Claudio Rossi

Antimicrobial activity against Helicobacter pylori strains and antioxidant properties of blackberry leaves (Rubus ulmifolius) and isolated compounds

Rubus spp. (Rosaceae) provide extracts used in traditional medicine as antimicrobial, anticonvulsant, muscle relaxant and radical scavenging agents. Resistance to antibiotics used to treat Helicobacter pylori infection as well as their poor availability in developing countries prompted us to test the antimicrobial activity of Rubus ulmifolius leaves and isolated polyphenols against two H. pylori strains with different virulence (CagA+ strain 10K and CagA(-) strain G21). The antioxidant activity (TEAC values) of the tested compounds ranged from 4.88 (gallic acid) to 1.60 (kaempferol), whilst the leaf extract gave a value of 0.12. All the isolated polyphenols as well as the leaf extract showed antibacterial activity against both of the H. pylori strains. The minimum bactericidal concentrations (MBCs) of the extract for H. pylori strains G21 and 10K, respectively, were 1200 microg/mL and 1500 microg/mL after 24h of exposure and 134 microg/mL and 270 microg/mL after 48 h exposure. Ellagic acid showed very low MBC values towards both of the H. pylori strains after 48 h (2 microg/mL and 10 microg/mL for strains G21 and 10K, respectively) and kaempferol toward G21 strain (MBC=6 microg/mL). A relationship between antimicrobial activity and antioxidant capacity was found only for H. pylori strain G21 CagA(-) strain.

Effect of trans-resveratrol on induced oxidative stress in human sperm and in rat germinal cells

Resveratrol is a phytoalexin with antioxidant properties. We evaluated resveratrol toxicity in swim-up selected human sperm and in rat spermatocytes and spermatids separated by the STAPUT method. Resveratrol antioxidant activity was tested against lipid peroxidation (LPO) induced by tert-butyl hydroperoxide. LPO was evaluated using the C11-BODIPY(581/591) probe and transmission electron microscopy in samples incubated with and without resveratrol. LD50 for human sperm and rat spermatids was 50 μM; spermatocytes were more sensitive to resveratrol cytotoxicity. Sperm motility increased progressively at 30 μM, 15 μM and 6 μM. 15 μM resveratrol acts against LPO, preserving sperm chromatin and plasma membranes. LPO were more marked in spermatocytes than in spermatids and the effect of resveratrol was more evident in spermatocytes. In this study, the scavenger properties of resveratrol were demonstrated in vitro in human sperm and rat germ cells, thus resveratrol could be added to the media used in assisted reproduction techniques and cryopreservation when oxidative stress is exacerbated.

Using Liposomes as Carriers for Polyphenolic Compounds: The Case of Trans-Resveratrol

Resveratrol (3,5,4′-trihydroxy-trans-stilbene) is a polyphenol found in various plants, especially in the skin of red grapes. The effect of resveratrol on human health is the topic of numerous studies. In fact this molecule has shown anti-cancer, anti-inflammatory, blood-sugar-lowering ability and beneficial cardiovascular effects. However, for many polyphenol compounds of natural origin bioavailability is limited by low solubility in biological fluids, as well as by rapid metabolization in vivo. Therefore, appropriate carriers are required to obtain efficient therapeutics along with low administration doses. Liposomes are excellent candidates for drug delivery purposes, due to their biocompatibility, wide choice of physico-chemical properties and easy preparation. In this paper liposome formulations made by a saturated phosphatidyl-choline (DPPC) and cholesterol (or its positively charged derivative DC-CHOL) were chosen to optimize the loading of a rigid hydrophobic molecule such as resveratrol. Plain and resveratrol loaded liposomes were characterized for size, surface charge and structural details by complementary techniques, i.e. Dynamic Light Scattering (DLS), Zeta potential and Small Angle X-ray Scattering (SAXS). Nuclear and Electron Spin magnetic resonances (NMR and ESR, respectively) were also used to gain information at the molecular scale. The obtained results allowed to give an account of loaded liposomes in which resveratrol interacted with the bilayer, being more deeply inserted in cationic liposomes than in zwitterionic liposomes. Relevant properties such as the mean size and the presence of oligolamellar structures were influenced by the loading of RESV guest molecules. The toxicity of all these systems was tested on stabilized cell lines (mouse fibroblast NIH-3T3 and human astrocytes U373-MG), showing that cell viability was not affected by the administration of liposomial resveratrol.

The Spatial Distribution of Optical Properties in the Ultraviolet and Visible in an Aquatic Ecosystem

Abstract In aquatic ecosystems, the UV and visible radiation environment is strongly influenced by variation in the chemical and physical parameters of the ecosystems. In shallow lakes, highly heterogeneous water characteristics produce a wide variety of optical environments. Such ecosystems require analysis approaches that consider a potential variability. In this study, 77 stations were used to characterize the optical properties of a shallow lake (open water surface 54 km2). The vertical attenuation of solar radiation at 305, 313, 320 and 340 nm and at photosynthetically active radiation was measured during the seasonal cycle. Dissolved organic matter (DOM), turbidity, fluorescence, pH, temperature, conductance and dissolved oxygen were simultaneously measured. The spatial variation of the extinction spectra of the dissolved fraction at each sampling station was also measured and analyzed between 270 and 400 nm. The spatial heterogeneity of the lake was examined by determining the distributions of the attenuation coefficients and biooptical parameters at high spatial resolution and describing the distributions in a series of maps. The methodology permitted a quantitative description of the interaction between solar radiation and aquatic ecosystems as well as a spatial classification of the dominating processes within the lake. This included the determination of the role played by DOM loading and changing chemical properties within the lake optical environment.

