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β-Thalassemia (β-thal) is considered rare in Central Europe. As in other malaria-free regions, the presence of β-thal in Central Europe reflects historical and recent immigration, and demographic changes that have influenced the genetic variability of the current populations living in this area. This study assesses the frequency and spectrum of mutations on the β-globin gene in Czech and Slovak subjects with clinical symptoms of thalassemia. The results of the initial part of this research were published more than two decades ago; the aim of this study was to update these original reports. During the period from 2002 to 2015, 400 cases from Czech and Slovak hematological centers were analyzed. Twenty-nine β-thal mutations, identified in 356 heterozygotes from 218 unrelated families, involve five unique mutations including a recently described insertion of a transposable L1 element into the β-globin gene. One mutation described here is reported for the first time. Most of the mutations were of Mediterranean origin and accounted for 82.0% of cases. All but one case studied were heterozygous carriers, manifesting β-thal minor, with rare exceptions represented by the rare (β(0)) codons 46/47 (+G) (HBB: c.142_142dupG) mutation associated with an α-globin gene quadruplication and by dominantly inherited β-thal with a more severe phenotype. One double heterozygous β-thal patient was a recent immigrant from Moldavia. The list of δβ-thal alleles (26 carriers, 16 families) contains Hb Lepore and two types of δβ(0)-thal deletions. In the past, genetic drift and migration as well as recent immigrations were responsible for the introduction of Mediterranean alleles, while several mutations described in single families were of local origin.

Tyrosine kinase inhibitors (TKI) have completely changed the prognosis of patients with Ph+ chronic myeloid leukemia (CML). The occurrence of a second malignancy (SM) in CML patients successfully treated with TKI may significantly affect their prognosis. In a retrospective study of 1,038 patients with CML treated at 10 centers in the Czech Republic and Slovakia between 2000 and 2009, SM was detected in 35 (3.37%) patients after TKI therapy was initiated. The median intervals from the diagnosis of CML and from the start of TKI therapy to the diagnosis of SM were 58 months (range 2 - 214) and 32 months (range 1 - 102), respectively. The observed age-standardized incidence of SM after the start of TKI therapy was 8.95 / 1,000 person-years. Comparison of the incidence of SM in CML patients with population data was performed only for patients from the Czech Republic. The age-standardized incidence rate of all malignant tumors except non-melanoma skin cancers was 6.76 (95% CI: 6.74; 6.78) / 1,000 person-years in 2000 - 2007 while the incidence rate of SM in 708 CML patients from the Czech Republic treated with TKI was 9.84 (95% CI: 6.20; 13.48) / 1,000 person-years, i.e. 1.5-fold higher, although the difference was statistically insignificant. The distribution of SM types in CML patients treated with TKI was similar to that in the age-standardized general Czech population. The median overall survival (OS) of patients treated with TKI who also developed SM (57 months) was shorter than the OS of patients treated with TKI but not suffering from SM (median OS not reached, log rank test p < 0.001. Prospective long-term population-based studies in CML patients treated with TKI as first-line therapy are needed to determine the relationship of SM to KTI therapy.

Interleukin-6 (IL-6) is an important pro-inflammatory mediator implicated in immune-mediated complications of allogeneic haematopoietic stem cell transplantation (aHSCT). In accord with previous reports, this preliminary study on 56 donor-recipient pairs revealed IL-6-174 single nucleotide polymorphisms as a risk factor for the development of acute graft-versus-host disease and decreased survival after aHSCT.

A novel HLA-B (human leukocyte antigen-B) allele, HLA-B*4442, was identified both in a Czech patient with leukaemia and in his mother. The presence of a novel allele was initially suspected because conflicting results were obtained by serological and DNA typing techniques. The HLA typing using the polymerase chain reaction-sequence-specific primers (PCR-SSP) at the two-digit level indicated an allele belonging to the HLA-B*44 group, whereas serological typing indicated HLA-B21. Typing with PCR-sequence-specific oligonucleotides (PCR-SSO) resulted in a unique reaction pattern that could not be assigned to a known allele, PCR-SSP typing at the four-digit level did not match any known B*44 allele, either. The sequencing-based typing of the HLA-B locus then revealed the novel B*4442 allele that is identical with B*4405 except a single C-->G nucleotide exchange at position 572. This exchange results in an amino acid substitution from serine to tryptophan at position 167 of the expressed HLA-B protein. The B21 serological reactivity of the novel B*4442 allele product was confirmed by employing an additional serological panel of typing sera. Our findings support previous reports claiming that serine at the position 167 in the alpha-2 domain of the HLA-B protein is a major determinant of the HLA-B44(12) serological epitope.

