In our previous Genome-wise Association Study we found that Cystic Fibrosis Transmem- brane Conductance Regulator gene (CFTR) is a candidate gene for sperm motility in fresh semen of Holstein-Friesian bulls. Since in cows thawed semen is commonly used for the artificial insem- ination (AI) we have decided to find out whether functional polymorphism within CFTR gene coding sequence is associated with selected parameters of thawed sperm, including their motility evaluated by computer-assisted sperm analysis (CASA), the activity of three antioxidant enzymes: glutathione peroxidase (GPx) catalase (CAT), superoxide dismutase (SOD), ATP con- tent and integrity of sperm membranes. One hundred twenty Holstein Friesian bulls kept in uni- form environmental conditions (one AI company) were included in the study. Significant associ- ations between genotypes of missense mutation within exon 11 of the CFTR gene (Met468Leu) and the activity of antioxidant enzymes and sperm mitochondrial function were revealed. No effect of CFTR genotypes on sperm motility was observed. Significant differences in CAT and SOD activity were found between AA and TT homozygous individuals. Bulls with TT genotype had the lowest activity of both antioxidant enzymes. The same bulls also showed the lowest num- ber of sperm with active mitochondria. Our results demonstrate that missense mutation Met468Leu within CFTR gene is associated with antioxidant enzyme activity and mitochondrial function of bovine thawed sperm without affecting their motility.
The aim of the present study was to determine the concentrations of glutathione (GSH), vitamin C, copper (Cu) and zinc (Zn) in the uterine tissues in diagnosis of canine pyometra. Fourteen samples of uterine tissues from female dogs with pyometra and twelve samples of healthy uteruses (control) were used. The concentrations of GSH and vitamin C were determined in the uterine tissue homogenates using spectrophotometric methods. The concentrations of Cu and Zn were measured using atomic absorption spectrometer. The results obtained showed the significantly lower (p<0.05) concentration of GSH and the trend towards lower concentration of vitamin C in the pyometra samples compared to the control. The concentrations of Cu and Zn were similar in the uterine tissues from female dogs with pyometra and those from healthy female dogs. The lower GSH and vitamin C concentrations in the uterine tissues of female dogs with pyometra indicate that the non-enzymatic antioxidant mechanisms are impaired in the uterus of dogs with pyometra. These findings suggest that the imbalance of oxidative-antioxidative can play an important role in pathogenesis of canine pyometra.
We used the Dpph method to assess in vitro the antiradical activity of extracts from the roots, leaves and fruits of six Rumex L. (dock) species. Data from preliminary screening indicated that all the tested extracts showed antioxidant properties. The degree of antiradical activity depended upon the plant part. Fruit extracts from R. hydrolapathum Huds., R. obtusifolius L. and R. confertus Willd. showed stronger antiradical properties than the other tested material. We also determined tannin content levels in the extracts and their correlation with antioxidant activity.
The influence of ambient solar UV-A or UV-B radiation on growth responses was investigated in three varieties of cotton (Gossypium hirsutum L.) after exclusion of solar UV-A/B radiation: JK-35, IH-63 and Khandwa-2. Cotton plants were grown from seeds in UV-exclusion chambers lined with selective UV filters to exclude either UV-B (280-315 nm) or UV-A/B (280-400 nm) from the solar spectrum under field conditions. Excluding UV-B and UV-A/B significantly increased plant height, leaf area and dry weight accumulation in all three varieties of cotton. The varieties differed considerably in their sensitivity to ambient UV-A/B. Khandwa-2 was most sensitive and JK-35 least sensitive to ambient solar UV. We monitored the activity of the antioxidant enzymes superoxide dismutase (SOD), ascorbic acid peroxidase (APX), glutathione reductase (GR) and guaiacol peroxidase (GPX), as well as the level of the antioxidant ascorbic acid (ASA), in primary leaves of the most UV-sensitive variety (Khandwa-2). The level of UV-B-absorbing substances was significantly decreased by exclusion of solar UV-B and UV-A/B. Exclusion of solar UV decreased the activity of all the antioxidant enzymes monitored and the level of ascorbic acid versus control plants (+UV-A/B) grown under filters transparent to solar UV. Reduction of the antioxidant defense after UV exclusion indicates that ambient solar UV exerts significant stress and induces some reactive oxygen species to accumulate, which in turn retards the growth and development of cotton plants. Ambient solar UV stresses cotton plants, shifting their metabolism towards defense against solar UV. Exclusion of solar UV eliminates the need for that defense and leads to enhancement of primary metabolism.
