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Ovarian cancer is one of the most common cancers in women. Due to its nonspecific symptoms, it is often diagnosed late. This reduces the possibility of successful therapy. The main treatments are surgery and chemotherapy. With late detection, complete surgical removal of the tumor is not possible. The use of chemotherapy is limited by side effects and the resistance of cancer. Therefore, new and promising treatments are being sought. There are studies on the usage of hormone therapy in ovarian cancer. Unfortunately, a small number of these studies provide insufficient evidence of effectiveness. Hence, additional targeted therapies in ovarian cancer, and especially in immunotherapy, are of interest. Immunotherapy uses antibodies directed against specific receptors on cells important in the development of cancer. The results of the latest research on targeted methods, collected in this work, are promising.

Introduction: Both microenvironmental signals from surrounding cells and changes in the genome of leukemic cells play essential role in the development of chronic lymphocytic leukemia. Nurse like cells (NLCs) are one of the important elements of the microenvironment of CLL cells. The key role in the interactions of leukemic cells with NLCs is played by chemokines, which may interfere with the programmed cell death process in the leukemic lymphocytes. The aim of our study was analysis of selected microenvironmental factors having potential impact on the leukemic cells survival, as well as their association with clinical, cytogenetic and molecular parameters. For this study we selected three types of molecules which can modulate microenvironment: chemokines IL-8 and CCL3 (which are classically secreted to extracellular matrix), soluble forms of adhesion molecules JAG1 and CD163 and secreted form of endogenous protein BIRC5. We assessed their expression in the serum of CLL patients as well as in medium of long term NLCs cultures. Methods: Long-term cell culture was prepared from mononuclear cells derived from the blood of 34 patients with CLL. Number of NLCs cells was evaluated, under a light inverted microscope. The concentration of IL-8, CCL3, sBIRC5, sCD163 and sJAG1 in culture medium and serum was assessed by enzyme-linked immunosorbent assays. Results: There were significant differences in the concentration of IL-8, sBIRC5, CCL3, sCD163 and sJAG1 between the patient's blood serum and the culture medium. The concentrations of IL-8, CCL3 and JAG1 were higher in the culture medium, which confirmed the role of the microenvironment in the production of these proteins. In addition, the concentration of CCL3 chemokine in both patient’s blood serum and in the culture medium correlated with the number of NLCs and with known prognostic factors in the course of CLL e.g. Rai stage, WBC, expression of ZAP-70, CD38 and CD5/19. Conclusion: The microenvironment of CLL cells, which includes NLCs, plays an important role in the pathogenesis of CLL. The CCL3 chemokine seems to be a good factor representing microenvironment of CLL cells. Chronic lymphocytic leukemia is a complex and very heterogeneous disease, therefore its progress should be considered both in the context of genetic changes and the interaction with microenvironmental cells.

Despite significant medical advances, cancer treatment is still associated with a high risk of side effects. The treatment is usually invasive and devastating and it affects the overall immunity of the whole organism, including the condition of the skin. In recent years there has been a growing interest in isoflavonoids, due to their wide range of biological and pharmacological activity, especially estrogen-like. It gives a broad perspective of their use as active ingredients of preparations, which eliminate skin lesions associated with oncological treatment. This article is an overview describing preclinical and clinical observations on the basis of available literature. It discusses the influence of genistein on skin health in women after breast cancer treatment. The overview focuses on studies conducted with genistein

Neutrophils to lymphocytes ratio (NLR) and platelets to lymphocytes ratio (PLR) are considered as laboratory markers of inflammation. They can be potentially useful in predicting the course of multiple neoplasms including selected hematological cancers. The aim of the study was to assess the value of NLR and PLR in predicting the effects of therapy and prognosis in multiple myeloma patients treated with thalidomide-based regimen. The study group consisted of 100 patients treated with the first line CTD (cyclophosphamide, thalidomide, and dexamethasone) chemotherapy. The NLR and PLR were calculated before treatment. High NLR was observed in patients with higher stage of the disease, with poor performance status, hypercalcemia, and high CRP. High PLR was associated with low BMI and high CRP. In patients with high NLR, significantly shorter PFS was observed (17 vs. 26 months, p = 0.0405). In addition, high values of NLR and PLR were associated with significantly shorter OS (38 vs. 79 months, p = 0.0010; 40 vs. 78 months, p = 0.0058). Summarizing, NLR and PLR have a significant independent prognostic value for multiple myeloma patients. Furthermore, the NLR can be a predictive marker for the outcome of thalidomide-based chemotherapy.

Chronic lymphocytic leukemia (CLL) is the most common form of leukemia in Western European countries. This disease occurs mainly in adult population at an age of over 50 years, more commonly in men. Many years of research have shown that CLL is a disease with a very diverse clinical course and variable sensitivity to treatment. Therefore, in each case of newly diagnosed CLL, the clinical stage, prognostic factors and individualized therapeutic goals should be precisely defined before the treatment initiation. Recent 50 years have been a period of significant development in the treatment of proliferative diseases of the hematopoietic and lymphatic systems. Along with dynamic development of molecular biology techniques, several new specific prognostic markers have been investigated and introduced, allowing better stratification of patients and more accurate direction of the therapy. A lot of effort has gone into searching for the next prognostic factors which allow to determine better probable life span and to implement the successful treatment. Furthermore, many preclinical and clinical studies continually extend the list of effective anticancer drugs, including these for CLL treatment. Increasingly, in addition to standard cytostatics, drugs with more selective mechanisms of action are being tested in monotherapy and new combinations. Considering the historical aspects, only a few anticancer drugs were available 50 years ago. In addition, they mainly presented the one mechanism of action directed at nucleic acids. Today, the list of effective drugs is significantly longer. In addition to typical cytostatics, it also contains drugs with selective mechanisms of action, affecting the cell life processes, their subcellular structures, signaling from outside and inside the cell, surface antigens, as well as drugs that act on the microenvironment of cancer cells and their vascularization. The aim of this paper was to show the historical background in the therapy of leukemia and summarize the current knowledge in CLL biology and treatments. The manuscript is focused on the clinical course of the disease, the main risk factors and mainly, on the available treatment strategies.