Chemical and optical phototransformation of dissolved organic matter

Dissolved organic matter represents the main reservoir of organic carbon in most aquatic ecosystems. In the present study, we determined the optical changes and the quantum yields of transient species formation for chromophoric dissolved organic matter (CDOM) samples undergoing photodegradation. The results show that the triplet states (3)CDOM* are potentially key players in CDOM photodegradation and that such transformations are strongly influenced by small differences in CDOM sources and sinks. In contrast, ·OH radicals are very unlikely to play a key role in phototransformation. These results represent an important first step in combining optical and transient species analyses to understand photodegradation processes of dissolved organic matter.

1H-13C selective NOE studies of the decapeptide gramicidin S

The cyclic decapeptide gramicidin S has been used as a model biopolymer to test the reliability of a structural method which is based on a relaxation analysis of heteronuclear selective NOEs. The observation of through-the-space dipolar couplings between intra- and inter residue amide protons and carbonyl carbons, perfectly consistent with the well established peptide solution conformation, confirms the effectiveness of this structural approach. As a corollary of the latter, carbonyl carbon resonances are unequivocally assigned. Moreover, a direct experimental proof of a Orn-NH2----Phe C = O hydrogen bonding is here given.

New perspectives in cell communication: Bioelectromagnetic interactions

This paper explores physical signalling in biological communications, the so-called biophysical pathways, and especially the role of electromagnetic signalling in cell-cell interactions. The experiments were designed to evaluate whether different cell populations physically interfere when incubated in separate Petri dishes placed in close proximity. Two different cell populations, immortalized mouse fibroblasts (NIH3T3) and adult human microvascular endothelial cells (HMVECad) were selected and seeded in separate polystyrene Petri dishes. Dishes seeded with NIH3T3 were then placed on top of those seeded with HMVECad at distances of 4mm and 11mm. A black filter was placed between dishes containing the two cell populations in another experiment, to prevent transmission of electromagnetic radiation between the two. Cell number and morphology of NIH3T3 and endothelial cells were found to be modified in dishes without the black filter, suggesting that specific signals emitted by the cells were transmitted through the polystyrene wall, affecting cell proliferation rate and morphology, even though the cells were growing in separate dishes.

Effect of quercetin, rutin, naringenin and epicatechin on lipid peroxidation induced in human sperm.

Quercetin, rutin, naringenin, epicatechin are flavonoids with diverse properties, including antioxidant potential. We evaluated, in vitro, the cytotoxicity of these flavonoids (20, 30, 50, 100, 200, 400 μM) in swim-up selected human sperm. Antioxidant activity was tested against tert-butylhydroperoxide induced lipid peroxidation using a C11-BODIPY(581/591) probe and transmission electron microscopy. A significant concentration-dependent effect on sperm viability (P<0.001) and motility (P<0.001) was observed. Lipid peroxidation was decreased in samples treated with 30 μM quercetin (P<0.01) and 30 μM rutin (P<0.05) versus samples incubated with tert-butylhydroperoxide alone. Naringenin (50-100 μM) showed a low protective effect and epicatechin (200 μM) was not efficacious. Transmission electron microscopy analysis confirmed the protective action of rutin and in particular quercetin on damages induced by lipid peroxidation. These results underlined the antioxidant properties of quercetin and rutin. A possible role of these compounds in the supplementation of media used during semen handling warrants attention and further studies.

Solution structure of hyaluronic acid oligomers by experimental and theoretical NMR, and molecular dynamics simulation.

The conformational properties of hyaluronic acid (HA) oligomers in aqueous solution were investigated by combining high‐resolution NMR experimental results, theoretical simulation of NMR two‐dimensional (2D) spectra by Complete Relaxation Matrix Analysis (CORMA), and molecular dynamics calculations. New experimental findings recorded for the tetra‐ and hexasaccharides enabled the stiffness of the HA and its viscoelastic properties to be interpreted. In particular, rotating frame nuclear Overhauser effect spectroscopy spectra provided new information about the arrangement of the glycosidic linkage. From 13C NMR relaxation the rotational correlation time (τc) were determined. The τc were employed in the calculation of geometrical constraints, by using the MARDIGRAS algorithm. Restrained simulated annealing and 1 ns of unrestrained molecular dynamic simulations were performed on the hexasaccharide in a box of 1215 water molecules.

Assessing the optical changes in dissolved organic matter in humic lakes by spectral slope distributions

The impact of photodegradation and mixing processes on the optical properties of dissolved organic matter (DOM) was examined using a distribution of absorption spectral slopes and fluorescence measurements in two Argentine lakes. By examining the variability of the absorption spectral slopes throughout the ultraviolet and visible wavelengths, it was possible to determine which wavelength intervals were most sensitive to dominant loss processes. For DOM photodegradation, results show that increases in the absorption spectral slope between 265 and 305 nm were highly sensitive to increased exposure to solar ultraviolet radiation. A slightly larger wavelength range (265-340 nm) was found to be influenced when both mixing and photodegradation processes were considered, in terms DOM residence time, DOM absorption and UV diffuse attenuation coefficients. This same interval of spectral slopes (265-340 nm) was found to highly correlate with changes in fluorescence emission/excitation in wavelengths that are typically associated with terrestrial humic-like DOM. The identification of specific wavelength intervals, rather than the use of standard wavelength intervals or ratios, improved our ability to identify the dominant dissolved organic matter (humic-like) and major loss mechanisms (photodegradation) in these lakes.