Frekvencija mikronukleusa u limfocitima periferne krvi osoba izloženih osiromašenom uranu

One of the negative environmental impacts of the last armed conflict in Bosnia and Herzegovina was the use of radioactive ammunition containing depleted uranium. The United Nations Environment Programme measurements detected higher radioactivity at several examined sites in Bosnia and Herzegovina. One of those places is in the area of Hadžiæi, close to Sarajevo. This research included an evaluation of genetic load in human lymphocytes due to exposure to depleted uranium. The study included individuals who were located in the area of Hadžići and who were directly exposed to depleted uranium. The control blood samples were taken from individuals who lived in West Herzegovina which is considered environmentally uncontaminated. The results of the micronucleus cytochalasin-B test in peripheral blood lymphocytes showed increased micronuclei frequencies in the exposed group.Jedan od genotoksina, prisutnih u okolišu kao posljedica ratnih djelovanja u Bosni i Hercegovini jest osiromašeni uran. Njegovo porijeklo veže se za upotrebu radioaktivne antitenkovske municije s osiromašenim uranom. UNEP-ova mjerenja otkrila su povećanu radioaktivnost na nekoliko ispitanih lokaliteta od kojih je jedan na području Hadžića, u blizini Sarajeva. Naše istraživanje obuhvatilo je evaluaciju genetičkog opterećenja u humanim limfocitima periferne krvi osoba koje su bile nastanjene na području Hadžića te bile direktno izložene osiromašenom uranu. Kao kontrola u istraživanju, uzeta je krv od osoba nastanjenih na području zapadne Hercegovine, koja se smatra ekološki nekontaminiranom. Korištena je metoda mikronukleus-citokalazin B testa, a frekvencije mikronukleusa ispitanika iz obje populacije međusobno su komparirane. Rezultati istraživanja pokazuju povećanu frekvenciju mikronukleusa među ispitanicima eksponirane populacije

Nova in vitro otkrića o citotoksičnosti halogeniranoga boroksina i deregulaciji gena povezanih sa staničnom smrću u stanicama GR-M melanoma

Anti-proliferative effects of halogenated boroxine – K2(B3O3F4OH) (HB) – have been confirmed in multiple cancer cell lines, including melanoma, but the exact mechanism of action is still unknown. This study aimed to determine its cytotoxic effects on human Caucasian melanoma (GR-M) cell growth in vitro as well as on the expression of cell death-related genes BCL-2, BECN1, DRAM1, and SQSTM1. GR-M and peripheral blood mononuclear (PBM) cells were treated with different HB concentrations and their growth inhibition and relative gene expression profiles were determined using the Alamar blue assay and real-time PCR. HB significantly inhibited cell growth of both GR-M and PBM cells but was even more effective in GR-M melanoma cells, as significant inhibition occurred at a lower HB concentration of 0.2 mg/mL. GR-M BCL-2 expression was significantly downregulated (P=0.001) at HB concentration of 0.4 mg/mL, which suggests that HB is a potent tumour growth inhibitor. At the same time, it upregulated BCL-2 expression in normal (PBM) cells, probably by activating protective mechanisms against induced cytotoxicity. In addition, all but the lowest HB concentrations significantly upregulated SQSTM1 (P=0.001) in GR-M cells. Upregulated BECN1 expression suggests early activation of autophagy at the lowest HB concentration in SQSTM1 cells and at all HB concentrations in PBM cells. Our findings clearly show HB-associated cell death and, along with previous cytotoxicity studies, reveal its promising anti-tumour potential.Antiproliferativni učinci halogeniranoga boroksina – K2(B3O3F4OH) (HB) – potvrđeni su u više staničnih linija raka, uključujući melanom, ali točan mehanizam djelovanja još uvijek nije poznat. Cilj ovoga istraživanja bio je utvrditi njegove citotoksične učinke na rast stanica ljudskoga melanoma (GR-M) in vitro, kao i na ekspresiju gena BCL-2, BECN1, DRAM1 i SQSTM1, povezanih sa staničnom smrću. GR-M melanom i mononuklearne stanice periferne krvi (PBM) tretirane su različitim koncentracijama HB-a, a njihova inhibicija rasta i relativni profili ekspresije gena određeni su Alamar blue testom i real-time PCR-om. HB je značajno inhibirao rast GR-M melanoma i PBM stanica, no u GR-M melanomu učinci su registrirani pri nižim koncentracijama HB-a. Ekspresija BCL-2 gena u GR-M melanomu bila je značajno smanjena (P=0,001) pri koncentraciji od 0,4 mg/mL, što sugerira da je HB snažan inhibitor rasta tumora. Istodobno, pojačao je ekspresiju BCL-2 u normalnim PBM stanicama, vjerojatno aktiviranjem zaštitnih mehanizama protiv inducirane citotoksičnosti. Osim toga, sve osim najniže koncentracije HB-a značajno su povećale ekspresiju SQSTM1 (P=0,001) u GR-M melanomu. Povećana ekspresija BECN1 u najnižoj koncentraciji HB-a u GR-M stanicama i pri svim koncentracijama u PBM stanicama sugerira ranu aktivaciju autofagije. Naša otkrića jasno pokazuju indukciju stanične smrti povezane s HB-om i zajedno s prethodnim studijama citotoksičnosti otkrivaju njegov obećavajući antitumorski potencijal

