Anti-Inflammatory, Antioxidant and Enzyme Inhibition Activities in Correlation with Mycochemical Profile of Selected Indigenous Ganoderma spp. from Balkan Region (Serbia).

The aim of this research work was to study the bioactivity potentials (antioxidant, anti-inflammatory, and enzyme inhibitory) of ethanol (EtOH), water (H2 O) and chloroform (CHCl3 ) extracts of G. applanatum, G. lucidum, G. pfeifferi and G. resinaceum as well as their mycochemical profile: the total content of phenolics (TP) and sugars (TS) and LC/MS/MS detection of phenolics. LC/MS/MS profile showed that p-hydroxybenzoic and protocatechuic acids were mostly found. The highest ABTS and DPPH activities were detected in polar G. applanatum extracts (159.84±0.59 mg TE/g d.w., IC50 =0.85±0.30 µg/mL, respectively), while G. resinaceum CHCl3 extract was the most potent in NO assay (IC50 =41.21±0.18-81.89±0.81 µg/mL). The highest TP and TS were generally determined in G. applanatum EtOH extracts. Enzyme inhibitory effects were determined in H2 O extracts. Generally, CHCl3 extracts showed the most powerful anti-inflammatory potential. These results suggest that analyzed species are a promising source of bioactive compounds and may be considered as candidates for new food supplements or drug formulations.

Bioactive Phenolic Compounds of Two Medicinal Mushroom Species Trametes versicolor and Stereum subtomentosum as Antioxidant and Antiproliferative Agents.

Medicinal mushrooms have tremendous potential in production of bioactive compounds with diverse bioactivities while the biochemical potential of some specific mushroom strains (autochthonous for the region) in production of specific bioactive agents may be of the main importance in a continuous search for novel strains with supreme activities all over the world. In this study, the ethanolic (EtOH) and water (H2 O) extracts of wild-growing polypore mushroom species were investigated: Trametes versicolor (L.) Lloyd and Stereum subtomentosum Pouzar. This study was designed to determine total phenol (TP), flavonoid (TF) and protein content (TPR) as well as LC/MS/MS phenolic profile related to in vitro antioxidant, antiproliferative (MTT assay) (AP) and DNA fragmentation properties. The H2 O extracts expressed better antioxidant scavenging potential than EtOH showing the highest activity for the T. versicolor (IC50 =5.6 µg/mL, IC50 =0.6 µg/mL for DPPH. and OH. , respectively) while O2 .- activity achieved the best activity for S. subtomentosum (IC50 =4.1 µg/mL). In contrary, the highest AP activity was obtained for the EtOH extracts of S. subtomentosum (IC50 =141.1 µg/mL). The EtOH extracts of both species showed the highest TP, TF and TPR content. Obtained results of DNA degradation indicate genotoxicity potential of the extracts at high concentration. The LC/MS/MS detection showed that the majority of analyzed extracts contained phenolic acids, p-hydroxybenzoic and protocatechuic acid. The obtained results suggest that analyzed medicinal mushroom species, T. versicolor and S. subtomentosum, could be of potential interest as new sources of strong natural antioxidants as well as antiproliferative agents in the future.

Chemical composition, nutritional profile and in vivo antioxidant properties of the cultivated mushroom Coprinus comatus.

This study investigated the chemical and nutritional profile and antioxidative properties of cultivated Coprinus comatus. Proximate analysis revealed that C. comatus is rich in carbohydrates, dietary fibres and proteins, and could also be a valuable source of phenolics. Additionally, fat content is low, consisting mainly of polyunsaturated and omega-3 fatty acids. Furthermore, the safety profile of C. comatus is satisfactory, with all elements of toxicological importance within the proposed limits. Oral treatment with C. comatus for 42 days improved the antioxidant capabilities and ameliorated carbon tetrachloride-induced liver damage in rats, marked by decreased serum aminotransferase levels and lipid peroxidation intensity. Glutathione concentrations increased in a dose-dependent manner. Histological morphometric and immunohistochemical analysis confirmed antioxidative and hepatoprotective potential. These findings imply that cultivated C. comatus could be considered a nutraceutical, having beneficial nutrient and therapeutic properties.

Antidiabetic effect of two different Ganoderma species tested in alloxan diabetic rats.

