Hyperglycemia is implicated both in micro-and macro-vascular complications in diabetes mellitus. ... more Hyperglycemia is implicated both in micro-and macro-vascular complications in diabetes mellitus. Pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal polypeptide (VIP) are two known nonclassic regulators of angiogenesis, although their biological role on endothelial cell proliferation remains poorly defined. In the present study we hypothesized that either peptides might play an inhibitory role on hyperglycemia-induced cell growth. To this end, we investigated the effect of both PACAP and VIP on cell proliferation in murine microvascular endothelial cells (H5V) cultured both under euglycemic and hyperglycemic conditions (5 and 25 mM glucose, respectively) for 24, 48 h, 7 and 15 days. Results demonstrated that high glucose treatment induced a time-dependent increase in cell viability after 48 h (p < 0.05), which was much more evident after 7 and 15 days (p < 0.001). Similar effects were observed in cell proliferation, although significant changes were obtained after prolonged exposures to high glucose (7 and 15 days; p < 0.001). The proliferative response to the glucose-enriched environment was correlated to changes in the expression of PAC1 and, to a minor extent, to VPAC2, but not VPAC1 receptors, as measured by quantitative real-time PCR. These results were further confirmed by Western blot and immunofluorescence analyses. Interestingly, 10 −7 M PACAP or VIP treatment significantly attenuated hyperglycemia-induced increase in cell viability and proliferation after 7 and 15 days. Taken together, our findings demonstrate that both PACAP and VIP peptides exert an inhibitory activity on hyperglycemia-induced endothelial cell proliferation, thus suggesting that the effect might be mediated by PAC1 and VPAC2 receptors.
Perturbation of oxidant/antioxidant cellular balance, induced by cellular metabolism and by exoge... more Perturbation of oxidant/antioxidant cellular balance, induced by cellular metabolism and by exogenous sources, causes delete- rious effects to proteins, lipids, and nucleic acids, leading to a condition named "oxidative stress" that is involved in several diseases, such as cancer, ischemia-reperfusion injury, and neu- rodegenerative disorders. Among the exogenous agents, both H2O2 and hyperthermia have been implicated in oxidative stress promotion
We consider here a reaction diffusion model which arises in the development of swarm colonies of ... more We consider here a reaction diffusion model which arises in the development of swarm colonies of the Proteus mirabilis. After having obtained the equivalence transformations admitted by the model we give a classification of two special subclasses of this last one.
Salvia verbenaca L. (syn. S. minore) is a perennial herb known in the traditional medicine of Sic... more Salvia verbenaca L. (syn. S. minore) is a perennial herb known in the traditional medicine of Sicily as "spaccapetri" and is used to resolve cases of kidney stones, chewing the fresh leaves or in decoction. The chemical composition of the essential oil obtained from aerial parts of S. verbenaca collected in Piano Battaglia (Sicily) on July 2009, was analyzed by GC and GC-MS. The oil was strongly characterized by fatty acids (39.5%) and carbonylic compounds (21.2%), with hexadecanoic acid (23.1%), (Z)-9-octadecenoic acid (11.1%) and benzaldehyde (7.3%) as the main constituents. The in vitro activity of the essential oil against some microorganisms in comparison with chloramphenicol by the broth dilution method was determined. The oil exhibited a good activity as inhibitor of growth of Gram + bacteria.
We report the synthesis, the antioxidant and the inhibitory activity (IC50) on metalloproteinases... more We report the synthesis, the antioxidant and the inhibitory activity (IC50) on metalloproteinases (MMPs) 3 and 13 of 2-benzo[d]isothiazolylimino-5-benzylidene-4-thiazolidinones. Their potential as protective agents in osteoarthritis (OA) was evaluated by biological assays on chondrocytes cultures, stimulated by IL-1β. The chondroprotective capability, related both to antioxidant activity and to inhibition of MMPs, was studied in vitro, by determining nitric oxide production and glycosaminoglycans release. Moreover, selected derivatives 1h and 1g was studied for nuclear factor kappaB (NF-κB) inhibition by Western Blot analysis and for MMP-3 protein release using ELISA test. The structure-activity relationship of tested compounds demonstrates a favorable effect of the para substitution on the 5-benzilydene ring. Compound 1g shows a potent and selective activity on MMP-3 versus MMP-13. Accordingly, 1g possesses high antioxidant effect, NO lowering and GAGs restoring capability and also...
