Barbara Tomasello

Response of cell cycle/stress-related protein expression and DNA damage upon treatment of CaCo2 cells with anthocyanins

Anthocyanins are a class of flavonoids, widely spread throughout the plant kingdom, exhibiting important antioxidant and anti-inflammatory actions as well as chemotherapeutic effects; nonetheless, little is known about the molecular mechanisms by which these activities are exerted. The present study is aimed at investigating molecular mechanisms involved in the chemotherapeutic effects induced by both cyanidin-3-O-beta glucopyranoside (CY3G) and its aglycon form, cyanidin chloride (CY), in human colon cancer cells (CaCo2). The effect on cell growth, reactive oxygen species (ROS) formation and cell cycle/stress proteins modification, including ataxia teleangectasia mutated protein (ATM), p53, p21, 8-oxoguanine DNA glycosylase (OGG1), 70 kDa heat shock protein (HSP70) and topoisomerase IIbeta, as well as on DNA fragmentation, was determined. CY and CY3G treatment affect cell growth and cell proliferation, this latter in a moderately dose-dependent way. Interestingly, ROS level is decreased by any concentration of CY and, only at the lowest concentration, by CY3G. Moreover, the two molecules exert their activities increasing ATM, topoisomerase II, HSP70 and p53 expression. The analysis of DNA fragmentation by Comet assay evidences: (1) a dose-dependent increase in DNA damage only after treatment with CY3G; (2) a more evident trend in the DNA fragmentation when the treatment is performed on agarose embedded cells (cellular atypical Comet); (3) a highly dose-dependent DNA fragmentation induced by CY when the treatment is carried out on agarose embedded naked DNA (acellular atypical Comet). The present findings substantiate a possible chemotherapeutic role of anthocyanins and suggest that CY and CY3G act on CaCo2 by different mechanisms, respectively, ROS-dependent and ROS-independent.

Preparation and optimization of PIT solid lipid nanoparticles via statistical factorial design

The objective of this study was the preparation, physico-chemical characterization and statistical optimization of cationic solid lipid nanoparticles (SLN) prepared by the PIT method as potential carrier for gene therapy, emphasizing the application of factorial design in such a kind of studies. The preliminary screening from a physico-chemical point of view on three cationic lipids (CTAB, DDAB and DOTAP), selected on the basis of their different chemical structure and increasing lipophilicity, allowed us to select SLN with DOTAP, due to its higher zeta potential and smaller particle size. Afterward, a 2(2) full factorial experimental design was developed in order to study the effects of two independent variables (amount of DOTAP and concentration of lipid matrix) and their interaction on mean particle size and zeta potential values. The factorial planning was validated by ANOVA analysis; the correspondence between the predicted values of size and zeta and those measured experimentally confirmed the validity of the design and the equation applied for its resolution. The factorial design showed a significant influence of the independent variables on the selected parameters; in particular, a higher effect of DOTAP was observed on zeta potential value. Different dilutions of the optimized SLN containing 7% w/w of cutina CP and 1% w/w of DOTAP, with size and zeta potential values respectively of 462.9 nm and 50.8 mV, were in vitro examined to evaluate the possible cytotoxicity on two models of cell cultures: human prostate cancer androgen-non-responsive DU-145 cells and primary cultures of rat astrocytes.

Assessment of environmental stress in Parablennius sanguinolentus (Pallas, 1814) of the Sicilian Ionian coast.

The blenny Parablennius sanguinolentus was selected as a useful bioindicator of environmental pollution. Chemical parameters in water and sediments from three different sampling sites along the Sicilian Ionian coast were determined and metal concentrations in fish muscle were measured. DNA fragmentation and oxidation in erythrocytes and hepatocytes was determined by the Comet assay and HSP70 expression levels were evaluated in the liver. The results show an increased level of chromium in sediments and high polycyclic aromatic hydrocarbon (PAH) concentrations in water at one site. The bioaccumulation of metals in muscle tissue shows high concentrations of lead in some samples. A high percentage of DNA damage in blood and liver cells, as well as high hepatic levels of HSP70, were found in all the sites. The results demonstrate the usefulness of an integrated chemical and biological approach for the determination of environmental stress.

