Mario Falchi

The introduction of the stilbene synthase gene enhances the natural antiradical activity of Lycopersicon esculentum mill

Tomato (Lycopersicon esculentum) is a vegetable rich in antioxidants, such as lycopene, lutein, and zeaxanthin. Their presence is responsible for the characteristic ability of this product to inhibit the formation of reactive oxygen species, including singlet oxygen. The grapes and wines derived from grapes also contain powerful antioxidants. The antioxidant effect is derived from the polyphenols such as resveratrol and proanthocyanidin. Resveratrol is phytoalexin that is synthesized via the activation of the gene, stilbene synthase (STS). We decided to determine if the introduction of this gene into Lycopersicon esculentum Mill could modify its antioxidant activity. Using Electronic Paramagnetic Resonance (EPR) spectroscopy, which permits the detection of antiradical activity, especially •OH (hydroxyl radical), we showed that the antioxidant activity of the products, into which the gene STS had been introduced, was almost double than that of natural products and that their activity was especially pronounced due to ripening. Moreover, resveratrol concentrations in modified tomatoes were much higher than that found in the individual fruit. In the isolated hearts subjected to ischemia/reperfusion, the rats fed with modified tomato exhibited better cardiac performance, reduced myocardial infarct size and decreased number of apoptotic cardiomyocytes, and reduced oxidative stress compared to unmodified tomato or resveratrol alone indicating superior cardioprotective abilities of modified tomatoes.

Cardioprotective properties of raw and cooked eggplant (Solanum melongena L)

Although eggplants are known to be part of a healthy diet, the effects of this fruit on cardioprotection are not known. The present study examined the role of raw and grilled eggplants on cardioprotection using an isolated perfusion heart model. The animals were fed freeze-dried products of either raw or grilled eggplants for 30 days. After 30 days, isolated working hearts were subjected to 30 min ischemia followed by 2 h of reperfusion. Left ventricular function was monitored, and myocardial infarct size and cardiomyocyte apoptosis were assessed. To determine the antioxidant function of eggplants, their DPPH scavenging ability were determined, and polyphenolic components, especially nasunin content, were determined. The chemical composition of raw and grilled eggplants were determined in order to examine whether grilling was associated with major changes in their composition. The results of this study demonstrated eggplants as containing potent cardioprotective compounds judging by their ability to increase left ventricular function, and reduce myocardial infarct size and cardiomyocyte apoptosis. However, there was no difference in cardioprotective ability between the raw and grilled products. The antioxidant vitamins, including vitamin A, vitamin C and β-carotene, were lower and some of the polyphenolic components, especially nasunin content, were higher in grilled eggplants, but they were unable to demonstrate better cardioprotective properties compared to the raw fruit.

Central antalgic activity of resveratrol

A single dose of resveratrol (25 μg/10μl) was injected directly into the right lateral cerebral ventricle (icv) of Wistar rats via an implanted cannula in order to study the analgesic properties of the compound. A control group of rats received 10 μl NaCl 0.9%. The lengthening of the time to reaction to painful stimuli was assessed in the radiant heat tail-flick latency time test. In this study, the response to painful stimuli of the animals treated with resveratrol had a bimodal profile with hypoalgesia or hyperalgesia. In the selected experimental conditions, resveratrol had a definite analgesic effect; the increase in time to reaction ranged from 100-120% (8 rats) to 600-700% (9 rats). In this experiment resveratrol exerts evident central antalgic effects in the majority of rats, which are related to the individual level of excitation and vigilance at baseline. Antinociceptive induced by resveratrol icv injection was maximal at 4-10 min and lasted no longer than 15 min. The effect of resveratrol to produce analgesia after a single icv injection may be interesting for preventing chronic pain.

