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Aim: This study aims to evaluate the diagnostic challenges and molecular aspects of pediatric gliomas, among the most common brain tumors in children, focusing on improving early detection and personalized treatment strategies., Methods: We conducted a comprehensive review of recent literature, examining current diagnostic techniques, including imaging and histopathological analysis, alongside molecular profiling methods such as next-generation sequencing (NGS) and methylation profiling., Results: The findings highlight significant diagnostic challenges due to the heterogeneity of pediatric gliomas. Molecular profiling has proven essential in identifying key genetic alterations, such as those in the H3F3A and MAPK pathway genes, offering insights into tumor behavior and therapeutic targets., Conclusions: Early and accurate diagnosis of pediatric gliomas is hampered by their molecular diversity. Integrating molecular diagnostics with traditional methods is crucial for enhancing diagnostic accuracy and guiding personalized treatment approaches.

Introduction: Plasma cell myeloma is a hematological malignancy with heterogeneous genomic landscape and diverse clinical course. Recurrent chromosomal and subchromosomal aberrations commonly occur in this entity and are associated with the pathogenesis and progression of the disease. The identification of these alterations aids genetic characterization, classification and prognostication of patients. Aim: Molecular cytogenetic investigations of plasma cell myeloma patients treated at the University of Pécs Clinical Center and János Balassa County Hospital of Tolna County, Szekszárd, between 2005 and 2018 were evaluated in our study. Method: 231 patients were screened for genetic aberrations using fluorescence in situ hybridization. Translocations involving the immunoglobulin heavy chain gene, losses of 1p and 17p chromosome arms, gains of 1q chromosome arm and unbalanced aberrations of chromosome 13 were investigated. Losses and gains of 1p, 1q, 5q, 12p, 13q, 16q and 17p chromosome arms were analyzed using multiplex ligation-dependent probe amplification in 42 patients. During the investigated period, 116 bone marrow karyotyping was also performed. Results: In total, 233 genetic aberrations were identified using our targeted approaches; the frequency of specific aberrations correlated with data of the recent literature. Concordance of results gained by fluorescence in situ hybridization and multiplex ligation-dependent probe amplification was 96.2% by analyzing the same chromosome arms. The latter technique revealed 21 additional genetic aberrations in 16/42 patient samples (38%) as compared to fluorescence in situ hybridization. Conclusions: Our results suggest that the combined application of the two molecular cytogenetic methods may facilitate a more detailed characterization of genetic aberrations of plasma cell myeloma patients in Hungary. Orv Hetil. 2019; 160(24): 944-951.

Introduction: Prognostic impact of the detection of minimal residual disease (MRD) in multiple myeloma (MM) has been confirmed in numerous studies., Aim: Retrospective examination of our patient database (107 newly diagnosed multiple myelomas between 2007 and 2017). Flow cytometry (FCM) was performed as MRD assessment., Method: MRD assessment was performed in 56 patients (median age: 68 years), after induction treatment of multiple myeloma. The treatment contained bortezomib in 91%, autologous haematopoetic stem cell transplantation (ASCT) was perfomed in 50%. MRD detection was performed on bone marrow samples, predominantly in our hospital (BD FACScan, 3 colour, panel: CD38, CD138, CD19, CD45, CD56, CD28, CD117, cyKappa, cyLambda, 100 000 events)., Statistical Analysis: SPSS 13.0., Results: The progression-free survival (PFS) and the overall survival (OS) were significantly longer in MRD negative (n = 22) patients (PFS: 54 months, OS: 79% after 5 years) than MRD positive patients (n = 34, PFS: 22 months, OS 21% after 5 years, p = 0.001). Patients achieving complete response (CR) (n = 29) have different PFS (MRD negative CR: 60 months, MRD positive CR: 21 months, p<0.001). Patents achiving MRD negative very good partial response (n = 5) have similar PFS (54 months) as patients with MRD negative CR. The longest PFS (68 months) was observed in MRD negative patients, after ASCT (n = 11), while the PFS was significantly (p<0.001) shorter in patients who were MRD positive after ASCT (n = 18, PFS: 25 months), similarly in MRD positive patients without ASCT (n = 15, PFS 21 months). Cox regression analysis (stage, cytogenetic risk, ASCT) confirmed that MRD is an independent prognostic factor of PFS and OS. We did not find significant relationship between MRD and stage, cytogenetic risk, number of treatment cycles, ASCT., Conclusions: The depth of response after induction treatment of MM is an independent predictor of survival. MRD assessment with FCM is recommended to define response. Consideration of maintenance treatment in MRD positive patients and eradication of MRD are also recommended. Orv Hetil. 2019; 160(13): 502-508.

