Insertion/deletion polymorphism of the angiotensin converting enzyme (ACE) gene is a key component of the Renin-Angiotensin-Aldosterone System (RAAS). It has been proposed as an independent factor for hypertension and other cardiovascular diseases. Consequently, it has been extensively studied in various populations. The aim of this study is to investigate I/D polymorphism of ACE gene and its connection to hypertension in population of Tuzla Canton (Bosnia & Herzegovina). The study included 60 hypertensive subjects and 60 healthy control subjects with no risk factors for hypertension. I/D polymorphism was genotyped by polymerase chain reaction followed by gel electrophoresis and data obtained were statistically analysed using Chi square test. Odd’s ratios were calculated with a 95% confidence interval. P-value < 0.05 was considered significant. Odd’s ratios were calculated with a 95% confidence interval. P-value <0.05 was considered significant. Higher frequency of genotype D/D and allele D was determined in subjects with hypertension compared to control subjects but there is no statistical significance (p>0.05). However, statistically significant association was found in compared groups of subjects with genotypes DD + ID, in regards to genotype I/I (p<0.05). The results indicate the conclusion that ACE I/D polymorphism cannot be considered the main risk factor for development of hypertension, but its influence should be investigated together with other genetic and acquired risk factors that are associated with hypertension. This research contributes to the on-going exploration of molecular-genetic associations with hypertension.

Introduction: Prenatal diagnostic is a diagnostic method which is used to prove the presence of chromosome changes, a large number of metabolic disorders and other morphological fetus abnormalities. Prenatal genetic testing mostly refers to the molecular genetic and cytogenetic methods used during pregnancy to diagnose genetic fetal conditions. Aim: To investigate the existence and incidence of cytogenetics abnormalities in fetuses. Material and Methods: The retrospective research is based on cytogenetic analysis of the 1510 amniotic fluid samples collected from pregnant women sent to the cytogenetic laboratory from January, 2012 to December, 2022. Results: The karyotype without visible structural and numerical changes was detected in 96.8% (1462/1510) cases. The fetal karyotype was abnormal in 3.2 % (48/1510) of the cases. Trisomy 21 was the most frequent chromosome aberration detected in 1.12% (17/1510) cases followed by pericentric inversion 9 (10/1510; 0.66%) and trisomy 18 (4/1510; 0.26%). Mosaics were detected in five cases (5/1510; 0.33%). Comparing the prevalence of chromosome abnormalities according to maternal age, we come to know the prevalence of chromosome aberrations in the group of females above age 35 (26/790; 17.2/1000) is higher than in the group of females under age 25 (7/95; 4.63/1000), but not significantly different (P= 0.09). Conclusion: Conventional cytogenetics maintains its role as a powerful diagnostic tool in detecting chromosomal changes during prenatal screening.

Chronic myeloid leukemia (CML) is a hematological disorder characterized by increased proliferation of the granulocytic cell lineage. We diagnose CML by presence of the breakpoint cluster region-abelson (BCR-ABL1) oncogene using the quantitative real-time polymerase chain reaction (QRT-PCR). This method provides an accurate and unambiguous follow-up of the treatment response to tyrosine kinase inhibitors (TKIs). This study aimed to determine the molecular response to the first and second generation of TKIs in first and second line of treatment using the QRT-PCR method. We conducted a retrospective study on 48 CML patients treated with the first and second generation of TKIs in first and second-line treatment. Treatment responses have been followed-up every 3 months using the QRT-PCR method. Patients were divided into three groups according to molecular responses to the first line of TKIs. Results obtained in this study showed that the first group of patients did not achieve major molecular response (MMR) in the first 18 months of TKI treatment. The second and third group of patients achieved MMR and deep molecular response (DMR) in the first 18 months of TKI treatment. These results indicate that patients with MMR and DMR in the first 18 months of TKIs treatment had a favourable clinical course of the disease. Inadequate molecular responses to the first line of TKIs can be improved with in increase of the dose of TKIs or by switching to other TKIs. Continuous and timely molecular monitoring of TKI’s response in CML patients provides a careful observation of the disease's course and a proper treatment approach.

Introduction: Since 1956 karyotype analysis becomes an essential part of routine medical diagnostics, and helped medical professionals investigate the origin of genetic abnormalities in many constitutional and cancer diseases. Karyotyping also provided more information in the monitoring of fertility problems. An inversion does not usually have a phenotypic effect, especially if it involves a heterochromatin area, such as 9qh. Chromosome 9 polymorphism, with breakpoints p11q13/p12q13, can be the cause of variant abnormal clinical conditions such as congenital abnormalities, hematological diseasesand also could have a connection with pregnancy loss and fertility failure. Methods: A retrospective study was conducted on 1784 cytogenetics examination results from peripheral blood samples in the period from January, 2012 to December, 2022. The patients, carriers inv(9) in their karyotype were highlighted for detailed analysis. Results: Among the 1784 patients, constitutional pericentric inversion inv(9)(p11q13) was found in 13 females (0,72%), while it was seen in 17 cases of males (0.95%). The total average amount of inv (9) in this study is 1.68%. The inv(9) population consists of 60% cases with infertility problems, 6,66% females who had spontaneous abortus and 33,33% were patientsreferred to our laboratory for other reasons. Conclusion: In this research, the prevalence of inv (9) in the population of patients of Northeast Bosnia and Herzegovina who had the reproductive failure is shown. We believe that these results will help in finding the key to the truth about the association of this chromosome polymorphism with some pathological conditions such as fertility problems.

