Alessandra Masci

Chemistry, stability and bioavailability of resveratrol.

Resveratrol is a bioactive polyphenol found in many vegetables. It is well known for its multiple pharmacological activities, such as anti-inflammatory, antioxidant, antimicrobial, anticancer, neuroprotective and cardioprotective effects. In vitro evidence of resveratrol efficacy is widespread, however, many concerns regarding its effectiveness in vivo arise from its poor stability in vitro and bioavailability following oral ingestion. This review focuses on the in vitro stability, with special focus on the photochemical stability of resveratrol, and on the therapeutic perspectives of this molecule due to its low bioavailability.

Evaluation of different extraction methods from pomegranate whole fruit or peels and the antioxidant and antiproliferative activity of the polyphenolic fraction

Pomegranate is a functional food of great interest, due to its multiple beneficial effects on human health. This fruit is rich in anthocyanins and ellagitannins, which exert a protective role towards degenerative diseases. The aim of the present work was to optimize the extraction procedure, from different parts of the fruit, to obtain extracts enriched in selected polyphenols while retaining biological activity. Whole fruits or peels of pomegranate cultivars, with different geographic origin, were subjected to several extraction methods. The obtained extracts were analyzed for polyphenolic content, evaluated for antioxidant capacity and tested for antiproliferative activity on human bladder cancer T24 cells. Two different extraction procedures, employing ethyl acetate as a solvent, were useful in obtaining extracts enriched in ellagic acid and/or punicalagins. Antioxidative and antiproliferative assays demonstrated that the antioxidant capability is directly related to the phenolic content, whereas the antiproliferative activity is to be mainly attributed to ellagic acid.

Antiviral activity of Lactobacillus brevis towards herpes simplex virus type 2: role of cell wall associated components.

The aim of this research was to study the mechanisms of Lactobacillus brevis antiviral activity towards HSV-2 and to identify the bacterial components responsible for the inhibiting effect. Bacterial extract and cell walls were prepared by lysozyme digestion of L. brevis cells untreated or treated with LiCl to remove S-layer proteins. Bacterial extract and cell wall fragments showed a dose dependent inhibitory effect on HSV-2 multiplication. In order to characterize the inhibitory activity of L. brevis, the bacterial extract was subjected to different physical and chemical treatments. The inhibitory activity was resistant to high temperature and proteases digestion and appeared to be associated with compounds with a molecular weight higher than 10 kDa. DNA, RNA and lipids isolated from bacterial cells were devoid of inhibitory effect. The antiviral activity of both bacterial extract and cell wall fragments obtained from L. brevis cells after the S-layer removal was significantly reduced compared to untreated cells suggesting that the inhibitory activity is likely due to a heat-resistant non-protein cell surface bacterial component.

5‐S‐cysteinyldopamine neurotoxicity: Influence on the expression of α‐synuclein and ERp57 in cellular and animal models of Parkinson's disease

Parkinsons disease (PD) is a progressive neurodegenerative disorder whose etiology is still unclear in spite of extensive investigations. It has been hypothesized that 5‐S‐cysteinyldopamine (CysDA), a catechol‐thioether metabolite of dopamine (DA), could be an endogenous parkinsonian neurotoxin. To gain further insight into its role in the neurodegenerative process, both CD1 mice and SH‐SY5Y neuroblastoma cells were treated with CysDA, and the data were compared with those obtained by the use of 6‐hydroxydopamine, a well‐known parkinsonian mimetic. Intrastriatal injection of CysDA in CD1 mice caused a long‐lasting depletion of DA, providing evidence of in vivo neurotoxicity of CysDA. Both in mice and in SH‐SY5Y cells, CysDA treatment induced extensive oxidative stress, as evidenced by protein carbonylation and glutathione depletion, and affected the expression of two proteins, α‐synuclein (α‐Syn) and ERp57, whose levels are modulated by oxidative insult. Real‐time PCR experiments support these findings, indicating an upregulation of both ERp57 and α‐Syn expression. α‐Syn aggregation was also found to be modulated by CysDA treatment. The present work provides a solid background sustaining the hypothesis that CysDA is involved in parkinsonian neurodegeneration by inducing extensive oxidative stress and protein aggregation.

Evaluation of processing effects on anthocyanin content and colour modifications of blueberry (Vaccinium spp.) extracts: Comparison between HPLC-DAD and CIELAB analyses

Colour is the first organoleptic property that consumers appreciate of a foodstuff. In blueberry (Vaccinium spp.) fruits, the anthocyanins are the principal pigments determining the colour as well as many of the beneficial effects attributed to this functional food. Commercial blueberry-derived products represent important sources of these healthy molecules all year round. In this study, blueberries were produced into purees comparing two homogenization methods and further heated following different thermal treatments. All the supernatants of the homogenates were monitored for pH. Then, the hydroalcoholic extracts of the same samples were characterized by CIELAB and HPLC-DAD analyses. These analytical techniques provide complementary information on fruit pigments content as a whole and on quali-quantitative profile of the single bioactive colorants. These data could be very interesting to know the best manufacturing procedure to prepare blueberry-derived products, well accepted by the consumers, while maintaining their healthy properties unaltered.

Inhibition of Amyloid Peptide Fragment Aβ25–35 Fibrillogenesis and Toxicity by N-Terminal β-Amino Acid-Containing Esapeptides: Is Taurine Moiety Essential for In Vivo Effects?

