Real-Time PCR and Droplet Digital PCR: two techniques for detection of the JAK2(V617F) mutation in Philadelphia-negative chronic myeloproliferative neoplasms.

INTRODUCTION: Philadelphia-negative chronic myeloproliferative neoplasms (MPNs) are clonal disorders that present JAK2(V617F) mutation in 50-95% of cases. The main objective of this study was the comparison of two PCR methods, real-time (qPCR) and droplet digital PCR (DD-PCR) for detection of the JAK2(V617F) mutation, to assess analytic sensitivity, specificity, and feasibility of the two methods. METHODS: Ninety-nine patients with MPN of 225 presenting the JAK2(V617F) mutation by qPCR have been evaluated by DD-PCR also. RESULTS: We demonstrated an absolute concordance in terms of specificity between the two methods, DD-PCR showing a higher sensitivity (half a log higher than qPCR). As expected, a progressive increase of mutant allele burden was observed from essential thrombocythemia (ET) to polycythemia vera (PV) and primary myelofibrosis (PMF) to secondary myelofibrosis (SMF). CONCLUSION: In conclusion, our study showed that DD-PCR could represent a new and promising technological evolution for detection of JAK2 mutation in MPNs.

The c.480C>G polymorphism of hOCT1 influences imatinib clearance in patients affected by chronic myeloid leukemia.

The aim of the study was to investigate any possible influence of polymorphisms of transmembrane transporters human organic cation transporter 1 (hOCT1), ABCB1, ABCG2 on imatinib pharmacokinetics in 33 men and 27 women (median age and range, 56 and 27-79 years, respectively) affected by chronic myeloid leukemia. A population pharmacokinetic analysis was performed to investigate imatinib disposition in every patient and the role of transporter polymorphisms. Results showed that the α1-acid glycoprotein and the c.480C>G genotype of hOCT1 had a significant effect on apparent drug clearance (CL/F) being responsible, respectively, for a 20% and 10% decrease in interindividual variability (IIV) of CL/F (from 50.1 up to 19.6%). Interestingly, 25 patients carrying at least one polymorphic c.480 G allele had a significant lower CL/F value with respect to the 35 c.480CC individuals (mean±s.d., 9.6±1.6 vs 12.1±2.3 l h(-1), respectively; P<0.001). In conclusion, the hOCT1 c.480C>G SNP may significantly influence imatinib pharmacokinetics, supporting further analyses in larger groups of patients.

Occurrence of Salmonella in swine in the Latium Region (central Italy) from 1980 to 1989: a retrospective study.

An extensive survey was carried out on 849 biological samples (dead animals, organs and viscera, faeces, and rectal swabs) from pigs in the Latium Region (Central Italy) throughout the years 1980-1989. In total, 46 of the samples (5.4%) were found to be bacteriologically positive for salmonellae. Among dead animals, typical gross lesions were observed in 6 clinically infected animals. As far as serotypes are concerned, 11 different ones were isolated, with a predominance of Salmonella typhimurium (26.1%), S. anatum (21.7%), S. bovis-morbificans (15.2%), and S. heidelberg (10.8%). According to Kauffmann-White's classification scheme, 39.1% of the isolated strains belonged to serogroup B, 13% to serogroup C1, 17.4% tot serogroup C2, 6.5% to serogroup D, and 23.9% to serogroup E1. In conclusion, the authors speculate that the relatively low Salmonella isolation frequency (5.4%) reported in this study was not dependent upon the cultural procedures used, since different enrichment and plating methods were used, but rather upon the great number of rural-type herds which were investigated over the decade compared to industrial-type herds. Other factors such as stress conditions, which are particularly pronounced in industrial-type herds, and overall climatic situations, which tend to be characterised by warm, dry and long summers in the Latium Region, are also considered in this respect.

[Treatment of thalassemia]. / Il trattamento della talassemia.

The management of Cooley's anemia today is based on: a careful transfusional therapy using international schedules with standardized parameters for evaluating its efficacy; prevention of iron overload improving the compliance for chelation therapy; blood transfusion with the optimal methods to prevent reactions mainly due to leukocytes; psychosocial and clinical approach of adolescent's problems.

[Critical evaluation of the causes of death in thalassemic subjects]. / Valutazione critica delle cause di morte nei soggetti talassemici.

We discuss 25 cases of death observed from 1971 to 1983 in a casistic of 155 patients with Thalassaemic Syndrome. Anemia as a cause of death is disappearing, new triggers are involved such as Yersinia Enterocolitica who can cause severe sepsis even in non-splenectomized patients. Iron overload appears to be the most severe complication in the second decade.

Early iron overload in beta-thalassaemia major: when to start chelation therapy?

Twenty-eight children with beta-thalassaemia major aged between 11 and 48 months were given intensive transfusions. Serum iron, transferrin saturation, serum ferritin, non-transferrin iron, and subcutaneous desferrioxamine-induced urinary iron excretion were measured. The results showed that even children with a limited number of transfusions had severe iron overload as indicated, in particular, by the raised serum ferritin levels and the high excretion rates after subcutaneous infusion of desferrioxamine. The desferrioxamine test was useful, even in very young children, in assessing response to chelation therapy thus enabling such treatment to be started early to prevent harm from iron overload.

Evaluation of the supertransfusion regimen in homozygous beta-thalassaemia children.

The maintenance of physiological levels of haemoglobin is beneficial in the management of patients with thalassaemia major since it leads to better tissue oxygenation, reduction of blood volume, and reduced intestinal absorption of iron. We have studied 11 patients with thalassaemia major while treating them for 4-12 months with a standard transfusion regimen (mean pre-transfusion Hb 10.2 g/dl) and then for a second period of 7-18 months with a supertransfusion regimen (mean pre-transfusion Hb 12.3 g/dl). Blood consumption was 16.71 +/- 2.0 ml/kg/month in the first period; it rose to 20.30 +/- 3.5 ml/kg/month in the first 5 months of the second period, and then returned to the values of the first period (16.53 +/0 2.0 ml/kg/month). There were no significant differences in blood consumption between the two transfusion regimens, after the 5-month equilibration period.