We previously showed that [Pt(O,O'-acac)(γ-acac)(DMS)] ([Pt(acac)2(DMS)]) exerted substantial cytotoxic effects in SH-SY5Y neuroblastoma cells, and decreased metalloproteases (MMPs) production and cells migration in MCF-7 breast cancer cells. The ubiquitously distributed sodium-hydrogen antiporter 1 (NHE1) is involved in motility and invasion of many solid tumours. The present study focuses on the effects of [Pt(acac)2(DMS)] in SH-SY5Y cell migration and also on the possibility that NHE1 may be involved in such effect. After sublethal [Pt(acac)2(DMS)] treatment cell migration was examined by wounding assay and cell invasion by transwell assay. NHE1 activity was measured in BCECF-loaded SH-SY5Y as the rate of Na+-dependent intracellular pH recovery in response to an acute acid pulse. Gelatin zymography for MMP-2/9 activities, Western blottings of MMPs, MAPKs, mTOR, S6 and PKCs and small interfering RNAs to PKC-ε/-δ mRNA were performed. Sublethal concentrations of [Pt(acac)2(DMS)] decreases NHE1 activity, inhibits cell migration and invasion and decreases expression and activity of MMP-2 and -9. [Pt(acac)2(DMS)] administered to SH-SY5Y cells provokes the increment of ROS, generated by NADPH oxidase, responsible for the PKC-ε and PKC-δ activation. Whilst PKC-δ activates p38/MAPK, responsible for the inhibition of MMP-2 and -9 secretion, PKC-ε activates a pathway made of ERK1/2, mTOR and S6K responsible for the inhibition of NHE1 activity and cell migration. In conclusion, we have shown a drastic impairment in tumour cell metastatization in response to inhibition of NHE1 and MMPs activities by [Pt(acac)2(DMS)] occurring through a novel mechanism mediated by PKC-δ/-ε activation.

The selectivity of [Pt(O,O’-acac)(γ-acac)(DMS)] stimulates a more detailed study aimed at pre-clinical investigation of its therapeutic potential in vivo. In this context, we employed a preclinical model based on the subcutaneous injection of MCF-7 breast cancer and ZL55 malignant pleural mesotelioma cell lines in SCID mice. Remarkably, [Pt(O,O’-acac)(γ- acac)(DMS)] stands out for higher anticancer activity than cisplatin toward both the murine tumor models examined, inducing up to 50% inhibition of tumor growth. We also demonstrated enhanced in vivo pharmacokinetics (PK), biodistribution and tolerability of [Pt(O,O’-acac)(γ-acac)(DMS)] when compared to cisplatin administered in Wistar rats. Altogether, these findings suggest that [Pt(O,O’-acac)(γ-acac)(DMS)] is a promising therapeutic agent for preventing growth of cancer, thus providing a solid starting point for its validation as a suitable candidate for further pharmacological testing.

In this study, we used the ovarian cancer cells Skov-3,a cell line resistant to cisplatin, in order to investigate whether [Pt(O,O’-acac)(γ-acac)(DMS)] was able to induce cell death. Skov-3 cells were treated with [Pt(O,O’-acac)(γ-acac)(DMS)] or with cisplatin and cytotoxicity tests were performed, together with western blotting of several proteins involved in apoptosis and autophagy, including LC-3 I/II and Beclin-1, considered important autophagic markers. The results obtained showed that [Pt(O,O’-acac)(γ-acac)(DMS)] induced a significant decrease in Skov-3 cell viability. We assessed the activation of both apoptotic and autophagic processes since western blotting analyses showed a time-dependent increment of the expression levels of autophagic markers Beclin-1 and LC-3, together with the proteolytic activation of caspase 9 and the degradation of PARP. The formation of autophagic vacuoles was detected by visualization of monodansylcadaverine (MDC) in Skov-3 cells incubated with [Pt(O,O’-acac)(gamma-acac)(DMS)]-treated cells, but not in cisplatin-treated cells. In addition, cisplatin caused apoptosis in Skov-3 cells since it activated caspases 7, 3 and 9 with consequent PARP proteolysis whilst it did not have any effect on LC-3II and Beclin-1 activation, thus showing the absence of autophagic processess. In conclusion, both [Pt(O,O’-acac)(gamma-acac)(DMS)] and cisplatin induced apoptotic Skov-3 cell death, but only [Pt(O,O’-acac)(gamma-acac)(DMS)] was able to induce autophagy.

Cisplatin is one of the most active chemotherapeutic agents used in the treatment of childhood and adult malignancies. Cisplatin induces cell death through different pathways. Despite its effectiveness, the continued clinical use of cisplatin is limited by onset of severe side effects (nephrotoxicity, ototoxicity and neurotoxicity) and drug resistance. Therefore, one of the main experimental oncology purpose is related to the search for new platinum-based drugs to create different types of adducts or more specific and effective subcellular targets. Thus, [Pt(O,O'-acac)(γ-acac)(DMS)], which reacts preferentially with protein thiols or thioether, was synthesized. In our research, different approaches were used to compare cisplatin and [Pt(O,O'-acac)(γ-acac)(DMS)] effects in B50 rat neuroblastoma cells. Our results, using immunocytochemical, cytometric and morphological techniques, showed that these compounds exert a cytostatic action and activate apoptosis with different pathways. Long-term effects demonstrated that [Pt(O,O'-acac)(γ-acac)(DMS)] exerts cytotoxic effects in neuronal B50 cell line not inducing drug resistance. Analysis was performed both to compare the ability of these platinum compounds to induce cell death and to investigate the intracellular mechanisms at the basis of their cytotoxicity.

The aim of this study was to compare antitumor activity of [Pt(O,O’-acac)(γ-acac)(DMS)] in epithelioid and sarcomatoid type of MPM. Thus, we employed the human cell line of epithelioid derivation ZL55, and the sarcomatoid cell line ZL34 in vitro and SCID mice. In epithelioid cells, [Pt(O,O’-acac)(γ-acac)(DMS)] was approximately 12-fold more cytotoxic than cisplatin after 24 h of incubation (IC50 were 3.9±0.11 μM for [Pt(O,O’-acac)(γ-acac)(DMS)] and 46.8±0.6 μM for cisplatin, n=6). Similarly, in sarcomatoid cells cisplatin was significantly less cytotoxic than [Pt(O,O’-acac)(γ-acac)(DMS)] (IC50 48.7±1.7 μM and ND n=4, for [Pt(O,O’-acac)(γ-acac)(DMS) and cisplatin, respectively). In addition, we employed a preclinical model based on the subcutaneous injection of ZL55 and ZL34 malignant pleural mesotelioma cell lines in SCID mice. Remarkably, [Pt(O,O’-acac)(γ-acac)(DMS)] stands out for higher anticancer activity than cisplatin toward both the murine tumor models examined, inducing up to 50% inhibition of tumor growth. Mice inoculated with MPM cells showed a statistically significant reduction of tumor volume at every time point in the [Pt(O,O’-acac)(γ-acac)(DMS)] groups compared with both not treated and cisplatin-treated mice (p < 0.05). In summary, our findings show that [Pt(O,O’-acac)(γ-acac)(DMS)] seems to be more potent than cisplatin in MPM, thus providing a solid starting point for its validation as a suitable candidate for further pharmacological testing.

[Pt(O,O′-acac)(γ-acac)(DMS)] can inhibit the metastatic diffusion of breast carcinoma

¹H Nuclear Magnetic Resonance (NMR) spectroscopy coupled with multivariate analysis has been applied in order to investigate metabolomic profiles of more than 200 extravirgin olive oils (EVOOs) collected in a period of over four years (2009-2012) from different geographic areas. In particular, commercially blended EVOO samples originating from different Italian regions (Tuscany, Sicily and Apulia), as well as European (Spain and Portugal) and non-European (Tunisia, Turkey, Chile and Australia) countries. Multivariate statistical analysis (Principal Component Analisys (PCA) and Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA)) applied on the NMR data revealed the existence of marked differences between Italian (in particular from Tuscany, Sicily and Apulia regions) and foreign (in particular Tunisian) EVOO samples. A possible correlation with available climate data has been also investigated. These results aim to develop a powerful NMR-based tool able to protect Italian olive oil productions.

The metabolomic profile of blue crab (Callinectes sapidus) captured in the Acquatina lagoon (SE Italy) was compared to an autochthonous (Eriphia verrucosa) and to a commercial crab species (Cancer pagurus). Both lipid and aqueous extracts of raw claw muscle were analyzed by 1H NMR spectroscopy and MVA (multivariate data analysis). Aqueous extracts were characterized by a higher inter-specific discriminating power compared to lipid fractions. Specifically, higher levels of glutamate, alanine and glycine characterized the aqueous extract of C. sapidus, while homarine, lactate, betaine and taurine characterized E. verrucosa and C. pagurus. On the other hand, only the signals of monounsaturated fatty acids distinguished the lipid profiles of the three crab species. These results support the commercial exploitation and the integration of the blue crab in human diet of European countries as an healthy and valuable seafood

In this work, we tested a NMR-based MVA method to assess the authenticity of EVOOs cultivar (from specific origin) declaration by using a total of over a 100 samples coming from different selected areas of Southern Italian Regions (Apulia and Calabria). We built a reference model, by performing unsupervised PCA and supervised PLS-DA, OPLS-DA analyses on the 1HNMR spectra of a 40 monovarietal EVOOs training set. These included 10 Coratina (the most popular Apulian cultivar) together with 10 Picholine, 10 Ogliarola Barese, and 10 Carolea samples (the most common Coratina bitterness smoothers cultivars used in Coratina-based blends). The training set originated PLS-DA reference model was profitably used to assess the class membership for a series of declared monocultivar EVOO samples and to obtain a clear indication for the composition of Coratina-based blends. By using the same reference model we analyzed in blind a test set formed by 40 unknown samples coming from the same areas and we predicted their class or blend composition affinity (later confirmed by suppliers as Coratina- or Coratina-based blends). This proposed method could be a simple and quick tool to assess, also for trading purpose, the cultivar (from given geographical areas) declaration of labeled EVOOs. Practical applications: EVOO is one of the most important element of the Mediterranean diet with its numerous health benefits due to the chemical composition (unsaturated fatty acids and minor components). For this high commercial value in the recent years, EVOO has been processed to adulteration process with seeds oils, refined pomace, and esterified oils with serious damage to its quality. In order to guarantee the quality of this important product, EU Regulation 182 of March 6, 2009, stated the obligatory labeling of EVOO samples with the geographical production area for all European countries. Surprisingly, this regulation lacks of an official methodology to assess the origin and the cultivar of EVOOs. A quick and reproducible method to check the label declaration of commercial EVOOs could be attractive for industry and ensure transparency for consumer choices. Work flow chart illustrating the procedures sequence to assess cultivars, from specific geographical areas, in EVOOs. A reference model is built and its prediction capability is tested with monovarietal and blend commercial extra virgin olive oil samples.

Coratina cultivar-based olives are very common among 100% Italian extra virgin olive oils (EVOOs). Often, the very spicy character of this cultivar, mostly due to the high polyphenols concentration, requires blending with other “sweetener” oils. In this work, monovarietal EVOO samples from the Coratina cultivar (Apulia, Italy) were investigated and compared with monovarietal EVOO from native or recently introduced Apulian (Italy) cultivars (Ogliarola Garganica, Ogliarola Barese, Cima di Mola, Peranzana, Picholine), from Calabria (Italy) (Carolea and Rossanese) and from other Mediterranean countries, such as Spain (Picual) and Greece (Kalamata and Koroneiki) by 1H NMR spectroscopy and multivariate analysis (principal component analysis (PCA)). In this regard, NMR signals could allow a first qualitative evaluation of the chemical composition of EVOO and, in particular, of its minor component content (phenols and aldehydes), an intrinsic behavior of EVOO taste, related to the cultivar and geographical origins. Moreover, this study offers an opportunity to address blended EVOOs tastes by using oils from a specific region or country of origin.

