miRNA profile of luminal breast cancer subtyptes in Slovak women.

OBJECTIVE: Aberrant expression of short, non-coding RNA molecules (miRNA) leads to breast cancer initiation, progression and metastasing. The miRNA expression level associates with imunohistochemical profile, histopathological parameters, clinical outcomes, prognoses and therapeutical response. The aim of this study was to analyse the whole spectrum of miRNA by microarray method and to define relevant miRNAs describing biological characteristics of luminal breast cancer subtypes. DESIGN: Cross-sectional study, basic research. SETTING: Biomedical center Martin, Jessenius Faculty of Medicine in Martin, Comenius University in Bratislava, Martin, Slovakia. METHODS: We analysed 16 tissue samples of Luminal A/B breast cancer types and 16 breast tissue samples without pathological findings. The microarray technology by Agilent was used to analyse 2549 miRNAs by SurePrint G3 Human miRNA kit v.21. The results were assessed by AgiMicroRNA Bioconductor library within Limma pack. RESULTS: The analyses of the lowest FDR p-value and the highest logFC value selected the oncomiR miR-182 as the most dominant with higher expression in cancer tissues than in normal tissues, followed by miR-21, miR342-3p/5p and miR-6826. The miR-4324 and cluster of miR-99a/let7c/miR-125b dominated in the group of miRNAs with lower expression in cancer tissues compared to normal tissues. CONCLUSION: The first results of this study complement biological characteristics of luminal breast cancer subptypes, represent basis for follow-up projects focused on the clarification of relevant signaling pathways and promise new and innovative breast cancer treatment based on the precise, tailored therapy by targeting specific miRNAs involved in the most important carcinogenesis mechanisms.

[Our experience with detection of JAK2 mutations in paraffin-embedded trephine bone marrow biopsies of patients with chronic myeloproliferative disorders]. / Nase skúsenosti s vysetrovaním JAK2 mutácií pacientov s myeloproliferatívnymi ochoreniami z trepanobioptického materiálu kostnej drene.

Polycythemia vera (PV), essential thrombocythemia (ET) and primary myelofibrosis (PMF) are Philadelphia chromosome-negative myeloproliferative neoplasms (MPN) characterized by JAK2 mutation. The exon 14 V617F mutation is present in almost all patients with PV and in approx. 60% of patients with ET and PMF. The importance of JAK2V617F in the differential diagnostic considerations is still unclear and here the BM morphology examination still represents an important diagnostic tool. In the WHO classification of Ph1-negative MPNs, the identification of JAK2 mutations represents a major diagnostic criterion of these diseases. Therefore we decided to implement the examination of JAK2V617F mutation in formalin-fixed paraffin-embedded biopsy specimens of patients with Ph1-negative MPN using allele-specific PCR. In addition, in all JAK2 V617F negative patients with PV we sequenced the whole JAK2 exon 12. Until now we examined up to 200 patients with clinically confirmed MPN and our results in all three categories PV, ET and PMF are in agreement with earlier published data. Paraffin embedded tissues represent a valuable source of DNA which can be used in the diagnostics of both JAK2 exon 12 and exon 14 mutations. It is of particular importance if the fresh material is not available and there is a clinical and/or research utility for the performance of PCR on archival bone marrow samples with PV, ET or PMF suspicion.

Eyelid edema as a first sign of lymphoma.

Chronic eyelid edema may be a symptom of different disease. The most common are autoimmune diseases such as orbital pseudotumor, vasculitis, sarcoidosis, or impaired vascular or lymphatic drainage. Rarely has it been reported as the sole manifestation of the lymphoma. Eyelid lymphoma is a special clinical entity in the spectrum of hematological malignancies. Here we present our clinical experience with eyelids lymphomas. First case is a 76-year-old female patient with bilateral edema of upper eyelid non-responding to anti-inflammatory therapy. Histological examination diagnosed mantle cells lymphoma. In the second case, 58-year-old patient was diagnosed with solitary unilateral tumor of the lower eyelid, where primary biopsy was ordered and diagnosis of MALT lymphoma was established after histological examination. In both cases, it was not solitary eyelid tumor, but systemic disease with multiple lymphadenopathy and bone marrow infiltration were found in follow-up examinations. Subsequently, patients care was given to the hemato-oncologist.

[Commentary on development of the prognostic factors concept in chronic lymphocytic leukaemia: the route from prognostic factors to therapy response predictors]. / Komentár k vývoju konceptu prognostických faktorov chronickej lymfocytovej leukémie: cesta od prognostických faktorov k prediktorom liecebnej odpovede.

Chronic lymphocytic leukaemia (CLL) is a lymphoproliferative disorder with variable clinical course. Determination of disease prognosis is based on the identification of different prognostic factors. The concept of CLL prognostic factors is still developing and has undergone several fundamental changes. Traditional (old) prognostic factors and staging systems are useful in describing the extent of the disease at any given moment, in determining clinical progression and in the identification of patients who need to start treatment. However, traditional prognostic factors are not sufficient for predicting a long-term prognosis because they are not able to identify potentially aggressive forms of CLL in the early stages. Nevertheless, clinical staging systems maintain their importance and in contrast to other traditional factors also their independent prognostic role. Otherwise, traditional prognostic factors play the role of disease activity descriptors rather than the role of actual prognostic factors. CLL risk profile determination is based on the identification of so-called new prognostic factors, the most relevant of which are chromosomal aberrations, TP53 gene mutations, mutational status of IgVH genes, ZAP-70 and CD38 expression. These factors are able to predict the prognosis already at the time of the initial diagnosis. In contrast to previous ideas, they are not incorporated into recommendations regarding indications for treatment. This is due to the risks associated with early treatment and the lack of data validated in prospective clinical trials demonstrating the justifiability of such procedure. In patients being treated, new prognostic factors may be useful for predicting the response to the therapy and some of them may directly influence the choice of treatment regime. New CLL treatment modalities have also raised the question of their influence on the prognostic and predictive power of new prognostic factors.