Antioxidant Activity of Calendula officinalis Extract: Inhibitory Effects on Chemiluminescence of Human Neutrophil Bursts and Electron Paramagnetic Resonance Spectroscopy

There is growing interest in natural chemical compounds from aromatic, spicy, medicinal and other plants with antioxidant properties in order to find new sources of compounds inactivating free radicals generated by metabolic pathways within body tissue and cells, mainly polymorphonuclear leukocytes (PMNs) whose overregulated recruitment and activation generate a large amount of reactive oxygen species (ROS) and reactive nitrogen species (RNS), leading to an imbalance of redox homeostasis and oxidative stress. The aim of this study was to examine whether a propylene glycol extract of Calendula officinalis interferes with ROS and RNS during the PMN respiratory bursts, and to establish the lowest concentration at which it still exerts antioxidant activity by means of luminol-amplified chemiluminescence. Electron paramagnetic resonance (EPR) spectroscopy was also used in order to confirm the activity of the C. officinalis extract. The C. officinalis extract exerted its anti-ROS and anti-RNS activity in a concentration-dependent manner, with significant effects being observed at even very low concentrations: 0.20 μg/ml without L-arginine, 0.10 μg/ml when L-arginine was added to the test with phorbol 12-myristate 13-acetate and 0.05 μg/ml when it was added to the test with N-formyl-methionyl-leucyl-phenylalanine. The EPR study confirmed these findings, 0.20 μg/ml being the lowest concentration of C. officinalis extract that significantly reduced 2,2-diphenyl-1-picrylhydrazyl. These findings are interesting for improving the antioxidant network and restoring the redox balance in human cells with plant-derived molecules as well as extending the possibility of antagonizing the oxidative stress generated in living organisms when the balance is in favor of free radicals as a result of the depletion of cell antioxidants.

Cardioprotection with the Parrodiene 2,4,6-Octatrienal and Its Potassium Salt through Activation of the Akt-Bcl-2 Survival Pathway

A study was undertaken to determine the cardioprotective effects of parrodienes prepared from the feather pigments of parrots (Ara macao). Adult male Sprague-Dawley rats were divided into three experimental groups and perfused with KHB buffer with or without treatment of 2,4,6-octatrienal (1) (50 mM) or its potassium salt (2) (50 mM). All hearts were then subjected to 30 min ischemia followed by a 2 h reperfusion. Ischemia/reperfusion resulted in a significant amount of tissue injury, cardiomyocyte apoptosis, and depression in hemodynamic functions. Parrodiene treatment prevented the development of myocardial injury after ischemia/reperfusion. Western blot analysis indicated that 1 and 2 reduced the oxidative stress, induced the expression of Bcl-2, and caused increased phosphorylation of Akt. These agents also reduced myocardial ischemic reperfusion injury presumably by reducing oxidative stress and activating the survival signal through the Akt-Bcl-2 pathway.

Free radical-scavenging activity of sulfurous water investigated by electron paramagnetic resonance (EPR) spectroscopy.

ABSTRACT The aim of the study was to explore the antiradical activity of sulfurous water, used for inhalatory therapy (characterized by the presence of sulfhydryl [HS]) by means of electron paramagnetic resonance (EPR) spectroscopy. The effects of sulfurous water corresponding to the concentrations from 16 down to 0.25 μg/mL of HS were tested by means of Fenton reaction (HO•), KO2–crown ether system (O2−•), and EPR of Tempol and of Fremys salt radical. All of these assays were made using natural sulfurous water or degassed sulfurous water (no detectable HS) or reconstituted sulfurous water (degassed plus NaHS). The free radicals were significantly inhibited by natural water with HS concentrations ranging from 16 to 1 μg/mL for HO•, Tempol, and Fremys salt, and O2−• was significantly inhibited from 16 and 2 μg/mL. The tests of degassed water did not reveal any significant differences from baseline values. The tests of reconstituted water led to significant results overlapping those obtained using natural water, thus confirming the importance of the presence of HS group (reductive activity). The positive effects of the activity of sulfurous thermal water is partially based on the patients’ subjective sense of well-being and partially on symptomatic (or general) clinical improvements that are sometimes difficult to quantify. These findings indicate that, in addition to their known mucolytic activity and trophic effects on respiratory mucosa, the HS groups in sulfurous water also have antioxidant activity that contributes to the waters therapeutic effects on upper and lower airway inflammatory diseases.

Free radical scavenging activity of erdosteine metabolite I investigated by electron paramagnetic resonance spectroscopy.

The aim of this study was to explore the antiradical activity of Met I (an active metabolite of erdosteine) containing a pharmacologically active sulphydryl group, by means of electron paramagnetic resonance (EPR) spectroscopy which has not previously been used to characterize the antiradical activity of Met I. The effects of concentrations of 20, 10, 5, 2.5, 1.25 and 0.625 µg/ml of Met I were tested against: (a) the Fenton reaction model system with EPR detection of HO·; (b) the KO2-crown ether system with EPR detection of O2–·; (c) the EPR assay based on the reduction of the Tempol radical, and (d) the EPR assay based on the reduction of Fremy’s salt radical. Our findings show that the intensity of 4 different free radicals was significantly reduced in the presence of Met I, thus indicating the presence of a termination reaction between the free radicals and Met I.

