An assessment of DNA integrity in erythrocytes using the alkaline comet assay was carried out to estimate the impacts of water pollution on Balkan loaches (Cobitis elongata Heckel et Kner, 1858) inhabiting the Sava and Kupa Rivers, rivers of varying different water quality. The amount of DNA damage in cells was estimated from three different parameters comet tail length as the extent of genetic material migration, tail intensity (% DNA in comet tail) and tail moment. The results suggest a genotoxicity of the aquatic environment in the Sava River and demonstrated significantly lower levels of DNA damage in fish captured from the Kupa River. This study confirmed that the comet assay, applied to fish erythrocytes, is a useful tool in determining potential genotoxicity of water pollutants. Although a good DNA damage pattern for Balkan loach was obtained, due to its global and regional conservation status, only restricted use of a small number of specimens per sampling site could be permitted.
The comet assay, or single-cell gel electrophoresis (SCGE), is
a sensitive, rapid, relatively simple and inexpensive method for
detecting DNA strand breaks in individual cells. It is used in
a broad variety of applications and as a tool to investigate DNA
damage and repair. The sensitivity and specificity of the assay
are greatly enhanced if the DNA incubated with an enzyme,
which recognizes a specific kind of DNA damage. This damage
induced by oxidative stress plays a pivotal role in many diseases
and in aging. This article is a critical review of the possible
application of the comet assay in some pathological states in
clinical practice. Most of the studies relate to evaluating the
response of an organism to chemotherapy or radiotherapy with
statistically significant evidence of DNA damage in patients. Other
useful applications have been demonstrated for patients with
heart or neurodegenerative diseases. Only a few studies have
been published on the use of this method in critically ill patients,
although its use would be appropriate. There are also other
scenarios where the comet assay could prove to be very useful in
the future, such as in predicting the likelihood of certain
pathological conditions.
Oxidative stress has been implicated to play a major role in aging and age-related diseases. In the present study, we investigated the effects of aging on the total antioxidant capacity, uric acid, lipid peroxidation, total sulfhydryl group content and damage to DNA in adult (6 months), old (15 months) and senescent (26 months) male Wistar rats. The antioxidant capacity, determined by phycoerythrin-based TRAP method (total peroxyl radical-trapping potential) was significantly decreased in the plasma and myocardium of old and senescent rats, whereas plasma level of uric acid was elevated in 26-month-old rats. Age-related decline in plasma and heart antioxidant capacity was accompanied by a significant loss in total sulfhydryl group content, increased lipid peroxidation and higher DNA damage in lymphocytes. Correlations between TRAP and oxidative damage to lipids, proteins and DNA suggest that the decline in antioxidant status may play an important role in age-related accumulation of cell damage caused by reactive oxygen species., M. Sivoňová, Z. Tatarková, Z. Ďuračková, D. Dobrota, J. Lehotský, T. Matáková, P. Kaplán., and Obsahuje bibliografii a bibliografické odkazy