This study was designed to define total protein, phenol and flavonoid content as well as LC-MS/MS phenolic profile related to antioxidant and antidiabetic activity of ethanolic (EtOH) and water extracts of G. pfeifferi and G. resinaceum. G. resinaceum water extract possessed the highest ability to scavenge DPPH˙ and O2˙-, while the EtOH extract of the same species showed better activity on NO˙ related to other extracts. The highest level of bioactive compounds was determined generally in EtOH extracts. Antidiabetic action was evaluated by the oral glucose tolerance test (OGTT) and histological examination of pancreas and liver in normoglycemic and alloxan-induced diabetic animals. Histological examination of pancreatic tissue demonstrated that G. pfeifferi extracts have protective effects. To conclude, analysed extracts could be considered as a promising candidate for further research with the aim to promote antidiabetic activity, which is for the first time reported for G. pfeifferi.

Professional Female Athletes Are at a Heightened Risk of Iron-Deficient Erythropoiesis Compared With Nonathletes.

This study primarily aimed to quantify and compare iron status in professional female athletes and nonathletes. Furthermore, this study also aimed to identify differences in iron status according to sporting discipline and explore the relationship between ferritin concentration and weekly training volume in professional athletes. A total of 152 participants were included in this study, including 85 athletes who were members of senior teams (handball, n = 24; volleyball, n = 36; soccer, n = 19; and judo, n = 6) involved at the highest level of competition and 67 nonathletes. A significantly greater proportion (p = .05) of athletes (27%) demonstrated iron-deficient erythropoiesis (IDE) compared with nonathletes (13%). There were nonsignificant differences (p > .05) in the prevalence of iron deficiency (ID; 49% vs. 46%) and iron deficiency anemia (IDA; 2% vs. 4%) between athletes and nonathletes. Similarly, the prevalence of ID, IDE, and IDA was not significantly different between sports (p > .05). Furthermore, training volume was negatively correlated with ferritin concentration in athletes (r: -.464, moderate, p < .001). Professional female athletes are at a heightened risk of IDE compared with nonathletes; therefore, they should be periodically screened for ID to reduce the deleterious effects on training and performance. The similar prevalence of ID, IDE, and IDA found across athletes competing in different sports suggests that overlaps exist between handball, volleyball, soccer, and judo athletes regarding risk of disturbance in iron metabolism.

Trifolium pratense L. as a potential natural antioxidant.

The essential oils of three different growth stages of Trifolium pratense L. (TP1, TP2 and TP3) were investigated by gas chromatography-mass spectrometry and tested for their antioxidant and antimicrobial activities. The highest content of volatile compounds was found in the essential oil sample TP1, where terpenes such as ß-myrcene (4.55%), p-cymene (3.59%), limonene (0.86%), tetrahydroionone (1.56%) were highlighted due to their biological activity. The antioxidant activity was determined by following the scavenging capacity of the essential oils for the free radicals DPPH·, NO· and O2·-, as well as effects of the investigated oils on lipid peroxidation (LP). In all three cases, the sample TP1 showed the best radical-capturing capacity for DPPH· (27.61±0.12 µg/mL), NO· (16.03±0.11 µg/mL), O2·- (16.62±0.29 µg/mL) and also had the best lipid peroxidation effects in the Fe2+/ascorbate induction system (9.35±0.11 µg/mL). Antimicrobial activity was evaluated against the following bacteria cultures: Escherichia coli (ATCC10526), Salmonella typhimurium (ATCC 14028), Staphylococcus aureus (ATCC 11632) and Bacillus cereus (ATCC 10876). None of the examined essential oil samples showed inhibitory effects on the tested bacterial strains.

Antioxidant profile of Trifolium pratense L.

In order to examine the antioxidant properties of five different extracts of Trifolium pratense L. (Leguminosae) leaves, various assays which measure free radical scavenging ability were carried out: 1,1-diphenyl-2-picrylhydrazyl, hydroxyl, superoxide anion and nitric oxide radical scavenger capacity tests and lipid peroxidation assay. In all of the tests, only the H2O and (to some extent) the EtOAc extracts showed a potent antioxidant effect compared with BHT and BHA, well-known synthetic antioxidants. In addition, in vivo experiments were conducted with antioxidant systems (activities of GSHPx, GSHR, Px, CAT, XOD, GSH content and intensity of LPx) in liver homogenate and blood of mice after their treatment with extracts of T. pratense leaves, or in combination with CCl4. Besides, in the extracts examined the total phenolic and flavonoid amounts were also determined, together with presence of the selected flavonoids: quercetin, luteolin, apigenin, naringenin and kaempferol, which were studied using a HPLC-DAD technique. HPLC-DAD analysis showed a noticeable content of natural products according to which the examined Trifolium pratense species could well be regarded as a promising new source of bioactive natural compounds, which can be used both as a food supplement and a remedy.