Preeclampsia and hypertension complicate several pregnancies. Identifying women at risk of developing these conditions is essential to establish potential treatment modalities. Biomarkers such as C19MC microRNA in pregnant patients wopuld assist in defining pregnancy surveillance and implementing interventions. This study sought to analyze circulating C19MC microRNA as an early marker of hypertension and preeclampsia in pregnant patients. A systematic review was undertaken using the following registers: disease registries, pregnancy registries, and pregnancy exposure registries, and the following databases: PubMed, CINAHL, Web of Science, Scopus, and EMBASE. The risk of bias was assessed using the Cochrane technique. From the 45 publications retrieved from the registers and databases, only 21 were included in the review after the removal of duplicates, screening, and eligibility evaluation. All 210 publications had a low risk of bias and illuminated the potential use of circulating C19MC microRNA as an early marker of hypertension and preeclampsia in pregnant patients. Therefore, it was concluded that C19MC microRNA can be used as an early marker of gestational preeclampsia and hypertension.

Pre-eclampsia is a placenta-related complication occurring in 2–10% of all pregnancies. miRNAs are a group of non-coding RNAs regulating gene expression. There is evidence that C19MC miRNAs are involved in the development of the placenta. Deregulation of chromosome 19 microRNA cluster (C19MC) miRNAs expression leads to impaired cell differentiation, abnormal trophoblast invasion and pathological angiogenesis, which can lead to the development of pre-eclampsia. Information was obtained through a review of articles available in PubMed Medline. Articles on the role of the C19MC miRNA in the development of pre-eclampsia published in 2009–2022 were analyzed. This review article summarizes the current data on the role of the C19MC miRNA in the development of pre-eclampsia. They indicate a significant increase in the expression of most C19MC miRNAs in placental tissue and a high level of circulating fractions in serum and plasma, both in the first and/or third trimester in women with PE. Only for miR-525-5p, low levels of plasma expression were noted in the first trimester, and in the placenta in the third trimester. The search for molecular factors indicating the development of pre-eclampsia before the onset of clinical symptoms seems to be a promising diagnostic route. Identifying women at risk of developing pre-eclampsia at the pre-symptomatic stage would avoid serious complications in both mothers and fetuses. We believe that miRNAs belonging to cluster C19MC could be promising biomarkers of pre-eclampsia development.

Malignant cells in chronic lymphocytic leukemia (CLL) show resistance to apoptosis, as well as to chemotherapy, which are related to deletions or mutations of TP53, high expression of MCL1 and BCL2 genes and other abnormalities. Thus, the main goal of the present study was to assess the impact of chlorambucil (CLB) combined with valproic acid (VPA), a known antiepileptic drug and histone deacetylation inhibitor, on apoptosis of the cells isolated from 17 patients with CLL. After incubation with CLB (17.5 µM) and VPA (0.5 mM), percentage of apoptosis, as well as expression of two TP53 target genes (p21 and HDM2) and two genes from Bcl-2 family (BCL2 and MCL1), were tested. As a result, an increased percentage of apoptosis was observed for CLL cells treated with CLB and VPA, and with CLB alone. Under the treatment with the drug combination, the expression of p21 gene was visibly higher than under the treatment with CLB alone. At the same time, the cultures under CLB treatment showed visibly higher expression of BCL2 than the cultures with VPA alone. Thus, the present study strongly suggests further investigations on the CLB and VPA combination in CLL treatment.

High performance liquid chromatography with ultra-violet detection (HPLC-UV) and gas chromatography–mass spectrometry (GC-MS) methods were developed and validated for the determination of chlorambucil (CLB) and valproic acid (VPA) in plasma, as a part of experiments on their anticancer activity in chronic lymphocytic leukemia (CLL). CLB was extracted from 250 µL of plasma with methanol, using simple protein precipitation and filtration. Chromatography was carried out on a LiChrospher 100 RP-18 end-capped column using a mobile phase consisting of acetonitrile, water and formic acid, and detection at 258 nm. The lowest limit of detection LLOQ was found to be 0.075 μg/mL, showing sufficient sensitivity in relation to therapeutic concentrations of CLB in plasma. The accuracy was from 94.13% to 101.12%, while the intra- and inter-batch precision was ≤9.46%. For quantitation of VPA, a sensitive GC-MS method was developed involving simple pre-column esterification with methanol and extraction with hexane. Chromatography was achieved on an HP-5MSUI column and monitored by MS with an electron impact ionization and selective ion monitoring mode. Using 250 µL of plasma, the LLOQ was found to be 0.075 μg/mL. The accuracy was from 94.96% to 109.12%, while the intra- and inter-batch precision was ≤6.69%. Thus, both methods fulfilled the requirements of FDA guidelines for the determination of drugs in biological materials.

Background: Immune modulatory factors like indoleamine 2,3-dioxygenase 1 (IDO1) generating kynurenine (Kyn) and receptor for advanced glycation end-products (RAGE) contribute to endometrial and cancer microenvironment. Using adequate experimental models is needed to learn about the significance of these molecular factors in endometrial biology. In this paper we study IDO1 activity and RAGE expression in the in vitro cultured primary human endometrial cells derived from cancerous and noncancerous tissue. Methods: The generated primary cell cultures from cancer and noncancerous endometrial tissues were characterized using immunofluorescence and Western Blot for expression of endometrial and cancer markers. IDO1 activity was studied by Kyn quantification with High Performance Liquid Chromatography with Diode Array Detector. Results: The primary cultures of endometrial cells were obtained with 80% success rate and no major genetic aberrations. The cells retained in vitro expression of markers (mucin MUC1 and HER2) or immunomodulatory factors (RAGE and IDO1). Increased Kyn secretion was associated with cancer endometrial cell culture in contrast to the control one. Conclusions: Primary endometrial cells express immune modulatory factors RAGE and IDO1 in vitro associated with cancer phenotype of endometrium.

