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Basic morphological and hematological parameters were studied in adult, sexually mature individuals of Carrasius gibelio living in two rivers with varying degrees and different types of anthropogenic pollution (less disrupted site, domestic sewage pollution and heavy metal pollution) in Southern Bulgaria. The study was conducted on 30 female fish, age 3+ from each site. The results show that in C. gibelio, which inhabit the site with heavy metal pollution, the morphological parameters BW and FCF have the lowest statistically significant values. Specimens of C. gibelio from the river site with domestic sewage pollution have the following hematological profiles: erythrocytosis, leukocytosis and hyperchromia, differential leukocyte count: neutrophilia, eosinophilia, monocytosis and lymphocytopenia. Red cell indices (MCH, MCHC and MCV) display the highest statistically significant values. The C. gibelio specimens from the site with heavy metal pollution had the following hematological profiles: erythrocytosis, leukocytopenia and hypоchromia, differential leukocyte count: neutrophilia, eosinophilia, monocytosis and lymphocytopenia. The lowest statistically significant values of red cell indices (MCH, MCHC and MCV) indicate anemic changes of microcytic and hypochromic type. The hematological parameters of C. gibelio are adequate biomarkers of the physiological state of animals and their habitats.

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This study investigated the anticancer effect of the anti-parasitic drug artemisinin in combination with two redox modulators: vitamin C and pro-vitamin K3 (C/K3) The experiments were conducted on leukaemia cells Jurkat. Cells were treated with either artemisinin or C/K3 alone and with all three compounds. Cell proliferation and viability were analysed using trypan blue stating and automated cell counting. The results showed that artemisinin (>10 mM) suppressed cell proliferation activity, but did not induce cell death up to 500 mM. The drug demonstrated a clear cytostatic effect at concentrations 250- 500 mM – Jurkat cells did not proliferate, but were alive. The combination C/K3 (200:2, 300:3 mM/mM) applied alone did not affect cell proliferation and viability. Vitamins C/K3 in concentration ratio 500:5 (μM/mM) decreased cell proliferation activity by ~10%. The triple combination artemisinin/C/K3 manifested synergistic anti-proliferative effects at all concentration ratios analysed. This synergistic effect increased with increasing C/K3 concentration. Based on literature data, it was assumed that the anti-proliferative effect of the triple combination was mediated by changes in the redox-homeostasis of cancer cells. The C/K3 redox system likely acted on cancer mitochondria and increased superoxide production and activation of pro-apoptotic signals, specific for cancer cells. On the other hand, artemisinin could generate hydroxyl radicals as a result of activation of Fenton reactions, depleting intracellular reducing equivalents. Both redox mechanisms lead to activation of signal pathways for induction of cancer cell death.

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