The green alga Caulerpa cylindracea is a non-autochthonous and invasive species that is severely affecting the native communities in the Mediterranean Sea. Recent researches show that the native edible fish Diplodus sargus actively feeds on this alga and cellular and physiological alterations have been related to the novel alimentary habits. The complex effects of such a trophic exposure to the invasive pest are still poorly understood. Here we report on the metabolic profiles of plasma from D. sargus individuals exposed to C. cylindracea along the southern Italian coast, using 1H NMR spectroscopy and multivariate analysis (Principal Component Analysis, PCA, Orthogonal Partial Least Square, PLS, and Orthogonal Partial Least Square Discriminant Analysis, OPLS-DA). Fish were sampled in two seasonal periods from three different locations, each characterized by a different degree of algal abundance. The levels of the algal bisindole alkaloid caulerpin, which is accumulated in the fish tissues, was used as an indicator of the trophic exposure to the seaweed and related to the plasma metabolic profiles. The profiles appeared clearly influenced by the sampling period beside the content of caulerpin, while the analyses also supported a moderate alteration of lipid and choline metabolism related to the Caulerpa-based diet.

An inverse linear relationship between 73Ge, 119Sn and 207Pb NMR chemical shifts and the overall sum of ionic radii of coordinated halido ligands has been discovered in tetrahedral [MXnY4−n] (M = Ge, Sn, Pb; 1 ≤ n ≤ 4; X, Y = Cl, Br, I) coordination compounds. This finding is consistent with a previously reported correlation found in octahedral, pentacoordinate and square planar platinum complexes. The effect of the coordinated halido ligands acting on the metal as shielding conducting rings is therefore confirmed also by 73Ge, 119Sn and 207Pb NMR spectroscopy.

Nutraceutics is a growing research field in which researchers study and attempt to improve the biological properties of metabolites in food. Wine is one of the most consumed products in the world and contains a plethora of molecules biologically relevant to human health. In this article, several polyphenols with potential antioxidant activity were measured in wines from Apulia, in Southeast Italy. Hydroxytyrosol, gallic and syringic acids, luteolin, quercetin, and trans-resveratrol were identified and quantified by HPLC. The amount of the analyzed metabolites in wines were largely dependent on their color, with red ones being the richest compared to white and rose wines. Gallic acid was the most abundant polyphenol, followed by syringic acid and luteolin. Nevertheless, significant amounts of hydroxytyrosol, quercetin, and trans-resveratrol were also found. The average concentration of polyphenols found in these wines could have potential health-promoting effects, especially if consumed in moderate quantities on a regular basis.

[Pt(O,O'-acac)(gamma-acac)(DMS)] (PtAcD) is able to induce apoptosis in various human cancer cells, including the cisplatin-resistant human breast carcinoma MCF-7 cells. Here, to confirm that PtAcD has the potentiality for therapeutic intervention, we studied its effects in primary cultured epithelial breast cells obtained from cancers and also from the corresponding histologically proven non-malignant tissue adjacent to the tumor. We demonstrated that PtAcD (1) is more cytotoxic in cancer than in normal breast cells; (2) activated NAD(P)H oxidase, leading to PKC-zeta and PKC-alpha tanslocations; (3) activated antiapoptotic pathways based on the PKC-alpha, ERK1/2 and Akt kinases; (4) activated PKC-zeta and, only in cancer cell PKC-delta, responsible for the sustained phosphorylation of p38 and JNK1/2, kinases both of which are involved in the mitochondrial apoptotic process. Moreover, crosstalk between ERK/Akt and JNK/p38 pathways affected cell death and survival in PtAcD-treated breast cell. In conclusion, this study adds and extends data that highlight the pharmacological potential of PtAcD as an anti breast cancer drug.

Abstract Immunoglobulin A nephropathy (IgAN) is the most common form of primary glomerulonephritis worldwide. Both one-dimensional NOESY and transverse-relaxation filter CPMG NMR spectra were recorded to investigate the urine metabolome of 24 IgAN patients and to detect altered metabolic profiles in comparison with 68 healthy matched controls. The spectral data were analyzed using multivariate statistical techniques. The analysis revealed that the NMR spectra of IgAN patients were statistically different from those of the controls (P = 4 × 10−7 for 1D-NOESY and P = 2 × 10−7 for CPMG). The robustness of the determined statistical model was confirmed by its predictive performance (for the 1D-NOESY dataset: sensitivity = 67 %, specificity = 95 %; for the CPMG dataset sensitivity = 60 %, specificity = 94 %). For the first time we found metabolites, including betaine and citrate, that are differentially modulated in IgAN patients compared to controls and that may be directly involved in the pathogenesis of IgAN. These metabolites may influence, directly or indirectly, the TNF-α, a regulating factor of the Th1/Th2 cell balance that is relevant in the pathology. The involvement of metabolites such as betaine and citrate in TNF-α regulation supports the power of the identified metabolic profiles to discern IgAN from controls.

[Pt(N-N)Cl(h1-CH2CH2OR)] complexes were generally considered to be indefinitely stable, both in solution and in the solid state. In a previous work we unexpectedly found that complex [Pt(Me2phen)Cl(1-CH2CH2OR)], 1, Me2phen = 2,9-dimethyl-1,10-phenanthroline, in solution, undergoes spontaneous decomposition to give vinyl-ether together with a Pt(II) hydride species. We demonstrated that this decomposition pathway is mediated by a H- b-shift process promoted by the unbearable steric hindrance introduced by Me2phen in the Pt(II) coordination plane. In this work we found that [Pt(Me2phen)Cl(h1-CH2CH2OR)] complexes can undergo H-D exchange on Me2phen Me’s, when dissolved in protic basic deuterated solvents. Moreover, we observed by 1H NMR that this exchange is considerably slowed down in the analogous complex [Pt(Me2bpy)Cl(h1-CH2CH2OR)], Me2bpy = 6,6’-dimethyl-2,2’-bipyridil, demonstrating the relevant effect of ligand flexibility. Increased acidity of the metal core in the formed T-shaped species reported in the Figure, could be responsible for the observed H-D exchange. An analogous H-D exchange process was observed and investigated by 1H NMR and LC-MS spectroscopy in the related complex [Pt(Me2phen)Cl2]. Further studies demonstrated that this phenomenon is considerably slowed down in the less sterically demanding complexes [Pt(Me2phen)(OR)2] and [Pt(Me2bpy)(OR)2], R = H, Me. It cannot be excluded a role of the H- b-shift process also in the catalysis of the observed H-D exchange process.

We used the human mesothelioma ZL55 cells, which are a reliable model for investigating the therapeutic potential of nucleotides in MPM clinical trials. In addition we attempted to potentiate the cytotoxic effects of cisplatin by ADP. In ZL55 cells ADP is able to block cellular proliferation in a time and dose dependent manner, but evaluating the major apoptotic markers, it is seen that the mechanism underlying this effect is not apoptosis. Cell cycle analysis revealed that inhibition of proliferation of ZL55 cells by ADP was mediated by block of cell cycle progression. Flow cytometry analysis of subconfluent ZL55 cell cultures showed that treatment with ADP (10–100μM) for 24 h caused a concentration dependent increase of cells in the G0/G1 phases. ADP also induced p53 protein levels and increased the levels of p53, p21 and p27 mRNA, whilst cisplatin did not. Cisplatin plus ADP were more efficacious than cisplatin alone in inducing apoptosis characterized by: (a) mitochondria depolarization, (b) increase of bax expression and its cytosol-to-mitochondria translocation and decrease of Bcl-2 expression, (c) activation of caspase-7 and -9 and cleavage of PARP-1. Therefore, the antiproliferative effect of ADP is due to the cell cycle block and increases cisplatin cytotoxicity.

The relevant adsorption of cis-[Pt(NH3)2(P2O7)](2-) (phosphaplatin) on hydroxyapatite nanocrystals (nHAP) was observed and studied in water suspension. Phosphaplatin cytotoxicity, which is very low for HeLa, MCF-7 and HS-5 cell lines could be enhanced, reaching that of cisplatin, by interaction with solid nHAP. This effect stems from nHAP ability to catalyze the phosphaplatin hydrolysis, producing the same hydrolytic species responsible for cisplatin antitumor activity.

In Puglia il Consorzio Interuniversitario di Ricerca in Chimica dei Metalli nei Sistemi Biologici ha supportato con uomini e mezzi diverse campagne di monitoraggio ambientale

Complexes of the type [PtCl(N-N)(eta(1)-CH2CH2OR)], N-N = diimine ligand, R = alkyl, were generally considered to be indefinitely stable, both in solution and in the solid state. Unexpectedly we found that complexes of the type [PtCl(Me(2)phen)(eta(1)-CH2CH2OR)], Me(2)phen 2,9-dimethyl-1,10-phenatroline, R = alkyl, undergo spontaneous decomposition, to give the corresponding vinyl-ether, CH2 = CHOR. Decomposition pathway studies suggest a pseudo-Wacker type mechanism (beta H- shift process) activated by sterical hindrance in the Pt(II) coordination plane, due to the Me(2)phen ligand sterically induced distortions in the Pt(II) coordination plane. A new useful synthetic pathway to access valuable and low toxic alkyl-vinyl-ethers is here reported.

Tumor growth, tumor cell proliferation and microvessel density, in a xenograft model of RCC, developed by injection of Caki-1 cells in BALB/c nude mice, were investigated. Exposure of the Caki-1 cells to cisplatin and to [Pt(O,O’-acac)(γ-acac)(DMS)] resulted in a dose-dependent inhibition of cell survival. [Pt(O,O′-acac)(γ-acac)(DMS)] was much more effective than cisplatin in the inhibition of tumor growth, proliferation and angiogenesis in vivo as well as migration, tube formation and MMP-1, MMP-2 and MMP-9 secretion of endothelial cells, in vitro. Wherease, cisplatin exerted a higher cytotoxic activity on Huvecs, but did not affect tube formation and migration of endothelial cells. In addition, treatment of xenograft mice with [Pt(O,O′-acac)(γ-acac)(DMS)] decreased, VEGF, MMP-1 and MMP-2 expression in tumors.The anti-angiogenic and anti-tumor activities of [Pt(O,O’-acac)(γ-acac)(DMS)] providing a solid starting point for its validation as a suitable candidate for further pharmacological testing.

The higher and selective cytotoxicity of [Pt(O,O′-acac)(γ-acac)(DMS)] towards cancer cell in both immortalized cell lines and in breast cancer cells in primary cultures, stimulated a pre-clinical study in order to evaluate its therapeutic potential in vivo. The efficacy of [Pt(O,O′-acac)(γ-acac)(DMS)] was assessed using a xenograft model of breast cancer developed by injection of MCF-7 cells in the flank of BALB/c nude mice. Treatment of solid tumor-bearing mice with [Pt(O,O′-acac)(γ-acac)(DMS)] induced up to 50% reduction of tumor mass compared to an average 10% inhibition recorded in cisplatin treated animals. Thus, chemotherapy with [Pt(O,O′-acac)(γ-acac)(DMS)] was much more effective than cisplatin. We also demonstrated enhanced in vivo pharmacokinetics, biodistribution, and tolerability of [Pt(O,O′-acac)(γ-acac)(DMS)] when compared to cisplatin administered in Wistar rats. Pharmacokinetics studies with [Pt(O,O′-acac)(γ-acac)(DMS)] revealed prolonged Pt persistence in systemic blood circulation and decreased nefrotoxicity and hepatotoxicity¬, a major target sites of cisplatin toxicity. Overall, [Pt(O,O′-acac)(γ-acac)(DMS)] turned out to be extremely promising in terms of greater in vivo anticancer activity, reduced nephrotoxicity and acute toxicity compared to cisplatin.

