Anica Horvat Knežević

Radioprotective Effects of Quercetin and Ethanolic Extract of Propolis in Gamma-Irradiated Mice

Radioprotective Effects of Quercetin and Ethanolic Extract of Propolis in Gamma-Irradiated Mice The aim of this study was to assess radioprotective effects of quercetin and the ethanolic extract of propolis (EEP) in CBA mice exposed to a single radiation dose 4 Gy (60Co). The mice were treated with 100 mg kg-1 quercetin or EEP a day for three consecutive days either before (pre-treatment) or after gamma-irradiation (therapy). Leukocyte count was determined in blood drawn from the tail vein, and DNA damage in leukocytes was assessed using the alkaline comet assay. Genotoxic effects of the test compunds were also evaluated in non-irradiated mice. The levels of radioprotection provided by both test compounds were compared with those established in mice that were given chemical radioprotector S-(2-Aminoethy1)isothiouronium bromide hydrobromide (AET). Mice that received pre-treatment were less sensitive to irradiation. Mice given the post-irradiation therapy showed a slight but not significant increase in total leukocyte count over irradiated negative control. Quercetin showed better protective properties than EEP in both pre-treatment and therapy, and activated a higher number of leukocytes in non-irradiated mice. The alkaline comet assay suggests that both natural compounds, especially when given as pre-treatment, protect against primary leukocyte DNA damage in mice. At tested concentrations, EEP and quercetin were not genotoxic to non-irradiated mice. AET, however, caused a slight but not significant increase in DNA damage. Although the results of this study show the radioprotective potential of the test compounds, further investigation is needed to clarify the underlying protection mechanisms. Primjena alkalnog kometnog testa u istraživanju radioprotektivnih učinaka alkoholnog ekstrakta propolisa i kvercetina na miševima ozračenim gama-zračenjem

Synergistic Effects of Irinotecan and Flavonoids on Ehrlich Ascites Tumour-Bearing Mice

Swiss albino mice were given Ehrlich ascites tumour cells (1 × 10(6)) intraperitoneally. For survival analysis and tumour growth analysis, the mice were administered quercetin and naringin (100 mg/kg) daily for 3 consecutive days, beginning on the third day after intraperitoneal (i.p.) injection of Ehrlich ascites tumour cells (1 × 10(6)). Irinotecan was administered ip at a dose of 50 mg/kg on days 1, 13 and 19. For the analysis of cell types and differential count of cells present in the peritoneal cavity, peripheral whole-blood leucocyte count and the comet assay, the mice were treated therapeutically with quercetin and naringin (100 mg/kg) and irinotecan (50 mg/kg) daily for 3 consecutive days beginning on third day after i.p. injection of Ehrlich ascites tumour cells (1 × 10(6)). We observed the synergistic anti-tumour effect expressed as the median survival time of mice treated with naringin in combination with irinotecan. All test components inhibited tumour growth and increased lifespan of mice except quercetin. The total number of cells present in the peritoneal cavity of mice significantly decreased in all treatments except quercetin. Single irinotecan and irinotecan combined with naringin had the highest DNA-damaging potential on peripheral blood leucocytes and lowest primary DNA damage, both in the kidney and liver cells as measured by the alkaline comet assay. Our results showed enhanced anti-tumour activity of irinotecan in combined treatment with flavonoids to reduce the deteriorating reaction of cytostatic drugs.

Addition of propolis to irinotecan therapy prolongs survival in ehrlich ascites tumor-bearing mice.

We investigated possible synergistic action of anticancer drug Irinotecan (IRI) combined with ethanolic (EEP) and water-soluble (WSDP) derivate of propolis on Swiss albino mice injected with Ehrlich ascites tumor (EAT). For survival analysis mice were administered WSDP and EEP (100 mg/kg) daily for 3 consecutive days, beginning on 3rd day after EAT cell (1×10⁶) injection. IRI was administered at a dose of 50 mg/kg on days 1, 13, and 19. We simultaneously studied peripheral white blood cell count, cell types washed from the peritoneal cavity, functional activity of macrophages from peritoneal cavity, and the level of primary DNA damage in leukocytes, kidney, and liver cells using the alkaline comet assay. Three out of 9 mice per group survived the entire duration of the experiment (90 days) in groups treated with IRI combined with WSDP and EEP. All test components increased survival of mice by 7.53% to 231.54%. Combined treatment with IRI and/or WSDP and EEP significantly decreased percentage of tumor cells in the peritoneal cavity as compared to nontreated EAT-injected mice. All treated animals had significantly higher percentage of neutrophils in the peritoneal cavity in comparison to nontreated EAT-injected mice. We observed significantly higher value of DNA damage in leukocytes of mice treated with IRI and combination of IRI and/or WSDP and EEP as compared to nontreated EAT-injected mice, while the same treatment decreased DNA damage in kidney. Our results showed that addition of propolis to IRI treatment enhanced antitumor activity of IRI and prolongs survival in EAT-bearing mice, which definitely deserve further studies to clarify the possible mechanisms of antitumor actions of combined herb-drug treatments.