Sporadic chromosome translocation frequencies in lymphocyte cultures – a retrospective study in a cohort of patients from Bosnia and Herzegovina

Aim Chromosome translocations are considered as one of the most severe forms of genome defects. Because of the clinical significance of chromosome translocations and scarce data on the incidence of sporadic translocations in population of Bosnia and Herzegovina, we aimed to report sporadic translocation frequencies in samples karyotyped in our laboratory. Methods The study group consisted of 108 samples. Whole blood was cultivated in complete medium for 72 hours with the thymidine application at 48th hour to synchronize the cell culture. Metaphases were arrested by colcemid 60 minutes before harvesting. Following hypotonic treatment, cells were fixed and cell suspension was dropped on coded slides. Dried slides were subjected to conventional GTG (G-banding with trypsin-Giemsa) banding and analyzed under 1000x magnification in the accordance with ISCN (International System for Human Cytogenetic Nomenclature) and E.C.A. Cytogenetic Guidelines and Quality Assurance. Results The incidence of all detected sporadic translocations was 27.81 x 10-4 per metaphase. The incidence of sporadic translocations involving chromosomes 7 and 14, being considered as the most frequent sporadic translocations of the human karyotype in phytohaemagglutinin (PHA) stimulated lymphocytes, was 15.89 x 10-4 per metaphase. The most frequent breakpoints were 7p21, 14q11 and 14q21. Other detected sporadic translocation breakpoints were: 1q25, 3p22, 7p13, 7q11.22, 7q33, 14q23 and 19q13.4. Conclusion Higher incidence of sporadic translocations compared to the similar studies was registered. Since potential explanations for this issue are smaller sample size and higher exposure of examined population to genotoxic agents, further monitoring of sporadic translocation incidences is recommended

Effects of dipotassium-trioxohydroxytetrafluorotriborate, K2[B3O3F4OH], on cell viability and gene expression of common human cancer drug targets in a melanoma cell line.

Recently it was found that dipotassium-trioxohydroxytetrafluorotriborate, K2(B3O3F4OH), is a potent and highly specific inhibitor of precancerous cell processes. We conducted gene expression profiling of human melanoma cells before and after treatment with two concentrations (0.1 and 1 mM) of this boron inorganic derivative in order to assess its effects on deregulation of genes associated with tumor pathways. Parallel trypan blue exclusion assay was performed to assess the cytotoxicity effects of this chemical. Treatment with K2(B3O3F4OH) induced a significant decrease of cell viability in melanoma cellline at both tested concentrations. Furthermore, these treatments caused deregulation of more than 30 genes known as common anti-tumor drug targets. IGF-1 and hTERT were found to be significantly downregulated and this result may imply potential use of K2(B3O3F4OH) as an inhibitor or human telomerase and insulin-like growth factor 1, both of which are associated with various tumor pathways

A retrospective analysis of spontaneous chromosomal aberrations in human lymphocyte cultures of individuals from Bosnia and Herzegovina

Spontaneous chromosomal aberrations are structural or numerical changes of chromosomes that occur naturally, without exposure to external genotoxic factors. They are not inherited, occur randomly in the karyotype, and do not have direct clinical significance. However, they can affect genomic instability and disease predisposition. They can result from DNA replication or repair processes errors, and typically are observed in cells that are actively dividing. Spontaneous chromosomal aberrations may arise due to the natural chromosomal instability and can be elevated in individuals exposed to mutagens. We analyzed frequencies of spontaneous chromosomal aberrations in 137 individuals subjected to karyotype analysis at the Laboratory for Cytogenetics and Genotoxicology, University of Sarajevo – Institute for Genetic Engineering and Biotechnology, during 2008-2023. Whole blood samples were cultivated for 72 hours with the thymidine added in the 48th hour. Metaphases were arrested by colcemid 60 minutes before harvesting. GTG banding was performed and slides were analyzed under 1000x magnification in accordance with An International System for Human Cytogenetic Nomenclature and E.C.A. Cytogenetic Guidelines and Quality Assurance. Constitutionally aberrant karyotypes were found in 2.92% of analysed individuals as well as altered karyotypes considered as normal chromosomal variants. In the total of 3092 analyzed metaphases, 20 spontaneous chromosomal aberrations were found in 13 individuals. This study contributes to the limited knowledge of the cytogenetic status of the Bosnian and Herzegovinian population. Further monitoring of spontaneous chromosomal aberrations incidences is recommended