Chemical and biological properties of the novel exopolysaccharide produced by a probiotic strain of Bifidobacterium longum

Bifidobacterium longum W11 is a commercialized probiotic that has an exopolysaccharide (EPS) layer covering its surface which could play a role in the beneficial properties attributed to the strain; thus, we have carried out chemical and biological analyses of this polymer. The eps cluster putatively involved in the polymer synthesis presented a unique structural organization not previously reported in bifidobacteria. B. longum W11 produced a complex polysaccharide blend with the main component composed of glucose and galactose. An exhaustive structural analysis identified two different repeating units: one linear [→6)-β-Galf-(1→3)-α-Galp-(1→] and one, more abundant, with the same backbone in which the β-Galf is 5-substituted by a β-Glcp unit. The antioxidant capability and the lack of toxicity of the whole EPS W11 mixture, as well as some functional characteristics of the producing strain, such as the in vitro resistance to gastrointestinal conditions and the adhesion of colonocytes, were also determined.

Synthesis of amphiphilic resveratrol lipoconjugates and evaluation of their anticancer activity towards neuroblastoma SH-SY5Y cell line

Resveratrol, a polyphenol present in grapes and other edible plants, possesses several important pharmacological activities, including anticancer activity. Nevertheless, its therapeutic use is still limited because of some unfavourable physicochemical and pharmacokinetic properties, mainly, poor cellular uptake and too rapid metabolism resulting in elimination from the body. To meet these drawbacks, some resveratrol conjugates would be useful, which would possess improved stability, uptake and bioavailability than the lead compound, and the ability to release it once it is internalized into the cell. In this paper we report a synthetic strategy which allowed us to obtain new amphiphilic resveratrol derivatives starting from different selectively protected resveratrol phosphoramidites or even from the resveratrol triphosphoramidite. Specifically, resveratrol was conjugated through phosphate bridge(s) to different lipophilic groups related to membrane lipids, such as cholesteryl or diacylglycero moieties. All the new lipoconjugates were tested towards human neuroblastoma SH-SY5Y cells and proved to be significantly more active than resveratrol, with a concentration-dependent activity.

ELIMED: a new hadron therapy concept based on laser driven ion beams

Laser accelerated proton beams have been proposed to be used in different research fields. A great interest has risen for the potential replacement of conventional accelerating machines with laser-based accelerators, and in particular for the development of new concepts of more compact and cheaper hadrontherapy centers. In this context the ELIMED (ELI MEDical applications) research project has been launched by INFN-LNS and ASCR-FZU researchers within the pan-European ELI-Beamlines facility framework. The ELIMED project aims to demonstrate the potential clinical applicability of optically accelerated proton beams and to realize a laser-accelerated ion transport beamline for multi-disciplinary user applications. In this framework the eye melanoma, as for instance the uveal melanoma normally treated with 62 MeV proton beams produced by standard accelerators, will be considered as a model system to demonstrate the potential clinical use of laser-driven protons in hadrontherapy, especially because of the limited constraints in terms of proton energy and irradiation geometry for this particular tumour treatment. Several challenges, starting from laser-target interaction and beam transport development up to dosimetry and radiobiology, need to be overcome in order to reach the ELIMED final goals. A crucial role will be played by the final design and realization of a transport beamline capable to provide ion beams with proper characteristics in terms of energy spectrum and angular distribution which will allow performing dosimetric tests and biological cell irradiation. A first prototype of the transport beamline has been already designed and other transport elements are under construction in order to perform a first experimental test with the TARANIS laser system by the end of 2013. A wide international collaboration among specialists of different disciplines like Physics, Biology, Chemistry, Medicine and medical doctors coming from Europe, Japan, and the US is growing up around the ELIMED project with the aim to work on the conceptual design, technical and experimental realization of this core beamline of the ELI Beamlines facility.