Biochemical and hematological profiles of a wild population of the nose-horned viper Vipera ammodytes (Serpentes: Viperidae) during autumn, with a morphological assessment of blood cells

BackgroundHematological and biochemical analyses of blood can be of great importance for studying the biology of species and determining the health status of animals in both the wild and captivity. In order to determine baseline ranges for the nose-horned viper Vipera ammodytes, we determined the blood cell morphology and measured 20 hematological and 5 biochemical variables from the blood samples taken from 31 (17 males and 14 females) healthy specimens of nose-horned viper caught in the wild at the beginning of autumn.ResultsMost of the measured parameters and blood cell morphology were similar to those reported for other reptile species. The males showed a significantly higher eosinophil count and higher mean cell hemoglobin (MCH) and MCH concentration values, while the females had a significantly higher mature erythrocyte count. The erythrocyte differential counts showed balanced hematopoiesis in this part of the annual cycle.ConclusionsWe suggest that these results can be used as a baseline for determining hematological values and blood cell morphology for V. ammodytes, as well as in further studies including monitoring and determination of the health status of this species, especially of animals kept in captivity for antiserum production. This is the most complete report regarding hematological parameters of this endangered snake species.

Synergism Between Propolis and Hyperthermal Intraperitoneal Chemotherapy with Cisplatin on Ehrlich Ascites Tumor in Mice

We investigated antitumor, genotoxic, chemopreventive, and immunostimulative effects of local chemoimmunotherapy and hyperthermal intraperitoneal chemotherapy (HIPEC) in a mouse-bearing Ehrlich ascites tumor (EAT). Mice were treated with water-soluble derivative of propolis (WSDP) at a dose of 50 mg kg(-1) , 7 and 3 days before implantation of EAT cells, whereas cisplatin (5 or 10 mg kg(-1) ) was injected 3 days after implantation of EAT cells at 37°C and 43°C. The following variables were analyzed: the total number of cells, differential count of the cells present in the peritoneal cavity, functional activity of macrophages, comet assay, and micronucleus assay. The combination of WSDP + CIS 5 mg kg(-1) at 37°C resulted in tumor growth inhibition and increased the survival of mice by additional 115.25%. WSDP with HIPEC increased the survival of mice by additional 160.3% as compared with HIPEC. WSDP reduced cisplatin toxic and genotoxic effect to normal cells without affecting cisplatin cytotoxicity on EAT cells. In addition, WSDP with HIPEC increased the cytotoxic actions of macrophages to tumor cells. Water-soluble derivative of propolis increases macrophage activity and sensitivity of tumor cells to HIPEC and reduces cisplatin toxicity to normal cells.

Brain Toxicokinetics of Prometryne in Mice

Brain Toxicokinetics of Prometryne in Mice Prometryne is a methylthio-s-triazine herbicide. Significant trace amounts are found in the environment, mainly in water, soil, and food plants. The aim of this study was to establish brain and blood prometryne levels after single oral dose (1 g kg-1) in adult male and female mice. Prometryne was measured using the GC/MS assay at 1, 2, 4, 8, and 24 h after prometryne administration. Peak brain and blood prometryne values were observed 1 h after administration and they decreased in a time-dependent manner. Male mice had consistently higher brain and blood prometryne levels than female mice. The observed prometryne kinetics was similar to that reported for the structurally related herbicide atrazine. Toksikokinetika Prometrina u Mozgu Miševa Prometrin je metiltio-s-triazinski herbicid. Značajne količine prometrina zaostaju u tragovima u okolišu, poglavito u vodi, tlu i biljkama koje rabimo za prehranu. Cilj je rada izmjeriti količinu prometrina koja se apsorbira u mozgu i krvi nakon primijenjene akutne oralne doze (1 g kg-1 tjelesne mase) u odraslih miševa obaju spolova. Razine prometrina u mozgu i krvi izmjerene su GC/MS-om tijekom 1., 2., 4., 8. i 24. sata nakon izlaganja. Utvrđeno je da je udio prometrina koji se zadržava u živčanom tkivu relativno nizak ali detektabilan u odnosu na koncentraciju u krvi i koncentraciju primijenjene doze. Najviše koncentracije u krvi i maseni udjeli u mozgu zabilježeni su tijekom 1. sata nakon izlaganja, a s vremenom izmjerene vrijednosti značajno opadaju. Uočena je značajna razlika između mužjaka i ženki pri čemu mužjaci imaju značajno više razine prometrina u mozgu i krvi nego ženke. Opisana toksikokinetika prometrina pokazuje sličnosti s otprije opisanom i poznatom toksikokinetikom strukturalno sličnog herbicida atrazina.

Sevoflurane and isoflurane genotoxicity in kidney cells of mice

Abstract The aim of this study was to evaluate the DNA damage and repair in kidney cells of Swiss albino mice after repeated exposure to sevoflurane and isoflurane and compare their detrimental effects. We used the alkaline comet assay to establish the genetic damage and measured three parameters: tail length, tail moment, and tail intensity of comets. These parameters were measured immediately after exposure to the above mentioned inhalation anaesthetics, two hours, six hours, and 24 hours later and were compared with the control group. Mean values of all three parameters were significantly higher in experimental groups compared to the control group. DNA damage in kidney cells of mice exposed to sevoflurane increased continuously before it reached its peak 24 hours after exposure. Isoflurane induced the highest DNA damage two hours after exposure. Levels of DNA damage recorded 24 h after cessation of exposure to both tested compounds suggest that sevoflurane was slightly more genotoxic than isoflurane to kidney cells of mice. According to these results, the currently used volatile anaesthetics sevoflurane and isoflurane are able to damage DNA in kidney cells of mice. Such findings suggest a possibility for similar outcomes in humans and that fact must be taken into account in everyday clinical practice.