Regeneration of peripheral nerves depends on the abilities of rejuvenating axons to migrate at th... more Regeneration of peripheral nerves depends on the abilities of rejuvenating axons to migrate at the injury site through cellular debris and altered extracellular matrix, and then grow along the residual distal nerve sheath conduit and reinnervate synaptic targets. Considerable evidence suggest that glial cells participate in this process, although the mechanisms remain to be clarified. In cell culture, regenerating neurites secrete PACAP, a peptide shown to induce the expression of the protease tissue plasminogen activator (tPA) in neural cell types. In the present studies, we tested the hypothesis that PACAP can stimulate peripheral glial cells to produce tPA. More specifically, we addressed whether or not PACAP promoted the expression and activity of tPA in the Schwann cell line RT4-D6P2T, which shares biochemical and physical properties with Schwann cells. We found that PACAP dose- and time-dependently stimulated tPA expression both at the mRNA and protein level. Such effect was m...
International journal of pharmaceutics, Jan 17, 2012
Physical protection of mucosa surface and reduction of inflammatory processes are currently consi... more Physical protection of mucosa surface and reduction of inflammatory processes are currently considered the main strategies in the treatment and prevention of mucosal diseases. However, the majority of models used to verify the activity of new mucoprotective agents are based on limiting instrumental assessment or the sacrifice of experimental animals. In this study, for the first time, some in vitro experimental methods using Caco-2 cell line are proposed as predicting in vivo behaviour and action of mucoprotective agents. To this purpose, hyaluronic acid and natural polysaccharides for their bioadhesive activity, hydrocortisone and natural polyphenols as anti-inflammatory agents have been chosen. The obtained results demonstrated that the techniques (Con A/o-pd assay and Franz cell system) of mucoadhesive evaluation on Caco-2 cells are useful to compare the activity of each experimental sample and to assess the adhesion time to the mucosal cell surface. Moreover, the reduction of in...
Zeitschrift für Naturforschung. C, Journal of biosciences
The stilbenoids E-resveratrol (E-3,5,4'-trihydroxystilbene, 1), E-3,5,4'-trimethoxystilbe... more The stilbenoids E-resveratrol (E-3,5,4'-trihydroxystilbene, 1), E-3,5,4'-trimethoxystilbene (2), E-3,4,4'-trimethoxystilbene (3) and E-3,4'-dimethoxy-5-hydroxystilbene (4) were converted by photoisomerization to their corresponding Z-isomers 5-8. Compounds 1-8 were subjected to antiproliferative activity bioassays towards a set of four different human cancer cell lines, namely DU-145 (androgen not responsive human prostate tumor), LNCaP (androgen responsive human prostate tumor), M-14 (human melanoma) and KB (human mouth epidermoid carcinoma). The methylated analogues of 1 are more active than the natural lead in the majority of bioassays. The most active compound was Z-3,5,4'-trimethoxystilbene (6), which showed against DU-145 and LNCaP cells GI50 values close to those of the anticancer drug vinorelbine; 6 resulted more active than its E-isomer 2 towards DU-145, LNCaP and especially KB cell lines. A number of methylated Z-isomers displayed a higher activity than...
The stilbenoid resveratrol (1) was subjected to regioselective acetylation catalysed by Candida a... more The stilbenoid resveratrol (1) was subjected to regioselective acetylation catalysed by Candida antarctica lipase (CAL) to obtain 4'-acetylresveratrol (2). CAL biocatalysed regioselective alcoholysis of 3,5,4'-triacetylresveratrol (3), 3,5,4'-tributanoylresveratrol (6), and 3, 4, 5'-trioctanoylresveratrol (9) afforded derivatives 4, 5, 7, 8, 10, and 11. Further resveratrol analogues (12-18) were obtained through methylation and hydrogenation reactions, whereas the 3,4,4'-trimethoxystilbene (19) was obtained by complete synthesis. Resveratrol and its lipophylic analogues were subjected to cell-growth inhibition bioassays towards DU-145 human prostate cancer cells. Compounds 2-19 showed cell-growth inhibition activity comparable to or higher than resveratrol (GI(50)=24.09 microM), displaying low or very low toxicity against non-tumorigenic human fibroblast cells. Comparison of the trimethoxy stilbenes 12 (GI(50)=2.92 microM) and 19 (GI(50)=25.39 microM) indicates t...