Szumna Klaudia, Piędel Faustyna, Jasielski Patryk, Habaj Kamila, Grosman Sylwia, Filip Agata. Cancer Stem Cells as a new promising approach of efficient oncological treatment - the review of literature. Journal of Education, Health and Sport. 2020;10(9):115‑120. eISSN 2391-8306. DOI http://dx.doi.org/10.12775/JEHS.2020.10.09.013 https://apcz.umk.pl/czasopisma/index.php/JEHS/article/view/JEHS.2020.10.09.013 https://zenodo.org/record/4016679 The journal has had 5 points in Ministry of Science and Higher Education parametric evaluation. § 8. 2) and § 12. 1. 2) 22.02.2019. © The Authors 2020; This article is published with open access at Licensee Open Journal Systems of Nicolaus Copernicus University in Torun, Poland Open Access. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author (s) and source are credited. This is an open access article licensed under the terms of the Creative Commons Attribution Non commercial license Share alike. (http://creativecommons.org/licenses/by-nc-sa/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. The authors declare that there is no conflict of interests regarding the publication of this paper. Received: 10.08.2020. Revised: 15.08.2020. Accepted: 06.09.2020. Cancer Stem Cells as a new promising approach of efficient oncological treatment - the review of literature Klaudia Szumna1, Faustyna Piędel1, Patryk Jasielski1, Kamila Habaj1, SylwiaGrosman1, Agata Filip2 Student's Association of Cancer Genetics in Department of Cancer Genetics with Cytogenetic Laboratory. Medical University of Lublin, Chodźki Street 19, 20-093 Lublin, Poland. Department of Cancer Genetics with Cytogenetic Laboratory. Medical University of Lublin, Chodźki Street 19, 20-093 Lublin, Poland. Klaudia Szumna; ORCID:0000-0002-0889-2627; klaudiadaria.szumna@gmail.com Faustyna Piędel; ORCID:0000-0002-8280-498X; faustyna.piedel@gmail.com Patryk Jasielski; ORCID: 0000-0002-0958-735X; patryk.jasielski111@gmail.com Sylwia Grosman; ORCID: 0000-0003-2126-6011; s.grosman@op.pl Kamila Habaj; ORCID: 0000-0002-1733-8046; habajkamila@gmail.com Agata Filip; ORCID: 0000-0001-7591-5887; A.filip@umlub.pl Summary: Introduction and purpose: For many years a lot of research projects have been conducted with the aim to discover special cells, which are responsible for a neoplasm development. One of the theories concerns cancer stem cells (CSCs). This theory presumes the existence of original, undifferentiated cancer cells. These cells are capable of transform to all types of cells in neoplasm mass. The purpose of this study was to review the currently available data about the theory of CSCs and especially potential usage of this theory in oncological treatment. Pubmed and Scopus databases from the last 10 years were searched for phrase “cancer stem cells”. A brief description of the state of knowledge: Impaired signaling pathways lead to an occurrence of CSCs which may contribute to the development of neoplasm eventually. It is proved, that gamma secretase inhibitors impede Notch signaling pathway, which leads to a decrease of tumour markers expression and consequently decline in vivo tumour growth. It is believed that epigenetic mechanisms, including DNA methylation, histones alteration and RNA orientation are pivotal regulators of CSCs. It was shown, that histone deacetylases (HDAC) modulate refractoriness on chemotherapy in haematological tumours. It is also evidenced that peculiar designed microRNA directly inhibit CD44 which suppresses prostatic cancer stem cells and its metastases. CSCs have high expression of programmed death-ligand 1 (PD-L1) including breast and colon cancer, which proclaims in favour of immunotherapy aimed on CSCs. Conclusions: Constantly growing knowledge about CSCs biology is creating new opportunities of detection, isolation and design of the therapies aimed on CSCs. Key words: Cancer stem cells, elective treatment, chemotherapy, remission.

Psiuk Dominika, Rocka Agata, Żak Klaudia, Tomczyk Żaklina, Filip Agata. The efficacy of HDAC inhibitors in neoplasm treatment. Journal of Education, Health and Sport. 2020;10(8):244-250. eISSN 2391-8306. DOI http://dx.doi.org/10.12775/JEHS.2020.10.08.028 https://apcz.umk.pl/czasopisma/index.php/JEHS/article/view/JEHS.2020.10.08.028 https://zenodo.org/record/3987520 The journal has had 5 points in Ministry of Science and Higher Education parametric evaluation. § 8. 2) and § 12. 1. 2) 22.02.2019. © The Authors 2020; This article is published with open access at Licensee Open Journal Systems of Nicolaus Copernicus University in Torun, Poland Open Access. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author (s) and source are credited. This is an open access article licensed under the terms of the Creative Commons Attribution Non commercial license Share alike. (http://creativecommons.org/licenses/by-nc-sa/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. The authors declare that there is no conflict of interests regarding the publication of this paper. Received: 01.08.2020. Revised: 05.08.2020. Accepted: 17.08.2020. The efficacy of HDAC inhibitors in neoplasm treatment Dominika Psiuk1, Agata Rocka1, Klaudia Żak1, Żaklina Tomczyk1, Agata Filip2 1Student Research Group, Department of Cancer Genetics, Medical University of Lublin, Radziwillowska 11, 20-080 Lublin, Poland, tel. +48 81 4486102 2Head of the Department of Cancer Genetics, Medical University of Lublin, Radziwillowska 11, 20-080 Lublin, Poland, tel. +48 81 4486102 Dominika Psiuk: ORCID:0000-0003-3319-3489; dominika.psiuk@gmail.com Agata Rocka: ORCID:0000-0003-4738-3160; agatarocka2@gmail.com Klaudia Żak: ORCID:0000-0003-2421-2553; zakklaudia3@gmail.com Żaklina Tomczyk: ORCID:0000-0002-0251-0448; zaklinka512@gmail.com Agata Filip: ORCID: 0000-0001-7591-5887; agata.filip@umlub.pl Abstract Introduction and purpose: Histone Deacetylases (HDACs) are associated with epigenetic regulation of gene expression. They are responsible of adequate action of not only histone proteins, but also crucial cell cycle proteins, such as p53, NF‐κB or alpha tubulin. Recent studies have shown the connection between expression of HDAC and carcinogenesis and impact of HDAC inhibitors on cancer therapy. The aim of the study was to review the recent studies on HDAC inhibitors efficiency in the treatment of hematologic neoplasms and solid tumors. It is a study review from 2010 to 2020 using the PubMed database and ClinicalTrials.gov. Description of state of knowledge: Most studies related to HDAC inhibitors impact in hematological neoplasms, such as multiple myeloma, Hodgkin lymphoma, T-cell lymphomas and acute myeloid leukemia. Among studied agents panobinostat, vorinostat and showed efficacy. Moreover, some HDACi demonstrated effectiveness in solid tumors treatment, foremost vorinostat in breast cancer and entinostat in non-small cell lung lymphoma. Summary: Including HDAC inhibitors in tumor treatment could be beneficial for patients, especially those with advanced, relapsed or refractory hematologic neoplasm. However, further studies are necessary to confirm their efficacy and to reveal side effects. Key words: HDAC, inhibitors, cancer, hematology

