Naturally occurring hydroxytyrosol: synthesis and anticancer potential.

Several epidemiological and animal studies have suggested that polyphenols, a group of secondary plant metabolites occurring mainly in the plant kingdom, may have a protective effect against some chronic degenerative diseases such as cancer. Polyphenols are part of the human diet, being present in vegetal food and beverages. Among them, an olive biophenol named hydroxytyrosol [2-(3,4- dihydroxyphenyl)ethanol, HTyr] has recently received particular attention because of its antioxidant, antiproliferative, pro-apoptotic, and anti-inflammatory activities, which have the potential to specifically counteract all cancer hallmarks, thus representing the expectant biological activities underlying the anti-tumor properties of this polyphenol. After a description of the synthetic procedures to prepare pure HTyr, this review takes into consideration the chemopreventive and chemotherapeutic potential of HTyr as the result of its antioxidant, antiproliferative and anti-inflammatory activities. In particular, the review is focused on the current knowledge of the main cellular and molecular mechanisms used by HTyr to affect carcinogenesis, highlighting the specific oncogenic and inflammatory signaling pathways potentially targeted by HTyr.

Induction of apoptosis in human pancreatic cancer cells by docosahexaenoic acid.

Polyunsaturated fatty acids have been indicated to induce anti-proliferative and/or apoptotic effects in various tumor cells. We showed that, at a 200- micro M concentration, both alpha-linoleic (18:2 n-6; LA) or docosahexaenoic (22:6 n-3; DHA) acid inhibited cell growth, while only DHA induced apoptosis in the human Paca-44 pancreatic cancer cell line. Investigating the mechanism underlying DHA-induced apoptosis, we showed that DHA induced a rapid and dramatic (>60%) intracellular depletion of reduced glutathione (GSH), without affecting oxidized glutathione (GSSG). Moreover, using two specific inhibitors of carrier-mediated GSH extrusion, cystathionine or methionine, we observed that GSH depletion occurred via an active GSH extrusion, and that inhibition of GSH efflux completely reversed apoptosis. These results provide the first evidence for a possible causative role of GSH depletion in DHA-induced apoptosis.

Th 1 cytokine production by peripheral blood mononuclear cells in X-linked adrenoleukodystrophy.

Cerebral adrenoleukodystrophy (ALD) and adrenomyeloneuropathy (AMN) are the two most frequent clinical phenotypes of the same genetic defect leading to the accumulation of very long chain fatty acids (VLCFA). Previous studies have suggested that inflammatory cytokines may play a role in the cerebral demyelination and in phenotype expression of the disease. We analyzed cytokine production by stimulated peripheral blood mononuclear cells (PBMC) from 17 patients (four asymptomatic subjects, eight AMN and five ALD). Our results show that lipopolysaccarides (LPS) stimulated PBMC from both symptomatic and asymptomatic patients have an increased production of IL-12 and TNFalpha compared to controls, while after phitoemoagglutinin (PHA) stimulation we observed a decreased production of IL-6 and IL-10. These data indicate that, following an immunological stimulus, PBMC from patients have an increased production of cytokines typical of a Th1 cell response which is able to promote the inflammatory process. This characteristic profile of cytokine production could be related to the biochemical defect and could have a role in central nervous system (CNS) pathogenesis.

Human intestinal epithelial cells express receptors for platelet-activating factor.

The intestinal epithelium produces and responds to cytokines and lipid mediators that play a key role in the induction and regulation of mucosal inflammation. The lipid mediator platelet-activating factor (PAF) can be produced and degraded by the human intestinal epithelium and is known to mediate a range of proinflammatory and other biological effects in the intestinal mucosa. In the studies herein, we assessed whether or not human intestinal epithelial cells express cell surface or intracellular PAF receptors (PAF-R), whether expression of these receptors can be regulated, and whether human intestinal epithelial cells respond to PAF. Several human colon epithelial cell lines (HT-29, Caco-2, T84, HCT-8, HCA-7, I407, and LS-174T) were shown by RT-PCR to constitutively express mRNA for PAF-R. In addition, PAF-R expression was demonstrated by immunoblot analysis and PAF-R was shown to be constitutively expressed on the cell surface of several of these cell lines, as assessed by flow cytometry. PAF-R expression by human colon epithelial cells was upregulated by stimulation with retinoic acid but not by stimulation with PAF, proinflammatory agonists (tumor necrosis factor-alpha, interleukin-1, interferon-gamma), or transforming growth factor-alpha. PAF-R on intestinal epithelial cells were functional, as PAF stimulation of the cells increased tyrosine phosphorylation of several cellular proteins, including proteins of 75 and 125 kDa, and this response was blocked by a PAF-R antagonist. Consistent with the findings using cell lines, PAF-R were also constitutively expressed by normal human colon and small intestinal epithelium in vivo, as shown by immunohistology. The constitutive and regulated expression of functional PAF-R by human intestinal epithelium suggests PAF produced by the intestinal epithelial cells or cells underlying the epithelium has autocrine or paracrine effects on intestinal epithelial cells.