70 KDa heat shock proteins (HSPs70), either as a constitutive or inducible form, are expressed at... more 70 KDa heat shock proteins (HSPs70), either as a constitutive or inducible form, are expressed at very high levels in malignant human tumors of various origin. In different cell types, they are known to play an antiapoptotic role. Resveratrol (3,5,4'-trihydroxy-trans-stilbene), a polyphenol present in red wine, grapes and other dietary and medicinal plants, has been shown to be active in inhibiting multistage carcinogenesis, inducing apoptotic cell death. With the present study, a possible relationship between HSP70 expression and cell death elicited by resveratrol in DU-145 cells, which mimic the late hormone-refractory stages of prostate carcinoma, was investigated. To this end, we treated DU-145 with different concentrations. (50, 100 and 200 microM) of resveratrol and cell viability, by tetrazolium salts assay (MTT) and membrane breakdown, by lactic dehydrogenase (LDH) release, were measured. The possible induction of oxidative stress was evidenced both by performing a fluor...
Experimental biology and medicine (Maywood, N.J.), 2003
Perturbation of oxidant/antioxidant cellular balance, induced by cellular metabolism and by exoge... more Perturbation of oxidant/antioxidant cellular balance, induced by cellular metabolism and by exogenous sources, causes deleterious effects to proteins, lipids, and nucleic acids, leading to a condition named "oxidative stress" that is involved in several diseases, such as cancer, ischemia-reperfusion injury, and neurodegenerative disorders. Among the exogenous agents, both H(2)O(2) and hyperthermia have been implicated in oxidative stress promotion linked with the activation of apoptotic or necrotic mechanisms of cell death. The goal of this work was to better understand the involvement of some stress-related proteins in adaptive responses mounted by human fibroblasts versus the oxidative stress differently induced by 42 degrees C hyperthermia or H(2)O(2.) The research was developed, switching off inducible nitric oxide synthase (iNOS) expression through antisense oligonucleotide transfection by studying the possible coregulation in the expression of HSP32 (also named HO-1)...
Ethanol intake is associated with increase in lipid peroxidation and formation of reactive oxygen... more Ethanol intake is associated with increase in lipid peroxidation and formation of reactive oxygen species in different cerebral areas, in neurons as well as in astrocytes. The latter's integrity is essential for the normal growth of neurons. In previous studies we observed, in different cerebral areas of both acutely and chronically ethanol-treated rats, correlation between ethanol-induced oxidative stress and the increased expression of HSP70 (70 kDa heat shock proteins), chaperonins having a protective and stabilizing effect on stress-induced cell injury. In this study we examined, in vitro, the role of HSP70 on chronically ethanol-treated rat astrocytes by transfection with an anti-HSP70 antisense oligonucleotide. The results show that treatment with ethanol, from 50 to 100 mmol/L, induces a dose-dependent increase in the production of reactive oxygen species and of HSP70 levels, together with an impairment of the respiratory chain activity and a decrease in cell viability. I...