Introduction: Many new prognostic factors established in recent years in chronic lymphocytic leukemia. May help predicting survival., Aims: The goal of the present study was to determine the frequency and the correlation of these novel prognostic factors in samples of 419 leukemia patients., Methods: The mutation status of the IgH gene was evaluated in 160 cases., Results: In 62% of cases, non-mutated IgH gene was found, the heavy chain family usage was different in mutated and non-mutated cases. The CD38 expression demonstrated 78% concordance with the mutation status, the ZAP-70 expression failed to show any correlation. Cytogenetic abnormalities were seen in 76% of cases, the most frequent were del(13q) (57%), trisomy 12 (15%), del(11q) (12%) and del(17p) (6%). 95% of cases with del(11q) harbored non-mutated, 74% of cases with del(13q) as the sole anomaly demonstrated mutated IgH genes., Conclusions: The parameters analysed are not independent of each other, utilization of them in the clinical routine needs careful planning.

The t(9;22) translocation, which results in a fusion protein with abnormal tyrosine-kinase activity, plays a central role in the pathogenesis of chronic myeloid leukemia. The selective inhibition of this chimeric protein with imatinib-mesylate is an efficient therapeutic option, haematologic and cytogenetic responses can be achieved in most of the patients. The primary goal of monitoring the disease is to assess the efficiency of the therapy, to highlight those patients, whose survival may be improved by modifying treatment. Three methods are widely used for the genetic monitoring of chronic myeloid leukemia. With karyotyping, the proliferating bone marrow cells can be evaluated. The use of fluorescent in situ hybridization makes the cytogenetic analysis of each cell within the sample possible. Real-time quantitative polymerase chain reaction is capable of quantifying residual leukemia far below the sensitivity of cytogenetics. The results of these three methods have different biological meanings, thus, for the interpretation of the results, the knowledge of the characteristics, the benefits and the draw-backs of the methods is required. The present study shows the most important characteristics of these methods based on the literature and data acquired from 1165 samples of 197 patients detected by the authors.

Introduction: Hepatitis C virus (HCV) infection may have an etiopathogenic role in development of B-cell non-Hodgkin lymphoma (NHL). Antiapoptotic effect of the HCV may display via activation of nuclear factor kappa B (NF-kappa B) transcription factor and can lead to development of carcinoma., Aims: 1. Activated NF-kappa B was examined in peripheral blood lymphocytes of patients with HCV infection or NHL, and HCV positive B-cell NHL. 2. NF-kappa B was detected in liver biopsies of patients with HCV positive hepatocellular carcinoma (HCC) and in lymphnodes of patients with HCV positive NHL., Patients and Methods: 1. Fifteen patients with chronic HCV infection, 13 patients with B-cell NHL (among them 4 HCV positive) and 10 healthy subjects were examined for the activity of NF-kappa B using electrophoretic mobility shift assay (EMSA). 2. Immunohistochemistry was performed in 8 liver biopsies from HCV positive patients and 2 HCC patients (1 HCV positive and 1 HCV negative) and 2 B-cell NHL patients (1 HCV positive and 1 HCV negative)., Results: 1. In the HCV negative control samples there was no shift detected, but we found NF-kappa B specific oligonucleotide-protein complexes in the lymphocyte extracts from all HCV positive patients and 10/13 (76.92%) in B-cell NHL. 2. NF-kappa B was detectable in 7/8 cases in liver biopsies from HCV positive patients. Strong reaction was detected in nuclei in HCV positive HCC and NHL patients (no reaction in HCV negative biopsies)., Conclusions: NF-kappa B activation was justified in both diseases, which can connect the development of both HCV infection and B-cell NHL. This factor activation may have role in hepato- and lymphomagenesis.