BACKGROUND AND PURPOSE It is thought that the mechanism of action of anticancer chemotherapeutic agents is mainly due to a direct inhibition of tumour cell proliferation. In tumour specimens, the endothelial cell proliferation rate increases, suggesting that the therapeutic effects of anticancer agents could also be attributed to inhibition of tumour angiogenesis. Hence, we investigated the potential effects of [Pt (O,O′-acac)(γ-acac)(DMS)] ([Pt(DMS)]), a new platinum drug for non-genomic targets, on human renal carcinoma and compared them with those of the well-established anticancer drug, cisplatin. EXPERIMENTAL APPROACH Tumour growth, tumour cell proliferation and microvessel density were investigated in a xenograft model of renal cell carcinoma, developed by injecting Caki-1 cells into BALB/c nude mice. The antiangiogenic potential of compounds was also investigated using HUVECs. KEY RESULTS Treatment of the Caki-1 cells with cisplatin or [Pt(DMS)] resulted in a dose-dependent inhibition of cell survival, but the cytotoxicity of [Pt(DMS)] was approximately fivefold greater than that of cisplatin. [Pt(DMS)] was much more effective than cisplatin at inhibiting tumour growth, proliferation and angiogenesis in vivo, as well as migration, tube formation and MMP1, MMP2 and MMP9 secretion of endothelial cells in vitro. Whereas, cisplatin exerted a greater cytotoxic effect on HUVECs, but did not affect tube formation or the migration of endothelial cells. In addition, treatment of the xenograft mice with [Pt(DMS)] decreased VEGF, MMP1 and MMP2 expressions in tumours. CONCLUSIONS AND IMPLICATIONS The antiangiogenic and antitumour activities of [Pt(DMS)] provide a solid starting point for its validation as a suitable candidate for further pharmacological testing.

Mesothelioma cancer cells have epithelioid or sarcomatoid morphology. The worst prognosis is associated with sarcomatoid phenotype and resistance to therapy is affected by cells heterogeneity. We recently showed that in ZL55 mesothelioma cell line of epithelioid origin [Pt(O,O'-acac)(γ-acac)(DMS)] (Ptac2S) has an antiproliferative effect in vitro and in vivo. Aim of this work was to extend the study on the effects of Ptac2S on ZL34 cell line, representative of sarcomatoid mesothelioma. ZL34 cells were used to assay the antitumor activity of Ptac2S in a mouse xenograft model in vivo. Then, both ZL34 and ZL55 cells were used in order to assess the involvement of p53 protein in (a) the processes underlying the sensitivity to chemotherapy and (b) the activation of various transduction proteins involved in apoptosis/survival processes. Ptac2S increases ZL34 cell death in vivo compared with cisplatin and, in vitro, Ptac2S was more efficacious than cisplatin in inducing apoptosis. In Ptac2S-treated ZL34 and ZL55 cells, p53 regulated gene products of apoptotic BAX and anti-apoptotic Bcl-2 proteins via transcriptional activation. Ptac2S activated PKC-δ and PKC-ε; their inhibition by PKC-siRNA decreased the apoptotic death of cells. PKC-δ was responsible for JNK1/2 activation that has a role in p53 activation. In addition, PKC-ε activation provoked phosphorylation of p38MAPK, concurring to apoptosis. In ZL34 cells, Ptac2S also activated PKC-α thus provoking ERK1/2 activation; inhibition of PKC-α, or ERK1/2, increased Ptac2S cytotoxicity. Results confirm that Ptac2S is a promising therapeutic agent for malignant mesothelioma, giving a substantial starting point for its further validation.

In invertebrate biomonitors of chemical pollution, emphasis has been generally given to mean accumulation patterns and how they reflect varying environmental levels of contamination. Intra-population variability, and how it relates with individual phenotypic traits, has received less attention. Here, a set of analytes including trace elements (B, Ba, Cd, Cr, Cu, Fe, Li, Mn, Ni, Pb, Sr, V, and Zn), macroelements (C, Ca, K, Mg, N, Na), and carbon and nitrogen stable isotopes (δ13C and δ15N) was measured in two populations of the crayfish Procambarus clarkii fromLake Trasimeno and Lake Bolsena (Central Italy). The influence of location, sex, body size, and condition factor was assessed; in addition, the analyte correlation profiles of the two populations were compared to verify their congruence. In general, significant inter-lake differences were observed in the concentration of both trace- and macroelements in crayfish tissues, generally mirroring the local chemistry of water and of benthic non-living matrices (sediment and plant detritus). Crayfish CN isotopic signatures excluded the occurrence of inter-lake variations in their omnivorous trophic habits. Correlation profiles varied considerably between the two populations in the nature and strength of bivariate relationships. However, Mantel tests and procrustean analyses indicated a general, significant congruence; C, N, and, to a lesser extent K, Li, Ni, Pb, and δ13C showed the highest procrustean residuals, suggesting that their associations with other analytes may be partially influenced by inter-population differences in growing phases. Our study indicates that the local geochemistry of the lacustrine environment influences the elemental fingerprint of Procambarus clarkii; the considerable inter-individual variability in the concentration of analytes, however, does not significantly reflect on their association, thus corroborating its effectiveness as an indicator species.

Aim of this study was to determine the content of bioactive phytochemicals in Capparis spinosa subsp. rupestris (syn. C. orientalis), a less investigated species of caper and compare the chemical profile of this species with that of other studied Capparis sp. and especially with the related cultigen C. spinosa subsp. spinosa. Chemical composition of seed oil and glucosinolates, as well as of glucosinolates and flavonoids from the aerial parts of the plant have been determined and data reported here. Oil from the plant seeds is rich in unsaturated and rare lipids such as cis-vaccenic acid; the main glucosinolate is glucocapperin. The aerial parts are characterized by rutin as the dominant flavonoid. The overall phytochemical data obtained from the analysis of C. spinosa subsp. rupestris indicate that this species represents a very rich source of bioactive compounds of nutraceutical relevance although the compositional profile does not differentiate this subspecies from C. spinosa subsp. spinosa.

Recently, with the advent of modern technologies, various marine organisms including algae are being studied as sources of natural substances effective on classical microorganisms and able to also combat the new trend of acquired resistance in microbes. In the present study the antimicrobial activity of the lipidic extract of the green seaweed Cladophora rupestris collected in a Mediterranean area, in two sampling periods (January and April), was assayed. The chemical characterization of the lipidic fractions was performed by gas-chromatography and multinuclear and multidimensional {NMR} spectroscopy. In the lipidic extract of C. rupestris collected in January an antibacterial activity against Enterococcus sp., Streptococcus agalactiae and Vibrio cholerae non-O1 was recorded; by contrast, bacterial inhibition was measured on several Vibrio species only in April. The fatty acid profile of C. rupestris lipidic extract, analyzed by gas chromatography, resulted mainly composed of palmitic, myristic, oleic, α linolenic, palmitoleic and linoleic acids. Moreover, since α-linolenic acid was the predominant ω3 fatty acid in April, we suggest its involvement in the antibacterial activity observed in this month, taking also into account that pure α-linolenic acid resulted effective towards some vibrios strains. C. rupestris fatty acid profile revealed also an interesting composition in polyunsaturated fatty acids in both the considered periods with the ω6/ω3 ratio lower than 1, leading to conclude that this macroalga may be employed as a natural source of ω3. Finally, the 1H {NMR} spectrum in {CDCl3} of algal lipid fractions showed the characteristic signals of saturated (SAFAs) and unsaturated fatty acids (UFAs) as well as other metabolites and a marked difference in free fatty acids (FFAs) content for the two examined algal lipid fractions. It is noteworthy that C. rupestris lipidic extracts show, by {NMR} spectroscopy, the signal pattern of polyhydroxybutyrate, a natural biocompatible and biodegradable polymer. In conclusion, on account of its antimicrobial activity, nutritional value and bioplastic content, C. rupestris lipidic extract can be considered a promising source for future biotechnological applications.

The series of complexes of formula [PtCl(h2-olefin)(N^N)]+, previously investigated for N^N = N,N,N0 ,N0-tetramethyl-ethylenediamine (Me4en), has been extended to the case of aromatic diimines 1,10-phenanthroline (phen) and 3,4,7,8-tetramethyl-1,10-phenanthroline (Me4phen) and to a variety of olefins (ethene, propene and styrene). The complexes have been prepared by reaction of the [PtCl3(h2-olefin)]-anions (K[PtCl3(h2-styrene)] reported here for the first time) with N^N in basic methanol. The initial [PtCl{h1-CH2-CH(R')-OMe}(N^N)] (R' = Me, Ph) complexes are formed in quantitative yield and as pure Markovnikov isomer. The reaction of the alkoxylic species with non coordinating acids, results in the quantitative formation of the desired cationic p-olefin complexes [PtCl(h2-olefin)(N^N)]+. The phenanthroline ligand confers peculiar properties to the new compounds. In particular, by reaction with triethylamine, [PtCl{h2-CH2=CH(Me)}(N^N)]+ species, undergo deprotonation of the olefin and formation of the dimeric species [{PtCl(N^N)}2(m-h1:h2-CH2CH=CH2)]+ which could be isolated and characterized. Interestingly such product in acetonitrile gives a disproportionation with precipitation of [PtCl2(phen)] and formation in solution of the new h3-allyl complex [Pt(h3-C3H5)(phen)]ClO4.

Cisplatin, cis-[Pt(NH3)2Cl2], was the first inorganic compound applied in clinics to treat a broad range of malignancies. It is able to bind DNA on the N7 positions of adjacent G/A residues, resulting in the cross-link lesions believed to be responsible for observed antitumor activity. Sometimes N7-metalated purines seem to be characterized by a relevant antitumor activity. This has led us to hypothesize a parallel mechanism of action of platinum drugs, based on free platinated purines, formed after drug administration and incorporated into DNA by DNA polymerases. In order to evaluate this possibility, as a key step to develop new drugs, we performed experiments focused on platinated nucleobases, i.e. [Pt(dien)(N7-G)] and cis-[Pt(NH3)2(py)(N7-G)], dien = diethylenetriamine, py = pyridine, G = 5’-dGTP, cell and mitochondrial uptake and processing. For the first time cell uptake and mobility mechanisms, related to plasmatic cell and/or mitochondrial membrane crossing, has been studied. The results of the present study suggest that nucleotide carriers can be actively implicated in the specific uptake of free cytoplasmic platinum bonded nucleotides. Moreover the possible insertion of metalated nucleobases into nuclear and/or mitochondrial new synthesized DNA/RNA, operated by DNA/RNA polymerases, has been evaluated.

Abstract Several chemotherapeutic drugs are known to cause neurotoxicity. Platinum-based agents in use or in clinical trials display neurotoxic potential accompanied by neurological complications; recent studies have identified a large number of behavioural issues in paediatric oncology patients. To understand the toxicity of platinum drugs at the molecular and cellular levels, this study compares the possible cytotoxic effects of an older platinum compound, cisplatin and a new platinum compound, [Pt(O,O′-acac)(γ-acac)(DMS)], on the {CNS} of postnatally developing rats, which is much more vulnerable to injury than the {CNS} of adult rats. Since several drugs interact with neurotransmitters during neuronal maturation, we performed immunostainings with antibodies raised against markers of glutamate and GABA, the major neurotransmitters in the cerebellum. After a single injection of cisplatin at postnatal day 10 (PD10), the labelling of Purkinje cells with the neurotransmitter markers evidenced alterations between {PD11} and PD30, i.e. atrophy of the dendrite tree, changes in the distribution of synaptic contacts of parallel and climbing fibres, delay in the elimination of transient synapses on cell soma and severely impaired pinceau formation at the axon hillock. After treatment with [Pt(O,O′-acac)(γ-acac)(DMS)], the sole relevant change concerned the timing of climbing fibres elimination; the transient synapses disappearance on the Purkinje cell soma was delayed in some cells; instead, the growth of Purkinje cell dendrite tree was normal as was the formation of inhibitory synaptic contacts on these neurons. These findings add new evidence not only on the lower neurotoxicity of [Pt(O,O′-acac)(γ-acac)(DMS)] vs cisplatin but also on the involvement of neurotransmitters and relative synaptic connections in the maturation of central nerve tissue.