Epitope identification is the basis of modern vaccine design. The present paper studied the super... more Epitope identification is the basis of modern vaccine design. The present paper studied the supermotif of the HLA-A3 superfamily, using comparative molecular similarity indices analysis (CoMSIA). Four alleles with high phenotype frequencies were used: A*1101, A*0301, A*3101 and A*6801. Five physicochemical properties-steric bulk, electrostatic potential, local hydrophobicity, hydrogen-bond donor and acceptor abilities-were considered and 'all fields' models were produced for each of the alleles. The models have a moderate level of predictivity and there is a good correlation between the data. A revised HLA-A3 supermotif was defined based on the comparison of favoured and disfavoured properties for each position of the MHC bound peptide. The present study demonstrated that CoMSIA is an effective tool for studying peptide-MHC interactions. #
Cellular and molecular biology (Noisy-le-Grand, France)
The in vitro biological response to fluoro-edenite (FE) fibres, an asbestos-like amphibole, was e... more The in vitro biological response to fluoro-edenite (FE) fibres, an asbestos-like amphibole, was evaluated in lung alveolar epithelial A549, mesothelial MeT-5A and monocyte-macrophage J774 cell lines. The mineral has been found in the vicinity of the town of Biancavilla (Catania, Sicily), where an abnormal incidence of mesothelioma has been documented. Cell motility, distribution of polymerized actin, and synthesis of vascular endothelial growth factor (VEGF) and of beta-catenin, critical parameters for tumour development, progression and survival, were investigated in A549 and MeT-5A cells exposed to 50 microg/ml FE fibres for 24 hr and 48 hr. The levels of cyclooxygenase (COX-2) and prostaglandin (PGE2), two molecules involved in cancer pathogenesis by affecting mitogenesis, cell adhesion, immune surveillance and apoptosis, were measured in J774 cells treated with FE fibres under the same experimental conditions. Finally, FE fibres were studied by SEM and EDS analysis to investigat...
International Journal of Developmental Neuroscience, 2000
The ether lipid 1-octadecyl-2-methyl-rac-glicero-3-phosphocholine (ET-18-OCH 3 ) is known to be s... more The ether lipid 1-octadecyl-2-methyl-rac-glicero-3-phosphocholine (ET-18-OCH 3 ) is known to be selectively cytotoxic toward several types of tumor cells, in which it seems to activate a process of apoptotic cell death. Moreover, the drug has been demonstrated to be active in normal cells too, particularly in rat astrocytes. In these cells at low dosage (from 1 to 6 mg/ml of medium) ET-18-OCH 3 stimulates maturation and protective responses, whereas at increasing dosages (from 8 to 20 mg/ml) it shows cytotoxic eects. The present study demonstrates that when ET-18-OCH 3 is added to astrocytes, it activates, in a timeand concentration-dependent manner, an oxidative process by increasing both the generation of reactive oxygen species (ROS), including nitric oxide, and lipid peroxidation. When there is a high ET-18-OCH 3 concentration or the time of treatment is prolonged, the increased oxidative condition seems to trigger DNA fragmentation (monitored by COMET assay) as well as loss in cell viability. These cytotoxic eects indicate that ROS may be considered, in our experimental model, as executioners of a program of cell death. In addition, ET-18-OCH 3 being a promising molecule in antitumor therapy, our data, while reinforcing the importance of monitoring the therapeutic drug dosage employed, also suggest that it may be useful to associate some antioxidants with antitumor treatments. 7
We have studied the in-vitro effects of liposomal formulations loaded with three oligopeptides (T... more We have studied the in-vitro effects of liposomal formulations loaded with three oligopeptides (Tp3, Tp4, Tp5), synthetic analogues corresponding to the active site of thymopoietin, on human thymic epithelial cells.
The aim of the present study was to investigate the effects of Levetiracetam, a new antiepileptic... more The aim of the present study was to investigate the effects of Levetiracetam, a new antiepileptic drug, on the synthesis of brain-derived neurotrophic factor (BDNF) and inducible nitric oxide synthase (iNOS) in rat cortical astrocyte cultures. The astrocytes were treated for 48 h with different concentrations of Levetiracetam and the expression of BDNF and iNOS was analyzed by immunostaining and immunoblotting analyses. We observed that Levetiracetam is able to stimulate expression of both BDNF and iNOS in a concentration-dependent manner on rat cortical astrocyte cultures. For the BDNF, this effect appears at very low concentrations (1 and 10 microgram/ml), while expression of iNOS appears only at higher dosages (50 microgram/ml). We conclude that Levetiracetam might exert neuroprotective effects, at least in part, via stimulation of neurotrophic factors, thus reducing the extent of inflammation and neuronal death under pathological conditions such as epilepsy.
