Eva Tušimová

Curcumin has protective and antioxidant properties on bull spermatozoa subjected to induced oxidative stress

Over the past decades, there has been an emphasis on assessment of the use of natural compounds in the prevention or repair of oxidative injury to spermatozoa. Curcumin (CUR) is a natural phenol with powerful antioxidant properties. The aim of the present study was to examine if CUR could reverse reactive oxygen species (ROS)-mediated alterations to the motility, viability and intracellular antioxidant profile of bull spermatozoa subjected to a prooxidant (i.e., ferrous ascorbate - FeAA). Spermatozoa were washed from recently collected semen samples, suspended in 2.9% sodium citrate and subjected to CUR treatment (5, 10, 25 and 50μmol/L) in the presence or absence of FeAA (150μmol/L FeSO4 and 750μmol/L ascorbic acid) during a 6h in vitro culture. Spermatozoa motility characteristics were assessed using the SpermVision computer-aided spermatozoa analysis (CASA) system. Cell viability was examined with the metabolic activity (MTT) assay, ROS generation was quantified using luminometry and the nitroblue-tetrazolium (NBT) test was used to quantify the intracellular superoxide formation. Cell lysates were prepared at the end of the culture to assess the intracellular activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) as well as the concentrations of glutathione (GSH) and malondialdehyde (MDA). Treatment with FeAA led to a reduced spermatozoa motility (P<0.001), viability (P<0.001) and decreased the antioxidant characteristics of the samples (P<0.001) but increased the ROS generation (P<0.001), superoxide production (P<0.001) and lipid peroxidation (P<0.001). The CUR treatment led to a preservation of spermatozoa motion (P<0.001), mitochondrial activity (P<0.001) and antioxidant characteristics (P<0.05 with SOD and GSH; P<0.01 with CAT and GPx), revealing the concentration range of 25-50μmol/L CUR to be the most effective for sustaining spermatozoa viability. Data from the present study suggest that CUR exhibits significant protective and ROS-scavenging characteristics which may prevent oxidative insults to spermatozoa and thus preserve the functional activity of male gametes.

Resveratrol offers protection to oxidative stress induced by ferrous ascorbate in bovine spermatozoa

Resveratrol (RES) is a natural polyphenol and phytoestrogen exhibiting cardioprotective, anticancer, antibacterial and vasorelaxing properties. It is also a powerful reactive oxygen species (ROS) scavenger and chelating agent. This study was designed to determine the efficiency of RES to reverse the ROS-mediated impairment of the motility, viability and intracellular antioxidant profile of bovine spermatozoa. Spermatozoa were washed out of fresh bovine semen, suspended in 2.9% sodium citrate and subjected to RES treatment (5, 10, 25 and 50 μmol L−1) in the presence or absence of a pro-oxidant, i.e., ferrous ascorbate (FeAA; 150 μmol L−1 FeSO4 and 750 μmol L−1 ascorbic acid) during a 6-h in vitro culture. Spermatozoa motion parameters were assessed using the SpermVision computer-aided sperm analysis (CASA) system. Cell viability was examined with the metabolic activity (MTT) assay, and the nitroblue-tetrazolium (NBT) test was applied to quantify the intracellular superoxide formation. Cell lysates were prepared at the end of the in vitro experiments in order to investigate the intracellular activity of superoxide dismutase (SOD), catalase (CAT), as well as the concentrations of glutathione (GSH) and malondialdehyde (MDA). FeAA treatment led to a reduced sperm motility (P < 0.001) and viability (P < 0.001), decreased the antioxidant parameters of the samples (P < 0.001 in case of SOD; P < 0.01 with respect to CAT; P < 0.05 in relation to GSH) but increased the superoxide production (P < 0.001) and lipid peroxidation (P < 0.001). RES supplementation resulted in a preservation of the spermatozoa vitality and antioxidant characteristics (P < 0.001 in case of SOD; P < 0.01 with respect to 25–50 μmol L−1 RES and P < 0.05 in relation to 10 μmol L−1 RES; P < 0.05 in case of GSH), with 50 μmol L−1 RES proving to be the most effective RES concentration. Our results suggest that RES possesses significant antioxidant properties that may prevent the deleterious effects caused by ROS to spermatozoa, and preserve the fertilization potential of male reproductive cells.

Blood concentration of copper, cadmium, zinc and lead in horses and its relation to hematological and biochemical parameters

Environmental pollution results in serious health hazards to animals and blood analysis serves as a good alternative for health status assessment. The target of this study was to analyze the concentration of selected metals in equine blood, to analyze the blood parameters and to find possible correlations. Blood samples were collected from the vena jugularis of healthy adult horses. The highest concentration of all elements was found in whole blood (Cu 3.84 ± 0.90 mg L−1; Cd = 0.81 ± 0.90 mg L−1; Zn 26.67 ± 14.12 mg L−1; Pb 9.33 ± 5.76 mg L−1). Higher concentrations of copper, cadmium, zinc and lead were detected in blood clots compared to blood sera (44.04%). A similar tendency was found for cadmium (50%), zinc (13.08%) and lead (46.02%), which showed generally higher concentrations in blood clots (cells). Correlation analysis proved some relations between analyzed elements. In blood clots there is a strong positive correlation between Cd – Pb (r = 0.93) and Zn – Pb (r = 0.71) was detected. For biochemical and hematological parameters mainly medium correlations were detected. Obtained results prove different correlations of analyzed elements in blood components as well as the effect on parameters of blood biochemical and hematological profiles.