ELIMED a new concept of hadrontherapy with laser-driven beams

ELIMED (Medical Applications at Extreme Light Infrastructure) is a task-force originally born by an idea of ELI-Beams (Prague, CZ)and INFN-LNS (Italian Institute for Nuclear Physics of Catania, I) researchers. It now involves other groups interested in the possibility to design and develop a new generation of hadrontherapy facilities using laser-accelerated ion beams. ELIMED main goal is to perform proof-of-principle experiments aimed to demonstrate that laser-accelerated high-energy proton beams (up to 70 MeV in the first phase) can be potentially used for the specific case of ocular proton therapy. For this purpose new devices for beam handling and transport will be developed as well as new methods for radiobiology and dosimetry. The involvement of INFN-LNS group takes advantage of the well-established expertise in dosimetry measurements and Monte Carlo calculations for medical physics, which has been achieved in several years of eye tumor treatments in the CATANA proton therapy facility. Recently, in the framework of an INFN activity, they have also designed, fabricated, calibrated and experimentally tested at PALS laser laboratory (Cz) a Thomson Parabola ion spectrometer with a wide acceptance and able to characterize laser-driven proton beams up to 20 MeV.

ELIMED: MEDICAL APPLICATION AT ELI-BEAMLINES. STATUS OF THE COLLABORATION AND FIRST RESULTS

ELI-Beamlines is one of the four pillars of the ELI (Extreme Light Infrastructure) pan-European project. It will be an ultrahigh-intensity, high repetition-rate, femtosecond laser facility whose main goal is to generate and apply high-brightness X-ray sources and accelerated charged particles. In particular, medical applications are treated by the ELIMED task force, which has been launched by collaboration between ELI and INFN researchers. ELIMED aims to demonstrate the clinical applicability of laser accelerated ions. In this article, the state of the ELIMED project and the first scientific results are reported. The design and realisation of a preliminary beam handling system and of an advanced spectrometer for diagnostics of high energy (multi-MeV) laser-accelerated ion beams will also be briefly presented.

BIOESOnet: A Tool for the Generation of Personalized Human Metabolic Pathways from 23andMe Exome Data.

The lowering of costs of whole exome sequencing (WES) services registered in the last two years has greatly increased the demand for managing different metabolic diseases, including autism spectrum disorders (ASD). WES allows the detection of a large part of exome single nucleotide polymorphisms (SNPs), whose expression can be in some cases modulated by epigenetics, life style and microbioma changes. However, such raw data usually needs to be manipulated in order to allow useful interpretation and analysis. We present BIOESOnet, a tool for the filtering and visualization of exome 23andMe raw data into a customized methylation pathway. The tool, available at: http://www.bionumeri.org/joomla/restricted-area/onecarbon-tool, enables a fast and extensive overview of possible mutations inside an extended metabolic pathway.

Biological activities of Teucrium flavum L., Teucrium fruticans L., and Teucrium siculum rafin crude extracts

Abstract The genus Teucrium (Lamiaceae) includes 300 species widespread all around the world, which are perennial herbs or shrubs commonly, named germanders. In Italy, Teucrium flavum L., Teucrium fruticans L., and Teucrium siculum Rafin are mostly present in Liguria, Sicily, and Sardegna. Teucrium species are characterized by mono and sesquiterpene hydrocarbons, flavonoids, fatty acid esters, and essential oils. Many species of this genus show antioxidant, antimicrobial, and antifungal activities, rendering them useful as natural preservative ingredients. In view of the interesting biological properties reported for Teucrium spp., in this study we determined the total phenol and flavonoid content of inflorescence extracts of T. flavum L., T. fruticans L., and T. siculum Rafin. In addition, we investigated the in vitro antioxidant and antibacterial activities of inflorescence extracts against pathogenic bacteria. Obtained results demonstrated that extracts had in vitro antioxidant activity and showed antimicrobial ability against Gram-positive and Gram-negative strains albeit with different effectiveness probably due to the different qualitative/quantitative composition of the extract also suggesting that these extracts might be useful in preventing several diseases in which oxidative stress may represent an important pathogenic mechanism.