Inhibitors of histone deacetylases (HDIs), by affecting the process of histone acetylation, lead to changes in chromatin condensation, and in consequence, to changes in the expression of numerous genes responsible for the cell cycle and differentiation. Therefore, they can be effective in the treatment of cancer. The antitumor activity for over 15 HDIs has been confirmed so far, and some of them have been approved in the USA and Europe, mainly in combination with cytostatics or radiotherapy. For several HDIs, large clinical trials are being carried out to estimate their effectiveness, in monotherapy and new combinations. Other synthetic and natural compounds with HDI activity are tested in preclinical studies. HDIs differ in terms of chemical structure, biological activity and specificity in relation to individual HDACs. The compounds active against classic HDACs (Zn2+-dependent metalloproteins) are usually characterized by the presence the Zn2+ binding group, the linker part and the capping group. Taking into account the type of Zn2+ binding group, classic HDIs are classified as short-chain fatty acids, hydroxamic acids, cyclic peptides and benzamides. In turn, the nicotinamide adenine dinucleotide-dependent SIRTs inhibitors are small molecules, mostly nicotinamide and β-naphthol derivatives. The presented paper summarizes the most important information regarding the use of HDIs as anticancer drugs, regarding their diversified chemical structure, activity against HDACs, additional therapeutic properties and side effects. The review was made taking into account the literature from the last five years (2013-2017).

Ginszt, M, Michalak-Wojnowska, M, Gawda, P, Wojcierowska-Litwin, M, Korszeń-Pilecka, I, Kusztelak, M, Muda, R, Filip, AA, and Majcher, P. ACTN3 genotype in professional sport climbers. J Strength Cond Res 32(5): 1311–1315, 2018—The functional RR genotype of the alpha-actinin-3 (ACTN3) gene has been reported to be associated with elite sprint/power athlete status. Although large and rapidly increasing number of studies have investigated the associations between the ACTN3 genotypes and athletic performance in various sport disciplines, there is a lack of studies on the genetic predisposition in sport climbing, which was selected to be part of the next Summer Olympic Games in Tokyo 2020 with three subdisciplines (“lead climbing,” “speed climbing,” and “bouldering”). The aim of the study is to determine the frequency distribution of ACTN3 genotypes and alleles in professional lead climbers and boulderers. 100 professional sport climbers from Poland, Russia, and Austria were divided into 2 equal groups: professional boulderers and professional lead climbers were involved in the study. ACTN3 allele frequencies and genotypes were compared with 100 sedentary controls. Genotypes were determined using polymerase chain reaction–restriction fragment length polymorphism method. The percent distribution of RR genotype in the boulderers was significantly higher than in lead climbers and controls (62 vs. 26%; 33%, respectively; χ2 = 17.230, p = 0.0017). The frequencies of ACTN3 R allele in boulderers differed significantly from lead climbers and controls (77 vs. 51%; 58%, respectively; χ2 = 15.721, p = 0.0004). The proportion of the ACTN3 RR genotype is significantly higher in boulderers than in lead climbers and may be related to the specific type of predisposition to this subdiscipline.

Introduction. Significant and accessible predictive factors for bortezomib treatment in plasma cell myeloma (PCM) are still lacking. TP53 codon 72 polymorphism (P72R) results in proline (P) or arginine (R) at 72 amino acid position, which causes synthesis of proteins with distinct functions. The aims of our study were to: 1) analyze whether this polymorphism is associated with an increased risk of PCM; 2) study whether the P72R polymorphism affects overall survival (OS) among PCM patients; 3) assess the possible association of the P72R polymorphism with sensitivity to bortezomib in cell cultures derived from PCM patients. Material and methods. Genomic DNA from newly diagnosed 59 patients (without IgVH gene rearrangements and TP53 deletions) and 50 healthy blood donors were analyzed by RFLP-PCR to identify TP53 polymorphism. Chromosomal aberrations were detected by use of cIg-FISH. The lymphocyte cell cultures from a subgroup of 40 PCM patients were treated with bortezomib (1, 2 and 4 nM). Results. The P allele of the P72R polymorphism was more common than the R allele in PMC patients compared to controls (39% vs. 24%), and the difference was significant (p = 0.02). The PP and PR genotypes (in combina­tion) were more frequent among cases than in controls (65% vs. 42%, OR = 2.32, p = 0.04). At the cell culture level and 2 nM bortezomib concentration the PP genotype was associated with higher necrosis rates (10.5%) compared to the PR genotype (5.7%, p = 0.006) or the RR genotype (6.3%, p = 0.02); however, no effect of genotypes was observed at bortezomib concentrations of 1 and 4 nM. The shortest OS (12 months) was observed in patients with the PP genotype compared to patients with the PR or RR genotypes (20 months) (p = 0.04). Conclusions. The results suggest that P72R polymorphisms may be associated with an increased PCM risk and may affect OS of PCM patients. However, we saw no consistent results of the polymorphism effect on apoptosis and necrosis in cell cultures derived from PCM patients. Further studies are need in this regard.

(1) Background: The aim of our study was to analyze the possible relationship of ABCB1 and CYP1A1 gene variants with susceptibility and outcome of multiple myeloma (MM), (2) Methods: Genomic DNA samples from 110 newly-diagnosed MM patients and 100 healthy blood donors were analyzed by methods-PCR-RFLP (for ABCB1 3435C &gt, T, CYP1A1 6235T &gt, C—m1), automated DNA sequencing (for ABCB1 1236C &gt, T, 2677G &gt, T/A) and allele-specific PCR (for CYP1A1 4889A &gt, G—m2), (3) Results: The genotypic frequencies of CYP1A1 4889A &gt, G variant were not in Hardy-Weinberg equilibrium for MM patients. The presence of m1 and m2 CYP1A1 alleles decreased the risk of MM—OR = 0.49 (p = 0.011) and OR = 0.27 (p = 0.0003), respectively. In turn, TT genotype (ABCB1 2677G &gt, T/A) increased the risk of this disease (p = 0.007). In the multivariate Cox analysis CT + TT genotypes (ABCB1 3435C &gt, T) were associated with decreased risk of death (HR = 0.29, p = 0.04). In log-rank test in patients with CT genotype (ABCB1 3435C &gt, T) was observed association of overall survival with the type of treatment, (4) Conclusions: Our findings suggest that T-alleles of ABCB1 2677G &gt, T/A and m1/m2 alleles of CYP1A1 affected the susceptibility of MM. Moreover, T-allele of ABCB1 3435C &gt, T might be independent positive prognostic factor in MM.