The evaluation of endocrine regulators after intramuscular and oral application of cyanogenic glycoside amygdalin in rabbits

Abstract The present in vivo study was designed to reveal whether amygdalin is able to cause changes in the endocrine profile and thus alter the key reproductive and physiological functions, using rabbits as a biological model. 40 adult female rabbits were randomly divided into five groups: the control group without any amygdalin administration, two experimental groups received a daily intramuscular injection of amygdalin at dose 0.6 and 3.0 mg kg-1 b.w. and other two groups were fed by crushed apricot seeds at dose 60 and 300 mg kg-1 b.w., mixed with commercial feed over the period of 28 days. The body weight of each experimental animal was recorded weekly during the whole study. Plasma levels of steroid (progesterone, 17β-estradiol, testosterone), thyroid (triiodothyronine, thyroxine, thyroid-stimulating hormone), as well as anterior pituitary (prolactin, luteinizing hormone) hormones were assessed by ELISA. Intramuscular application of amygdalin did not affect (P ≥ 0.05) the plasma levels of none of the endocrine regulators selected. Oral form of amygdalin did not induce significant changes in the plasma levels of examined hormones either. Similarly, no clinically noticeable changes in the average body weight of experimental animals were observed. Our findings indicate that intramuscular and oral application of amygdalin did not significantly affect the plasma levels of studied endocrine regulators in experimental rabbits. In this in vivo study, no obvious beneficial or negative effects of amygdalin on the physiological functions of female rabbits were demonstrated. Short-term intake of amygdalin at the recommended doses does not represent risk for animal health.

Antioxidant status of rabbits after treatment with epicatechin and patulin

Abstract The aim of this study was to assess antioxidant status by measuring of antioxidant parameters (SOD – superoxide dismutase, GPx – glutatione peroxidase, CAT – catalase, Trolox, TAC – total antioxidant capacity, bilirubin, albumin) and parameters associated with oxidative stress (MDA – malondialdehyde, MDA Adduct – malondialdehyde adduct, H2O2 – hydrogen peroxide) in rabbit blood after five weeks exposure to epicatechin alone or in combination with patulin in vivo. We investigated the protective role of epicatechin against possible negative effect of patulin in rabbits. Adult female rabbits (n = 32) were used in the experiment. Animals were divided into eight groups with four rabbits in each group. In the experiment there were two control groups: one without addition of patulin and epicatechin and one with addition of patulin (10 μg kg−1). Rabbits in the experimental groups received three times a week intramuscular injection of low dose (10 μg kg−1), medium dose (100 μg kg−1) or high dose (1000 μg kg−1) of epicatechin alone or in combination with low dose (10 μg kg−1) of patulin. Patulin alone in low dose significantly (P < 0.05) affected total antioxidant status in comparison with the experimental groups with the highest dose of epicatechin alone (1000 μg kg−1) and in combination with patulin. SOD was negatively affected by patulin. Enzymatic activity of this enzyme was significantly lower (P < 0.05) in the experimental group with patulin in comparison with the experimental group receiving the lowest concentration of epicatechin (10 μg kg−1). Patulin can induce oxidative stress in rabbits in low dose and catechin could have possible protective effect. Our results showed that patulin negatively affected TAC, and SOD in rabbit blood. Further research needs to be focused on the generation of data dealing with antioxidant effects.

Blood plasma levels of anterior pituitary hormones of rabbits after apricot seed exposure in vivo

The present study describes possible changes in plasma levels of anterior pituitary hormones induced by bitter apricot (Prunus armeniaca L.) seeds in young female rabbits in vivo. Prunus armeniaca L. is an important medicinal edible plant species commonly known as “apricot”. The apricot is a member of the Rosaceae and subfamily Prunoideae. It is one of the most delicious and commercially traded fruits in the world. Apricot kernel is the inner part of the seed of the apricot fruit. The kernel is used to produce oil and other chemicals used for medicinal purposes. The seeds are potentially useful in human nutrition and for treatment several diseases especially cancer. In the present study apricot seeds were mixed with feed at different doses 0, 60, 300, 420 mg*kg-1 of body weight. ELISA was used to determine the levels of follicle stimulating hormone (FSH), luteinizing hormone (LH) and prolactin (PRL). 58- days application of apricot seeds did not affect the concentration (P≥0.05) of PRL, LH in blood plasma. Significant (P≤0.01) inhibition of FSH levels induced by the seeds was found at the dose of 420 mg*kg-1 but not at 60 and 300 mg*kg-1 of body weight. These results are suggesting that the natural substances present in apricot seeds may be involved in mechanisms of ovarian folliculogenesis.

