Search

A new high pressure liquid chromatography (HPLC) method has been used to measure urinary sugar levels for the intestinal permeability test with cellobiose and mannitol (C/M test). Urinary specimens have been prepared by simple filtration through a Millipore membrane. The method is highly sensitive (minimal detectable concentration of urinary sugars = 0.01 mg/ml) and reproducible (coefficient variation between samples = 0.47% for cellobiose and 0.25% for mannitol). By this method a C/M test has been performed in a large series of gastroenterological patients. High values of mean urinary C/M percentage recovery ratio (C/M%) were found in 30 children with active gluten-sensitive enteropathy compared with controls (0.42 +/- 0.66 versus 0.014 +/- 0.005). In 44 treated celiacs and 34 children with chronic nonspecific diarrhea the mean C/M%s were 0.027 +/- 0.018 and 0.021 +/- 0.012, respectively. The results of this study confirm that the C/M test is a valuable investigation in the diagnostic studies of children with chronic diarrhea. The simple HPLC method described for the determination of urinary sugar levels should allow a wider diffusion of this test.

Human milk has always been the reference parameter for the preparation of commercial formulae. The advent of more advanced technologies has enabled increasingly precise information on the composition of human milk to be obtained. Our knowledge in the field of carbohydrates also has improved considerably in the last few years following the pioneering studies of Montreuil (1, 2). From a quantitative point of view it has been demonstrated that in the different phases of lactation, in addition to lactose, human milk contains a consistent amount of oligosaccharides (about 20 g/1 in colostrum and 10–13 g/l in mature milk) (3, 4), whereas monosaccharides make up only about 1% of the total carbohydrates (4). More than 100 different types of oligosaccharides have been identified so far in human milk (5–7), mostly tri-octasaccharides (8). From a biochemical point of view, oligosaccharides are constituted by glucose, galactose, N-acetyl-glucosamine, fucose and sialic acid, and present a linear or branching structure (5). Little is known yet about their physiological role or metabolic fate (9); nevertheless, it has been demonstrated that, in addition to their nutritional function, they participate also in the regulation of the intestinal ecosystem, encouraging the growth of bifid flora (10) and contributing to the defense mechanisms against pathogens in various organs (11–13).

Abstract 2494 Introduction. Acute myeloid leukemia (AML) is an heterogeneous disease with known specific recurrent genetic aberrations. The continuous and increasing identification of new genetic mutations has permitted to identify new subgroups with different prognosis. In the present work we evaluated the incidence of rare genetic abnormalities in pediatric AML such as del(4)(q12)FIP1L1-PDGFRA, t(16;21)(p11;q22)FUS-ERG, t(8;16)(p11;p13)MOZ-CBP, t(11;17)(q23;q12–21)MLL-AF17, t(4;11)(q35;q23)MLL-ArgB2, t(5;11)(q35;p15.5)NUP98-NSD1, t(3;5)(q25;q34)NPM1-MLF1, and MLLPTD. Methods. We selected 306 patients with AML other than acute promyelocytic leukemia, negative for known recurrent genetic abnormalities involving MLL, CBF-beta and FLT3 genes. RNA was extracted from fresh bone marrow at diagnosis, and multiplex RT-PCR was employed. Sequencing by Sanger method was applied to all positive cases to characterize breakpoints of fusion. The Kaplan-Meier method was used for estimating the probability of event-free survival (EFS). Results. We identified one patient each positive for t(16;21)(p11;q22)FUS-ERG, t(11;17)(q23;q12–21)MLL-AF17, and t(4;11)(q35;q23)MLL-ArgB2, respectively, this suggesting that these rearrangements are rare in pediatric AML. 2/306 patients had del(4)(q12)FIP1L1/PDGFRA, and 4/306 the t(8;16)(p11;p13)MOZ-CBP; both these anomalies should be investigated in larger cohorts for definining their prognostic value. Interestingly 6/306 (2%) patients had the t(3;5)(q25;q34)NPM1-MLF1, 6/306 (2%) the MLLPTD, and 8/306 (2.6%) were found to carry the t(5;11)(q35;p15.5)NUP98-NSD1. Since the t(5;11) fusion was recently associated to FLT3ITD, we enlarged the screening to 42 de novo AML harbouring FLT3ITD mutation enrolled in the AIEOP-LAM 2002 protocol finding that 6 of them (14%) had the NUP98-NSD1 fusion gene. We documented a poor EFS for patients with t(5;11)NUP98-NDS1 (n=12) as compared to patients negative for molecular lesions and enrolled in the LAM 2002-AIEOP protocol (25% vs 53.1% at 3 years, p Conclusions. We provide evidence that NUP98-NSD1 may be considered a recurrent translocation in pediatric AML with poor prognosis. Being cryptic to conventional karyotyping, we confirmed the need of using molecular approaches for a proper identification of this anomaly. We also suggest that the NUP98-NSD1 fusion gene be considered for a better evaluation of the FLT3ITD+ patients. Disclosures: No relevant conflicts of interest to declare.