Advances in genotypic technologies enable identification of possible associations between genetic variants of certain genes and increased risk of developing plaque psoriasis or psoriatic arthritis. The aim of the study was to analyze the NOTCH3 (6746T>C) (rs1044009) and PSMA6 (8C>G) (rs1048990) polymorphisms and their role in genetic susceptibility to psoriasis. The study included 158 psoriatic patients and 100 healthy controls. The frequencies of the NOTCH3 genotypes differed between the psoriatic patients and controls (p=0.050). No differences were found in the distribution of PSMA6 genotypes and alleles between the psoriatic patients and controls. The studied psoriatic patients presented a higher frequency of the CC genotype of PSMA6 compared to the healthy controls (8.8% vs 2%, respectively). Psoriatic arthritis was more frequent among patients with the CC genotype of PSMA6 (p=0.059). Simultaneous CC homozygosity of NOTCH3 and PSMA6 was significantly more commonly observed in the studied psoriatic patients than in the controls (p=0.032). The obtained data suggest that genetic variants of NOTCH3 (6746T>C) and PSMA6 (-8C>G) genes may play significant roles in psoriatic patients. Further studies are necessary to unequivocally determine their role as genetic risk factors of psoriasis development.

Introduction. NOTCH pathway and TP53 protein are involved in the development of fibrosis and autoimmune disorders, respectively. The aim of this study was to evaluate the role of single nucleotide polymorphisms (SNPs) of NOTCH3 and TP53 genes and serum anti-TP53 antibodies with the susceptibility, clinical subset of systemic sclerosis (SSc), and clinical profile of SSc patient, particularly with lung involvement and disease activity. Objects and Methods. 124 white Polish SSc patients (101 with limited cutaneous SSc–lcSSc, and 23 with diffuse cutaneous SSc–dcSSc) and 100 healthy individuals were included in the study. Patients were assessed for the presence of autoantibodies and interstitial lung disease. Two SNPs at position 6746 of NOTCH3 gene (C/T alleles) and 215 of the TP53 gene (P/R alleles) were genotyped by PCR-restriction fragment length polymorphism. Serum levels of anti-TP53 antibodies were analyzed by means of ELISA. Results. The genotypic frequencies of the NOTCH3 gene for SSc patients diverged significantly from Hardy–Weinberg equilibrium (p=0.03; χ2 = 4.63). There was no significant difference between SSc patients and the control population in allele frequencies of both SNPs. The CT + CC genotypes of NOTCH3 influenced the susceptibility to SSc (OR = 1.85, p=0.04), including dcSSc (OR = 3.43, p=0.04), and active form of SSc (OR = 5.46, p<0.001). The PR + RR genotypes of the TP53 gene were associated only with dcSSc susceptibility (OR = 3.30, p=0.034). The levels of anti-TP53 antibodies were not related to studied SNPs and clinical parameters of SSc including the presence of specific antibodies and interstitial lung disease. Conclusion. The CT + CC genotypes of NOTCH3 gene and PR + RR genotypes of the TP53 gene increased the risk of dcSSc development. Moreover, genotypes of CT + CC were associated with the active form of SSc suggesting the role of the NOTCH pathway in the pathogenesis of this disease.

Introduction. Multiple myeloma (MM) is a hematological malignancy characterized by genetic variety. The 3020insC variant of the NOD2/CARD15 gene results in the upregulation of proinflammatory cytokines. Chronic inflammation and abnormal function of the proteasome system may lead to MM development. The polymorphism (-8C>G) in the PSMA6 gene affects proteasome activity. The aim of our study was to analyze the possible relationship of NOD/CARD15 and PSMA6 genes with the risk of development and outcome of MM, as well as the sensitivity to bortezomib (proteasome inhibitor) in cell cultures derived from MM patients. Objects and Methods. Genomic DNA from 100 newly diagnosed MM patients and 100 healthy blood donors was analyzed by methods such as PCR-RFLP (for PSMA6 genotyping) and automated DNA sequencing (for NOD2/CARD15 genotyping). In a subgroup of 50 MM patients, nucleated bone marrow cells were treated with bortezomib in vitro. Results. Patients with PSMA6 CG+GG genotypes had higher chances for progressive disease (OR=5.0, 95% CI 1.07-23.16, p=0.05), shorter overall survival taking into account the type of treatment (p=0.039), and increased risk of death due to MM at the level of tendency (OR=4.74, 95% CI 1.02-21.97, p=0.06). The presence of NOD2/CARD15 3020insC decreased the risk of renal dysfunction in MM (OR=0.23, 95% CI 0.07-0.74, p=0.009). The analyzed changes in NOD2/CARD15 and PSMA6 genes did not impact the MM risk. In an in vitro study, bortezomib increased the number of apoptotic cells at 8 nM and 12 nM between wild-type and 3030insC variants of NOD2/CARD15 (p=0.018 and p=0.03, respectively). Conclusion. The presented results suggest a possible impact of PSMA6 CG+GG genotypes on the MM outcome and the association of the NOD2/CARD15 variant with bortezomib in vitro sensitivity.

Oxidative stress and systemic inflammation are closely linked with increased risk of cancer development. Tumor necrosis factor alpha (TNF-α) is one of the pro-inflammatory cytokines. Glutathione S-transferases (GSTs) are enzymes involved in oxidative stress handling. Polymorphisms of genes encoding mentioned molecules may potentially influence the risk and the outcome in neoplastic diseases. Multiple myeloma (MM) is a hematological malignancy characterized by clonal, atypical plasma cell proliferation. In the present study we investigated the association of deletion polymorphisms in GSTT1/GSTM1 genes and single nucleotide polymorphisms (SNPs) in the TNF-α gene at positions −308/−238 with the risk and outcome in MM and sensitivity to bortezomib under in vitro conditions. One hundred newly diagnosed MM patients and 100 healthy blood donors were genotyped by means of multiplex PCR (for GSTs) and PCR-RFLP (for TNF-α). In a subgroup of 50 MM patients, bone marrow cells were treated with bortezomib in vitro. Patients with −238GA+AA or GSTT1-null genotypes had 2.0 (p = 0.002) or 2.29 (p = 0.013) fold increased risk of MM. The interaction effects and risk of MM were observed in GSTT1/GSTM1-null (OR = 2.82, p = 0.018), −308/−238GA+AA (OR = 5.63, p < 0.001), as well as in all combinations of −308 with GSTs. The −308/−238GA+AA genotypes in comparison to GG were associated with earlier MM onset−61.14 vs. 66.86 years (p = 0.009) and 61.72 vs. 66.52 years (p = 0.035), respectively. Patients with GSTM1-present had shorter progression-free-survival (15.17 vs. 26.81 months, p = 0.003) and overall-survival (22.79 vs. 34.81 months, p = 0.039) compared with GSTM1-null. We did not observe relationship between response rate and studied polymorphisms. The in vitro study revealed significantly higher number of apoptotic cells at 12 nM of bortezomib in GSTT1-present, GSTM1-null/present, −308GG and −238GG/GA+AA genotypes. Our findings comprise large analysis of studied polymorphisms in MM.