The products obtained by forcing the reaction with nucleosides (guanosine, Guo, and adenosine, Ado) of potential anticancer drugs for nongenomic targets [PtCl(O,O'-acac)(L)] (L = dimethyl sulfoxide, DMSO; dimethyl sulfide, DMS), closely related to their very powerful organometallic analogues [Pt(O,O'-acac)(γ-acac)(L)], have been studied. [PtCl(O,O'-acac)(L)] and [Pt(O,O'-acac)(γ-acac)(L)] complexes were reported unreactive toward nucleobases. Aquo species [Pt(O,O'-acac)H2O(L)]+, obtained from [PtCl(O,O'-acac)(L)] by Ag+ driven coordinated Cl– removal, gave access to [Pt(O,O'- acac)(L)(nucleoside)]+ ([Pt(O,O'-acac)(DMSO)(Guo)]+, [Pt(O,O'-acac)(DMS)(Guo)]+, [Pt(O,O'-acac)(DMSO)(Ado)]+). The effect of the chelate oxygen donor acac (with respect to a chelate diammine), the role of the sulfur ligand (DMSO, DMS), and the influence of the purinic nucleoside itself on the coordinated Guo or Ado dynamic motions in [Pt(O,O'- acac)(L)(nucleoside)]+ complexes have been investigated by NMR spectroscopy. Interestingly, a slow rotation of nucleobase around the Pt–N(7) bond with formation of two rotamers was observed already at room temperature only in the case of [Pt(O,O'- acac)(DMSO)(Guo)]+. On the other hand, no hindered rotation at room temperature was detected in the analogous [Pt(O,O'-acac)(DMS)(Guo)]+ and [Pt(O,O'-acac)(DMSO)(Ado)]+ complexes. Data suggest that rotation of the nucleoside in [Pt(O,O'-acac)(L)(nucleoside)]+ is very different with respect to the analogous [Pt(diammine)(L)(nucleoside)]2+ systems, due to specific interactions between the acac chelate ligand, the DMSO, and the nucleobase

There is increasing interest in development of methods for wine characterization to guarantee traceability and product safety. Metabolomics represents a valuable tool to trace the metabolic fingerprint of complex mixtures. Wines from 22 pure varieties (autochthonous from Southern Italy and grown in the same area of Apulia) were genetically identified and metabolically characterized using both NMR and HPLC-MS. NMR associated with chemometrics was used to identify amino acid (proline, alanine, and tyrosine), organic acid (malic, lactic, acetic, and succinic acids), alcohol (2-phenylethanol), sugar (sucrose), and polyphenol (gallic acid) contents. Minor components of wines were determined using HPLC-MS analyses. Wines showed potential health-promoting effects with good amounts of phenolic compounds. In particular, consistent levels of trans-resveratrol and flavonoids were found in Negro Amaro, Uva di Troia, Ancellotta, and Montepulciano wines.

Porous silicon is a nano material in which pores with different sizes, densities and depths are infiltrated in conventional silicon imparting it augmented properties including biodegradability, biocompatibility, photoluminescence. Here, we realized porous silicon substrates in which the pore size and the fractal dimension were varied over a significant range. We loaded the described substrates with a PtCl(O, O′ − acac)(DMSO) antitumor drug and determined its release profile as a function of pore size over time up to 15 days. We observed that the efficacy of delivery augments with the pore size moving from small (∼ 8 nm, efficiency of delivery ∼ 0.2) to large (∼ 55 nm, efficiency of delivery ∼ 0.7). Then, we verified the adhesion of MCF-7 breast cancer cells on the described substrates with and without the administration of the antitumor drug. This permitted to decouple and understand the coincidental effects of nano-topography and a controlled dosage of drugs on cell adhesion and growth. While large pore sizes guarantee elevated drug dosages, large fractal dimensions boost cell adhesion on a surface. For the particular case of tumor cells and the delivery of an anti-tumor drug, substrates with a small fractal dimension and large pore size hamper cell growth. The competition between nano-topography and a controlled dosage of drugs may either accelerate or block the adhesion of cells on a nanostructured surface, for applications in tissue engineering, regenerative medicine, personalized lab-on-a-chips, and the rational design of implantable drug delivery systems.

The proximate composition and element contents of claw muscle tissue of Atlantic blue crabs (Callinectes sapidus) were compared with the native warty crab (Eriphia verrucosa) and the commercially edible crab (Cancer pagurus). The scope of the analysis was to profile the chemical characteristics and nutritive value of the three crab species. Elemental fingerprints showed significant inter-specific differences, whereas non-significant variations in the moisture and ash contents were observed. In the blue crab, protein content was significantly lower than in the other two species, while its carbon content resulted lower than that characterizing only the warty crab. Among micro-elements, Ba, Cr, Cu, Li, Mn, Ni, and Pb showed extremely low concentrations and negligible among-species differences. Significant inter-specific differences were observed for Na, Sr, V, Ba, Cd and Zn; in particular, cadmium and zinc were characterized in the blue crab by concentrations significantly lower than in the other two species. The analysis of the available literature on the three species indicated a general lack of comparable information on their elemental composition. The need to implement extended elemental fingerprinting techniques for shellfish quality assessment is discussed, in view of other complementary profiling methods such as NMR-based metabolomics.

Extra-virgin olive oil (EVOO) is among the basic constituents of the Mediterranean diet. Its nutraceutical properties are due mainly, but not only, to a plethora of molecules with antioxidant activity known as biophenols. In this article, several biophenols were measured in EVOOs from South Apulia, Italy. Hydroxytyrosol, tyrosol and their conjugated structures to elenolic acid in different forms were identified and quantified by high performance liquid chromatography (HPLC) together with lignans, luteolin and α-tocopherol. The concentration of the analyzed metabolites was quite high in all thecultivarsstudied, but it was still possible to discriminate them through multivariate statistical analysis (MVA). Furthermore, principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) were also exploited for determining variances among samples depending on the interval time between harvesting and milling, on the age of the olive trees, and on the area where the olive trees were grown.

Comparative studies on the effects of the platinum complexes in use or in clinical trials are carried out in order to discover differences in the neurotoxic potential and the reversibility of neurotoxicity. In this paper, we summarized the current literature on neurotoxicity and chemoresistance of cisplatin (cisPt) and discussed our recent efforts on the interference of cisPt and a new platinum compound [Pt(O,O-acac)(γ-acac)(DMS)] (PtAcacDMS), with high specific reactivity with sulphur ligands instead of nucleobases as cisPt, on some crucial events of rat postnatal cerebellum development. The acute effects of drug treatments on cell proliferation and death in the external granular layer and granule cell migration and the late effects on the dendrite growth of Purkinje cells were evaluated. Together with the demonstrated antineoplastic effectiveness in vitro, compared with cisPt, data suggest a lower neurotoxicity of PtAcacDMS, in spite of its presence in the brain that involves considerations on the blood brain barrier permeability.

Programmed cell death is regulated by prototypes of a large family of Bcl-2-like proteins such as Bax and Bcl-2. A neuroprotective role for cellular prion protein (PrPc) on programmed cell death has been reported, although the cytosolic accumulation of PrPc correlates with toxicity and death of some neurons by apoptosis. In order to understand the signalling function of PrPc in promoting survival or suppressing cell death, we analyzed the expression and co-localization of PrPc, Bax and Bcl-2 proteins in the developing cerebellum of rats treated at PD10 (postnatal day 10) with the chemotherapeutic drug cisplatin (cisPt) or the new platinum (Pt) compound [Pt(O,O0-acac)(g-acac)(DMS)] (PtAcacDMS). Differences in the expression of PrPc, Bax and Bcl-2 were found in proliferating cells and immature Purkinje neurons. One day after administration (PD11), cisPt markedly increased the apoptosis of the proliferating cells of the EGL (external granular layer); at the same time, several apoptotic bodies with strong Bax immunoreactivity were noticed. After PtAcacDMS, changes in PrPc and apoptotic proteins, with respect to the controls, were found but Bax immunopositive apoptotic bodies were not detectable, which could mean that apoptotic cell death of proliferating cells is preserved. Co-localization was clearly detected in the Purkinje cell population and may explain better the mechanisms by which PrPc and the apoptotic proteins function, and particularly the role of PrPc. Considering the reactivity of Purkinje neurons to these proteins at PD11 and Pd17, at least PrPc expression increased after cisPt and PtAcacDMS treatments or, if PrPc decreased, balanced itself with Bcl-2. The noteworthiness of this finding is that it emphasizes that most of the post-mitotic Purkinje cells need to be rescued, otherwise they undergo degeneration and are not replaced. Based on the effects of both Pt compounds on Purkinje cell differentiation, it should be emphasized that PrPc, together with the synergistic action of the co-localized anti-apoptotic protein, acts as a neuroprotective protein countering cytotoxicity in the postnatal critical phases of cerebellum development.

BACKGROUND AND PURPOSE: The aim of this study was to determine whether [platinum (Pt)(O,O'-acetylacetonate (acac))(γ-acac)(dimethylsulphide (DMS))] is differentially cytotoxic in normal and cancer cells, and to measure comparative levels of cytotoxicity compared with cisplatin in the same cells. EXPERIMENTAL APPROACH: We performed experiments on cancerous and normal epithelial breast cells in primary culture obtained from the same patients. The apoptotic effects [Pt(O,O'-acac)(γ-acac)(DMS)] and cisplatin in cancerous and normal breast cells were compared. KEY RESULTS: Cancer cells were more sensitive to [Pt(O,O'-acac)(γ-acac)(DMS)] (IC50 = 5.22 ± 1.2 μmol·L(-1)) than normal cells (IC50 = 116.9 ± 8.8 μmol·L(-1)). However, the difference was less strong when cisplatin was used (IC50 = 96.0 ± 6.9 and 61.9 ± 6.1 μmol·L(-1) for cancer and normal cells respectively). Both compounds caused reactive oxygen species (ROS) production with different mechanisms: [Pt(O,O'-acac)(γ-acac)(DMS)] quickly activated NAD(P)H oxidase while cisplatin caused a slower formation of mitochondrial ROS. Cisplatin and [Pt(O,O'-acac)(γ-acac)(DMS)] caused activation of caspases, proteolysis of PARP and modulation of Bcl-2, Bax and Bid. [Pt(O,O'-acac)(γ-acac)(DMS)] also caused leakage of cytochrome c from the mitochondria. Overall, these processes proceeded more quickly in cells treated with [Pt(O,O'-acac)(γ-acac)(DMS)] compared with cisplatin. [Pt(O,O'-acac)(γ-acac)(DMS)] effects were faster and quantitatively greater in cancer than in normal cells. [Pt(O,O'-acac)(γ-acac)(DMS)] caused a fast decrease of mitochondrial membrane potential, especially in cancer cells. CONCLUSIONS AND IMPLICATIONS: [Pt(O,O'-acac)(γ-acac)(DMS)] was specific to breast cancer cells in primary culture, and this observation makes this compound potentially more interesting than cisplatin.

Background Recently, a novel concept ‘bio-effectors’ rose on to describe a group of products that are able to improve plant performance rather than fertilizers. Agro-food processing residues and by-products potentially represent important sources of bio-effectors but they are currently not properly taken in consideration. To fulfill this gap, in these study, three food processing by-products: (i) brewers' spent grain, (ii) fennel processing residues, and (iii) lemon processing residues were chosen as bio-effector candidates. Raw materials were chemically characterized, and green extraction methodology was optimized by using water, ethanol, and their mixture based on the extraction yields. Aqueous extracts were used for seed germination bioassays on Lepidium sativum seeds to evaluate their potential bioactivities. Thereafter, the extracts were chemically characterized and metabolites were detected by 1D and 2D NMR spectroscopy. Results Results are summarized as follows: (i) raw materials showed an interesting nutritional content; (ii) aqueous extraction resulted higher yield more than other used solvent; (iii) at high solvent extraction ratio, aqueous extracts were not phytotoxic but enhanced seed germination and root elongation; (iv) all aqueous extracts are differently rich in nutrients, amino-acids, sugars, and other low molecular weight molecule compounds. Conclusions This study confirmed that efficient and simple recovery of bioactive compounds other than nutrients from agro-food processing by-products appear to be the new frontier in their valorization.