Introduction: Besides cardiovascular, malignant diseases are one of the leading causes of death in Bosnia and Herzegovina. At the top of this list are hematological diseases. This research aimed to identify cytogenetic and molecular biomarkers in patients treated for different types of hematological neoplasms. Methods: The retrospective study included 1600 samples of patients with different hematological diseases in the period from January 2006 to May 2022. The Polymerase Chain Reaction (RT-PCR) method was used to determine the presence of genetic rearrangements and to confirm the findings of conventional cytogenetic analysis. Results: Chromosomal aberrations were found in 739 (46,18%) patients. Using the RT-PCR technique, positive cases were increased by 1,5%. The BCR-ABL fusion gene was present in e14-a2 transcript form in 73% of samples, e13-a2 isoform in 21%, e1-a2 in 2%, while e14-a2/e1-a2 transcript coexpression was present in a percentage of 4% of the samples. The PML-RARa fusion gene was found in the form of bcr 1 transcripts in 21%, bcr2 32% and bcr3 59% of the samples. In twelve cases A type of the CBFB-MYH11 fusion transcript was detected. The MLL-AF4 fusion was found in only one case. Conclusion: The obtained percentages of frequency of individual molecular gene isoforms are in accordance with the results of most other researchers. This refers to the Balkan population and the Caucasian ethnic group.

Cytogenetic testing plays a major role in the diagnosis of different types of lymphadenopathies, assessment of survival prognosis, but also in the selection of adequate therapeutic strategies. Reports on aggressive head and neck lymphomas combining (cyto)genetics with pathology are rare, also lacking in Bosnia and Herzegovina. The aim of this retrospective study was to provide all chromosome aberrations data recorded in the group of patients diagnosed with malignant head and neck lymphadenopathy, and to analyze advantages and disadvantages of bone marrow (BM) cytogenetics analysis. Out of 819 patients who underwent cytogenetic analysis of BM in five years’ time spread, chromosomal abnormalities were analysed in 54 karyotypes of patients with clinically suspected head and neck lymphadenopathy. We recorded 66,6% Non-Hodgkin lymphoma, 26% Hodgkin lymphoma, 3,7% Acute lymphoblastic leukemia and 3,7% Chronic lymphocytic leukemia. Chromosomal abnormalities in the karyotype were detected in 32 (59.2%) of a total of 54 patients. A normal karyotype was observed in 14 (26%) patients. In 8 (14.8%) subjects, it was not possible to perform cytogenetic analysis. The results of this research are representative in a term of the karyotype characteristics of patients with head and neck lymphoma. This is the first work of its kind in Bosnia and Herzegovina and will continue through a multicenter study in order to characterise the diagnostic and prognostic significance of cytogenetic abnormalities in lymphoma patients.

As we know, the Philadelphia chromosome (Ph) is a highly specific marker for chronic myeloid leukemia (CML). This hematological disease is characterised by the formation of the BCR/ABL1 fusion gene, usually with typical translocation pattern including 9q34 and 22q11. In this paper we describe a 55 years old female patient with typical clinical and haematological signs of CML and a chromosome 9 differing from that which normally participates in translocation t(9;22). The karyotype of this Ph positive patient is characterised by pericentric inv(9)(p13q34) of the der(9)t(9;22)(q34;q11). Reverse transcriptase-polymerase chain reaction revealed a e14a2 type of BCR/ABL1 fusion transcript. As a consequence of this unusual translocation, FISH also found the separation of the ABL1/BCR1 fusion gene on chromosome 9. On reviewing the literature, to date only 10 Ph-positive leukemia patients have been noticed to have pericentric inversion inv(9)(p22q34)der(9)t(9;22)(q34;q11). No one case has been described with pericentric inversion inv(9)(p13q34) of the der(9)t(9;22)(q34;q11). This indicate that pericentric inv(9)(p13q34) of the der(9)t(9;22)(q34;q11) is a novel, rare, chromosomal abnormality in Phpositive CML.

