Analysis of Lyso-Globotriaosylsphingosine in Dried Blood Spots

Recently, lyso-globotriaosylsphingosine (lyso-Gb3) was found to be elevated in plasma of treatment naive male patients and some female patients with Fabry Disease (FD). This study tested whether lyso-Gb3 could be analyzed in dried blood spots (DBS) from filter cards and whether concentrations are elevated in newborn infants with FD. Lyso-Gb3 concentrations were analyzed in DBS following extraction using a novel HPLC-mass spectrometry (MS)/MS method. Lyso-Gb3 levels in DBS were above the lower limit of quantitation (0.28 ng/mL) in 5/17 newborn FD infants (16 males; range: 1.02-8.81 ng/mL), but in none of the newborn controls, in all 13 patients (4 males) with classic FD (range: 2.06-54.1 ng/mL), in 125/159 Taiwanese individuals with symptomatic or asymptomatic FD who carry the late onset alpha-galactosidase A (GLA) mutation c.936+919G>A (IVS4+919G>A) (3.75+/-0.69 ng/mL; range: 0.418-3.97 ng/mL) and in 20/29 healthy controls (0.77+/-0.24 ng/mL; range: 0.507-1.4 ng/mL). The HPLC-MS/MS method for analysis of lyso-Gb3 is robust and yields reproducible results in DBS in patients with FD. However, concentrations of lyso-Gb3 were below the limit of quantitation in most newborn infants with FD rendering this approach not suitable for newborn screening. In addition, most females with the late onset mutation have undetectable lyso-Gb3 concentrations.

Bcl-2 oncoprotein and sphingolipids blood levels in childhood lymphoma and leukemia

In this work, BCL-2 oncoprotein and sphingolipids levels were determined in blood of 60 children including 27 children with non- Hodgkin's lymphoma[NHL], 20 children with acute lymphoblastic leukemia [ALL] and 13 children with acute monoblastic leukemia [AML]. The study also included 11 healthy controls with comparable ages to patients. The study revealed significant increase in BCL-2 oncoprotein levels in patients compared with the controls; where levels in cases with AML were lower than cases with ALL or NHL. All patients had BCL-2 levels higher than cut-off levels i.e. 100% sensitivity. Children with enlarged liver and/or spleen showed higher BCL-2 oncoprotein levels compared with those without this enlargement. There was significant positive correlation between BCL-2 oncoprotein levels and splenic size. The levels of sphingolipids were significantly increased in patients compared with controls. Patients with ALL and NHL had significantly higher levels than cases with AML. Among cases with lymphoma and leukemia, 53.3% had levels above the cut-off levels. Children with liver and/or splenic enlargement had higher sphingolipid levels than children without enlargement. Sphingolipid levels correlated significantly in a positive manner with the size of the liver and spleen