Fibrosis identified in the bone marrow biopsies of patients with essential thrombocythemia: its incidence and significance for the differential diagnostic considerations.

Myelofibrosis (MF) may develop in all types of myeloproliferative disorders and its identification is of clinical relevance. Typical bone marrow (BM) morphology of patients with essential thrombocythemia (ET) shows either "normal" amount or "a slight increase" of reticulin fibers, but the published data differ in relation to the applied MF definition and ET diagnostic criterias. The aim of this study was to evaluate retrospectivelly MF in BM biopsies of 30 cases in which the diagnosis of ET was confirmed also clinically by local hematologists. In 7 of the patients not only primary but also sequential biopsy was available. The MF grade and extent were evaluated semiquantitativelly in archival slides stained by Gömöri silver impregnation. The analysis was based on the European clinicopathological criteria 2004 (ECP) defining a) normal bone marrow fibrosis (MF0), b) slight reticulin fibrosis (MF1), c) advanced reticulin and initial collagen fibrosis (MF2) and d) advanced collagen fibrosis (MF3). Generally, in majority of the biopsies MF0 (n = 6) or MF1 (n = 25, 18x focal and 7x diffuse) was found. More advanced MF2 was much less common as it was present in 6 biopsies (5x focal and 1x diffuse). In relation to the actual time of BM biopsy during course of the disease, the introductory biopsies done at the time of diagnosis (n = 18) showed 3x MF0, 14x MF1 and 1x MF2. The biopsies performed after a long time of patients observations (n = 12) showed 3x MF0, 7x MF1 and 2x MF2. In 5 of 7 sequential biopsies the progress of MF was evident, but 4 of these patients were treated by cytoreductive therapy. We conclude that the BM of patients with ET in initial phase shows either MF0 or focal slight increase of reticulin fibers (MF1). In addition, the long course of the disease and/or applied therapy may lead to more developed MF and more advanced MF stages (diffuse MF1 or MF2). Therefore their finding in the BM biopsies examined in the later phases of the disease should not exclude the diagnosis of ET.

Heart rate recovery after exercise: relations to heart rate variability and complexity.

Physical exercise is associated with parasympathetic withdrawal and increased sympathetic activity resulting in heart rate increase. The rate of post-exercise cardiodeceleration is used as an index of cardiac vagal reactivation. Analysis of heart rate variability (HRV) and complexity can provide useful information about autonomic control of the cardiovascular system. The aim of the present study was to ascertain the association between heart rate decrease after exercise and HRV parameters. Heart rate was monitored in 17 healthy male subjects (mean age: 20 years) during the pre-exercise phase (25 min supine, 5 min standing), during exercise (8 min of the step test with an ascending frequency corresponding to 70% of individual maximal power output) and during the recovery phase (30 min supine). HRV analysis in the time and frequency domains and evaluation of a newly developed complexity measure - sample entropy - were performed on selected segments of heart rate time series. During recovery, heart rate decreased gradually but did not attain pre-exercise values within 30 min after exercise. On the other hand, HRV gradually increased, but did not regain rest values during the study period. Heart rate complexity was slightly reduced after exercise and attained rest values after 30-min recovery. The rate of cardiodeceleration did not correlate with pre-exercise HRV parameters, but positively correlated with HRV measures and sample entropy obtained from the early phases of recovery. In conclusion, the cardiodeceleration rate is independent of HRV measures during the rest period but it is related to early post-exercise recovery HRV measures, confirming a parasympathetic contribution to this phase.

Heart rate recovery after exercise: relations to heart rate variability and complexity

Physical exercise is associated with parasympathetic withdrawal and increased sympathetic activity resulting in heart rate increase. The rate of post-exercise cardiodeceleration is used as an index of cardiac vagal reactivation. Analysis of heart rate variability (HRV) and complexity can provide useful information about autonomic control of the cardiovascular system. The aim of the present study was to ascertain the association between heart rate decrease after exercise and HRV parameters. Heart rate was monitored in 17 healthy male subjects (mean age: 20 years) during the pre-exercise phase (25 min supine, 5 min standing), during exercise (8 min of the step test with an ascending frequency corresponding to 70 percent of individual maximal power output) and during the recovery phase (30 min supine). HRV analysis in the time and frequency domains and evaluation of a newly developed complexity measure - sample entropy - were performed on selected segments of heart rate time series. During recovery, heart rate decreased gradually but did not attain pre-exercise values within 30 min after exercise. On the other hand, HRV gradually increased, but did not regain rest values during the study period. Heart rate complexity was slightly reduced after exercise and attained rest values after 30-min recovery. The rate of cardiodeceleration did not correlate with pre-exercise HRV parameters, but positively correlated with HRV measures and sample entropy obtained from the early phases of recovery. In conclusion, the cardiodeceleration rate is independent of HRV measures during the rest period but it is related to early post-exercise recovery HRV measures, confirming a parasympathetic contribution to this phase