Unlabelled: Hepatitis C virus (HCV) has cytopathogenic effect not only on hepatocytes, however on salivary glands, monocytes of peripheral blood and lymphoid cells, may explain the systemic manifestations of the infection. HCV activates B and T-cells, modifies the immune response, causes lymphoproliferation, leading the development of B-cell non-Hodgkin's lymphoma (NHL). In the majority of B-cell NHLs immunoglobulin heavy chain (IgH) and light chain (IgL) genes are rearranged and expressed on cell surface in the early stage of the ontogenesis. The analyses of IgH rearrangement prove the clonality of lymphoproliferative disorders giving a powerful approach to the B-cell origin identification of cell proliferation., Patients and Methods: IgH gene rearrangements were examined from the sera of 57 chronic HCV infected patients and 11 HCV-positive cryoglobulinemic patients as well., Results: Cryoglobulinemia was detected in 20% of all chronic hepatitis C virus infected patients and IgH rearrangement was observed in 10.29% (7/68), 4/7 patients (57.14%) suffered from cryoglobulinemia., Conclusions: These results support the hypothesis that IgH gene rearrangement in HCV positive patients can indicate the lymphoproliferative disorder in the HCV infection released B-cell proliferation and lymphoma development.

Oncogenesis is a multifactorial process in which environmental, genetical and infectious factors may be of importance. Specific viruses are supposed to have etiological role in about 15% of human tumors. Recently the B-cell proliferation inducing effect of the hepatotropic and lymphotropic hepatitis-C virus (HCV) came into the limelight based on the high prevalence of HCV positivity in B-cell non-Hodgkin's lymphoma (NHL) patients. The aim of the authors was to establish the prevalence of HCV infection in NHL patients. Paralelly the HBV, CMV and EBV markers, and the alterations of the humoral immune response (immunoglobulins, cryoglobulins, rheumatoid factor) were determined. 42 patients (24 male, 18 female; the mean age: 54.1 years, range 22-80 years) classified as 16 indolent (low risk), and 25 aggressive (intermediate risk) NHL and one with very aggressive Burkitt's lymphoma, according to the modified REAL classification were examined. Enzyme-linked immunosorbent assay (ELISA) for HBsAg and anti-HCV, HBsAg, anti EBV, anti CMV, furthermore polymerase chain reaction (PCR) for HCV-RNA were used. Anti-HCV was found in 6/42 NHL patients (14.3%), while anti-HCV and/or HCV-RNA PCR positivity revealed on overall HCV infection in 10/42 (23.8%) patients. None of them were HBsAg positive. Our findings support the hypothesis, that HCV might have an aetiological role in the lymphoproliferation leading to B-cell NHL.

The authors have treated 38 patients with chronic phase chronic myeloid leukemia in their single center in the last five years. Conventional chemotherapy provided about 40-50% hematological response, interferon-alpha seems to be more effective, complete hematological remission occurred in 65%. Interphase cytogenetics and fluorescein in situ hybridisation technique was used to measure the cytogenetic response. They observed complete cytogenetic remission in two cases (8%), major response in 11 (39%), minor response in 4 (15%) and minimal response in 4 cases (15%). Interferon-alpha is an effective, well-tolerated medicine in the treatment of chronic myeloid leukemia.