Does apricot seeds consumption cause changes in human urine

Natural substances, such as amygdalin, used in alternative medicine gained high popularity. Common people as well as patients with different diseases have almost unlimited access to various natural supplements. To protect human health, it is very important to study effect of these substances. Amygdalin is a cyanogenic glucoside derived from seeds of rosaceous plants, for example seeds of bitter almonds ( Prunus dulcis ), or apricot, cherry, apple, peach, plum, etc. It is a natural product that owns antitumor activity, it has also been used for the treatment of asthma, bronchitis, emphysema, leprosy and diabetes and produces a kind of antitussive and antiasthmatic effects. The present in vivo study was designed to reveal whether amygdalin in apricot seeds has got an effect on human urine composition, pH value and urine associated health status after six weeks of oral administration. The study group finally consisted of 34 healthy adult volunteers (21 females and 13 males). All participants were asked to consume 60 mg.kg -1 body weight of bitter apricot seeds daily (approximately 3.0 mg.kg -1 of amygdalin) during 6 weeks. During the experiment, three urine collections were carried out (first collection - at the beginning of the experiment; second collection - after 21 days; third collection - after 42 days). Quantification of urine calcium (Ca), magnesium (Mg), phosphorus (P), sodium (Na), potassium (K), chlorides (Cl - ), urea and pH value after apricot seeds supplementation was performed. Statistical analysis of variance showed, that consumption of bitter apricot seeds during 42 days had a significant ( p <0.01) effect on amount of calcium excreted in urine, though this decrease shifted its level from elevated mean value in control collection into normal physiological range. Significant changes were observed in urea ( p <0.05) and phosphorus ( p <0.01) levels in urine after apricot seed ingestion, but gender was also considered to be a source of their variation.

Oral and intramuscular application of cyanogenic glycoside amygdalin did not induce changes in haematological profile of male rabbits

Amygdalin is a cyanogenic glycoside initially obtained from the seeds of bitter almonds. It is composed of one molecule of benzaldehyde, two molecules of glucose and one molecule of hydrocyanic acid. Various ways of amygdalin application play a different role in recipient organism. Intravenous infusion of amygdalin produced neither cyanidemia nor signs of toxicity, but oral administration resulted in significant blood cyanide levels. The present in vivo study was designed to reveal whether amygdalin is able to cause changes in the haematological profile and thus alter the physiological functions, using rabbits as a biological model. Adult male rabbits (n = 20) were randomly divided into five groups: the control group without any amygdalin administration, two experimental groups received a daily intramuscular injection of amygdalin at a dose 0.6 and 3.0 mg.kg -1 b.w. and other two groups were fed by crushed apricot seeds at dose 60 and 300 mg. kg -1 b.w., mixed with commercial feed over the period of 14 days. After two weeks, haematological parameters in whole blood were analysed (WBC - total white blood cell count, LYM - lymphocytes count, MID - medium size cell count, GRA - granulocytes count, RBC - red blood cell count, HGB - haemoglobin, HCT - haematocrit, MCV - mean corpuscular volume, MCH - mean corpuscular hemoglobin, MCHC - mean corpuscular hemoglobin concentration, RDWc - red cell distribution width, PLT - platelet count, PCT - platelet percentage, MPV - mean platelet volume, PDWc - platelet distribution width) using haematology analyser Abacus junior VET. Our findings indicate that intramuscular and oral application of amygdalin for two weeks did not significantly affect the haematology parameters in experimental animals. In this study, no obvious beneficial or negative effects of amygdalin administration on the blood of male rabbits were observed.

Human Urine Alterations Caused by Apricot Seeds Consumption

Abstract Amygdalin is a cyanogenic glucoside derived from various plants seeds. It is a natural product that owns antitumor activity, it has also been used for the treatment of asthma, bronchitis, emphysema, leprosy and diabetes and produces antitussive and antiasthmatic effects. The present in vivo study was designed to reveal whether amygdalin in apricot seeds has got an effect on human urine composition, pH value and urine associated health status after twelve weeks of oral administration. The study group finally consisted of 12 healthy adult volunteers. All participants consumed 60 mg/kg of body weight of bitter apricot seeds daily (approximately 3.0 mg/kg of amygdalin) during 12 weeks. Two urine collections were carried out (at the beginning of the experiment and after 84 days). Quantification of urine calcium, magnesium, phosphorus, sodium, potassium, chlorides, urea and pH value after apricot seeds supplementation was performed. Paired t-test showed, that consumption of bitter apricot seeds during 84 days had a significant (P < 0.01) effect on amount of calcium excreted in urine, though this decrease shifted its level from elevated mean value in control collection into normal physiological range. No significant changes were observed in other parameters after apricot seed ingestion.