j o u r n a l h o m e p a g e : w w w . e l s e v i e r . c o m / l o c a t e / c h e m b i o i n... more j o u r n a l h o m e p a g e : w w w . e l s e v i e r . c o m / l o c a t e / c h e m b i o i n t Corrigendum Corrigendum to "Genista sessilifolia DC. and Genista tinctoria L. inhibit UV light and nitric oxide-induced DNA damage and human melanoma cell growth" [
Hyperglycemia is implicated both in micro-and macro-vascular complications in diabetes mellitus. ... more Hyperglycemia is implicated both in micro-and macro-vascular complications in diabetes mellitus. Pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal polypeptide (VIP) are two known nonclassic regulators of angiogenesis, although their biological role on endothelial cell proliferation remains poorly defined. In the present study we hypothesized that either peptides might play an inhibitory role on hyperglycemia-induced cell growth. To this end, we investigated the effect of both PACAP and VIP on cell proliferation in murine microvascular endothelial cells (H5V) cultured both under euglycemic and hyperglycemic conditions (5 and 25 mM glucose, respectively) for 24, 48 h, 7 and 15 days. Results demonstrated that high glucose treatment induced a time-dependent increase in cell viability after 48 h (p < 0.05), which was much more evident after 7 and 15 days (p < 0.001). Similar effects were observed in cell proliferation, although significant changes were obtained after prolonged exposures to high glucose (7 and 15 days; p < 0.001). The proliferative response to the glucose-enriched environment was correlated to changes in the expression of PAC1 and, to a minor extent, to VPAC2, but not VPAC1 receptors, as measured by quantitative real-time PCR. These results were further confirmed by Western blot and immunofluorescence analyses. Interestingly, 10 −7 M PACAP or VIP treatment significantly attenuated hyperglycemia-induced increase in cell viability and proliferation after 7 and 15 days. Taken together, our findings demonstrate that both PACAP and VIP peptides exert an inhibitory activity on hyperglycemia-induced endothelial cell proliferation, thus suggesting that the effect might be mediated by PAC1 and VPAC2 receptors.
Perturbation of oxidant/antioxidant cellular balance, induced by cellular metabolism and by exoge... more Perturbation of oxidant/antioxidant cellular balance, induced by cellular metabolism and by exogenous sources, causes delete- rious effects to proteins, lipids, and nucleic acids, leading to a condition named "oxidative stress" that is involved in several diseases, such as cancer, ischemia-reperfusion injury, and neu- rodegenerative disorders. Among the exogenous agents, both H2O2 and hyperthermia have been implicated in oxidative stress promotion
We consider here a reaction diffusion model which arises in the development of swarm colonies of ... more We consider here a reaction diffusion model which arises in the development of swarm colonies of the Proteus mirabilis. After having obtained the equivalence transformations admitted by the model we give a classification of two special subclasses of this last one.
Salvia verbenaca L. (syn. S. minore) is a perennial herb known in the traditional medicine of Sic... more Salvia verbenaca L. (syn. S. minore) is a perennial herb known in the traditional medicine of Sicily as "spaccapetri" and is used to resolve cases of kidney stones, chewing the fresh leaves or in decoction. The chemical composition of the essential oil obtained from aerial parts of S. verbenaca collected in Piano Battaglia (Sicily) on July 2009, was analyzed by GC and GC-MS. The oil was strongly characterized by fatty acids (39.5%) and carbonylic compounds (21.2%), with hexadecanoic acid (23.1%), (Z)-9-octadecenoic acid (11.1%) and benzaldehyde (7.3%) as the main constituents. The in vitro activity of the essential oil against some microorganisms in comparison with chloramphenicol by the broth dilution method was determined. The oil exhibited a good activity as inhibitor of growth of Gram + bacteria.
We report the synthesis, the antioxidant and the inhibitory activity (IC50) on metalloproteinases... more We report the synthesis, the antioxidant and the inhibitory activity (IC50) on metalloproteinases (MMPs) 3 and 13 of 2-benzo[d]isothiazolylimino-5-benzylidene-4-thiazolidinones. Their potential as protective agents in osteoarthritis (OA) was evaluated by biological assays on chondrocytes cultures, stimulated by IL-1β. The chondroprotective capability, related both to antioxidant activity and to inhibition of MMPs, was studied in vitro, by determining nitric oxide production and glycosaminoglycans release. Moreover, selected derivatives 1h and 1g was studied for nuclear factor kappaB (NF-κB) inhibition by Western Blot analysis and for MMP-3 protein release using ELISA test. The structure-activity relationship of tested compounds demonstrates a favorable effect of the para substitution on the 5-benzilydene ring. Compound 1g shows a potent and selective activity on MMP-3 versus MMP-13. Accordingly, 1g possesses high antioxidant effect, NO lowering and GAGs restoring capability and also...
