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.
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 Hadzići (target of NATO air strikes), and 28 inhabitants of Posusje (not exposed to war-related activities) were treated as sample of control population. The mean of chromosome aberration frequencies for the population from Hadzići 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.
Paracetamol is a common analgesic and antipyretic drug. It has been recognized as one of the most ordinary medications taken in overdoses. We examined the possible genotoxic effects of high paracetamol concentrations expected to occur after overdose. Paracetamol was added to the cultures at the beginning of the cultivation period. Separate cultures for three tested concentrations of paracetamol (50 μg/mL, 100 μg/mL, and 200 μg/mL) were set. Effects of paracetamol were evaluated by micronucleus cytokinesis-block assay, chromosome aberration analysis, and nuclear division index. Results demonstrate that paracetamol concentration of 200 μg/mL expresses certain genotoxic effects in human peripheral blood lymphocytes.
During the 1992-1995 siege, as well as after the war activities, citizens of Sarajevo were most probably exposed to various potential genotoxic agents. The effects of those potential genotoxins were evaluated by micronucleus-cytokinesis blocked assay. The study included 30 individuals who resided in the area of Sarajevo during the war and the postwar period. Point bi-serial coefficient analysis did not reveal any relationship between the frequencies of binuclear cells with micronuclei as well as total number of micronuclei and smoking habits or gender. Simple linear regression revealed statistically significant positive correlation between the age and micronuclei formation. Due to the war related environmental contamination more extensive study is recommended.
Standard molecular techniques, with only a slight modification, are very useful in obtaining and interpreting the final results in the field of forensic genetic. Data obtained through such analysis are highly reliable and can be used as a very powerful tool that produces valuable results. However, success and swiftness of DNA typing of biological evidence either that found at a crime scene or used in disputed paternity testing, depends on the optimization of numerous factors. One of the most important and critical phases that ensures reliability of the whole procedure is the choice of the most suitable volume for the amplification protocol. Buccal swabs were collected from volunteers. DNA was extracted by Qiagen Dnaeasy Tissue Kit. PowerPlex 16 kit was used to simultaneously amplify 15 STR loci by PCR. Amplification was carried out as described previously. The tested total working reaction volumes were 5, 10 and 25 microl. The PCR amplification was carried out in PE Gene Amp PCR System Thermal Cycler (ABI, Foster City, CA). Amplification products were analyzed on an ABI PRISM 377 instrument (ABI, Foster City, CA) in 5% bis-acrilamide gel. Amplification was generally successful for all the tested reaction volumes. Lower partial to complete DNA profiles ratio, the quality of obtained STR profiles, significantly reduced amount of reaction's components give advantage to 5 microl reaction volume over other two tested volumes in this case.
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 Hadzići, 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 Hadzići 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.
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