Chemoenzymatic Synthesis and Some Biological Properties of O-phosphoryl Derivatives of (E)-resveratrol

3- O-, 3,5-di- O- and 4′- O-phosphoryl derivatives of ( E)-resveratrol have been obtained following a chemoenzymatic strategy. Variedly acylated resveratrol derivatives have been obtained first by exploiting regioselective properties of Pseudomonas cepacea or Candida antarctica lipases in organic solvents. Each acyl-resveratrol was then phosphorylated by the phosphoramidite chemistry protocol and in sequence freed of protective groups, affording the desired O-phosphoryl derivative. Following UV-absorption spectroscopic investigation on the interaction of the newly synthesized compounds with DNA, 4′- O-phosphorylresveratrol exhibited the best binding affinity. As a result of cytotoxicity tests, 3- O-phosphorylresveratrol was more active than resveratrol against DU 145 prostate cancer cells

Rosmarinus officinalis extract inhibits human melanoma cell growth.

Rosmarinus officinalis L. is receiving increasing attention due to its anti-inflammatory and antioxidative constituents. Our recent studies showed that R. officinalis extract, containing 31.7 % of carnosic acid, was able to counteract the deleterious effects of UV-R, by protecting plasmid DNA from hydroxyl radicals generated by UV-A. In this work, we evaluated the effects of this extract on pBR322 DNA cleavage induced by nitric oxide, and the growth inhibitory activity against two human melanoma cell lines, M14 and A375. The extract showed a protective effect on plasmid DNA damage, and at concentrations of 10-80 μg/mL was able to reduce significantly (p<0.001) the growth (MTT assay) of both melanoma cell lines. In addition, our results indicate that apoptotic cell demise is induced in M14 and A375 cells. No statistically significant increase in LDH release was observed in melanoma cells, correlated to a fragmentation of genomic DNA, determined by COMET assay

Potential Anticancer Activity against Human Epithelial Cancer Cells of Peumus Boldus Leaf Extract

The potential in vitro antineoplastic effect has been studied of a methanolic extract of leaves of Peumus boldus Molina (Monimiaceae) on two human cancer epithelial cell lines, DU-145 cells (androgen-insensitive prostate cancer cells) and KB cells (oral squamous carcinoma cells). Our findings show that this extract exhibited comparable effects on the cancer cells examined as judged by IC50 values (5.07±0.4 μg/mL and 5.28±0.5 μg/mL in DU-145 and KB cells, respectively). In addition, with respect to genomic DNA damage, determined by Comet assay, the results obtained show a high fragmentation of DNA, not correlated to lactic dehydrogenase (LDH) release, a marker of membrane breakdown, in both cell lines treated with the extract at 5–20 μg/mL concentrations. Taken together, our experimental evidence may justify further investigation of the chemopreventive and chemotherapeutic potential of this natural drug

Mesenchymal stem cells from adipose tissue differentiated in chondrocytes into three-dimensional clusters termed chondrocyte “nodules” express lubricin

Lubricin is recognized to have an important role in preventing cartilage wear and synovial cell adhesion and proliferation. This study focused on isolation, cultivation, characterization of mesenchymal stem cells (MSCs) from adipose tissue and on their differentiation into chondrocytes through the NH ChondroDiff Medium. The main aim was to investigate some biochemical markers, such as collagen type I and II and lubricin verify the possibility to suggest of employing autologous three-dimensional clusters termed chondrocyte “nodules” in cartilage repair. Three-dimensional chondrocyte “nodules” were assessed with histology (haematoxylin and eosin), histochemistry (Alcian blue and Safranin-O/fast green) and Hoechst 33258 staining. Collagen type I, II and lubricin expression was determined by immunohistochemistry, immunofluorescence and Western blot. The results showed that, compared to control cartilage and monolayer chondrocytes showing just collagen type I, chondrocytes from MSCs (CD44, CD90 and CD105 positive; CD45, CD14 and CD34 negative) of adipose tissue grown in “nodules”, at 24 days, were able to express lubricin, collagen type I, and II, indicative of hyaline cartilage formation. Based on the function of lubricin in the joint cavity and disease and as a potential therapeutic agent, our results suggest the possibility of applying autologous cell transplantation from adipose tissue differentiated in chondrocyte “nodules”

Dehydroxymethylepoxyquinomicin Inhibits Expression and Production of Inflammatory Mediators in Interleukin-1β β β β β-induced Human Chondrocytes