We report the synthesis and fibrillogenesis inhibiting activity of the new peptide derivatives 1–4, related to the pentapeptide Ac‐LPFFD‐NH2 (iAβ5p), proposed by Soto and co‐workers and widely recognized as one of the most active β‐sheet breaker agents. The Aβ25–35 fragment of the parent full‐length Aβ1–42 was used as fibrillogenesis model. The activity of peptide derivatives 1–4 was tested in vitro by thioflavin T binding assay, far UV CD spectroscopy, and scanning electron microscopy. Their ability to hinder the toxic effect of Aβ25–35in vivo was studied by monitoring the viability of human SH‐SY5Y neuroblastoma cells and the prevention of superoxide anion radical release from BV2 microglial cells. The results point to a favourable role in the fibrillogenesis inhibitory activity of the sulphonamide junction for compounds 1 and 2, containing an N,N‐dimethyltaurine and a taurine amino‐terminal moiety, respectively. Furthermore, compounds 1 and 2 show a significant protective effect on cell viability, rescuing the cells from the toxicity exerted by Aβ25–35 treatment.

Neuroprotective Effect of Brassica oleracea Sprouts Crude Juice in a Cellular Model of Alzheimer's Disease.

β-Amyloid peptide (Aβ) aberrant production and aggregation are major factors implicated in the pathogenesis of Alzheimers disease (AD), causing neuronal death via oxidative stress. Several studies have highlighted the importance of polyphenolic antioxidant compounds in the treatment of AD, but complex food matrices, characterized by a different relative content of these phytochemicals, have been neglected. In the present study, we analyzed the protective effect on SH-SY5Y cells treated with the fragment Aβ 25–35 by two crude juices of broccoli sprouts containing different amounts of phenolic compounds as a result of different growth conditions. Both juices protected against Aβ-induced cytotoxicity and apoptotic cell death as evidenced by cell viability, nuclear chromatin condensation, and apoptotic body formation measurements. These effects were mediated by the modulation of the mitochondrial function and of the HSP70 gene transcription and expression. Furthermore, the juices upregulated the intracellular glutathione content and mRNA levels or activity of antioxidant enzymes such as heme oxygenase-1, thioredoxin, thioredoxin reductase, and NAD(P)H:quinone oxidoreductase 1 via activation of NF-E2-related factor 2 (Nrf2). Although the effects of the two juices were similar, the juice enriched in phenolic compounds showed a greater efficacy in inducing the activation of the Nrf2 signalling pathway.

Use of Probiotics for Dermal Applications

The concept of probiotic bacteria is considerably evolving. Clinical and experimental researches extensively document that beyond probiotic capacity to influence positively the intestinal functions, they can exert their benefits at the skin level thanks to their peculiar properties. Indeed, scientific and evidence-based reports strengthen the assumption that certain probiotics can contribute to modulate cutaneous microflora, lipid barrier, and skin immune system, leading to the preservation of the skin homeostasis. In this chapter, the most relevant evidences available from scientific literature as well as registered patents have been summarized in relation to actual or potential topical applications of probiotics in the field of dermatology. Altogether the evidences reported in this review afford the possibility of designing new strategies based on a topical approach for the prevention and treatment of cutaneous disorders.

Biological Effects of MC2050, a Quinazoline-Based PARP-1 Inhibitor, in Human Neuroblastoma and EBV-Positive Burkitt's Lymphoma Cells

Recently, many effortswere dedicated to the development of new highly selectiveand less toxic PARP-1 inhibitors. Most of these compounds actas competitive inhibitors by blocking the active site of theenzyme, although some benzamides have also been shown tohave additional effects, such as inhibition of the binding ofPARP-1 to DNA.

LPS, Oleuropein and Blueberry extracts affect the survival, morphology and Phosphoinositide signalling in stimulated human endothelial cells

Endothelial cells (EC) act as leading actors in angiogenesis. Understanding the complex network of signal transduction pathways which regulate angiogenesis might offer insights in the regulation of normal and pathological events, including tumours, vascular, inflammatory and immune diseases. The effects of olive oil and of Blueberry extracts upon the phosphoinositide (PI)-specific phospholipase C (PLC) enzymes were evaluated both in quiescent and inflammatory stimulated human umbilical vein EC (HUVEC) using molecular biology (multiliquid bioanalysis) and immunofluorescence techniques. Oleuropein significantly increased the number of surviving HUVEC compared to untreated controls, suggesting that it favours the survival and proliferation of EC. Our results suggest that Oleuropein might be useful to induce EC proliferation, an important event during angiogenesis, with special regard to wound healing. Blueberry extracts increased the number of surviving HUVEC, although the comparison to untreated controls did not result statistically significant. Lipopolysaccharide (LPS) administration significantly reduced the number of live HUVEC. LPS can also modify the expression of selected PLC genes. Adding Blueberry extracts to LPS treated HUVEC cultures did not significantly modify the variations of PLC expression induced by LPS. Oleuropein increased or reduced the expression of PLC genes, and statistically significant results were identified for selected PLC isoforms. Oleuropein also modified the effects of LPS upon PLC genes’ expression. Thus, our results corroborate the hypothesis that Oleuropein owns anti-inflammatory activity. The intracellular localization of PLC enzymes was modified by the different treatments we used. Podosome-like structures were observed in differently LPS treated HUVEC.