ABSTRACT
River water in Kosovo is exposed to various discharges from industrial and agricultural activities as well as to urban wastewater. Rivers Sitnica and Drenica are among the most affected ones and water samples drawn from these rivers show the presence of various toxic substances. Genotoxic effects are seen in fish living in these rivers indicating a cytotoxic and mutagenic potential of the river water. Aiming at substantiating these observations, we assessed cyto- and genotoxic effects of water samples collected at different locations from Drenica and Sitnica river. Samples drawn from Lake Badovc served for comparison. To address seasonal effects, samples were collected at different seasons/time points during the period summer 2016 - spring 2018. The water samples were analyzed employing primary rat hepatocytes as reliable in vitro cell model for the assessment of cytotoxic effects (mitotic arrest and cell death) and DNA damage/genotoxicity (micronucleus assay, Comet assay). The results do not account for significant effects associated with specific locations but demonstrate seasonal differences of the genotoxic potential of the water samples collected along both rivers, which are accompanied by a limited cytotoxic potential. Our data provide substantial support to earlier observations and strongly warrant the need for continuous chemical as well as biological monitoring of the river water in Kosovo, focusing on an improved toxicant profiling of the river water and investigations addressing the observed seasonal variations.
ABSTRACT
The alkaline comet assay, or single cell gel electrophoresis, is one of the most popular methods for assessing DNA damage in human population. One of the open issues concerning this assay is the identification of those factors that can explain the large inter-individual and inter-laboratory variation. International collaborative initiatives such as the hCOMET project - a COST Action launched in 2016 - represent a valuable tool to meet this challenge. The aims of hCOMET were to establish reference values for the level of DNA damage in humans, to investigate the effect of host factors, lifestyle and exposure to genotoxic agents, and to compare different sources of assay variability. A database of 19,320 subjects was generated, pooling data from 105 studies run by 44 laboratories in 26 countries between 1999 and 2019. A mixed random effect log-linear model, in parallel with a classic meta-analysis, was applied to take into account the extensive heterogeneity of data, due to descriptor, specimen and protocol variability. As a result of this analysis interquartile intervals of DNA strand breaks (which includes alkali-labile sites) were reported for tail intensity, tail length, and tail moment (comet assay descriptors). A small variation by age was reported in some datasets, suggesting higher DNA damage in oldest age-classes, while no effect could be shown for sex or smoking habit, although the lack of data on heavy smokers has still to be considered. Finally, highly significant differences in DNA damage were found for most exposures investigated in specific studies. In conclusion, these data, which confirm that DNA damage measured by the comet assay is an excellent biomarker of exposure in several conditions, may contribute to improving the quality of study design and to the standardization of results of the comet assay in human populations.
Subject(s)
Comet Assay/methods , Biomarkers/blood , DNA Damage/genetics , DNA Damage/physiology , HumansABSTRACT
Being exposed to untreated urban and industrial water, the rivers Drenica and Sitnica are considered to be the most polluted ones in the Kosovo. Our previous investigations on the cyto- and genotoxic potential of water samples from these rivers evaluated with primary rat hepatocyte cultures indicated a risk for the health of aquatic organisms. In order to assess the genotoxic risk to aquatic organisms, we therefore performed a two year study (2016-2017) on roach (Rutilus rutilus) from these rivers. Specimens were collected at three locations along the Drenica river and two locations along the Sitnica river, and the genotoxicity was evaluated by the micronucleus as well as the Comet assay (DNA damage) in erythrocytes. The frequencies of micronucleated cells were determined for samples collected in four seasons, whereas the Comet assay was employed on samples collected in five seasons during the two-year period. The data obtained revealed an increase of the frequency of micronucleated erythrocytes from Rutilus rutilus collected at most sampling locations and from both rivers at all seasons investigated. Significant differences to the control (lake Badovc) were found in summer 2016 and spring 2017 samples. When comparing the seasons, the summer 2016 samples were most genotoxic, followed by spring 2017 and autumn 2016. With regard to the Comet assay data, a similar but more prominent "response" was observed. Another important observation is that micronucleus rates as well as DNA damage levels were significantly higher in samples collected in 2016 compared to the respective seasons in 2017. Altogether, the "response" obtained with both markers confirmed a genotoxic risk for fish due the pollution of these rivers. Since there were, however, seasonal and annual variations of the genotoxicity levels further in depth studies have to be carried out addressing the nature of these changes.
Subject(s)
Cyprinidae/physiology , Environmental Monitoring , Water Pollutants, Chemical/toxicity , Animals , Comet Assay , Kosovo , Micronucleus Tests , Risk Assessment , Rivers/chemistry , Seasons , Water Pollutants, Chemical/analysisABSTRACT
The actual stage of the development of Kosovo is characterized by the concerning levels of environmental pollution and the serious health problems attributed to the emission of pollutants into air, soil and water. In this context, river pollution is one of the main threats due to the discharge of untreated urban and industrial waste waters that affect the quality of surface and ground water. In addition, urban and agricultural discharges are affecting the river water quality. In this article, we are presenting data on the cyto- and genotoxic potential of water samples from three rivers (Sitnica, Drenica and Lepenci) in the Kosovo as determined in the cultures of primary rat hepatocytes. Sitnica and Drenica (as the most important Sitnica tributary) drain into the Black Sea, whereas the Lepenci river drains into the Aegean Sea. These rivers are polluted mainly by industry in the Kosovo together with municipal discharges. The results of this study show that the samples have primarily a cytotoxic potential by causing necrotic cell death that was not accompanied by any increase of the rate of micronucleated cells as an indicator for a genotoxic potential. The different effects in 2 consecutive years can be attributed to variations in physico-chemical parameters such as water levels, intake of pollutants, and so on, indicating the need for continuous monitoring and risk assessment.
