapy was given. Each time, about 5 ml of saliva was collected with sterile tubes, then preserved with aprotinin, centrifuged (2000?rpm for 10 minutes) and then stored at -80 degrees Celsius. In the salivary samples, the levels of immunoglobulin A (with use of Human IgA Platinum ELISA BMS by eBioscience) and total protein (with use of TP0300-1KT-total protein kit by SIGMA, Germany) were tested. The levels of immunoglobulin A and total protein in blood were also checked PETCM in a clinically-certified diagnostic laboratory using a Cobas INTEGRA device (Roche). All symptoms that occurred during therapy were recorded. Adverse events (AEs) were assessed and graded with the use of the Common Terminology Criteria for Adverse Events (version 4.0).[13] 2.4. Statistical evaluation The distribution of the quantitative variables was assessed using the Shapiro-Wilk test. nonparametric statistics were used to compare the levels of repeated measurements and non-parametric analysis was applied for between time-point comparisons. All analyses were performed using Statistical Software, version 10.0.A (Stat soft) and a em P /em -value .05 was considered statistically significant. 3.?Results The salivary Ig A level correlated with serum IgA level at diagnosis of ALL. The Spearman correlation rank for the relationship was em r /em ?=?0.28 and em PETCM P /em ?=?.031 (Fig. ?(Fig.22). Open in a separate windows Physique 2 The relationship between levels of immunoglobulin A in blood and saliva, independently of the time point measured ( em r /em ?=?0.28; CI?=?95% and em P /em ?=?.031). During the examined period, the level of salivary IgA decreased from 10.7+/?4.8 to 9.6+/?6.4 between days 1 and 8, and to PETCM 5.7+/?3.9?(ng/mL) on day 22 ( em P /em ?=?.04); (Table ?(Table2).2). This was even more pronounced when the level of salivary immunoglobulin A was normalized with the level of salivary total protein ratio, as the figures decreased from 6,6+/? 3,2 around the first day, to 3,8+/-1,2?(ng/mg) around the last PETCM day of the study ( em P /em ?=?.02); (Table ?(Table22). Table 2 Changes in Mouse monoclonal to OCT4 salivary and serum proteins during remission induction treatment of ALL children. Open in a separate windows In saliva, the significant reduction of immunoglobulin A level was not associated with any changes in total protein, which remained constant during the first 22 days of the therapy. In blood, the total protein content decreased from 6.2+/-0.4 to 5.1+/-0.3?g/dL between days 1 and 22 ( em P /em ?=?.01). The level of serum IgA also decreased, but not significantly. A correlation between lymphocyte count and serum IgA level was observed ( em r /em ?=?0.54; em p /em ? ?.03); however, no correlation was found between lymphocyte count and salivary IgA level ( em r /em ?=?0.06; em P /em ?=?.33); (Fig. ?(Fig.3A3A and B). Total lymphocyte count (per microliter) decreased considerably between day 1 and day 8, and day 22 (2.12+/?0.8 vs 0.41?+/?0.1 vs 1.08?+/0.5; em P /em ? ?.002). More detailed data is given in Table ?Table22. Open in a separate window Physique 3 a, b The significant correlation between lymphocyte PETCM count and serum IgA level ( em r /em ?=?0.54; em P /em ? ?.03); (panel a), and the lack of correlation between lymphocyte count and salivary IgA level ( em r /em ?=??0.06; em P /em ?=?.33); (panel b). No cases of mucositis were noted before day 8 of treatment; however, 4 children were found to be suffering from oral mucosal inflammation until day 22. Three cases of mucositis were graded as 1 (moderate form) according to the common terminology criteria for adverse events criteria while 1 case was graded as 3 (severe). One of 3 patients with mucositis graded as 1.