Furthermore, the girls’ erythrocyte count was elevated whereas reticulocytosis count was within the reference values for all the patients The 17-year old met the diagnostic criteria according to the WHO for polycythemia in man. Hyperbilirubinemia was also noted in the oldest boy and a hepatology consultation was recommended. Iron concentration and transferrin saturation exceeded

the norm in all the children, while ferritin concentration, transaminases, creatinine and erythropoietin levels remained within the reference ranges. The coagulation profile, CRP and capillary blood gas tests were also within the norm. HBV and HCV infection was excluded and so were mononucleosis, Belinostat in vitro cytomegalovirus and toxoplasmosis protozoan. The bone marrow biopsy performed in the oldest boy and the girl revealed no deviations from the norm. Molecular studies on the oldest boy excluded a V617F mutation. The characteristic biochemical parameters of the patients are listed in Table I. On the basis of diagnostic indications learn more given in previous publications [13], molecular diagnosis of type 1 hemochromatosis (HFE mutation) was performed on the children. Diagnostic materials – DNA isolated from 200 μl of whole blood collected in EDTA using High Pure PCR Template Preparation Kit (Roche) reagents. DNA fragment consisting of 354 base pairs, which include the H63D and S65C HFE gene region, and a DNA fragment consisting of 276 base pairs, which encompass the C282Y HFE gene

region, which were amplified using multiplex Real-Time PCR. The genotype identification was based on melting curve analysis, using HybProbe probes, based on the specific melting points (Tm): Tm for a normal H63 and S65 HFE genotype = 57 °C, using the 530 nm channel; mutant HFE 63D Tm = 65.5 °C, mutant HFE 65 °C Tm = 52 °C, normal HFE C282 Tm = 56.5 °C, using the 640 nm channel; mutant HFE 282Y Tm = 62 °C. The presence of HFE mutations was confirmed in

all the patients. In the oldest boy a His/Asp phenotype at position 63 of the HFE protein (heterozygous for the HFE gene at position 187, C/G, H63D), the aminophylline 16-year old had a Cys/Tyr phenotype at position 282 of the HFE protein (heterozygous for the HFE gene at position 845, G/A, C282Y), whereas the girl was diagnosed with a homozygous mutation in the HFE 282Y (Tyr/Tyr phenotype, homozygous for the HFE gene at position 845, A/A). All the patients remain under clinical observation in the department. Their hemoglobin concentration and erythrocyte count are comparable with previous tests. For a preliminary assessment of the hematopoietic system, the primary diagnostic tool is a full blood count. Elevated hemoglobin concentrations, as opposed to anemia, are rarely observed during childhood [1] and [2]. In the 3 children mentioned above, elevated levels of hemoglobin were found in full blood counts performed without any specific medical indication in an outpatient setting. The children did not report any complaints or infections.