Regeneration of peripheral nerves depends on the abilities of rejuvenating axons to migrate at th... more Regeneration of peripheral nerves depends on the abilities of rejuvenating axons to migrate at the injury site through cellular debris and altered extracellular matrix, and then grow along the residual distal nerve sheath conduit and reinnervate synaptic targets. Considerable evidence suggest that glial cells participate in this process, although the mechanisms remain to be clarified. In cell culture, regenerating neurites secrete PACAP, a peptide shown to induce the expression of the protease tissue plasminogen activator (tPA) in neural cell types. In the present studies, we tested the hypothesis that PACAP can stimulate peripheral glial cells to produce tPA. More specifically, we addressed whether or not PACAP promoted the expression and activity of tPA in the Schwann cell line RT4-D6P2T, which shares biochemical and physical properties with Schwann cells. We found that PACAP dose- and time-dependently stimulated tPA expression both at the mRNA and protein level. Such effect was m...
International journal of pharmaceutics, Jan 17, 2012
Physical protection of mucosa surface and reduction of inflammatory processes are currently consi... more Physical protection of mucosa surface and reduction of inflammatory processes are currently considered the main strategies in the treatment and prevention of mucosal diseases. However, the majority of models used to verify the activity of new mucoprotective agents are based on limiting instrumental assessment or the sacrifice of experimental animals. In this study, for the first time, some in vitro experimental methods using Caco-2 cell line are proposed as predicting in vivo behaviour and action of mucoprotective agents. To this purpose, hyaluronic acid and natural polysaccharides for their bioadhesive activity, hydrocortisone and natural polyphenols as anti-inflammatory agents have been chosen. The obtained results demonstrated that the techniques (Con A/o-pd assay and Franz cell system) of mucoadhesive evaluation on Caco-2 cells are useful to compare the activity of each experimental sample and to assess the adhesion time to the mucosal cell surface. Moreover, the reduction of in...
Zeitschrift für Naturforschung. C, Journal of biosciences
The stilbenoids E-resveratrol (E-3,5,4'-trihydroxystilbene, 1), E-3,5,4'-trimethoxystilbe... more The stilbenoids E-resveratrol (E-3,5,4'-trihydroxystilbene, 1), E-3,5,4'-trimethoxystilbene (2), E-3,4,4'-trimethoxystilbene (3) and E-3,4'-dimethoxy-5-hydroxystilbene (4) were converted by photoisomerization to their corresponding Z-isomers 5-8. Compounds 1-8 were subjected to antiproliferative activity bioassays towards a set of four different human cancer cell lines, namely DU-145 (androgen not responsive human prostate tumor), LNCaP (androgen responsive human prostate tumor), M-14 (human melanoma) and KB (human mouth epidermoid carcinoma). The methylated analogues of 1 are more active than the natural lead in the majority of bioassays. The most active compound was Z-3,5,4'-trimethoxystilbene (6), which showed against DU-145 and LNCaP cells GI50 values close to those of the anticancer drug vinorelbine; 6 resulted more active than its E-isomer 2 towards DU-145, LNCaP and especially KB cell lines. A number of methylated Z-isomers displayed a higher activity than...
The stilbenoid resveratrol (1) was subjected to regioselective acetylation catalysed by Candida a... more The stilbenoid resveratrol (1) was subjected to regioselective acetylation catalysed by Candida antarctica lipase (CAL) to obtain 4'-acetylresveratrol (2). CAL biocatalysed regioselective alcoholysis of 3,5,4'-triacetylresveratrol (3), 3,5,4'-tributanoylresveratrol (6), and 3, 4, 5'-trioctanoylresveratrol (9) afforded derivatives 4, 5, 7, 8, 10, and 11. Further resveratrol analogues (12-18) were obtained through methylation and hydrogenation reactions, whereas the 3,4,4'-trimethoxystilbene (19) was obtained by complete synthesis. Resveratrol and its lipophylic analogues were subjected to cell-growth inhibition bioassays towards DU-145 human prostate cancer cells. Compounds 2-19 showed cell-growth inhibition activity comparable to or higher than resveratrol (GI(50)=24.09 microM), displaying low or very low toxicity against non-tumorigenic human fibroblast cells. Comparison of the trimethoxy stilbenes 12 (GI(50)=2.92 microM) and 19 (GI(50)=25.39 microM) indicates t...