Effects of St John's wort (Hypericum perforatum L.) extracts on epileptogenesis.

The purpose of this study was to investigate the effects of treatment with water, n-butanol and ether extracts of Hypercom perforatum L. on epileptogenesis in rabbits. Animals from the control group received solvent-ethanol, and the kindling model of epilepsy was used. Epileptic focus was induced in Chinchilla rabbits by stimulation of the hippocampus. The following parameters were determined: the minimum current strength necessary to induce after-discharge (AD) - discharges appearing after cessation of stimulation; AD duration; the number of stimulations necessary to induce spontaneous kindling; and the latency time for the development of full kindling. The results obtained indicate that epileptogenesis is influenced by Hypericum perforatum L. extract treatment. Animals treated with an ether extract of Hypericum perforatum L. required significantly weaker minimum current strengths for the development of epileptogenic focus, and displayed longer AD times, while the number of electro-stimulations necessary for full kindling was less. In contrast, animals treated with water and n-butanol extracts required increased electro-stimulations for the development of epileptic discharge, and displayed shortened AD durations versus controls.

Antioxidant capacity of Ocimum basilicum L. and Origanum vulgare L. extracts.

The antioxidant properties of five different extracts (Et2O, CHCl3, EtOAc, n-BuOH, and H2O) of Ocimum basilicum L. and Origanum vulgare L. were studied. Antioxidant activity was assessed in six different model systems. Free radical scavenging capacity (RSC) was evaluated by measuring the scavenging capacity of extracts on DPPH, NO, O2•⁻ and OH radical, as well as on hydrogen peroxide (H2O2). In addition, the protective effects on lipid peroxidation in liposomes (LPx) were evaluated by TBA-assay using the Fe²âº/ascorbate induction system. The amount of total phenolic compounds and content of total flavonoids was also determined. EtOAc, n-BuOH and H2O extracts of O. basilicum and O. vulgare expressed very strong scavenger activity. Furthermore, the mentioned extracts showed notable inhibition of LPx. On the other hand, Et2O and CHCl3 extracts showed much weaker effect in the neutralization of DPPH, NO and O2•⁻ radicals and the neutralization of H2O2. When examining the production of OH radicals and inhibition of LPx, the Et2O and CHCl3 extracts showed weak prooxidative properties. The observed differences in antioxidant activity could be partially explained by the levels of phenolics and flavonoids in the investigated O. basilicum and O. vulgare extracts.

Antioxidant activities of Melittis melissophyllum L. (Lamiaceae).

Extracts of Melittis melissophyllum leaves in ether, chloroform, ethyl acetate, n-butanol and water were evaporated to dryness and dissolved in 50% ethanol to make 10% (w/v) solutions. The potential protective action of the extracts was assessed by the corresponding in vitro and in vivo tests. In the in vitro experiments extracts were tested as potential scavengers of free radicals (DPPH, O2·â», NO, and OH radicals), as well as inhibitors of liposomal peroxidation (LPx). The results obtained show that all extracts (exept n-BuOH extract) are good scavengers of radicals and reduce LPx intensity in liposomes, which points to their protective (antioxidant) activity. In vivo experiments were concerned with antioxidant systems (activities of GSHPx, GSHR, Px, CAT, XOD, GSH content and intensity of LPx) in liver homogenate and blood-hemolysate of experimental animals after their treatment with extracts of M. melissophyllum leaves, or in combination with CCl4. On the basis of the results obtained it can be concluded that the examined extracts have protective (antioxidative) effect and this antioxidative behaviour is more pronounced in liver than in blood-hemolysate. The reason is probably the fact that liver contains other enzymatic systems, which can also participate in the antioxidative mechanism. Of all the extracts the H2O one showed the highest protective activity.

Thermodynamics of fatty acid degradation.

In this communication, the correlation of some thermodynamic properties of structurally similar biological compounds with items such as the number of carbon atoms or the number of its characteristic groups, such as phosphates, was applied to enzyme thermodynamics, to advance the thermodynamics of fatty acid degradation. It was shown that these concepts apply equally well for all three major thermodynamic functions: Gibbs energy of formation, enthalpy of formation, and entropy of formation. First, the standard transformed Gibbs energies of formation, Δ(f)G'(o), were calculated for the major portion of saturated fatty acids at 25 °C and pH 5, pH 7, and pH 9. Second, a total ATP yield was calculated for complete combustion of major saturated fatty acids to CO(2) and water. Third, the standard transformed Gibbs energies of reaction, Δ(r)G'(o), were calculated for enzyme reactions catalyzing complete ß-oxidation of butyryl-CoA to acetyl-CoA at 25 °C and pH 7.