Introduction: The treatment response and outcome in acute myeloid leukaemia (AML) is heterogeneous. Aim: To analyze the prognostic parameters of AML at presentation. Methods: The total sample of 44 AML patients was analyzed on the basis of age <55 and ≥55 years, sex, WBC count <50x10/9/l and ≥50x10/9/l, the Hb concentration <100 g/l and ≥100 g/l, PLT count <100x10/9/l and ≥100x10/9/l, Karnofsky score <60% and >60%, cytogenetics, CD56 expression, morphological type and types of treatment (standard and reduced induction chemotherapy, high–dose chemotherapy/stem cell transplantation – autologous and HLA matched, related, allogeneic, together and separately). Results: The age <55 years, Karnofsky score >60% and standard induction chemotherapy statistically correlated with the higher complete remission (CR) rates, longer relapse free survival (RFS), lower relapse rate (RR), and longer overall survival (OS) (p<0.01). The difference in terms of CR and RR between the sexes were not statistically significant (p<0.05), however women had statistically lower OS comparing to men (9.71±4.54 months vs. 38.03±9.17 months) (p<0.01). WBC count ≥ 50x10/9/l and the Hb concentration <100 g/l statistically correlated with shorter OS (p<0.05), while the WBC count ≥50x10/9/l statistically correlated with shorter RFS (p<0.05). The PLT count <100x10/9/l and ≥100x10/9/l was not found as prognostically significant for CR, RR, RFS, and OS (p<0.05). In comparison to the standard induction chemotherapy, both types of high dose chemotherapy/stem cell transplantation (HDT/SCT) (10/22), together and separately, resulted in longer RFS, lower RR, and longer OS (p<0.05). The frequency of cytogenetic risk was intermediate 81.6%, unfavorable 13.2%, and favorable 5.3%, respectively. CD56 + expression statistically correlated with the lower PLT count, higher RR, shorter RFS, and shorter OS (p<0.05). Statistical analysis of the cytogenetic risk and morphological types of AML were not possible due to the small number of patients in stratified groups. Conclusions: Female sex, the WBC count >50x10/9/l, the concentration of Hb <100 g/l, and CD56 + expression, at presentation of AML, should be considered as parameters of adverse risk, especially in latter decisions considering post-remission treatment with HDT/SCT.

Molecular monitoring of chronic myeloid leukemia patients using robust BCR-ABL1 tests standardized to the International Scale (IS) is key to proper disease management, especially when treatment cessation is considered. Most laboratories currently use a time-consuming sample exchange process with reference laboratories for IS calibration. A World Health Organization (WHO) BCR-ABL1 reference panel was developed (MR 1 – MR 4 ), but access to the material is limited. In this study, we describe the development of the fi rst cell-based secondary reference panel that is traceable to and faithfully replicates the WHO panel, with an additional MR 4.5 level. The secondary panel was calibrated to IS using digital PCR with ABL1 , BCR and GUSB as reference genes and evaluated by 44 laboratories worldwide. Interestingly, we found that 4 40% of BCR-ABL1 assays showed signs of inadequate optimization such as poor linearity and suboptimal PCR ef fi ciency. Nonetheless, when optimized sample inputs were used, 4 60% demonstrated satisfactory IS accuracy, precision and/or MR 4.5 sensitivity, and 58% obtained IS conversion factors from the secondary reference concordant with their current values. Correlation analysis indicated no signi fi cant alterations in %BCR-ABL1 results caused by different assay con fi gurations. More assays achieved good precision and/or sensitivity than IS accuracy, indicating the need for better IS calibration mechanisms.

Molecular monitoring of chronic myeloid leukemia patients using robust BCR-ABL1 tests standardized to the International Scale (IS) is key to proper disease management, especially when treatment cessation is considered. Most laboratories currently utilize a time-consuming sample exchange process with reference laboratories for IS calibration. A World Health Organization (WHO) BCR-ABL1 reference panel was developed (MR 1 -MR 4 ), but access to the material is limited. In this study, we describe the development of the first cell-based secondary reference panel that’s traceable to and faithfully replicates the WHO panel, with an additional MR 4.5 level. The secondary panel was calibrated to IS using digital PCR with ABL1 , BCR , and GUSB as reference genes and evaluated by 44 laboratories worldwide. Interestingly, we found that > 40% of BCR-ABL1 assays showed signs of inadequate optimization such as poor linearity and suboptimal PCR efficiency. Nonetheless, when optimized sample inputs were used, > 60% demonstrated satisfactory IS accuracy, precision and/or MR 4.5 sensitivity, and 58% obtained IS conversion factors from the secondary reference concordant with their current ones. Correlation analysis indicated no significant alterations in %BCR-ABL1 results caused by different assay configurations. More assays achieved good precision and/or sensitivity than IS accuracy, indicating the need for better IS calibration mechanisms. 78% (14 of 18) at MR 3 obtained results with ≥ 0.2 log difference. Overall, these results showed that some BCR-ABL1 tests were non-linear and might therefore yield statistically different %BCR-ABL1 results against different sample inputs. To mitigate the risk of inaccurate %BCR-ABL1 measurements from using an