70 KDa heat shock proteins (HSPs70), either as a constitutive or inducible form, are expressed at... more 70 KDa heat shock proteins (HSPs70), either as a constitutive or inducible form, are expressed at very high levels in malignant human tumors of various origin. In different cell types, they are known to play an antiapoptotic role. Resveratrol (3,5,4'-trihydroxy-trans-stilbene), a polyphenol present in red wine, grapes and other dietary and medicinal plants, has been shown to be active in inhibiting multistage carcinogenesis, inducing apoptotic cell death. With the present study, a possible relationship between HSP70 expression and cell death elicited by resveratrol in DU-145 cells, which mimic the late hormone-refractory stages of prostate carcinoma, was investigated. To this end, we treated DU-145 with different concentrations. (50, 100 and 200 microM) of resveratrol and cell viability, by tetrazolium salts assay (MTT) and membrane breakdown, by lactic dehydrogenase (LDH) release, were measured. The possible induction of oxidative stress was evidenced both by performing a fluor...
Experimental biology and medicine (Maywood, N.J.), 2003
Perturbation of oxidant/antioxidant cellular balance, induced by cellular metabolism and by exoge... more Perturbation of oxidant/antioxidant cellular balance, induced by cellular metabolism and by exogenous sources, causes deleterious effects to proteins, lipids, and nucleic acids, leading to a condition named "oxidative stress" that is involved in several diseases, such as cancer, ischemia-reperfusion injury, and neurodegenerative disorders. Among the exogenous agents, both H(2)O(2) and hyperthermia have been implicated in oxidative stress promotion linked with the activation of apoptotic or necrotic mechanisms of cell death. The goal of this work was to better understand the involvement of some stress-related proteins in adaptive responses mounted by human fibroblasts versus the oxidative stress differently induced by 42 degrees C hyperthermia or H(2)O(2.) The research was developed, switching off inducible nitric oxide synthase (iNOS) expression through antisense oligonucleotide transfection by studying the possible coregulation in the expression of HSP32 (also named HO-1)...
Ethanol intake is associated with increase in lipid peroxidation and formation of reactive oxygen... more Ethanol intake is associated with increase in lipid peroxidation and formation of reactive oxygen species in different cerebral areas, in neurons as well as in astrocytes. The latter's integrity is essential for the normal growth of neurons. In previous studies we observed, in different cerebral areas of both acutely and chronically ethanol-treated rats, correlation between ethanol-induced oxidative stress and the increased expression of HSP70 (70 kDa heat shock proteins), chaperonins having a protective and stabilizing effect on stress-induced cell injury. In this study we examined, in vitro, the role of HSP70 on chronically ethanol-treated rat astrocytes by transfection with an anti-HSP70 antisense oligonucleotide. The results show that treatment with ethanol, from 50 to 100 mmol/L, induces a dose-dependent increase in the production of reactive oxygen species and of HSP70 levels, together with an impairment of the respiratory chain activity and a decrease in cell viability. I...
Epitope identification is the basis of modern vaccine design. The present paper studied the super... more Epitope identification is the basis of modern vaccine design. The present paper studied the supermotif of the HLA-A3 superfamily, using comparative molecular similarity indices analysis (CoMSIA). Four alleles with high phenotype frequencies were used: A*1101, A*0301, A*3101 and A*6801. Five physicochemical properties-steric bulk, electrostatic potential, local hydrophobicity, hydrogen-bond donor and acceptor abilities-were considered and 'all fields' models were produced for each of the alleles. The models have a moderate level of predictivity and there is a good correlation between the data. A revised HLA-A3 supermotif was defined based on the comparison of favoured and disfavoured properties for each position of the MHC bound peptide. The present study demonstrated that CoMSIA is an effective tool for studying peptide-MHC interactions. #
Cellular and molecular biology (Noisy-le-Grand, France)
The in vitro biological response to fluoro-edenite (FE) fibres, an asbestos-like amphibole, was e... more The in vitro biological response to fluoro-edenite (FE) fibres, an asbestos-like amphibole, was evaluated in lung alveolar epithelial A549, mesothelial MeT-5A and monocyte-macrophage J774 cell lines. The mineral has been found in the vicinity of the town of Biancavilla (Catania, Sicily), where an abnormal incidence of mesothelioma has been documented. Cell motility, distribution of polymerized actin, and synthesis of vascular endothelial growth factor (VEGF) and of beta-catenin, critical parameters for tumour development, progression and survival, were investigated in A549 and MeT-5A cells exposed to 50 microg/ml FE fibres for 24 hr and 48 hr. The levels of cyclooxygenase (COX-2) and prostaglandin (PGE2), two molecules involved in cancer pathogenesis by affecting mitogenesis, cell adhesion, immune surveillance and apoptosis, were measured in J774 cells treated with FE fibres under the same experimental conditions. Finally, FE fibres were studied by SEM and EDS analysis to investigat...