A novel human leukocyte antigen-B (HLA-B) allele, B*420502, was identified in a patient with leukemia (Caucasoid, Czech ancestry) and his mother during intrafamily search for the hematopoietic stem cell donor. The novel allele was initially detected by HLA typing at low resolution using both sequence specific primers and sequence specific oligonucleotides techniques that resulted in unique reaction patterns. The alleles of the HLA-B locus were separated by the haplotype-specific extraction technique. Sequencing of those alleles revealed a novel allele, B*420502, that is identical with B*420501 except a T-->G exchange (synonymous mutation) at position 618.

Introduction: The aim of this study was the retrospective assessment of therapy results and the changing prognosis of multiple myeloma (MM) patients from the Middle and North Moravia region of the Czech Republic during the last 40 years., Patients and Methods: The analyzed group of 562 patients was gathered in the years 1959-2000, mean age 63 (28-91) years, M/F ratio 1.1 : 1.0. Overall survival (OS) curves were plotted according to the method of Kaplan and Meier and compared using the log rank test (P<0.05)., Results: During the long assessed period, a significant change in prognosis and therapy response was achieved (P=0.0000). The 'first turning point' (1963-1975) characterized by unsystematic melphalan and prednisone therapy (MP), led to significant improvement of OS (8-19 months, P=0.0031) in comparison with the period of plain symptomatic therapy (1959-1963) and the increase in 3-year survival rate from 4 to 23% of patients. The 'second turning point' (1976-1980), characterized by introduction of the systematic conventional polychemotherapy (VMP, VMCP, VBAP, VCAP) and complex supportive treatment, was associated with subsequent improvement of therapeutic response (OS 40 months, P=0.0000; 3- or 10-year survival was seen in 55 and 5.5% of patients). Nevertheless, the results of the next 20 years (regimens VB(C)MCP, VAD, Cy-VAD and CIDEX) are a little disappointing with insignificant progress. In the whole group of 295 patients from the years 1976 to 1995, divided into 5-year periods, remission (R< or =25% of the initial M-protein level) was achieved in 10-24% of cases, the OS median was 32-44 months, and 5 (respectively 10)-year survival improved from 25 to 36% and from 5.5 to 16.5% of patients. The 'third turning point' represented years 1996-2000 marked with the introduction of high dose (HD)-therapy with ASCT support leading in a group of 31 patients (< or =65 years) to remission in 71%, to complete remission in 32% and to significant improvement of OS (P=0.0037) and 5-year survival in 91% of the patients. The improvement of therapy results in this 'third turning point' was demonstrated in comparison with the 1976-1995 group with conventional therapy with a group of patients from 1996 to 2000 (both conventional and HD therapy with ASCT), characterized by 36% of remissions and 5-year survival of 57% of the patients (P=0.030). HD therapy with ASCT achieved in the years 1996-2000 had better therapy results (R-71%, 5-year survival 91%) than that achieved in two comparable groups of patients (1991-1995 and 1996-2000), fulfilling all criteria of HD therapy with ASCT but treated with conventional therapy only (R - 24 and 32%, 5-year survival 46 and 68% of the patients)., Conclusion: The retrospective analysis showed that the meaningful change in overall prognosis of MM patients achieved in Moravia is in accord with worldwide trends in the course of 1959-2000 years, which was achieved due to the gradual application of the modern innovated systems of complex therapy.

We have identified through sequencing of amplified DNA a GCC-->GAC mutation in codon 115 of the beta-globin gene in a mother and daughter of a small Czech family. This base change was confirmed by hybridization with a 32P-labeled specific oligonucleotide probe and by gene mapping because it creates a new Ava II site. The mutation results in an Ala-->Asp replacement at beta 115(G17); this beta chain is severely unstable and could not be identified either as chain or as hemoglobin variant by isoelectrofocusing and various high performance liquid chromatography methods. Stability tests were mildly positive in freshly prepared lysates, but an unstable hemoglobin could not be detected in older lysates with these methods. Its presence results in a dominant type of beta-thalassemia in the two heterozygotes, with moderate anemia, reticulocytosis, nucleated red cells, target cells, and other red cell changes, Heinz body formation, and splenomegaly; the oldest of the two patients was splenectomized. Both subjects had a marked increase in fetal hemoglobin synthesis.