In a prospective survey clinical haematological and molecular biological data of 31 patients with chronic myelogenous leukaemia (chronic phase, CML) observed in their haematological outpatient department were analyzed. During 1996 and 1999 a regular follow-up of the Philadelphia chromosome level in treated patients was performed with molecular biological techniques, i.e. with reverse transcription polymerase chain reaction (RT-PCR) and with fluorescence in situ hybridization (FISH) methods. During the follow-up period of 33 months from the 31 patients with CML (16 males, 15 females) 25 ones were treated with human recombinant interferon-alpha (IFN), however, six others were getting only hydroxyurea (HU). During this period in three patients allogen bone marrow transplantation was performed and seven ones expired (six out of them of blastic crisis). The quality of therapeutic response at cytogenetical level was determined by decrease of the bcr-abl level due to the treatment (non-responders, major, minor and complete cytogenetic remission groups). In nine patients (five from the IFN-group, four from the HU-group) achieved no cytogenetic therapeutic response (non-responders, 29%). However, in 13 patients a minor (bcr-abl range of 60-30%), and in nine patients a maior/complete (bcr-abl of 30-10%) cytogenetic response (42% and 29%, respectively) were detected. Moreover, the quality of cytogenetic response correlated with the haematological remission. The maior/complete cytogenetic response was durable in eigth patients. The improvement of the overall survival of patients with CML, the postpone of the fatal accelerated-blastic phase could be expected only from the early introduced, in individually adjusted and given in maximally tolerated dosage of interferon (3-5 million UI/m2/day). The qualitative (RT-PCR) and quantitative (FISH) detections of the Philadelphia chromosome are reliably reproducible up-to-date molecular biological methods getting relevant results, which could be very helpful in the planning, monitoring, in setting of optimal dosage of the interferon therapy of patients with CML, in addition in the judgement of the effectiveness of the therapy, in the reduction of adverse effects, as well as in forecasting of the cytogenetic progression.

The authors have monitored the treatment of chronic myeloid leukaemia by simultaneous investigation of the residual tumour mass as well as the expression of the bcr-abl chimera. The first parameter was obtained by calculating the bcr-abl rearrangement positive cells as determined by fluorescent in situ hybridisation (FISH). The other value was determined by a quantitative (competitive) reverse transcription polymerase chain reaction (Q-RT-PCR). To avoid any inaccuracy associated with the variable yield of RNA isolation and reverse transcription, respectively, the amount of chimera mRNA was related to that of abl 2-3 transcript as internal control. The threshold for the sensitivity of the Q-RT-PCR procedure was 16 bcr-abl mRNA molecule/microgram RNA. The lowest relative concentration was represented by detecting 1 bcr-abl mRNA molecule out of 12,600 abl 2-3 transcripts. The analysis of peripheral blood samples of 33 untreated patients indicated that the amount of chimera mRNA changes between 80 and 1,000,000 molecules per microgram RNA and this parameter correlated hardly (r = -44) with the tumour load. Data from blood samples of 48 treated patients indicated that this four order of magnitude difference in the expression persists in that patient population which exhibited on average major cytogenetic and also in those without any cytogenetic response. In the individual patients not only correlated changes of residual tumour mass and chimera expression, but mainly independent changes of these two parameters were observed.

The authors investigated by reverse transcription-polymerase chain reaction the incidence of the t(12;21)(p13;q22) translocation among 130 pediatric acute lymphoblastic leukemia registered by the Hungarian Pediatric Oncology Workgroup. The distribution of this translocation was analysed in the ploidy categories as defined by the flow cytometric DNA analysis and interphase cytogenetics. The incidence of the translocation proved to 19%, the positive patients' age ranged between 2 and 14 with an average of 5.8 years. Ninety percent of the leukemic patients harbouring the t(12;21) exhibited the precursor B-cell phenotype, 10% coexpressed myeloid markers. Coexistence of the t(12;21) with the m-bcr type of Philadelphia translocation was not observed. Ninety five percent of the t(12;21) positive children was diploid by flow cytometry whereas the same figure proved to be 58% using interphase cytogenetics. This difference was due to the hypo- and pseudodiploidy undetectable by flow cytometry but revealed by interphase cytogenetics. The authors conclude that the t(12;21) positive patients which seemed to be homogeneous at gross DNA level were markedly heterogeneous with interphase cytogenetics and this might explain the inconsistent data in the literature in connection with prognostic significance of the t(12;21).