Artemisia annua tea has been proven to be a very effective treatment for malaria in various clinical trials, but to date its efficacy has not been investigated in vitro. A study was therefore performed to evaluate the effects of A. annua tea on Plasmodium falciparum cultures in vitro. The concentration of artemisinin in the herbal tea preparation was also determined. The herbal tea extract was tested against chloroquine (CQ)-sensitive D10 and CQ-resistant W2 strains of P. falciparum using the parasite lactate dehydrogenase assay. Quantification of artemisinin in the extract of leaves of A. annua was performed using proton nuclear magnetic resonance (1H-NMR). Results of the in vitro tests were consistent with the clinical efficacy of A. annua tea [50% inhibitory concentration (IC50) for strain D10=1.11±0.21μg/ml; IC50 for strain W2=0.88±0.35μg/ml]. The concentration of artemisinin in A. annua tea (0.18±0.02% of dry weight) was far too low to be responsible for the antimalarial activity. The artemisinin present in the tea is probably co-solubilised with other ingredients, some of which also have antimalarial activity and act synergistically with it. These compounds also merit further research to determine whether their presence hinders the development of parasite resistance compared with pure artemisinin.

Extraction methods are critical in the production and success of an extra virgin olive oil ready for consumption. The first step of olive oil extraction procedure produces an olive paste, which is subsequently processed by separation of the liquid phases (oil and vegetable water) from the pomace. Profiles of Apulian extra virgin olive oils and the deuterated chloroform extracts of its paste and pomace were studied by 1D and 2D NMR (NMR) spectroscopy without separation and pre-treatment of samples. In order to study the metabolic profiles variation in the paste and pomace extracts with respect to the corresponding EVOOs, multivariate statistical analyses (PCA and OPLS DA) of the 1H NMR data were performed. As expected, a different minor components profile, in particular for the presence of higher content of secoiridoids, lignans and phenolic compounds in olive paste and pomace extracts with respect to the EVOOs, was observed. Practical applications: Amount of extractable oil from olive paste and pomace after the production processes of EVOO was measured by 1H NMR spectroscopy. Identification of metabolites in olive paste and pomace was performed by 1D and 2D NMR spectroscopy without separation and pre-treatment of samples. The chemical composition of major and minor components (in particular secoiridoids, lignans, and phenolic compounds) present in olive paste and pomace chloroform extracts could be used for extraction process trimming purposes as well as in fingerprint analyses for cultivar and/or geographical origin certification of olive oil production. Moreover, determination of the entire metabolic content of different materials of the olive oil production process could be exploited in other applications such as recycling on agricultural land, studies of the health benefits on human nutrition and alternative dietary lipid source in animals. In fact, it is worth noting that the olive paste and pomace extracts could be even healthier than other vegetable oils. The extraction processes are critical in the extra virgin olive oil production. The chemical composition of EVOOs and its paste and pomace extracts have been investigated by Nuclear Magnetic Resonance Spectroscopy (NMR).

An inverse linear relationship between the experimentally observed (195)Pt NMR signals and the overall sum of coordinated halido ligands' ionic radii was discovered in Pt(ii) and Pt(iv) complexes. The reduction of (195)Pt NMR frequencies parallels the increase of coordinated halido ligands' ionic radii sum. This suggests that each halido ligand may act as a conducting ring whose induced electric current shields the (195)Pt NMR signals proportionally to the ionic radius of the coordinated halido ligand.

Glucosinolates (GLSs) from a variety of Brassica oleracea, known locally as "mugnolo" and widely distributed in southern Italy, were studied. It was found that "mugnolo" inflorescences are characterized by the presence of the aliphatic GLSs glucoraphanin (1.79 mu mol/g), glucoiberin, glucoerucin, and sinigrin, of the aromatic GLSs glucobarberin (0.56 mu mol/g) and gluconasturtin, and of the indole GLSs glucobrassicin (3.51 mu mol/g), neoglucobrassicin, 4-methoxyglucobrassicin, and 4-hydroxyglucobrassicin. Indole GLSs were predominant, while aliphatic and aromatic GLS were found in lower quantities. The metabolic profile of "mugnolo" leaves was also studied. It was found that 4-hydroxyglucobrassicin (0.13 mu mol/g) and glucoraphanin (0.11 mu mol/g)are the predominant GLSs. Vegetables of the Brassicaceae family are seldom consumed raw; therefore we also analyzed the GLS profile of "mugnolo" after cooking in water. The results showed variations in the GLSs content with a fall in concentration of 50%.

Multivariate analysis of H-1 NMR data has been used for the characterization of 12 blended olive oils commercially available in the U. S. as Italian products. Chemometric methods such as unsupervised Principal Component Analysis (PCA) allowed good discrimination and gave some affinity indications for the U. S. market olive oils compared to other single cultivars of extra virgin olive oil such as Coratina and Ogliarola from Apulia, one of Italy's leading olive oil producers, Picual (Spain), Kalamata (Greece) and Sfax (Tunisia). The olive oils commercially available as Italian products in the U. S. market clustered into 3 groups. Among them only the first (7 samples) and the second group (2 samples) showed PCA ranges similar to European references. Two oils of the third group (3 samples) were more similar to Tunisian references. In conclusion, our study revealed that most EVOO (extra virgin olive oils) tested were closer to Greek (in particular) and Spanish olive oils than Apulia EVOO. The PCA loadings disclose the components responsible for the discrimination as unsaturated (oleic, linoleic, linolenic) and saturated fatty acids. All are of great importance because of their nutritional value and differential effects on the oxidative stability of oils. It is evident that this approach has the potential to reveal the origin of EVOO, although the results support the need for a larger database, including EVOO from other Italian regions.

Nine hundred extra virgin olive oils (EVOO) were extracted from individual olive trees of four olive cultivars (Coratina, Cima di Mola, Ogliarola, Peranzana), originating from the provinces of Bari and Foggia (Apulia region, Southern Italy) and collected during two consecutive harvesting seasons (2013/14 and 2014/15). Following genetic identification of individual olive trees, a detailed Apulian EVOO NMR database was built using 900 oils samples obtained from 900 cultivar certified single trees. A study on the olive oil lipid profile was carried out by statistical multivariate analysis (Principal Component Analysis, PCA, Partial Least-Squares Discriminant Analysis, PLS-DA, Orthogonal Partial Least-Squares Discriminant Analysis, OPLS-DA). Influence of cultivar and weather conditions, such as the summer rainfall, on the oil metabolic profile have been evaluated. Mahalanobis distances and J2 criterion have been measured to assess the quality of resulting scores clusters for each cultivar in the two harvesting campaigns. The four studied cultivars showed non homogeneous behavior. Notwithstanding the geographical spread and the wide number of samples, Coratina showed a consistent behavior of its metabolic profile in the two considered harvests. Among the other three Peranzana showed the second more consistent behavior, while Cima di Mola and Ogliarola having the biggest change over the two years.

The present work analyzed three hard-bottom and two soft-bottom species of sabellid polychaetes to determine the content of several heavy metals in their branchial crown and body. The highest concentrations of heavy metals were recorded in the hard-bottom species Branchiomma bairdi, a recent Mediterranean introduction. Differences in the metal concentrations were most notable in the high trace metal levels of the branchial crown for all the studied species. Statistical analysis showed that the Mediterranean hard-bottom species were similar each other in their heavy metal content in the body as well as in the branchial crown and appeared separated from all the other species. Arsenic and vanadium hyperaccumulation in the branchial crowns of the considered sabellid species probably acts as a deterrent for predation. The observed differences among the examined species were discussed not only at the light of habitat colonization but also in terms of the phylogeny.

The effects of a Pt-based drug on human mesothelioma cells NCI-H28 and H2373

A series of pentacoordinate d8 PtII complexes, of the type [PtCl2(η2-CH2=CH2)(Men∩NN)], where Men∩NN = bis-nitrogen ligand, with a variable number of Me groups (i.e., 2,2′-bipyridyl; 1,10-phenanthroline; 6-methyl-2,2′-bipyridyl; 2,9-dimethyl-1,10-phenanthroline; 2,9-dimethyl-1,10-phenanthroline; N,N′-trimethyl-ethylenediamine; N,N,N′,N′-tetramethyl-ethylenediamine; N,N,N′,N′-tetramethyl-1,2-diaminocyclohexane) was studied. The compounds are characterized by variable steric hindrance, due to the variable number (n) of Me substituents, on or ortho to the N-donors of aliphatic diamines or aromatic diimines, respectively. This approach was developed to investigate the interaction of substituents with the metal coordination sphere. With this aim, we analyzed the NMR properties of the considered complexes, with respect to modulation of the metal electron density (195Pt NMR signal frequency) by alkyl groups close to the N-donors. 1H and 13C NMR analysis of the η2-olefin signals has revealed, for each kind of bis-nitrogen ligand, a positive or negative chemical shift variation that is proportional to the number of Me groups geminal or vicinal to the N-donors. Interestingly, the 1JPt,C values increase by approximately 45 Hz for each additional Me on the series of diamine or diimine bis-nitrogen ligands. A rationale for the stability changes observed in such pentacoordinate complexes is suggested, based on the NMR spectroscopic data analysis.

Aiming to contribute to the design of new antitumoral drugs, we synthesized new hydrophilic Pt(II) complexes of general formula [PtCl2(N,N0)] containing nitrogen bidentate amine–imine and di-imine ligands derived from glucose. Some chemical properties were discussed. The X-ray molecular structure of [PtCl2(a-D-glucopyranoside-methyl-6-deoxy-6(2-(methylimino)methyl)pyridine) (D) was reported. [Pt Cl2(b-D-glucopyranosylimine-N-(2-pyridinylmethyl))] (A), which is well-soluble both in organic solvents and in water, was tested for cytotoxicity.

Malignant pleural mesothelioma (MPM) is an aggressive malignancy highly resistant to chemotherapy. There is an urgent need for effective therapy inasmuch as resistance, intrinsic and acquired, to conventional therapies is common. Among Pt(II) antitumor drugs, [Pt(O,O'-acac)(γ-acac)(DMS)] (Ptac2S) has recently attracted considerable attention due to its strong in vitro and in vivo antiproliferative activity and reduced toxicity. The purpose of this study was to examine the efficacy of Ptac2S treatment in MPM. We employed the ZL55 human mesothelioma cell line in vitro and in a murine xenograft model in vivo, to test the antitumor activity of Ptac2S. Cytotoxicity assays and Western blottings of different apoptosis and survival proteins were thus performed. Ptac2S increases MPM cell death in vitro and in vivo compared with cisplatin. Ptac2S was more efficacious than cisplatin also in inducing apoptosis characterized by: (a) mitochondria depolarization, (b) increase of bax expression and its cytosol-to-mitochondria translocation and decrease of Bcl-2 expression, (c) activation of caspase-7 and -9. Ptac2S activated full-length PKC-δ and generated a PKC-δ fragment. Full-length PKC-δ translocated to the nucleus and membrane, whilst PKC-δ fragment concentrated to mitochondria. Ptac2S was also responsible for the PKC-ε activation that provoked phosphorylation of p38. Both PKC-δ and PKC-ε inhibition (by PKC-siRNA) reduced the apoptotic death of ZL55 cells. Altogether, our results confirm that Ptac2S is a promising therapeutic agent for malignant mesothelioma, providing a solid starting point for its validation as a suitable candidate for further pharmacological testing.

In this chapter, we discuss several immunohistochemical methods to monitor apoptosis and autophagy in two models, the cultured rat neuroblastoma cells and the developing rat cerebellum . Apoptosis and autophagy have been implicated in many physiological and pathological processes. Apoptotic cell death is characterized by membrane blebbing and nuclear chromatin condensation /fragmentation , alteration of cytoskeleton components and cytoplasmic organelles , and formation of membrane-enveloped apoptotic bodies that are rapidly phagocytosed by macrophages . Autophagy is a sequential set of events including double membrane formation and elongation, vesicle maturation , and finally delivery of the targeted materials to the lysosome . Platinum compounds can induce different apoptotic pathways in neural cell cultures and alter the normal balance between the formation and degradation of cellular proteins . These compounds also affect cell death by apoptosis and autophagy in the proliferating and differentiating cells of rat cerebellum in vivo .