Antiproliferative and genotoxic potential of xanthen-3-one derivatives

Twelve previously synthesized, biologically active 2,6,7-trihydroxyxanthen-3-one derivatives were evaluated in vitro for antiproliferative activity. Compounds were screened against HeLa, SW620, HepG2 and A549 tumor cell lines. Compound with the trifluormethyl group on C-4\u27 position of the phenyl ring showed the best inhibitory activity towards HeLa and A549 tumor cells with IC50 of 0.7 µmol L–1 4.1 µmol L–1, resp. Compound with chlorine and fluorine substituents on aryl ring showed the best antiproliferative activity against SW620 with IC50 of 4.1 µmol L–1 and against HepG2 tumor cell line with IC50 of 4.2 µmol L–1. Analyses of cytotoxic and genotoxic potential of the trifluormethyl derivative were performed with cytokinesis-block micronucleus cytome assay in human lymphocyte culture and revealed no genotoxic and cytotoxic effects. The most potent compounds were subjected to molecular docking simulations in order to analyse bindings to molecular targets and, at the same time, further support the results of experimental cytotoxic tests. Docking studies showed sites of importance in forming hydrogen bonds of the most potent compounds with targets of interest

IN VITRO STUDY OF THE ANTI-PROLIFERATIVE EFFECTS OF DIPOTASSIUM-TRIOXOHYDROXYTETRAFLUOROTRIBORATE ON THE H520 NON-SMALL CELL LUNG CANCER CELL LINE

Non-small cell lung cancer has been shown to be resistant to many forms of chemotherapy and is amongst the deadliest cancers worldwide. The anti-proliferative effects of halogenated boroxine - K2(B3O3F4OH), have been confirmed in multiple cancer cell lines including melanoma and breast cancer. The potential for this chemical treatment on non-small cell lung cancer cells was studied and the lower threshold concentration with the clear biological effect of halogenated boroxine, was determined. Appling MTT assays and the relative gene expression analysis of two genes of interest, RRBP1 and PER1, novel knowledge on the biological potential of halogenated boroxine (HB) was gained, but did not lead to biological explanations of the mechanisms of halogenated boroxine activity. The results of MTT assay showed a significant HB effect on non-small cell lung cancer in concentration of 0.5 mg/mL, while relative expression levels of RRBP1 and PER1 did not significantly differ regardless the concentration applied

Complementarity of Standard Cytogenetic Assays

Standard cytogenetic assays used in genotoxicology usually include chromosome aberrations analysis and micronucleus cytokinesis-block assay. Both tests originate on standard protocol for lymphocyte culture and can be used as complement or substitute to each other. Aim of this study was to evaluate complementarities between results of chromosome aberration analysis assay and results of micronucleus cytokinesis-block assay in representative sample of inhabitants from Bosnia and Herzegovina. The aim was achieved by calculating Pearson correlation coefficient and simple linear regression

Complementarity of Standard Cytogenetic Assays

Standard cytogenetic assays used in genotoxicology usually include chromosome aberrations analysis and micronucleus cytokinesis-block assay. Both tests originate on standard protocol for lymphocyte culture and can be used as complement or substitute to each other. Aim of this study was to evaluate complementarities between results of chromosome aberration analysis assay and results of micronucleus cytokinesis-block assay in representative sample of inhabitants from Bosnia and Herzegovina. The aim was achieved by calculating Pearson correlation coefficient and simple linear regression

Chromosome Aberrations as Bioindicators of Environmental Genotoxicity

Due to the exposure to various potentially genotoxic xenobiotics, derived from recent war activities such as NATO air strikes with antitank ammunition containing depleted uranium, we have evaluated chromosome aberrations in 84 peripheral blood samples from three local populations. One population sample included 30 individuals who lived in the Sarajevo area during and after the war (exposed to potential genotoxins), second population was presented with 26 employees of the tank repair facility in Hadžići (target of NATO air strikes), and 28 inhabitants of Posušje (not exposed to war-related activities) were treated as sample of control population. The mean of chromosome aberration frequencies for the population from Hadžići was significantly higher than the frequencies for the two other populations. Point bi-serial coefficient analysis did not reveal any relationship between the frequencies of chromosome aberrations and smoking habits or gender. Results suggest that depleted uranium could be a risk factor for human health