Amelioration of Myocardial Ischemic Reperfusion Injury with Calendula Officinalis

Calendula officinalis of family Asteraceae, also known as marigold, has been widely used from time immemorial in Indian and Arabic cultures as an anti-inflammatory agent to treat minor skin wound and infections, burns, bee stings, sunburn and cancer. At a relatively high dose, calendula can lower blood pressure and cholesterol. Since inflammatory responses are behind many cardiac diseases, we sought to evaluate if calendula could be cardioprotective against ischemic heart disease Two groups of hearts were used: the treated rat hearts were perfused with calendula solution at 50 mM in KHB buffer (in mM: sodium chloride 118, potassium chloride 4.7, calcium chloride 1.7, sodium bicarbonate 25, potassium biphosphate 0.36, magnesium sulfate 1.2, and glucose 10) for 15 min prior to subjecting the heart to ischemia, while the control group was perfused with the buffer only. Calendula achieved cardioprotection by stimulating left ventricular developed pressure and aortic flow as well as by reducing myocardial infarct size and cardiomyocyte apoptosis. Cardioprotection appears to be achieved by changing ischemia reperfusion-mediated death signal into a survival signal by modulating antioxidant and anti-inflammatory pathways as evidenced by the activation of Akt and Bcl2 and depression of TNFα. The results further strengthen the concept of using natural products in degeneration diseases like ischemic heart disease.

Diclofenac-Choline Antioxidant Activity Investigated by means of Luminol Amplified Chemiluminescence of Human Neutrophil Bursts and Electron Paramagnetic Resonance Spectroscopy.

A new diclofenac salt called diclofenac-choline (DC) has recently been proposed for the symptomatic treatment of oropharyngeal inflammatory processes and pain because its greater water solubility allows the use of high concentrations, which are useful when the contact time between the drug and the oropharyngeal mucosa is brief, as in the case of mouthwashes or spray formulations. The antioxidant activity of DC has not yet been investigated, and so the aim was to use luminol-amplified-chemiluminescence (LACL) to verify whether various concentrations of DC (1.48, 0.74 and 0.37 mg/mL for incubation times of 2, 4 and 8 min) interfere with oxygen and nitrogen radicals during the course of human neutrophils respiratory bursts; electron paramagnetic resonance (EPR) spectroscopy was used to investigate its direct antiradical (scavenger) activity. The EPR findings showed that DC has concentration-dependent scavenging activity against the ABTS, the DPPH, and the hydroxyl radicals, but no activity on superoxide anion, as has been previously reported in the case of other NSAIDs. LACL revealed an inhibitory effect that was statistically significant after only 2 min of incubation, and similar after 4 and 8 min. The effects on the peroxynitrite radical paralleled those observed in the previous test. High concentrations and short incubation times showed that there is no interference on PMN viability, and so the inhibitory findings must be attributed to the effect of the drug. The anti-inflammatory effects of DC cannot be attributed solely to the inhibition of prostaglandin synthesis, but its effects on free radicals and neutrophil bursts suggest that they may contribute to its final therapeutic effect.

Intracerebroventricular capsaicin influences the body weight increasing of rats

Adult rats were treated for ten days with capsaicin or with NaCl 0.9% directly injected into the lateral cerebral ventricles through a surgically implanted cannula. A third group of rats was implanted with the same cannula but did not receive any treatment. The food intake and the body weight were recorded for at least six weeks after stopping the treatment. The animals were always kept at constant ambient temperature of 22 °C. The body weight of the capsaicin-treated group was reduced by the treatment, and showed a regular but lower degree of recovery trend than the control groups after the treatment period. In fact the capsaicin treated animals never reached the body weight of the controls. Nevertheless, food intake did not significantly vary after the capsaicin treatment. On the basis of these and previous findings, we can assume that capsaicin injected into the cerebral ventricles to rats kept at constant ambient temperature can acts on hypothalamic neurons, but a permanent action on metabolic pathways can not be excluded.