Results from numerous pre-clinical studies suggest that a well known anticonvulsant drug valproic acid (VPA) and other short-chain fatty acids (SCFAs) cause significant inhibition of cancer cell proliferation by modulating multiple signaling pathways. First of all, they act as histone deacetylase (HDAC) inhibitors (HDIs), being involved in the epigenetic regulation of gene expression. Afterward, VPA is shown to induce apoptosis and cell differentiation, as well as regulate Notch signaling. Moreover, it up-regulates the expression of certain G protein-coupled receptors (GPCRs), which are involved in various signaling pathways associated with cancer. As a consequence, some pre-clinical and clinical trials were carried out to estimate anticancer effectiveness of VPA, in monotherapy and in new drug combinations, while other SCFAs were tested in pre-clinical studies. The present manuscript summarizes the most important information from the literature about their potent anticancer activities to show some future perspectives related to epigenetic therapy.

Chronic lymphocytic leukemia (CLL) is one of the most frequent lymphoproliferative diseases. CLL is characterized by unusual heterogeneity, which probably reflects its biological and genetic lack of homogeneity. Clonal chromosome aberrations belong to the most important prognostic and predictive factors in CLL. This research was aimed at observing clonal evolution in CLL at the chromosomal level, and assessing its clinical significance in relation to selected prognostic factors. The study involved 72 untreated patients with CLL. The preliminary investigations using cytogenetic banding analysis (CBA) and FISH were performed at the time of diagnosis, and again after about 24 months to observe clonal changes (clonal evolution). In addition, other parameters were evaluated, i.e., the expression of ZAP-70 kinase, CD38 antigen, and the mutation statuses of IGVH and NOTCH1 genes. Classic prognostic factors, i.e., categorized ZAP70 and CD38 expressions as well as mutations in IGVH and NOTCH1 genes did not influence the course of clonal evolution in the examined group of patients. Clonal evolution was detected in 45.8% of patients by means of CBA, and in 19.4% patients with FISH. Analysis of chromosomal aberrations in the examined group of patients showed that the incidence of 17p deletions and translocations in karyotypes has a negative impact on overall survival. CE was found to be a risk factor for the occurrence of disease progression (OR = 2.22). Our observations indicate that combined CBA and FISH are the most optimal techniques for monitoring clonal evolution in the course of CLL.

Introduction: The insertion (I allele) deletion (D allele) polymorphism of ACE gene (rs4646994) may influence the etiopathogenesis of multiple myeloma (MM). ACE gene is expressed in bone marrow cells and encodes angiotensin converting enzyme (ACE). It converts angiotensin I to active peptide angiotensin II, which stimulates proliferation of hematopoietic stem cells. This suggests possible association of ACE I/D gene polymorphism with MM. The aim of our study was to check possible impact of this polymorphism on risk of development and outcome of MM, as well as, sensitivity to bortezomib in cell cultures derived from MM patients. Objects and Methods: Genomic DNA from 98 newly diagnosed MM patients and 100 healthy blood donors were analyzed by PCR method. Chromosomal aberrations were detected by use of cIg-FISH. In a subgroup of 40 MM patients nucleated bone marrow cells were treated with bortezomib in vitro. Results: The Hardy-Weinberg equilibrium test showed that genotypic frequencies diverged significantly from the equilibrium. The differences between I and D allele frequencies in control and study population were significant (p = 0.046). We observed the association between DD genotype and more than 2-fold risk of MM - OR = 2.69; p < 0.0001. We did not detect any significant differences among studied genotypes regarding clinical and laboratory parameters. Moreover, we did not observe the association between survival of MM patients and I/D genotypes. Bortezomib increased number of apoptotic and necrotic cells, but the only statistically significant differences were observed in the number of viable cells at 1 nM between ID and DD genotypes (p = 0.026). Conclusion: Presented results confirmed the significant relationship between ACE (I/D) polymorphism and risk of MM development. We did not observe the association of ACE I/D polymorphism with disease outcome and bortezomib in vitro sensitivity.

Introduction : Breast cancer (BC) in young women of Podkarpackie province accounts for approximately 11.0% of all diagnosed breast tumors. Aim of the study was to assess the trends in incidence of BC among women younger than 44 from Podkarpackie in the years 2002–2011. Material and methods: 614 cases of malignant BC and 26 cases of cancer in situ were analyzed. The crude and the standardized incidence ratios were estimated; the percentage of histopathologically confirmed cancer cases and the percentage share of registered malignant breast tumors were calculated. The analysis of incidence in individual counties was also presented, as well as the stages of clinical advancement at diagnosis and the methods of treatment. Results : The number of registered cases at 2011 was 73, and it was 37.7% higher as compared to 2002. During the period analyzed, the increase in the crude and the standardized cancer incidence ratios was observed. The percentage share of BC in the examined group increased of 5.4% in 2011. Significant variation in incidence among different counties was observed. The incidence ratios ranged from 65.8 to 93.1/100 000. BC in young women most commonly was diagnosed as locally advanced and over 70% of patients were radically treated. Conclusions : Even though the progress in diagnostics and treatment has been made, BC in young women is diagnosed later than it should be and at considerably advanced stage. It is relevant to propagate the knowledge among women and health professionals to emphasize that BC may affect young women.