We assessed the therapeutic potential of [Pt(O,O’-acac)(γ-acac)(DMS)] in vivo using a xenograft model of breast cancer developed by injection of MCF-7 cells in the flank of BALB/c nude mice. Solid tumor-bearing mice showed up to 50% reduction of tumor mass after [Pt(O,O’-acac)(γ-acac)(DMS)] treatment, whereas an average 10% inhibition was recorded in cisplatin treated animals. Moreover, enhanced in vivo pharmacokinetics (PK), biodistribution and tolerability for [Pt(O,O’-acac)(γ-acac)(DMS)] administered Wistar rats have been observed with respect to cisplatin treated.

The reactivity with acetylene of [PtX2(Me2phen)] (X = Cl, Br, I) complexes has been investigated. Whereas the chlorido species [PtCl2(Me2phen)] exhibits negligible reactivity at short reaction times, the bromido and iodido species [PtBr2(Me2phen)] and [PtI2(Me2phen)] lead initially to formation of Pt(II) five-coordinate complexes, [PtX2(η2-CHuCH)(Me2phen)], that evolve to four-coordinate alkenyl complexes of the type [PtX(η1 -E-CHvCHX)(Me2phen)]. The alkenyl complexes, in the presence of excess acetylene, establish an equilibrium with the five-coordinate alkyne–alkenyl species [PtX(η1-E-CHvCHX)(η2-CHuCH)(Me2phen)] (X = Br, I). The π-bonded acetylene can be exchanged with free olefins or CuO, affording the new alkene–alkenyl or carbonyl–alkenyl complexes [PtX(η1-E-CHvCHX)(η2-olefin)(Me2phen)] and [PtX(η1-E-CHvCHX)(CuO)(Me2phen)]. The five-coordinate geometry of the alkyne–alkenyl and alkene–alkenyl complexes was assessed from NMR data and is fully consistent with that of a previously determined X-ray structure of [PtBr(η1-E-CHvCHBr)(η2-CH2vCH2)(Me2phen)].

The reactivity of [PtX2(Me2phen)] complexes (X = Cl, Br, I; Me2phen = 2,9-dimethyl-1,10-phenanthroline) with terminal alkynes has been investigated. Although the dichlorido species [PtCl2(Me2phen)] exhibits negligible reactivity, the bromido and iodido derivatives lead in short time to the formation of five-coordinate Pt(II) complexes of the type [PtX2(Me2phen)(η2-CHuCR)] (X = Br, I; R = Ph, n-Bu), in equilibrium with the starting reagents. Similar to analogous complexes with simple acetylene, the five coordinate species can also undergo dissociation of an halido ligand and formation of the transient square-planar cationic species [PtX(Me2phen)(η2-CHuCR)]+. This latter can further evolve to give an unusual, sparingly soluble square planar product where the former terminal alkyne is converted into a :CvC(H)(R) moiety with the α-carbon bridging the Pt(II) core with one of the two N-donors of coordinated Me2phen. The final product [PtX2{κ2-N,C-(Z)-N̲1–N10–C̲vC(H)(R)}] (N1–N10 = 2,9-dimethyl-1,10-phenanthroline; X = Br, I) contains a Pt–N–C–C–N–C six-membered chelate ring in a square planar Pt(II) coordination environment.

The Radio Frequency Identification (RFID) is a wireless technology that is becoming more and more important as auto-identification solution for many application scenarios. The adoption of this innovative technology in the pharmaceutical sector promises to solve several problems related to tracing and tracking systems at item level. Unfortunately, there are still some barriers limiting the large-scale deployment of RFID technologies. One of these is related to very interesting research topics on the evaluation of potential effects of electromagnetic fields on drugs. In detail, this work aimed to analyze the impact of UHF RFID devices, used in tracing systems, on the molecular structure and potency of a commercial human insulin preparation, ActrapidTM. In order to investigate possible induced alterations of molecular structure, the Reverse Phase-High Pressure Liquid Chromatography and the Nuclear Magnetic Resonance spectroscopy have been mainly used in the experimental protocol. To obtain some indications about drug performance, in vitro cell proliferation assays have been also conducted. The experimental results, achieved by a protocol combining an accurate structural analysis on 5 min to 24 h irradiated drug samples with functional in vitro assays, have shown that the electromagnetic field generated by UHF RFID devices does not cause significant effects on ActrapidTM insulin. These findings are strongly encouraging the use of RFID-based technologies for item-level tracing systems in the pharmaceutical supply chain.

The experiments here reported evidence on the importance of the residual charge of a nucleotide derivative, for the adsorption on nHAP (hydroxyapatite nanocrystals), in water solution.We found that the simple presence of phosphates on the nucleotide derivative does not guarantee adsorption on nHAP. On the other hand, we demonstrated that a cationic or neutral charge on a nucleotide derivative produces a strongly reduced chemical adsorption (chemisorption) whereas, in the presence of a net negative charge, relevant adsorption on nHAP is observed. The number of phosphates can only modulate the adsorption efficiency of a molecule provided that this latter bears an overall negative charge. The neutral zwitterionic nucleotide Pt(II) complexes, bearing negatively charged phosphates, are unable to give stable chemisorption. Previous considerations are important to model the binding ability of phosphate bearing nucleotide derivatives or molecules on hydroxyapatite. The findings reported in the present paper could be relevant in bone tissue targeting or nHAP mediated drug delivery.

Cisplatin, cis-diamminedichloroplatinum(II), is one of the most widely used antitumor drugs in cancer chemotherapy, as a critical component against a broad range of malignancies. Platinum anticancer drugs are generally known to target DNA, where they can bind to the N7 of purine bases. Cisplatin, as other bifunctional agents, is able to bind to adjacent purines, resulting in crosslink lesions believed to be responsible for the observed antitumor activity. N7-metalated purines, in some cases, seem to be characterized by a relevant antitumor activity. This has led us to hypothesize a cisplatin parallel mechanism of action: based on free platinated purines formation, directly in tissues, after drug administration. In order to evaluate this possible mechanism, as a key path to develop new drugs, we performed a series of experiments focused on platinated nucleobases cell and mitochondrial uptake and processing. For the first time cell uptake and mobility mechanisms, related to plasmatic cell and/or mitochondrial membrane crossing, has been studied. Moreover the possible insertion of metalated nucleobases into nuclear and/or mitochondrial new synthesized DNA/RNA, operated by DNA/RNA polymerases was evaluated. At this scope model metalated nucleosides/nucleotides, with nitrogen carrier ligands, have been synthesized, isolated and characterized. The possible development of new drugs based on this new rational drug design will be discussed.

To get further insight in the reaction of nucleophilic substitution upon changing the ligand trans to a eta(2)-olefin, the reactivity of some monoanionic platinum(II) complexes (trans-[PtCl2X(eta(2)-C2H4)](-), X = Cl-, 1, OH-, 2, and CH2NO2-, 3) towards pyridines with different steric hindrance (py, 4-Mepy, and 2,6-Me(2)py) has been tested. All crystallographic (2 and 3 reported for the first time) and spectroscopic data are in accord with a platinum-olefin interaction decreasing in the order 2 > 1 > 3, paralleling the decreasing electronegativity of the donor atom (O > Cl > C). Not only the platinum-olefin bond but also the bond between platinum and the ligand trans to the olefin appear to be strongest in 2 (Pt-O distance at the lower limit for this type of bond). In the reaction with py, the ligand trans to the olefin is displaced in 1 and 2. Moreover the reaction is in equilibrium in the case of sterically hindered 2,6-Me2py, the equilibrium being shifted moderately or prevalently toward the reagents in the case of 1 and 2, respectively. In the case of 3, the reaction with pyridines leads to substitution of the olefin instead of the carbanion. This is in accord with the observation that carbanions strongly weaken the trans Pt-olefin bond.

Mono-varietal extra virgin olive oils were micro-extracted from drupes that were selectively collected from 28 trees distributed in five different Southern Italian Apulian areas. Nuclear Magnetic Resonance (NMR) profiles of these oil samples were correlated to the genetic (young green material) and soil (samples collected within the foliage projection) data of the tree of origin. Genetic analysis, performed on the samples using SSRs (Simple Sequence Repeats) by 9 microsatellite loci, confirmed the specific cultivar assignment (among Cima di Mola, Coratina, Ogliarola, and Oliva Rossa cultivars). Chemometric methods applied to 1H-NMR spectroscopic data were used for cultivar and geographical origin discrimination of the studied extra virgin olive oils. Linear Discriminant Analysis (LDA) afforded a high reliability degree for discriminating cultivars (almost 90% of prediction ability), and a good assigning ability for the geographical origin (Ogliarola and Coratina samples used as subsets). Soil analyses were performed for each tree. Regression analysis was applied to soil composition in order to correlate available nutrients and total metals with the content of fatty acids and minor components present in monovarietal extra virgin olive oils. In the case of oleic and linoleic fatty acids, and for some terpenes, B, Cr, Mn, Zn were found to give significant correlations. Zn and Mn were the most significant trace elements for all the correlations found (p < 0.01). The results obtained (genetic, spectroscopic and soil analyses) are discussed as a multidisciplinary approach for setting up a strategy for a cultivar and/or geographic origin certification committed database construction.

In this work, we assessed the capacity of RNA polymerases to use platinated ribonucleotides as substrates for RNA synthesis by testing the incorporation of the model compound [Pt(dien)(N7-5'-GTP)] (dien=diethylenetriamine; GTP=5'-guanosine triphosphate) into a natural RNA sequence. The yield of in vitro transcription operated by T7 RNA polymerase, on the LacZ (Escherichia coli gene encoding for β-galactosidase) sequence, decreases progressively with decreasing the concentration of natural GTP, in favor of the platinated nucleotide, [Pt(dien)(N7-5'-GTP)]. Comparison of the T7 RNA polymerase transcription activities for [Pt(dien)(N7-5'-GTP)] compound incorporation reaction test, with respect to the effect of a decreasing concentration of natural GTP, showed no major differences. A specific inhibitory effect of compound [Pt(dien)(N7-5'-GTP)] (which may pair the complementary base on the DNA strand, without being incorporated in the RNA by the T7 RNA polymerase) was evidenced. Our findings therefore suggest that RNA polymerases, unlike DNA polymerases, are unable to incorporate N7-platinated nucleotides into newly synthesized nucleic acids. In this respect, specifically designed N7-platinated nucleotides based compounds could be used in alternative to the classical platinum based drugs. This approach may offer a possible strategy to target specifically DNA, without affecting RNA, and is potentially able to better modulate pharmacological activity.

Carbohydrate polimeric microcapsules were assembled using a LbL approach onto a CaCO3 core. The microcapsules were used to delivery the anticancer drug cisplatin into HeLa and MCF-7 cancer cell lines. Drug encapsulation, measured by ICP spectroscopy, was around 50% of the charging solution. Fluorimetric measurements showed an efficient cellular uptake of polysacchardic microcapsules in both cell lines. The drug-loaded capsules demonstrated a better efficiency against cell viability than the free drug. Specifically, the amount of platinum reaching genomic DNA was measured, showing that encapsulation improves the nuclear delivery of the drug for both cell lines.