Subject(s)
Hepatocytes/drug effects , Rivers/chemistry , Water Pollutants, Chemical/toxicity , Animals , Female , Kosovo , Mutagenicity Tests , Rats , Rats, Inbred F344 , Toxicity TestsABSTRACT
ß-Carotene has been shown to increase the risk of developing lung cancer in smokers and asbestos workers in two large scale trails, the Beta-Carotene and Retinol Efficacy Trial (CARET) and the Alpha-Tocopherol Beta-carotene Cancer Prevention Trial (ATBC). Based on this observation, it was proposed that genotoxic oxidative breakdown products may cause this effect. In support of this assumption, increased levels of sister chromatid exchanges, micronuclei, and chromosomal aberrations were found in primary hepatocyte cultures treated with a mixture of cleavage products (CPs) and the major product apo-8'carotenal. However, because these findings cannot directly be transferred to the lung due to the exceptional biotransformation capacity of the liver, potential genotoxic and cytotoxic effects of ß-carotene under oxidative stress and its CPs were investigated in primary pneumocyte type II cells. The results indicate that increased concentrations of ß-carotene in the presence of the redox cycling quinone dimethoxynaphthoquinone (DMNQ) exhibit a cytotoxic potential, as evidenced by an increase of apoptotic cells and loss of cell density at concentrations > 10 µM. On the other hand, the analysis of micronucleated cells gave no clear picture due to the cytotoxicity related reduction of mitotic cells. Last, although CPs induced significant levels of DNA strand breaks even at concentrations ≥ 1 µM and 5 µM, respectively, ß-carotene in the presence of DMNQ did not cause DNA damage. Instead, ß-carotene appeared to act as an antioxidant. These findings are in contrast with what was demonstrated for primary hepatocytes and may reflect different sensitivities to and different metabolism of ß-carotene in the two cell types.
ABSTRACT
BACKGROUND: Host genetic factors may impact susceptibility to infection. A small number of studies have investigated the association between factors such as ABO blood groups and selected phenotypes on the incidence and severity of H1N1 infections with inconclusive results. METHODS: Using data from the Clinic of Infectious Diseases - University Clinical Centre Prishtina and based on the examination of 125 patients hospitalized with H1N1 in the period 2009-2014, the frequency of blood groups from ABO and Rhesus (Rh) systems as phenotypical markers were evaluated. In addition, other phenotypes such as ear lobe free/ear lobe attached, normal chin/cleft chin, tongue roller/non roller, hand clasping right thumb over/hand clasping left thumb over, right-handed/left-handed, dark eyes/light eyes were also analyzed. The data obtained from the 125 hospitalized patients were compared with the data from the Kosovar population (n = 2000) as a reference group. RESULTS: A total of 303 patients with H1N1 were hospitalized in the period 2009-2015. Blood group and phenotype data available from 125 hospitalized H1N1 patients showed significant differences in the frequencies of the blood groups from Rh system as well as in two (out of six) phenotypes of the selected morphological traits compared to reference groups. CONCLUSIONS: The findings from this preliminary study indicate that these Rh system and phenotype differences may be linked to H1N1 susceptibility and may guide identification of risk groups and populations.
Subject(s)
Genetic Predisposition to Disease/epidemiology , Genetic Predisposition to Disease/genetics , Influenza A Virus, H1N1 Subtype , Influenza, Human/epidemiology , Influenza, Human/genetics , Adult , Female , Humans , Incidence , Kosovo/epidemiology , Male , Phenotype , Retrospective Studies , Young AdultABSTRACT
Reports on the state of the environment in Kosovo have emphasized that river and ground water quality is affected by pollution from untreated urban water as well as the waste water from the industry. One of the main contributors to this pollution is located in Obiliq (coal power plants). Prishtina-the capital city of Kosovo-is heavily influenced too. Furthermore, the pollutants combined together with those from heavy traffic are dissolved in Prishtina runoff water, which is discharged into the creek entering the river Sitnica together with urban waste water. The available data show the complex pollution with excessive quantities of nitrites, suspended materials, organic compounds, detergents, heavy metals, polychlorinated biphenyls, etc. In this study, the cytotoxic and genotoxic potential of water samples taken at these sites was tested in primary rat hepatocytes. The results obtained indicate that water samples collected in Prishtina and Obiliq had a significant cytotoxic potential in primary rat hepatocyte cultures even when diluted to 1 %. The increased cytotoxicity, however, was not accompanied by an increased genotoxicity as measured by the percentage of micronucleated cells. Further investigations addressing the chemical composition of the samples and the identification of the toxicants responsible for the cytotoxic effects found will be carried out in a next step.