Toxicity and physiological alterations were determined in Pseudococcus viburni nymphs treated with Artemisia annua methanolic extract. The leaf dipping bioassay showed LC50 values of 0.287% and 0.194% 24 and 48 hours post-exposure. Activities of general esterases were significantly higher in the control nymphs than in those which had been treated except for the 48 h time interval using α-naphtyl acetate. The activity of glutathione S-transferase using CDNB (1-chloro-2,4-dinitrobenzene) in the control nymphs, was significantly higher than in the control at both time intervals while no significant difference was observed after 24 h in addition to the higher enzymatic activity in the treated nymphs after 48 h. All three aminotransferases were significantly more active in the control nymphs except for time intervals of 24 h for γ-glutamyl transferase and 48 h for alanine aminotransferase. Higher activities of lactate dehydrogenase, acid- and alkaline phosphatase were found in the control nymphs than in treated nymphs for all time intervals. Activities of the enzymes involved in the antioxidant system including catalase, peroxidase, superoxide dismutase, ascorbate peroxidase and glucose-6-phosphate dehydrogenase was increased in the treated nymphs compared to the control. Results of the current study demonstrated toxic effects of A. annua methanolic extract on P. viburni nymphs causing mortality and physiological turbulences.
The chemical composition and bioactivity of a water/methanol extract prepared from aerial parts of Circaea lutetiana were determined. HPLC-DAD-MS3 analysis revealed the presence of 14 different compounds comprising phenolic acids, ellagitannins and flavonoids. Antioxidant assays showed the extract's strong scavenging activity towards DPPH (SC50 33.1±3.1 μg/ml), O2 - (SC50 4.0±2.3 μg/ml) and H2O2 (SC50 below 2 μg/ml). Enzyme-based studies revealed that Circaea lutetiana extract inhibits the activity of hyaluronidase (IC50 13.3±2.4 μg/ml) and lipoxygenase (IC50 44.7±1.4 μg/ml). These results support some traditional uses of Circaea lutetiana.
We examined the effects of feeding by the polyphagous insect Coccus hesperidum on its host plant Nephrolepis biserrata under different intensities of infestation. As an effect of scale insect feeding there were significant changes in the values of parameters reflecting the state of cell membranes. N. biserrata plants reacted to the biotic stress by increasing guaiacol peroxidase activity and decreasing catalase activity. Our data show that these processes play key roles in plant tolerance mechanisms, here the fern’s response to insect feeding. The observed complex reaction of N. biserrata testifies to actively proceeding, complex and very often contrasting mechanisms triggered with the aim of neutralizing the effects of biotic stress and enabling normal cell functioning in plants attacked by scale insects
I n t r o d u c t i o n: Interactions between oral microbiota and systemic diseases have been suggested. We aimed to examine the composition of oral microbiota with reference to antioxidative defense and its correlation with clinical state in Crohn’s disease (CD) in comparison to ulcerative colitis (UC). Ma t e r i a l s a n d Me t h o d s: Smears were taken from the buccal and tongue mucosa of patients with CD, UC and controls, and cultured with classical microbiology methods. Bacterial colonies were identified using matrix-assisted laser desorption/ionization (MALDI) with a time-of-flight analyzer (TOF). Blood morphology and C-reactive protein (CRP) were analyzed in the hospital laboratory. Antioxidative defense potential (FRAP) was determined using spectrophotometry in saliva and serum. R e s u l t s: Oral microbiota in CD patients were characterized by lower diversity in terms of the isolated bacteria species compared to UC and this correlated with reduced FRAP in the oral cavity and intensified systemic infl ammation. Oral microbiota composition in CD did not depend on the applied treatment. In CD patients, a negative correlation was observed between the FRAP value in saliva and serum and the CRP value in serum. Individual differences in the composition of oral microbiota suggest that different bacteria species may be involved in the induction of oxidative stress associated with a weakening of antioxidative defense in the oral cavity, manifested by ongoing systemic inflammation. C o n c l u s i o n s: Analysis of both the state of the microbiota and antioxidative defense of the oral cavity, as well as their referencing to systemic inflammation may potentially prove helpful in routine diagnostic applications and in aiding a better understanding of CD and UC pathogenesis associated with oral microbiota.
Polygonum orientale with beautiful red flowers can be found as one dominant species in the vicinity of most water bodies and wetlands in China. However, its phytoremediation potential has not been sufficiently explored because little is known about its resistance to inorganic or organic pollutants. We investigated P. orientale response to low and moderate levels of phenol stress (≤ 80 mg L-1). Endpoints included phenol tolerance of P. orientale and the removal of the pollutant, antioxidant enzyme activities, damage to the cell membrane, osmotic regulators and photosynthetic pigments. In plant leaves, phenol stress significantly increased the activities of peroxidase (POD) and catalase (CAT), as well as the contents of proline, soluble sugars and carotenoids, whereas superoxide dismutase (SOD), H2O2 and electrolyte leakage (EL) levels remained unaltered. On the other hand, there were significant decreases of soluble protein and chlorophyll contents. We demonstrated that, in combination with phenol tolerance and its removal, P. orientale has efficient protection mechanisms against phenol-induced oxidative damage (≤ 80 mg L-1). We propose that P. orientale could be used as an alternative and interesting material in the phytoremediation of phenol.