Introduction Acute myeloid leukemia (AML) is a genetically heterogeneous disease at both the cytogenetic and molecular levels. In AML cells many chromosomal aberrations are observed, some of them being characteristic of a particular subtype of patients, and others being less significant. Besides chromosomal abnormalities, the leukemic cells can have a variety of mutations involving individual genes. The aim of this work was to investigate the frequencies of molecular alterations with the focus on FLT3-ITD and NPM1 mutations in AML patients of different age groups living in a southeastern region of Poland. Material and methods The study group comprised 50 consecutive AML patients. We analyzed bone marrow samples by conventional cytogenetics. Cytogenetic evaluation in selected cases was complemented by the FISH technique. The internal tandem mutation in the FLT3 gene was identified using polymerase chain reaction (PCR), and the NPM1 mutation was assessed by direct nucleotide sequencing. Results The studies using classical cytogenetics showed chromosomal aberrations in 32 (64%) patients. In 18 cases no changes in the karyotype were found by conventional karyotyping. FLT3-ITD mutation was detected in 4 (8%) patients and mutation of NPM1 in 3 patients with AML (6%). Conclusions The incidence of both mutations in our study group was lower than described elsewhere. We have confirmed that FLT3-ITD occurred more commonly in older patients and it was associated with shorter overall survival. By contrast, mutation of exon 12 of the NPM1 gene seems to be a good prognostic factor in AML patients with normal karyotype.

In Western countries the majority of gastric cancers (GC) are usually diagnosed in advanced stages reporting a 5-year survival rate of only 26%. The Lauren classification of GC was most widely used in clinical practice since it reflects GC morphology, epidemiology, tumor biology, clinical management and outcome. Despite the initial promise of individualizing antitumor treatment, the management of GC still remains relatively broad and general. Apart from clinical staging, molecular profiling enables targeting of the identified underlying alterations, rather than histology. In contrast to breast carcinoma, molecular classification of GC does not yet imply treatment modality. Molecular classifications of GC and their therapeutic implications are therefore extensively studied. The current proposed molecular divisions of GC come from three different parts of the world where different standard treatment modalities for advanced GC are recommended. Wider use of GC molecular subtyping may solve problems, such as susceptibility to novel systemic therapy regimens or selection of patients for aggressive surgery and targeted adjuvant/conversion therapy. In any case, the rapid entry of novel molecular targeted therapies into routine oncology practice clearly underscores the urgent need for clinicians to be aware of these new possibilities.

The Dopamine receptor D4 gene (DRD4) has been previously linked to financial risk-taking propensity. Past works demonstrated that individuals with a specific variant of the DRD4 gene (7R+) are more risk-seeking than people without it (7R−). The most prominent explanation for this effect is the fact that 7R+ individuals are less sensitive to dopamine and thus seek more stimulation to generate “normal” dopaminergic activity and feel pleasure. However, results about this relationship have not been conclusive, and some revealed a lack of the relationship. In the current work, we tested if those unclear results might be explained by the motivation that underlies the risk-taking activity; i.e., if people take risks to feel excitement or if they take risk to obtain a specific goal. In our study we tested the differences in risk-taking between 7R+ and 7R− among people who are experienced in financial risk-taking (113 investors) and non-experienced financial decision makers (104 non-investors). We measured risk-taking propensity with the Holt-Laury test and the Stimulating-Instrumental Risk Inventory. Moreover, we asked investors about their motivations for engaging in investment activity. Our study is the next one to report a lack of differences in risk-taking between 7R+ and 7R− individuals. As well, our results did not indicate any differences between the 7R+ and 7R− investors in motivation to engage in investment activity. We only observed that risk-taking propensity was higher among investors than non-investors and this was noticed for all measures. More research is needed to better understand the genetic foundations of risk-taking, which could answer the question about the substantial variation in the domain of risky financial decisions.

Jedną z grup lekow immunomodulacyjnych, modyfikujących dzialanie ukladu odpornościowego, są przeciwciala monoklonalne. Negatywne receptory, takie jak CTLA-4 i PD-1, obnizają poziom stymulacji komorki, a w opisywanych przypadkach — hamują aktywacje i proliferacje limfocytow T. Fizjologiczną funkcją tych receptorow jest eliminacja autoreaktywnych komorek odpornościowych, ktore nie zostaly zniszczone w mechanizmie tolerancji centralnej, oraz ochrona tkanek organizmu zmienionych zapalnie. Nowotwory, poprzez ekspresje ligandow dla negatywnych receptorow, unikają rozpoznania przez uklad immunologiczny. Dzialanie przeciwcial obejmuje blokowanie receptorow CTLA-4 i PD-1, przywracając limfocytom zdolnośc rozpoznawania i reagowania na antygeny nowotworowe. Dotychczasowe badania przeprowadzone nad lekami anty-CTLA, anty-PD-1 i anty-PD-L1 wykazują umiarkowaną skutecznośc w leczeniu niektorych zaawansowanych nowotworow, czesto opornych na leczenie standardowe, polączoną z trwalą remisją guza. Szczegolnie korzystne okazalo sie lączenie dwoch grup lekow z efektem synergistycznym. Pomimo zachecających wynikow dotychczasowych badan wciąz przed naukowcami stoją wyzwania, takie jak znalezienie obiektywnych markerow szacujących korzyści leczenia danego pacjenta, zmniejszenie liczby dzialan niepoządanych oraz normalizacja cen nowych lekow.

The Dopamine receptor D4 gene (DRD4) has been previously linked to financial risk-taking propensity. Past works demonstrated that individuals with a specific variant of the DRD4 gene (7R+) are more risk-seeking than people without it (7R−). The most prominent explanation for this effect is the fact that 7R+ individuals are less sensitive to dopamine and thus seek more stimulation to generate “normal” dopaminergic activity and feel pleasure. However, results about this relationship have not been conclusive, and some revealed a lack of the relationship. In the current work, we tested if those unclear results might be explained by the motivation that underlies the risk-taking activity; i.e., if people take risks to feel excitement or if they take risk to obtain a specific goal. In our study we tested the differences in risk-taking between 7R+ and 7R− among people who are experienced in financial risk-taking (113 investors) and non-experienced financial decision makers (104 non-investors). We measured risk-taking propensity with the Holt-Laury test and the Stimulating-Instrumental Risk Inventory. Moreover, we asked investors about their motivations for engaging in investment activity. Our study is the next one to report a lack of differences in risk-taking between 7R+ and 7R− individuals. As well, our results did not indicate any differences between the 7R+ and 7R− investors in motivation to engage in investment activity. We only observed that risk-taking propensity was higher among investors than non-investors and this was noticed for all measures. More research is needed to better understand the genetic foundations of risk-taking, which could answer the question about the substantial variation in the domain of risky financial decisions.