Antioxidant activities of celery and parsley juices in rats treated with doxorubicin.

We have examined the influence of diluted pure celery and parsley leaf and root juices and their combinations with doxorubicin on the antioxidant status [as measured by the content of reduced glutathione (GSH) and ferric reducing antioxidant power (FRAP)] in liver homogenate and hemolysate and on the contents of cytochrome P450 in liver homogenate. It was found that doxorubicin significantly decreased the content of reduced glutathione and the total antioxidative status (FRAP) in liver homogenate and hemolysate, while celery and parsley juices alone and in combination with doxorubicin had different actions. Doxorubicin and celery juice had no effect on content of cytochrome P450. However, in combination with doxorubicin, parsley root juice significant increased, and parsley leaves juice decreased the cytochrome P450 content (compared to doxorubicin treated animals). Only parsley root juice significantly increased the content of cytochrome P450.

Anti-oxidative activity of an aqueous suspension of commercial preparation of the mushroom Coprinus comatus.

In this study the effects of an aqueous suspension of a commercial preparation of the mushroom Coprinus comatus on oxidative stress induced in rats by alloxane and carbon tetrachloride was examined. The effects were estimated from changes in the biochemical parameters (xanthine oxidase, glutathione peroxidase and catalase activity, reduced glutathione content, and extent of lipid peroxidation) of liver homogenate as well as histological changes in the liver of the rats treated with alloxane and carbon tetrachloride. Two screening doses of alloxane sufficient to induce diabetes in rats did not have any significant effect on the examined biochemical parameters of liver homogenate or on the cytoarchitectonics of liver cross-sections. Treatment with carbon tetrachloride resulted in a significant increase in the intensity of lipid peroxidation and peroxydasis activity, as well as with decrease in catalase activity. Certain changes in liver cross sections were detected, such is lymphocyte infiltration of dilated sinusoid capillaries. Administration of Coprinus comatus suspension thus showed antioxidative potential, evidenced by an increase of antioxidative status of liver homogenate and prevention of histological changes in liver cross sections.

In vitro and in vivo effects of Laurus nobilis L. leaf extracts.

The in vitro and in vivo antioxidant activities of different extracts of laurel leaves were studied. Free radical scavenging capacity (RSC) was evaluated measuring the scavenging activity on the DPPH, NO, O(2)(.-) and OH radicals. The effects on lipid peroxidation (LP) were also evaluated. Experimental results indicate that ethyl acetate extract of leaves has exhibited the largest RSC capacity in neutralization of DPPH, NO, O(2)(.-) and OH radicals. The same result was obtained in investigation of extracts impact on LP. The in vivo effects were evaluated on some antioxidant systems (activities of GSHPx, LPx, Px, CAT and XOD, and GSH content) in the mice liver and blood-hemolysate after treatment with the examined laurel extracts, or in combination with carbon tetrachloride (CCl(4)). On the basis of the results obtained it can be concluded that the examined extracts exhibited a certain protective effect, which is more pronounced on the liver than on blood-hemolysate parameters. The results obtained indicate toxicity of CCl(4), probably due to the radicals involved in its metabolism. Combined treatments with CCl(4) and the examined extracts showed both positive and negative synergism. Based on the experimental results, the strongest protective effect was shown by the EtOAc extract.

Inhibitory effect of Ginkgo biloba extract on the tonus of the small intestine and the colon of rabbits.

Ginkgo biloba is widely used in folk medicine. Patients very often use the plant preparation with no concern for purity. They also tend to increase the dosage by themselves and this may result in certain insufficiently researched acute effects. Due to this extremely widespread application, the aim of this work is an examination of the possible acute effects of Ginkgo bilobaon the motility of the small and the large intestine of rabbits. capital TE, Cyrilliche effects of Gingium - a standardized ginkgo biloba extract (GBE) [one milliliter preparation contained 8.8-10.8 mg ginkgo flavonol glycoside and 2.0-2.8 mg lactone ring-containing terpenes (ginkgolides and bilobalides)], on the tonus of isolated segments of the ileum and the colon of rabbits were examined. The experiments were carried out on isolated bowel incisions according to the Magnus method. Data was registered by physiography (Narco-Bio-System). Our results show that GBE (0.006 g/L, - 0.06 g/L) concentration-dependently reduces the tonus of the ileum and the colon of rabbits. Apart from that, GBE reduces the increase of the tonus of the ileum caused by acetylcholine (ACh), but does not change colon tonus intensified by ACh. This indicates that the effects of the used extract in the ileum are predominantly achieved through cholinergic mechanisms, while the relaxant effects in the colon are achieved in some other way.