The present research was carried out to determine the effects of a nuclear factor-kappaB (NF-kappaB) inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ), derivative of the antibiotic epoxyquinomicin C, on normal human chondrocytes treated with interleukin-1beta (IL-1beta). This is a cell model particularly useful to reproduce the mechanisms involved in degenerative arthropathies, where oxidative-inflammatory stress determines a progressive destruction of the articular cartilaginous tissue. The expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and inter-cellular adhesion molecule (ICAM)-1 was evaluated through Western blot analysis. The release of chemokines like monocyte chemoattractant protein-1 (MCP-1), regulated upon normal activation T-cell expressed and secreted (RANTES), and interleukin-8 (IL-8) were determined by ELISA assays. DHMEQ acts as a potent inhibitor of iNOS and COX-2 gene expression while also suppressing the production of nitrite in human chondrocytes. In addition, DHMEQ induces a significant dose-dependent decrease in ICAM expression, MCP-1, RANTES, and IL-8 release. DHMEQ helps to decrease the expression and production of pro-inflammatory mediators in IL-1beta-induced chondrocytes. DHMEQ may become a therapeutic agent for treatment of chondro-degenerative diseases

Adaption of Lung Fibroblasts to Fluoro-Edenite Fibers: Evaluation of Molecular and Physiological Dynamics.

Background/Aims: The fluoro-edenite fibrous amphibole was identified as an environmental pollutant associate to risk of carcinogenicity. In Sicily (Italy), it represents a public health issue because fluoro-edenite fibers are present in the soil of Biancavilla, a town located on the south-west slopes of the volcano Etna. Since the relationship between exposure to fluoro-edenite and the onset of lung disorders have been documented, in vitro studies were performed to clarify the mechanisms of damage, but most aspects remain unknown. Here, we focus on the effects of mineral fibers in a primary culture of lung fibroblasts. We supposed that the cells react to fluoro-edenite exposure by establishing a process of adaption that could modify their metabolic activity, their proliferation, and their physiological functions, as the production of extracellular matrix (ECM) components. Methods: To verify our hypothesis, we used immunofluorescence, cell proliferation, senescence, apoptosis, scratch, Western blot, Reverse transcription-polymerase chain reaction (RT-PCR), and evaluation of extracellular matrix components assays. Results: Results demonstrated that lung fibroblasts react to fluoro-edenite by a down-regulation of mitochondrial activity, a reduction of cell growth and migration, and a resistance to apoptosis. These elements suggested the induction of a premature senescent phenotype that was confirmed by senescence-associated beta-galactosidase (SA-β-Gal) activity, and by the analysis of ECM elements. We found an unbalance of collagens ratio, and changes in matrix metalloproteinase3 production and release. Conclusion: Our data suggest that fluoro-edenite-induced senescence of lung fibroblasts could be an early and underestimated step that may drive fibroblasts toward a fibrotic and carcinogenic phenotype

Induction of tissue plasminogen activator (tPA) by pituitary adenylate cyclase-activating polypeptide (PACAP) in Schwann cell-like cultures

Peripheral nerve regeneration is dependent on the ability of regenerating neurites to migrate through cellular debris and altered extracellular matrix at the injury site, grow along the residual distal nerve sheath conduit, and reinnervate synaptic targets. In cell culture, growth cones of regenerating axons secrete PACAP, a peptide known to induce the expression of the protease tPA1. Here we tested the hypothesis that PACAP might also stimulate peripheral glial cells to release tPA to participate in nerve regeneration. More specifically, we addressed whether PACAP promoted the release and expression of tPA in the Schwann cell-like culture RT4-D6P2T, which shares biochemical and physical properties with Schwann cells2. We found that PACAP dose- and time-dependently stimulated tPA expression. Maxadilan, a potent PACAP receptor agonist, mimicked the effect of PACAP, whereas VIP, a PACAP-related peptide, produced only a moderate response. PACAP ability to stimulate tPA expression seemed to be dependent on the Akt/CREB signaling cascade, as inhibition using the PI3K/Akt blocker wortmannin or the TrkA/B inhibitor K252a both significantly dampened PACAP-evoked tPA expression. A similar effect was obtained in cells treated with the PACAP/VIP receptor antagonist PACAP6-38. We conclude that PACAP through the Akt/CREB intracellular pathway, acts as a potent inducer of tPA expression and release in Schwann-cell like cultures