W niniejszej pracy przedstawiono zalezności pomiedzy zmianami w ekspresji poszczegolnych mikroRNA a powsta­waniem i rozwojem wybranych nowotworow. Dokonano przeglądu doniesien na temat uzyteczności badan nad ekspresją mikroRNA, ktore w przyszlości mogą byc wartościowymi i poządanymi przez diagnostow i lekarzy wskaź­nikami prognostycznymi i predykcyjnymi. Mogą sie one stac podwaliną do opracowania nowych metod leczniczych z wykorzystaniem antysensownych miRNA (antagomiry) czy lekow mających na celu kompensacje ilości cząsteczek w przypadku delecji lub uszkodzen genow dla konkretnych mikroRNA. Daloby to potencjalną mozliwośc regulacji ekspresji genow o znaczeniu strategicznym w procesach związanych z powstawaniem i rozwojem nowotworow.

Steatocystoma multiplex (SM) is an unusual benign disorder of the pilosebaceous duct characterized by multiple cysts with little or no nail and hair involvement. We report a 30-year-old woman with multiple cystic nodules located on the neck, axillae and forearms as well as patchy scalp alopecia. Histopathological examination of the lesions was diagnostic of SM. Trichoscopy revealed pili torti and pili canaliculi. This patient represents an unusual clinical presentation of SM because of the presence of hair abnormalities.

Chronic lymphocytic leukemia (CLL) mainly affects people older than 60 years. Accumulation of morphologically mature but dysfunctional B-lymphocytes in the bone marrow, lymph nodes and peripheral blood is a characteristic feature of this disease. Chromosomal aberrations are observed in lymphocytes of most CLL patients. Typical alterations include deletions of 13q14 and 11q, trisomy 12, and deletions of 17p. Altered expression of the genes located within involved regions, i.e. microRNA15/16 (13q14.3), ATM (11q22-q23) or TP53 (17p13) may be associated with the development and progression of the disease. Cryptic mutations may also contribute to leukemogenesis. Among others, they affect TP53, NOTCH1, SF3B1 and BIRC genes. CLL is a disease with heterogeneous course. There are two clinical forms – indolent and aggressive. The former is characterized by long time to first treatment and demise usually occurs because of coexisting diseases or is associated with leukemia-dependent immunodeficiency. Rapid clinical course and short overall survival, sometimes in spite of appropriate treatment implementation, is typical for aggressive form of CLL. For patients with this form, the moment of treatment initiation and the choice of first-line therapy are especially important, and depend inter alia on prognostic and predictive factors. Established poor prognostic factors in CLL include chromosomal aberrations, i.e., deletion of 17p or 11q, high ZAP-70 kinase expression, mutations/deletions of TP53, and lack of mutation of immunoglobulin heavy chain variable region genes (IgVH). In this paper we tried to point out the importance of some of the prognostic and predictive factors used routinely in the diagnostic management of CLL. Prognostic and predictive potential of microRNA expression level and recently described cryptic changes in the TP53, NOTCH1, SF3B1 and BIRC3 have also been presented.

B-cell chronic lymphocytic leukemia (B-CLL) is one of the most common hematologic malignancies in Western countries. Accumulation of leukemic lymphocytes in peripheral blood, bone marrow and secondary lymphatic organs of CLL patients is due to decreased apoptosis rather than to increased proliferation. The former is driven by signals from a specific microenvironment, created by stromal cells of mesenchymal origin, follicular dendritic cells, T lymphocytes and others. Nurse-like cells (NLCs) were first described to differentiate from peripheral blood mononuclear cells of CLL patients in vitro , then they have been also found in proliferation centers of their lymphatic tissues. Like tumor-associated macrophages (TAMs) in solid tumors, nurse-like cells promote survival of CLL lymphocytes. NLC gene expression patterns suggest their similarity to TAMs and differ between patients depending on ZAP70 protein expression status. NLC number in vitro corresponds with CD14 expressing cell count and beta-2-microglobulin serum level, and positively correlates with leukemic lymphocyte viability. As NLCs strongly express genes for adhesion molecules and secrete chemokines of antiapoptotic activity, they should be considered as a target for anti-microenvironment therapy of this incurable disease.

MicroRNAs (miRNAs) are a group of small molecules, about twenty nucleotides in length. They are involved in the regulation of gene expression mainly at a posttranscriptional level. This function depends on their complementarity to the 3'UTR regions of mRNAs. MicroRNAs are essential for proper development and functioning of the organism. They are so important because of their participation in such processes as angiogenesis, apoptosis, cell cycle control and oncogenesis. Over thirty percent of human genes are controlled by miRNAs. This indicates the great importance of these molecules. Alterations of numerous cellular processes can be caused by the dysregulation of miRNA expression. Such disturbances are observed in cancer cells, and signatures of microRNA expression are specific to particular types of cancer. It is suggested that miRNAs may serve as diagnostic and prognostic factors in oncologic and hematooncologic disorders. The expression of specific miRNAs can indicate benign or aggressive course of disease. The overall survival or time to treatment are also possible to estimate based on the microRNA expression profile. Knowledge about changes in miRNA expression observed in leukemia patients may enable the selection of appropriate individual therapy. Recent reports indicate that various hematooncologic disorders may be well characterized by microRNAs circulating in plasma or serum. It is of great potential importance, considering the availability of material for analysis, simplicity of diagnostic procedures and shortening of time required to conduct them.

B-cell chronic lymphocytic leukemia (B-CLL) is one of the most common leukemias of the elderly. To date, although many prognostic factors are known, none are universal or easily accessible thus allowing for the stratification of patients to slow-go and aggressive-course groups. Recent studies have identified new recurrent mutations in CLL cells, including mutation of the gene encoding one of the spliceosome subunits, SF3B1, mutation or rearrangement of NOTCH1, a gene of well-known tumorigenesis association, and disruption of BIRC3, a member of the inhibitors of apoptosis (IAP) family. This article presents the current state-of-the-art findings concerning the prognostic significance of these new alterations, as well as an explanation of the mechanisms underlying their biological impact on CLL lymphocytes.

Conventional cytogenetics is invariably one of the most important methods used in diagnostics of chronic lymphoproliferations. It complements fluorescence in situ hybridization (FISH) and molecular analysis. Presence of particular chromosomal alterations in chronic lymphocytic leukemia enables patients' stratification into appropriate cytogenetic risk groups and influences treatment decisions. In other non-Hodgkin lymphomas cytogenetic analyses are employed also in minimal residual disease assessment.As lymphocytes in chronic lymphoid malignancies are characterized by low proliferation rate in vitro, it is critical to induce their division in the culture properly. Here, we describe methods of lymphocyte isolation from patient's samples, conditions of cell culture, and the most commonly used mitogens for B- and T-lymphocytes in hemato-oncologic analyses.