Liquid chromatography/tandem mass spectrometry study of anti-inflammatory activity of plantain (Plantago L.) species.

To evaluate anti-inflammatory activity of selected Plantago species (P. lanceolata L. and P. major L.) an optimized in vitro test for determination of cyclooxygenase-1 (COX-1) and 12-lipoxygenase (12-LOX) inhibition potency was undertaken. By using intact cell system (platelets) as a source of COX-1 and 12-LOX enzymes and highly sensitive and specific LC-MS/MS technique for detection of main arachidonic acid metabolites formed by COX-1 and 12-LOX, this test provides efficient method for evaluation of anti-inflammatory potential of plant extracts and isolated compounds. Our results validated the well-known COX-1 inhibitory activity of P. lanceolata and P. major methanol extracts (concentration required for 50% inhibition (IC(50)) was 2.00 and 0.65 mg/ml, respectively). Furthermore, 12-LOX inhibitory activity of examined extracts was reported for the first time (IC(50)=0.75 and 1.73 mg/ml for P. lanceolata and P. major, respectively). Although renowned inhibitors, such as acetylsalicylic acid and quercetin showed higher activity, this study verifies P. lanceolata and P. major as considerable anti-inflammatory agents.

Antioxidant effects of some drugs on immobilization stress combined with cold restraint stress.

The aim of this work was to investigate the effect on antioxidant potential of some commonly used drugs (morphine, tramadol, bromocriptine, haloperidol and azithromycin) on immobilization stress (IS) combined with cold restraint stress (CRS) in the rat. After the drug treatment the animals were kept immobilized in the cold chamber at 4+/-0.3 degrees C for 3 hours and then decapitaed and the livers were extracted. The following parameters were determined in the liver homogenate: content of reduced glutathione, activities of catalase, xanthine oxidase, glutathione reductase, glutathione peroxidase, peroxidase, and lipid peroxidation intensity. A battery of biochemical assays was used and the resulting data were statistically analyzed. Combined stress exhibited a prooxidative action (increased catalase activity, lowered content of reduced glutathione). Significantly enhanced catalase activity that was observed in all groups compared to the control indicates that the primary reactive oxygen species (ROS) metabolite is hydrogen peroxide, which decomposes very rapidly (very high catalase activity), thus hindering formation of OH radicals as the most toxic ROS. None of the tested drugs showed a protective effect on combined IS and CRS. The intensity of lipid peroxidation did not change either in the combined stress or under additional influence of the drugs. Probably, under our experimental conditions, the time was not sufficiently long to observe damage of lipid membrane by ROS.

Effect of aroma and magnum hops extracts and paracetamol on antioxidant liver parameters in mice.

Hop varieties that are mainly grown in Serbia are Magnum (German variety) and Aroma, which is grown only in the Vojvodina region. About the use of hops extracts as an auxiliary remedy there are divergent opinions. Our findings indicate that extracts of Magnum and Aroma varieties, among the others, enhance and prolong the analgesic action of paracetamol. For this reason we undertook a study of the effects of these extracts alone and in combination with paracetamol, along with the action of paracetamol alone, on the activity of the antioxidant enzymes GSHPx, CAT, Px, XOD, GSHR, glutathione content, LPx intensity, as well as activities of AST and ALT. Paracetamol in the dose applied exerted no influence on the investigated parameters and neither did ethanolic extract of Magnum variety. On the other hand, ethanolic extract of Aroma hops caused a significant reduction of GSH content. In combination with paracetamol, extracts of both Magnum and Aroma varieties reduced significantly the LPx intensity, activities of CAT and GSHPx, as well as GSH content in the liver homogenate. A significant increase in the AST value with respect to control was also observed. These findings indicate the disturbance in the functioning of hepatocytes, probably by decelarating metabolism and elimination of paracetamol.

Protective effects of celery juice in treatments with Doxorubicin.

The aim of this work was to investigate possible protective effect of celery juice in doxorubicin treatment. The following biochemical parameters were determined: content of reduced glutathione, activities of catalase, xanthine oxidase, glutathione peroxidase, peroxidase, and lipid peroxidation intensity in liver homogenate and blood hemolysate. We examined influence of diluted pure celery leaves and roots juices and their combinations with doxorubicine on analyzed biochemical parameters. Celery roots and leaves juices influenced the examined biochemical parameters and showed protective effects when applied with doxorubicine.