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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 the cyto- and genotoxic effects of water samples collected at different locations from the Drenica and Sitnica rivers. Samples drawn from Lake Badovc served for comparison. To address seasonal effects, samples were collected at different seasons/time points during the period of summer 2016—spring 2018. The water samples were analyzed employing primary rat hepatocytes as a reliable in vitro cell model for the assessment of cytotoxic effects (mitotic arrest and cell death) and DNA damage/genotoxicity (micronucleus assay and 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 improved toxicant profiling of the river water and investigations addressing the observed seasonal variations. [ABSTRACT FROM AUTHOR]

The available monitoring data from Kosovo Environmental Protection Agency show concerning levels of the air pollution in Prishtina - the capital city of Kosovo and the most populated city in the country. Due to the air pollutants emitted mostly from the heavy traffic and the coal-fired power plants located in the vicinity, the residents of urban area in Prishtina are exposed to unhealthy air. It is reported that the situation worsens during the autumn and winter months due to more frequent smog episodes. Based on the concerns raised, the aim of the study was to assess the eventual genotoxic effects of air pollution among residents of Prishtina during the autumn and winter months. For this purpose, 29 healthy female non-smoker students (aged 20-26) were involved in this preliminary study. The first sampling of buccal cells took place beginning of November 2019 whereas the second sampling took place by the end of January 2020. Buccal cell samples were analyzed for the frequency of micronuclei and the obtained data demonstrate increased genotoxicity in a sample population as an effect of the exposure to increased air pollution levels in Prishtina urban area during the autumn and winter period. On the other hand, these preliminary data clearly indicate the need for continuing with bio-monitoring studies by extending the timeframe and increasing the number of seasons under investigation. [ABSTRACT FROM AUTHOR]

Background/Aims: Trypan blue is routinely used in cell culture experiments to distinguish between dead cells, which are labelled by trypan blue, and viable cells, which are apparently free of any staining. The assumption that trypan blue labelling is restricted to dead cells derives from the observation that rupture of the plasma membrane correlates with intense trypan blue staining. However, decades ago, trypan blue has been used to trace fluid uptake by viable macrophage-like cells in animals. These studies contributed to the concept of the reticuloendothelial system in vertebrates. Trypan blue itself does not show a fluorescence signal, but trypan blue-labelled proteins do. Therefore, intracellular localization of trypan blue-labelled proteins could give a clue to the entrance pathway of the dye in viable cells. Methods: We used fluorescence microscopy to visualize trypan blue positive structures and to evaluate whether the bactericide, silver, enhances cellular trypan blue uptake in the brain macrophage-like cell line, BV-2. The pattern of chromatin condensation, visualized by DAPI staining, was used to identify the cell death pathway. Results: We observed that silver nitrate at elevated concentrations (≥ 10 µM) induced in most cells a necrotic cell death pathway. Necrotic cells, identified by pycnotic nuclei, showed an intense and homogenous trypan blue staining. Apoptotic cells, characterized by crescent-like nuclear chromatin condensations, were not labelled by trypan blue. At lower silver nitrate concentrations, most cells were viable, but they showed trypan blue labelling. Viable cells showed a cell-type specific distribution of heterochromatin and revealed a perinuclear accumulation of bright trypan blue-labelled vesicles and, occasionally, a faint homogenous trypan blue labelling of the cytoplasm and nucleus. Amiloride, which prevents macropinocytosis by blocking the Na+ / H+ exchange, suppressed perinuclear accumulation of dye-labelled vesicles. Swelling of cells in a hypotonic solution induced an intense intracellular accumulation of trypan blue. Cells exposed to a hypotonic solution in the presence of 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB), which blocks volume-regulated ion channels, prevented labelling of the cytoplasm and nucleus but did not affect labelling of perinuclear vesicles. Conclusion: In viable cells trypan blue-labelled vesicles indicate trypan blue uptake by macropinocytosis and trypan blue-labelled cytosol could indicate a further entry pathway for the dye, like activated volume-regulated channels. Accordingly, fluorescence microscopic analysis of trypan blue-labelled cells allows not only a discrimination between necrotic and apoptotic cell death pathway but also a discrimination between the mode of trypan blue uptake in viable cells – via pinocytosis or via activated volume-regulated ion channels – in the same preparation at the single cell level. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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