Influence of thermal and dietary stress on immune response of rabbits.

With the aim of analyzing the effects of prolonged thermal stress or food intake reduction on lymphoid cell proliferation and antibody synthesis, New Zealand White (NZW) male rabbits, both immunized and nonimmunized with Mycobacterium tuberculosis, were kept in individual cages for 24 d at controlled climatic conditions. Both immunized and nonimmunized rabbits were divided into two experimental groups and one control group. The thermal-stressed rabbits (TS) were exposed to a room temperature of 33.5 degrees C, and dietary restricted rabbits (DR) were pair-fed on the basis of the average feed intake of the TS groups and maintained at a room temperature of 18.0 degrees C. The control group (Ctr) was maintained at 18.0 degrees C and was given ad libitum access to feed. All rabbits were maintained at relative humidity 62 +/- 5%. Sera and peripheral blood mononuclear cells (PBMC) were isolated from blood samples collected on d 0, 6, 12, and 24. Sera were used for determining total proteins and immunoglobulins (Ig) specific or not to mycobacterial antigens. Antibodies to heat-shock protein (HSP) were also determined. The PBMC were used to measure cell proliferation and in vitro Ig synthesis. Both experiments in vivo and in vitro suggest that thermal stress and dietary restriction severely affect the immune cell functions. In fact, both stress treatments decreased the capacity of PBMC to proliferate and inhibited the differentiation of B lymphocytes in antibody-secreting cells. However, a recovery of immune cell functions was only observed in vivo after 12 d of treatment, suggesting that other defensive mechanisms may come into play in vivo. Sera collected from both TS and DR rabbits after 24 d presented antibodies to HSP70, suggesting that the analysis of anti-HSP antibodies could represent a useful indicator to reveal the effects of different stress effectors regardless of the nature of the stress.

Differential effect of alpha-tocopherol and ascorbate on oxidative injury induced in immune cells by thermal stress.

As immune cells are often subjected to hyperthermia that can easily occur either after intense and/or prolonged exercise or during defense against pathogens, in this paper we analysed whether superoxide anion production occurred in lymphocytes exposed to high temperature and, consequently, if antioxidants could exert any protective function. The results demonstrated that an increase of superoxide anion was induced in rabbit lymphocytes exposed to 42 degrees C for 1h, although cell viability was no affected. However, suppression of either Pokeweed mitogen (PWM)-driven cell proliferation, or immunoglobulin production or IL-2 synthesis was observed. To evaluate the capacity of antioxidants to restore the immune suppressed responses, two vitamins, alpha-tocopherol and ascorbic acid, were added to PWM-stimulated cultures following heat treatment. The data demonstrated that alpha-tocopherol was able to totally abrogate the inhibitory effects mediated by thermal stress, while ascorbic acid did not give any protective results.

The gene encoding for MC56 determinant (drug-sensitivity marker) is located on the short arm of human chromosome 11.

A panel of mouse x human B- and T-cell hybrids was analyzed for the expression of MC56 determinant which marks the drug-sensitive state of CEM cells. Karyotypic and phenotypic analyses of the tested clones showed that the expression of MC56 determinant correlated to the presence of human chromosome 11 and segregated concordantly to the epitopes recognized by monoclonal antibodies in the CD44 cluster. By using a particular class of interspecific rodent x human-cell hybrids in which the human genome counterpart is represented in the different clones only by human chromosome 11 or its fragments, we showed that the gene encoding for MC56 determinant is located on the region p13-pter of the short arm of chromosome 11. Therefore, the hypothesized homology between the drug-sensitivity marker MC56 and the CD44 determinant is supported also by gene mapping studies.