Platinum compounds cause significant clinical neurotoxicity. Several studies highlight neurological complications especially in paediatric oncology patients with Central Nervous System (CNS) and non-CNS malignancies. To understand the toxicity mechanisms of platinum drugs at cellular and molecular levels in the immature brain, which appears more vulnerable to injury than in the adult one, we compared the effects in vivo of the most used platinum compounds, i.e., cisdichlorodiammineplatinum (cisplatin, cisPt), and the new [Pt(O,O′-acac)(γ-acac)(DMS)] (PtAcacDMS). As models of developing brain areas, we have chosen the cerebellum and hippocampus dentate gyrus. Both areas show the neurogenesis events, from proliferation to differentiation and synaptogenesis, and therefore allow comparing the action of platinum compounds with DNA and non-DNA targets. Here, we focused on the changes in the intracellular calcium homeostasis within CNS architecture, using two immunohistochemical markers, the calcium buffer protein Calbindin and Plasma Membrane Calcium ATPase. From the comparison of the cisPt and PtAcacDMS effects, it emerges how essential the equilibrium and synergy between CB and PMCA1 is or how important the presence of at least one of them is to warrant the morphology and function of nervous tissue and limit neuroarchitecture damages, depending on the peculiar and intrinsic properties of the developing CNS areas.

In this work it is described a new synthetic pathway for the synthesis of bis-cresolate Pt(II) complexes with (N-N) dinitrogen ligands. In particular, we were able, for the first time, to isolate, in the solid state, both anti-[Pt(N-N)(4-Me-C6H4O)2], N-N = 1,10-phenanthroline, (Head to Tail, HT) and syn-[Pt(N-N)(4-Me-C6H4O)2], N-N = 2,2’-bipyridile, (Head to Head, HH) rotamers, which were also characterized by single crystal X-ray diffraction

The reactivity of the Zeise’s anion with bidentate dinitrogen ligands (N-N) has been the main subject of several studies on the Pt(II) coordination and organometallic chemistry. The chemistry of Pt(II) organometallic ketonyl derivatives it is recognized to be important for many potential applications. For these reasons we exploit the possible extension of Zeise’s anion reactivity, in basic media, to the general synthesis, of organometallic ketonyl derivatives of the type [PtCl{h1-CH2C(O)R}(N-N)], even in the case of aliphatic diamines and sterically hindered diimine ligands. In these conditions we observed the formation of methastable intermediate complexes of the type trans-[PtCl2{h1-CH2C(O)R}(h2-C2H4)]-. Further reaction with dinitrogen ligands (N-N) gives [PtCl{h1-CH2C(O)R}(N-N)] complexes. In particular this new simple pathway results interesting from the synthetic point of view for the possibility to synthesize ketonyl complexes with several ketones and dinitrogen ligands not suitable for other methods.

In the [ABrnIm] (A = C, Si, Ge, Sn; n + m = 4) compounds, with the heavier halides bonded to the central IV group elements, the experimental 13C, 29Si, 73Ge and 119Sn NMR chemical shifts of the central atoms were found to be strictly linearly proportional to the bonded halides ionic radii overall sum Σ(rh). Based on this, calibration lines relating the chemical shifts to Σ(rh) could be built for the considered subgroup of [ABrnIm] compounds. Using such calibration lines we could calculate the equivalent NMR radius, NMRrH–A, attributable to each of the bonded hydrogens in [AH4] species, according to the overall NMR shielding produced on the central A atom. Interestingly, the calculated NMRrH–A value resulted to be almost constant in all [AH4] examined systems (A = 13C, 29Si, 73Ge, 119Sn) with an average NMRrH–A value equal to 194.6 ± 1.6 pm. Based on this approach, we could calculate the 207Pb NMR chemical shift of the unstable [PbH4] complex using the value of 192.7 pm calculated for NMRrH–Sn in the stable closest hydride [SnH4]. The obtained unprecedented NMR value is in accord with the 207Pb NMR chemical shift estimation, independently calculated for [PbH4] from the [SnH4] data, using the Pb/Sn chemical shift correlation defined in the Mitchell equation.

The properties of three novel Platinum(II) compounds toward the insulin-degrading enzyme (IDE) enzymatic activity have been investigated under physiological conditions. The rationale of this study resides on previous observations that these compounds, specifically designed and synthesized by some of us, induce apoptosis in various cancer cell lines, whereas IDE has been proposed as a putative oncogene involved in neuroblastoma onset and progression. Two of these compounds, namely [PtCl(O,O′-acac)(DMSO)] and [Pt(O,O′-acac)(γ-acac)(DMS)], display a modulatory behavior, wherefore activation or inhibition of IDE activity occurs over different concentration ranges (suggesting the existence of two binding sites on the enzyme). On the other hand, [Pt(O,O′-acac)(γ-acac)(DMSO)] shows a typical competitive inhibitory pattern, characterized by a meaningful affinity constant (K i = 0.95 ± 0.21 μM). Although all three compounds induce cell death in neuroblastoma SHSY5Y cells at concentrations exceeding 2 μM, the two modulators facilitate cells’ proliferation at concentrations ≤ 1.5 μM, whereas the competitive inhibitor [Pt(O,O′-acac)(γ-acac)(DMSO)] only shows a pro-apoptotic activity at all investigated concentrations. These features render the [Pt(O,O′-acac)(γ-acac)(DMSO)] a promising “lead compound” for the synthesis of IDE-specific inhibitors (not characterized yet) with therapeutic potentiality.

Synthesis and characterization of the new pentacoordinate [PtCl2(eta(2)-C2H4)(Mebpy)] and square planar [PtCl2(Mebpy)] complexes both containing the asymmetrically hindered nitrogen donor chelate Mebpy = 6-Methyl-2,2'-bipyridil are reported. By nucleophilic addition of MeO- to the coordinated ethene of [PtCl2(eta(2)-C2H4)(Mebpy)], syn-[PtCl(eta(1)-CH2CH2OMe)(Mebpy)] and anti-[PtCl(eta(1)-CH2CH2OMe)(Mebpy)] complexes have been obtained as the kinetic and thermodynamic product, respectively. The mechanism for selective formation of both stereoisomers is also reported. This synthetic route opens new perspectives for the stereospecific synthesis of syn- and anti-square planar organometallic complexes.

Strategies to control diffusion of malaria needs to account for the increase of resistance of the parasite to the conventional antimalarial drugs. It has been proposed that a traditional aqueous preparation from Artemisia annua, with a low content of the active compound, artemisinin, may reduce the risk of resistance of the protozoa and be relatively more effective in the treatment of the disease. The solubility properties of the molecule have been the matter of concern about the therapeutic usefulness of herbal teas from A. annua. The present study aimed at analysing the chemical profile of a tea infusion from A. annua. Tea from A. annua was prepared through infusion of the plant aerial parts in water for 1, 24 and 48&#xa0;h. Content of artemisinin was determined by HPLC-ELSD. Overall chemical characterization of the extracts was carried out by a combination of metabolomic techniques. The artemisinin content varied only slightly in the three different extracts (about 0.12%). A series of mono-caffeoyl- and mono-feruloyl-quinic acids, di-caffeoyl- and di-feruloyl-quinic acids was identified as main components of the tea infusion, together with some flavonoids. Reconstitution of the same extracts in less polar or apolar solvents resulted in a different composition with no phenolics and a much lower concentration of artemisinin.

PON 254/Ric. Potenziamento del “Centro Ricerche per la Salute dell’Uomo e dell’Ambiente” Cod. PONa3_00334;Consorzio Interuniversitario di Ricerca in Chimica dei Metalli nei Sistemi Biologici (CIRCMSB), Bari (Italy).

The item-level traceability is a very important requirement for many practical application scenarios, where it needs to guarantee perfect transparency for products flow along the whole supply chain. Among these, the pharmaceutical distribution is a very interesting scenario, characterized by many challenges, where, the Radio Frequency Identification (RFID) technology will play a very important role. Unfortunately, there are still some technical barriers that are retarding the deployment of these innovative technologies in large-scale. For the pharmaceutical supply chain, there have been concerns raised regarding the potential effects on the quality of drugs due to electromagnetic fields exposure. This work aimed to evaluate potential effects of tracing RFID systems on the molecular structure of biological drugs. In particular, some samples of a commercial human insulin preparation have been exposed for different periods to electromagnetic fields generated by RFID devices. In order to evaluate possible alterations on the molecular structure, the following diagnostic techniques were used: High Pressure Liquid Chromatography (HPLC) and Nuclear Magnetic Resonance (NMR). HPLC analysis demonstrated that there is are no differences between the RFID exposed samples and the control. On the contrary, a first and partial NMR analysis detected some changes on the insulin molecule spectra after one hour of exposition to the electromagnetic field. Unfortunately, this approach did not allow us to verify possible damages on the protein because of presence of expicients and low drug concentration. Further investigations, e.g. in vitro functional analysis, are required.

Abstract The influence of temperature and heating rate on the yield and properties of biochar derived from pyrolysis of solid olive mill waste (pomace) was investigated. Three pyrolysis temperatures (430 ± 10 °C, 480 ± 10 °C and 530 ± 10 °C) and 3 heating rates (25 °C min−1, 35 °C min−1 and 45 °C min−1) were studied. The biochar production was carried out using a vertical downdraft gasifier. Increasing the pyrolysis temperature, and/or the heating rate, the biochar yield lowered, the C content and biochar aromaticity increased and the surface functional groups were reduced. The highest biochar yield was obtained by low pyrolysis temperature (430 ± 10 °C) and low heating rate (25 °C min−1). This biochar is characterized by a high heating value (31 MJ/kg) that makes it a possible fuel candidate and, in the meantime, due to its high concentration in C (70.2%–84.1%), low electrical conductivity (0.28 dS m−1–0.47 dS m−1) and the lack of phytotoxicity it is suitable for amendment in agricultural soils and for long term carbon sequestration.

We used ZL55 mesothelioma cell line in order to study the cytotoxicity of cisplatin and the cellular pathways eventually ending to the apoptotic process. To this end, ZL55 cells were treated with increasing doses (1-200 μM) and for different incubation times (1-72 hours) with cisplatin and then the number of viable cells was evaluated by the MTT test. The cleavage patterns of caspase-3, -7 and -9, and Poly-ADP ribose polymerase (PARP) by cisplatin were assessed by Western methods. The results showed here suggest that activation of PKC-δ is an essential part of the apoptotic program provoked by cisplatin in mesothelioma cells and that PKC-δ transduces a pro-apoptotic signal in these cells.

A. annua was usually used to prepare a tea and if it contains effective amounts of artemisinin, it might be used today as a self-reliant treatment of malaria [1]. Artemisia annua tea has proven itself to be a very effective treatment for malaria in various clinical trials, but to date, his efficacy has not been investigated in vitro. Therefore, we have carried out a study for the evaluation of effects of A. annua tea on Plasmodium falciparum cultures in vitro. We also determined the concentration of artemisinin in herbal tea preparation. The compound was tested against chloroquine-sensitive D10 and chloroquine-resistant W2 strains of P. falciparum using the parasite lactate dehydrogenase assay [2]. The quantification of artemisinin in the extract of leaves of A. annua was obtained using an 1H NMR method. The in vitro tests conducted in this study confirm the clinical efficacy demonstrated by the tea of A. annua in vivo on both chloroquine-sensitive D10 and chloroquine-resistant W2 strains. The concentration of artemisinin in A. annua tea is lower with respect to that of pure artemisinin responsible for the same antimalarial activity. The artemisinin present in the tea is probably co-solubilised with other ingredients, some of which may also have antimalarial activity and act synergistically with it. The presence of other active ingredients suggests that A. annua is a natural artemisinin combination therapy. These compounds also merit further research, to see whether their presence hinders the development of parasite resistance compared to pure artemisinin [3]. [1] C.H.Blanke, G.B.Naisabha, M.B.Balema, G.M.Mbaruku, L.Heide and M.S.Muller, Trop. Doct., 38, 2008, 113–6. [2] M.T.Makler, J.M.Ries, J.A.Williams, J.E.Bamcroft, R.C.Piper, B.L.Gibbins, et al, Am.J.Trop.Med.Hyg., 48(6), 1993, 739-41. [3] Work supported by Regione Puglia Progetto Strategico PS70.