International Journal of Developmental Neuroscience, 2000
The ether lipid 1-octadecyl-2-methyl-rac-glicero-3-phosphocholine (ET-18-OCH 3 ) is known to be s... more The ether lipid 1-octadecyl-2-methyl-rac-glicero-3-phosphocholine (ET-18-OCH 3 ) is known to be selectively cytotoxic toward several types of tumor cells, in which it seems to activate a process of apoptotic cell death. Moreover, the drug has been demonstrated to be active in normal cells too, particularly in rat astrocytes. In these cells at low dosage (from 1 to 6 mg/ml of medium) ET-18-OCH 3 stimulates maturation and protective responses, whereas at increasing dosages (from 8 to 20 mg/ml) it shows cytotoxic eects. The present study demonstrates that when ET-18-OCH 3 is added to astrocytes, it activates, in a timeand concentration-dependent manner, an oxidative process by increasing both the generation of reactive oxygen species (ROS), including nitric oxide, and lipid peroxidation. When there is a high ET-18-OCH 3 concentration or the time of treatment is prolonged, the increased oxidative condition seems to trigger DNA fragmentation (monitored by COMET assay) as well as loss in cell viability. These cytotoxic eects indicate that ROS may be considered, in our experimental model, as executioners of a program of cell death. In addition, ET-18-OCH 3 being a promising molecule in antitumor therapy, our data, while reinforcing the importance of monitoring the therapeutic drug dosage employed, also suggest that it may be useful to associate some antioxidants with antitumor treatments. 7
We have studied the in-vitro effects of liposomal formulations loaded with three oligopeptides (T... more We have studied the in-vitro effects of liposomal formulations loaded with three oligopeptides (Tp3, Tp4, Tp5), synthetic analogues corresponding to the active site of thymopoietin, on human thymic epithelial cells.
The aim of the present study was to investigate the effects of Levetiracetam, a new antiepileptic... more The aim of the present study was to investigate the effects of Levetiracetam, a new antiepileptic drug, on the synthesis of brain-derived neurotrophic factor (BDNF) and inducible nitric oxide synthase (iNOS) in rat cortical astrocyte cultures. The astrocytes were treated for 48 h with different concentrations of Levetiracetam and the expression of BDNF and iNOS was analyzed by immunostaining and immunoblotting analyses. We observed that Levetiracetam is able to stimulate expression of both BDNF and iNOS in a concentration-dependent manner on rat cortical astrocyte cultures. For the BDNF, this effect appears at very low concentrations (1 and 10 microgram/ml), while expression of iNOS appears only at higher dosages (50 microgram/ml). We conclude that Levetiracetam might exert neuroprotective effects, at least in part, via stimulation of neurotrophic factors, thus reducing the extent of inflammation and neuronal death under pathological conditions such as epilepsy.
j o u r n a l h o m e p a g e : w w w . e l s e v i e r . c o m / l o c a t e / c h e m b i o i n... more j o u r n a l h o m e p a g e : w w w . e l s e v i e r . c o m / l o c a t e / c h e m b i o i n t Corrigendum Corrigendum to "Genista sessilifolia DC. and Genista tinctoria L. inhibit UV light and nitric oxide-induced DNA damage and human melanoma cell growth" [
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Papers by Venera Cardile