Subject(s)
Environmental Monitoring/methods , Groundwater/chemistry , Mutagens/analysis , Rivers/chemistry , Water Pollutants, Chemical/analysis , Animals , Cell Survival/drug effects , Cities , Female , Hepatocytes/drug effects , Hepatocytes/pathology , Kosovo , Metals, Heavy/analysis , Metals, Heavy/toxicity , Micronuclei, Chromosome-Defective/chemically induced , Mutagens/toxicity , Polychlorinated Biphenyls/analysis , Polychlorinated Biphenyls/toxicity , Rats , Rats, Inbred F344 , Wastewater/chemistry , Water Pollutants, Chemical/toxicityABSTRACT
It has been reported by the Ministry of Environment in Kosova that particle emissions from one of the units of the coal-fired power plants (Kosova A) in Kastriot/Obiliq were exceeding the European standard by some 74 times. Besides the particle emission, there is also release of sulphur dioxide, mono-nitrogen oxide (NOx), carbon monoxide, carbon dioxide, organic compounds and heavy metals. In addition, there is also release of heavy metals and organic compounds from a nearby solid waste dumpsite. Together, they are considered to be responsible for the increased health problems of the population living in the vicinity.To study the genetic effects of these emissions we focused on the genetic load, that is, recessive mutations that affect the fitness of their carriers, of exposed wild living Drosophila melanogaster The effects of ash from the dumpsite on the other hand were investigated upon feeding the ash with the nutrient medium. Our results revealed that the D. melanogaster population from the Kastriot/Obiliq area carries a high genetic load of 54.7%. Drosophila fed with the nutrient medium containing ash in a concentration of 1% carried a genetic load of 37.1%, whilst increasing concentrations (2% and 3% of ash) led to higher genetic loads of 68.7% and 67.4%, respectively.
Subject(s)
Drosophila melanogaster/genetics , Environmental Pollutants/toxicity , Genetic Load , Power Plants , Animals , Carbon Dioxide/toxicity , Carbon Monoxide/toxicity , Chromosomes, Insect/genetics , Drosophila melanogaster/drug effects , Environmental Monitoring , Female , Industrial Waste/adverse effects , Kosovo , Male , Metals, Heavy/toxicity , Nitrogen Oxides/toxicity , Particulate Matter/toxicity , Sulfur Dioxide/toxicityABSTRACT
The aim of this study was to investigate the distribution of specific phenotypes in patients with lung diseases as well as their eventual association with the risk of developing lung diseases. For this purpose 2777 patients with lung diseases and 2778 healthy individuals from all over Kosova were examined for the appearance of the following selected phenotypes: ear lobe free (ELF)/ear lobe attached, normal chin (NC)/cleft chin, tongue roller (TR)/non roller, hand clasping right thumb over (HC)/hand clasping left thumb over, righthanded (RH)/lefthanded. In addition, the blood group from ABO system and the presence or absence of the Rhesus factor asphenotypical markers were observed. The results obtained show significant differences between control and lung disease patients for NC (p ≤ 0.05) and TR (p ≤ 0.005) as well as for blood groups AB (p ≤ 0.05) and O (p ≤ 0.005). These results point to eventually increased levels of genetic load as a result of the increased homozygosity in some gene loci causing an increased frequency of some recessive phenotypes in patients with lung diseases. Together with the specific associations observed, these preliminary findings could serve as a basis for further in depth investigations with respect to the types of lung diseases, occupational exposure and dietary habits, and thus is expected to contribute to an understanding of predispositions and susceptibility to lung diseases.
Subject(s)
Genetic Association Studies , Genetic Predisposition to Disease , Lung Diseases/genetics , Female , Humans , Male , PhenotypeABSTRACT
When we investigated the genotoxicity of beta-carotene cleavage products (CPs) in primary rat hepatocytes stimulated to proliferate, we observed dose-dependent increases of chromosomal aberrations, sister chromatid exchanges and micronuclei. In contrast to other genotoxic substances, however, this increased genotoxicity was not accompanied by increased cytotoxicity. As a consequence we observed metaphases showing massive chromosomal damage, indicating inhibition of apoptosis by CPs enabling these cells to proceed in the cell cycle. Since proliferative stimulation by growth factors may support this effect, the in vitro toxicological effects of CPs were studied on proliferatively quiescent primary rat hepatocytes. A significant increase of both apoptosis and necrosis was found. Supplementation with antioxidants did not significantly lower the level of apoptosis, while the level of necrosis was significantly reduced by Trolox and N-acetylcysteine at all concentrations tested as well as ascorbic acid (50 microM) and a combination of Trolox (50 microM) and ascorbic acid (50 microM). These observations indicate that a) the cytotoxic potential in combination with the genotoxic potential of CPs may promote the initiation of cells due to compensatory cell division in exposed tissues and may aggravate inflammatory processes under chronic exposure, and b) the applied antioxidants may protect from cytotoxicity primarily via the detoxification of aldehydic beta-carotene cleavage products.