We investigated the antioxidant defense mechanism, metal uptake and lipid peroxidation (LPO) levels at different leaf positions in Mentha piperita L. grown in Mn2+-deficient and control conditions. Under manganese deficiency the activity of superoxide dismutase (SOD), catalase (CAT) and guaiacol peroxidase (GuaPOX) and the content of ascorbate, chlorophyll, and carotenoid under Mn2+ deficiency were significantly lower than in the control for all leaf positions. SOD activity correlated positively with Mn2+ uptake. Fe2+ uptake was inhibited by Mn2+ deficiency. During early stages of Mn2+ deficiency, M. piperita leaves showed relatively more antioxidant activity and lower LPO. Towards the final stages of the treatment period, comparatively lower SOD, CAT and GuaPOX activity and higher LPO levels accelerated the senescence process.
We used DPPH scavenging assays to study the antioxidant activity of three native Polish species of blackberry leaves (Rubus kuleszae Ziel., R. fabrimontanus (Sprib.) Sprib. and R. capitulatus Utsch.). All the studied extracts (methanolic, water, methanolic-water) showed high DPPH free radical scavenging activity (IC50 450.0-186.0 μg/ml). The most effective of the studied species was Rubus kuleszae. Total content of phenolic compounds (70.50-136.04 mg GAE/g) and phenolic acids (14.70-38.26 mg CAE/g) was determined spectrophotometrically. Antioxidant activity correlated positively with total content of phenolic compounds and phenolic acids.
Hylocereus undatus flower is commonly used as food or for medicinal purposes in south China. To study its antioxidant activity and mechanism we used antioxidant and chemical assays to compare two commercial samples from different locations (Shenjing, Qixing). The difference in antioxidant levels corresponded with differences in chemical content (including total phenolics, total flavonoids, kaempferol and quercetin) between Shenjing and Qixing. The antioxidant ability of H. undatus flower seems attributable to total phenolics (mainly total flavonoids). Kaempferol is one of the main bioactive components. H. undatus flower exerts its antioxidant effects through metal chelation and radical scavenging via hydrogen atom (H•) and electron (e) donation.
Two fungal strains, isolated from Livingston Island, Antarctica (Penicillium commune 161, psychrotolerant and Aspergillus glaucus 363, mesophilic) were investigated for a relationship between growth temperature and oxidative stress response. Cultivation at temperatures below - (10 and 15°C and 10 and 20°C for P. commune and A. glaucus, respectively) and above (25°C and 30°C for P. commune and A. glaucus, respectively) the optimum caused significant difference in growth and glucose uptake in comparison with the control cultures. Enhanced level of reserve carbohydrates (glycogen and trehalose) was determined under cultivation at different temperatures from the optimal one. While the highest content of trehalose was found in the exponential phase, glycogen accumulation was observed in the stationary phase when growth conditions deteriorate. The growth at temperature below- and above-optimum caused strain-dependent changes in two antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). While SOD activity in the psychrotolerant strain increases with decreasing of growth temperature, the mesophilic A. glaucus demonstrated marked reduction of it at below- and above-optimal temperature. Decreasing trend of CAT activity was observed in both strains below the optimal temperature indicating a lack of antioxidant protection from this enzyme under the cold stress conditions.
This work presents a comparative analysis of the phenolic composition (UHPLC-PDA-ESI-MS3, HPLC-PDAfingerprint, UV-spectrophotometric methods) and antioxidant activity (DPPH, FRAP) of leaf samples from two vegetation seasons of a medicinal and dietary plant Sorbus domestica growing in its natural habitat (Croatia, C) and cultivated in Poland (P). The samples from both sources were rich in structurally diverse polyphenols (44 analytes; P: 73.4–76.6 and C: 98.3–106.7 mg GAE/g dry leaves) including the dominating flavan-3-ols and flavonoids. The greatest qualitative and quantitative differences were observed for flavonoids (P: 14.3–20.3%; C: 27.5–34.1% of polyphenols) – in the Polish samples flavonoid diglycosides predominated, in the Croatian samples the contents of both monoglycosides and diglycosides were similar. In the case of dry methanolic extracts, despite the higher extraction efficiency obtained for the Croatian samples (32–36% vs 23–24%), the quality of the extracts was comparable, both in terms of the total phenolic content (P: 269.4–280.0; C: 297.6–304.4; mg GAE/g dry extract) and antioxidant activity parameters (DPPH, EC50, μg/mL. P: 10.5–10.9, C: 10.0–10.3; and FRAP, mmol Fe2+/g, P: 6.64–7.13, C: 7.06–7.11). As a result, the study confirmed the influence of environmental conditions on the phenolic profile and antioxidant capacity of S. domestica leaves, as well as showed that despite some differences, plant materials from both Poland and Croatia might be suitable for production of natural health products.