Nowadays great deal of research is physiological field is conducted on experimental animals and there is a lot of criticism from the wide public on methods used. Therefore, recently there is a lot of effort focused on the welfare of the animals. Main aim of this study is to determine the effect of experimental sample collection method on the selected parameters of stress. In the experiment two sample collections of rabbit blood from marginal ear vein were realized – first using standard method with one person fixing the animal and other collecting the blood using gently fixating the animal. In the second groups experimental method of inserting the experimental animal into a sack and further collection in dark was realized. During the experiment the levels of cortisol – main stress indicator in organism and other health parameters of animals including mineral profile and haematological parameters were observed. Our results show no significant changes in levels of cortisol but also a decreasing tendency in the sample from the second (dark) collection. Haematological parameters were generally in the reference values and any significant changes except levels of lymphocytes and percent of lymphocytes which shown significant increase in the second collection period were found. Also the levels of mean corpuscular haemoglobin and percent of neutrophils unveiled a significant decrease in values. Values of mineral profile parameters have indicated no significant changes except the levels of phosphorus. Based on the result we can state that the experimental sample collection has no effect on blood parameters of the animals but we spectated a statistically insignificant decrease in the levels of cortisol which can suggest that the dark collection is possibly less stressful to the animals.
The present review summarizes the current knowledge concerning provenance, properties, physiological and therapeutic actions of elderberry and the bioactive molecules present in the plant, with emphasis on their action on female reproduction. Elderberry or black elder (Sambucus nigra L.) attracts attention due to its easy cultivation and high availability of bioactive compounds. Most of the available data concerning black elder’s therapeutic action are focused on its effects such as activation of immune processes and anti-inflammatory processes (cytokine production, etc.) and regulation of hormones and their receptors in cancer cells. The effects of elderberry on reproduction have been poorly investigated so far. Nevertheless, conducted studies so far demonstrate the stimulatory influence of black elder extract and its constituents, such as rutin, anthocyanins and agglutinins, on the viability and steroidogenesis of healthy ovarian cells as well as their ability to promote apoptosis and reduce the viability and proliferation of ovarian cancer cells. Furthermore, the action of black elder extract and its constituent biomolecules, such as anthocyanins and lectins, on embryogenesis and the embryonal estradiol-estradiol receptor system have also been reported. The available information, despite limitations, suggest the applicability of black elder constituents for improvement of reproductive processes in animal biotechnology, animal production and assisted reproduction, as well as for prevention and treatment of reproductive disorders (including cancer) in veterinary and human medicine.
Pomegranate fruit (Punica granatum L.) is rich in antioxidants with a content of bioactive substances with high medicinal value. Punicalagin, a polyphenol from pomegranate fruit, has been studied for its antioxidant, anti-proliferative and anti-cancer activities. Ovarian cancer is one of the most common cancers in the female reproductive organs and with high rate of lethality. While it is confirmed that pomegranate has significant beneficial effects on several types of cancer, there are few detailed reports on epithelial ovarian cancer. In accordance with the potential health-promoting effects of pomegranate, the aim of our study was to examine the in vitro effect of punicalagin and pomegranate peel extract at the different concentrations (12.5, 25, 50, 100, and 200 µg/mL) for 24 h on the human ovarian granulosa cell line HGL5 and human ovarian carcinoma cell line OVCAR-3. For this experiment, the ethanol extract from lyophilized pomegranate peel was prepared. The metabolic activity was determined by AlamarBlueTM cell viability assay, the secretion of steroid hormones was assayed by the ELISA method. The results showed a significant (P≤0.001) decrease in the viability of HGL5 cells after the addition of the highest concentration of punicalagin (200 µg/mL). The number of viable OVCAR-3 cells was not significantly (P≥0.05) affected compared to the control. On the other hand, the concentrations 25, 50, 100, and 200 µg/mL of pomegranate peel extract led to a significant decrease in the viability of OVCAR-3 cells but did not cause any significant (P≥0.05) changes in the viability of HGL5. Although our studies revealed an increase in the release of 17β-estradiol levels by HGL5 cells after punicalagin treatment at the concentration 50 (P≤0.01) and 100 (P≤0.05) µg/mL, progesterone secretion was not significantly (P≥0.05) affected. Also, the release of 17ß-estradiol was significantly increased after the supplementation of pomegranate peel extract at the concentrations 50 (P≤0.01), 100, and 200 (P≤0.001) µg/mL. Furthermore, the levels of progesterone were significantly (P≤0.05) decreased at concentrations 12.5, 25, 50, and 100 µg/mL. In conclusion, pomegranate phytonutrients might be a promising modulator of secretion of steroid hormones and it might serve to be a potential chemoprotective agent, reducing viability of ovarian cancer cells.
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