In this work, we studied the reactivity of the Zeise’s salt, K[PtCl3(h2-C2H4)], with ketones in the presence of strong bases. In these conditions we demonstrated, by NMR, the formation of metastable intermediate complexes of the type trans-[PtCl2{h1-CH2C(O)R}(h2-C2H4)]-. Further reaction with dinitrogen ligands (N-N) gives [PtCl{h1-CH2C(O)R}(N-N)] complexes. This reactivity of the Zeise’s anion can be exploited for the first general synthesis, in a one pot reaction of s-carbon Pt(II) ketonyl derivatives with aliphatic or aromatic dinitrogen ligands (even if sterically hindered).

The Zeise’s salt, K[PtCl3(h2-C2H4)], is a paradigmatic example of the olefin to metal bond and still an important starting material for the synthesis of platinum-based organometallic compounds. The great reactivity of Zeise’s anion, essentially related to the lability of the chlorido ligand trans to the olefin, which can be easily substituted by any ligand having a reasonable affinity for platinum. To get further insights in the reaction of nucleophilic substitution upon changing the ligand trans to the h2-olefin, we tested the reactivity of three monoanionic platinum(II) substrates (Zeise’s anion itself, [PtCl3(h2-C2H4)]-, 1, trans-[PtCl2(OH)(h2-C2H4)], 2, and trans-[PtCl2(h1-CH2NO2)(h2-C2H4)]-, 3), towards aromatic imines with different steric requirements (pyridine, 4-methylpyridine, and 2,6-dimethylpyridine). We also performed a X-ray crystal structure characterization of the tetraphenylphosphonium salts of 2 and 3. Our data have highlighted the nature of the Pt-Cl Pt-OH, and Pt-C s bonds. Cl– is the only one of the three trans-to-olefin ligands which can have a h-acceptor capacity towards the metal. The s-donor capacity of the ligands can be ranked in the order: CH2NO2 C- >> Cl– > OH–. The carbanion has an unexpectedly high weakening effect on the bond between platinum and the trans-olefin. As a consequence the olefin is displaced in preference not only by another olefin but also by h-donors such as pyridines. In the light of the present results, the series ranking the trans-effect, which were set up in the 1980’s, needs to be reconsidered in view of the importance for synthetic chemistry of trans-directing ligands.

Renal cell carcinoma (RCC) is a heterogeneous cancer often showing late symptoms. Until now, some candidate protein markers have been proposed for its diagnosis. Metabolomics approaches have been applied, predominantly using Mass Spectrometry (MS), while Nuclear Magnetic Resonance (NMR)-based studies remain limited. There is no study about RCC integrating NMR-based metabolomics with transcriptomics. In this work, 1H-NMR spectroscopy combined with multivariate statistics was applied on urine samples, collected from 40 patients with clear cell RCC (ccRCC) before nephrectomy and 29 healthy controls; nine out of 40 patients also provided samples one-month after nephrectomy. We observed increases of creatine, alanine, lactate and pyruvate, and decreases of hippurate, citrate, and betaine in all ccRCC patients. A network analysis connected most of these metabolites with glomerular injury, renal inflammation and renal necrosis/cell death. Interestingly, intersecting metabolites with transcriptomic data from CD133+/CD24+ tumoral renal stem cells isolated from ccRCC patients, we found that both genes and metabolites differentially regulated in ccRCC patients belonged to HIF-α signaling, methionine and choline degradation, and acetyl-CoA biosynthesis. Moreover, when comparing urinary metabolome of ccRCC patients after nephrectomy, some processes, such as the glomerular injury, renal hypertrophy, renal necrosis/cell death and renal proliferation, were no more represented.

Nuclear Magnetic Resonance (NMR) spectroscopy is emerging as a powerful technique in olive oil fingerprinting, but its analytical robustness has to be proved. Here, we report a comparative study between two laboratories on olive oil (1)H NMR fingerprinting, aiming to demonstrate the robustness of NMR-based metabolomics in generating comparable data sets for cultivar classification. Sample preparation and data acquisition were performed independently in two laboratories, equipped with different resolution spectrometers (400 and 500 MHz), using two identical sets of mono-varietal olive oils. Partial Least Squares (PLS)-based techniques were applied to compare the data sets produced by the two laboratories. Despite differences in spectrum baseline, and in intensity and shape of peaks, the amount of shared information was significant (almost 70%) and related to cultivar (same metabolites discriminated between cultivars). In conclusion, regardless of the variability due to operator and machine, the data sets from the two participating units were comparable for the purpose of classification.

In this work, we show by a simple empirical approach that a linear relationship between observed 195Pt NMR frequencies and the overall sum of the ionic radii of the coordinated halido ligands [Σ(rh)] exists in square-planar PtII complexes of the type [PtXnY4–n]2– (1 ≤ n≤4; X, Y = Cl, Br, I). Another finding was that such square-planar complexes could be empirically described as octahedral complexes, with the two lobes of the 5dz2 orbital above and below the coordination plane acting as two pseudo-halido ligands, each showing a constant apparent radius of around 207 pm. According to our approach, the overall apparent radius of around 415 pm produces constant 195Pt NMR shielding for all [PtXnY4–n]2– complexes of about 10450 ppm. This result is 1) consistent with the theoretically calculated overall 5d shell lone-pair shielding observed in square-planar PtII with respect to octahedral PtIV complexes and 2) almost coincident with the already measured chemical shift anisotropy (CSA) of the K2[PtCl4] complex both in solution and in the solid state (single crystal).

[Pt(O,O′-acac)(γ-acac)(DMS)] change the expression of 84 proteins as assessed by a proteomic approach in SH-SY5Y cells

BACKGROUND AND PURPOSE: We showed previously that a new Pt(II) complex ([Pt(O,O'-acac)(gamma-acac)(DMS)]) exerted high and fast apoptotic processes in MCF-7 cells. The objective of this study was to investigate the hypothesis that [Pt(O,O'-acac)(gamma-acac)(DMS)] is also able to exert anoikis and alter the migration ability of MCF-7 cells, and to show some of the signalling events leading to these alterations. EXPERIMENTAL APPROACH: Cells were treated with sublethal doses of [Pt(O,O'-acac)(gamma-acac)(DMS)], and the efficiency of colony initiation and anchorage-independent growth was assayed; cell migration was examined by in vitro culture wounding assay. Gelatin zymography for MMP-2 and -9 activities, Western blottings of MMPs, MAPKs, Src, PKC-epsilon and FAK, after [Pt(O,O'-acac)(gamma-acac)(DMS)] treatment, were also performed. KEY RESULTS: Sub-cytotoxic drug concentrations decreased the: (i) anchorage-dependent and -independent growth; (ii) migration ability; and (iii) expression and activity of MMP-2 and MMP-9. [Pt(O,O'-acac)(gamma-acac)(DMS)] provoked the generation of reactive oxygen species (ROS), and the activation of p38MAPK, Src and PKC-epsilon. p38MAPK phosphorylation, cell anoikis and migration due to [Pt(O,O'-acac)(gamma-acac)(DMS)] were blocked by PKC-epsilon inhibition. Furthermore, Src inhibition blocked the [Pt(O,O'-acac)(gamma-acac)(DMS)]-provoked activation of PKC-epsilon, while ROS generation blockage inhibited the activation of Src, and also the decrement of phosphorylated FAK observed in detached [Pt(O,O'-acac)(gamma-acac)(DMS)]-treated cells. CONCLUSIONS AND IMPLICATIONS: Sublethal concentrations of [Pt(O,O'-acac)(gamma-acac)(DMS)] induced anoikis and prevented events leading to metastasis via alterations in cell migration, anchorage independency, stromal interactions and MMP activity. Hence, [Pt(O,O'-acac)(gamma-acac)(DMS)] may be a promising therapeutic agent for preventing growth and metastasis of breast cancer.

A smart nanocarrier system for cancer therapy, based on a recently developed technique for preparing pure nanometric calcium carbonate (CaCO3), was studied. Different approaches were used to obtain sustained release of cisplatin: at first, pure CaCO3 nanoparticles were evaluated as carriers, then the nanoparticles were functionalized with polymer or silanes, and finally they were employed as a substrate to build layer by layer (LbL) self-assembled polyelectrolyte nanocapsules. Loading efficiency and release kinetics were measured. The best loadings were obtained with the LbL nanocapsules, allowing for high loading efficiency and the possibility of controlling the release rate of the drug. The behavior of all the carriers was evaluated on four neoplastic cell lines, representative of different types of neoplastic disease, namely MCF-7 (breast cancer), SKOV-3 (ovarian cancer), HeLa (cervical cancer) and CACO-2 (human epithelial colorectal adenocarcinoma). Negligible cytotoxicity of the nanoparticles, functionalized nanoparticles, and nanocapsules was observed in experiments with all cell lines. Nanocapsules were functionalized with fluorescein isothiocyanate (FITC) in order to track their kinetic of internalization and localization in the cell line by confocal laser scanning microscopy (CLSM). The cytotoxicity of the loaded capsules was evaluated, showing cell survival rates close to those expected for non-encapsulated cisplatin at the same nominal concentration.

The new b-diketonate compounds ([Pt(O,O’-acac)(gamma-acac)(NH3)] and [Pt(O,O’-acac)(g-acac)(py)]) were chemical characterized by NMR spectroscopy and preliminary cytotoxic assays of [Pt(O,O’-acac)(g-acac)(NH3)] on cisplatin resistant MCF-7 breast cancer cell line were performed. Interestingly, [Pt(O,O’-acac)(gamma-acac)(NH3)] resulted in dose dependent inhibition of cell survival comparable to [Pt(O,O’-acac)(γ-acac)(DMS) and approximately 90-fold greater than that observed for cisplatin.

In the present study, we aimed to investigate the anti-angiogenesis effects of [Pt(O,O’-acac)(γ-acac)(DMS)]. [Pt(O,O′-acac)(γ-acac)(DMS)] significantly inhibited human umbilical vein endothelial cell (HUVEC) proliferation. Then, the ex vivo rat aortic ring capillary-network sprouting has been carried out. It was observed that after one week of treatment under regular growth conditions, [Pt(O,O′-acac)(γ-acac)(DMS)] potently inhibited the sprouting as well as capillary-network formation from rat aortic ring in a dose-dependent manner. Compared to control 0,5 and 1 μM doses of [Pt(O,O′-acac)(γ- acac)(DMS)] suppressed capillary-network formation by 68 to 100%, respectively. In addition, in mouse xenograft model of breast cancer, treatment of mice with [Pt(O,O′-acac)(γ-acac)(DMS)], decrease MMP-1 content in tumours. These observations demonstrated that [Pt(O,O′- acac)(γ-acac)(DMS)] could inhibit angiogenesis, which warrants further studies in other in vivo models.

In the field of experimental oncology, many efforts are being carried out to search new platinum-based drugs overcoming the CNS toxicity and drug resistance. One of the adopted strategies is the synthesis of platinum compounds able to form Pt-DNA adducts different from the cisplatin ones or to react with other subcellular targets. In this context a novel Pt(II) complex, [Pt(O,O'-acac)(γ-acac)(DMS)](PtAcacDMS), was synthesized which reacts preferentially with protein thiols or thioethers. In this work we investigated the in vivo effects of cisplatin and PtAcacDMS on normal development. Moreover, to verify the dose-dependence of the effects, different groups of animals were treated with 5 μg/g or 10 μg/g body weight of cisPt and PtAcacDMS. We have focused our attention on the cerebellum because it provides a useful model system to evaluate the outcomes of perinatal treatment with chemotherapeutic agents on key CNS developmental processes such as neural cells proliferation, migration and differentiation. We have demonstrated the ability of both cisPt and PtAcacDMS to reach the brain tissue once injected. The brain platinum content after PtAcacDMS treatment was notably higher (approximately 4-fold as much) than after cisPt. The platinum accumulation in the brain was still considerable 7 days after PtAcacDMS administration. However, compared with cisplatin, PtAcacDMS induces less severe changes on fundamental events of neuroarchitecture development, such as no high apoptotic events, less altered granule cell migration and Purkinje cell dendrite growth, suggesting a low neurotoxicity of this new Pt complex for normal CNS. The mild damages could be attributable to the different subcellular target of this compound as well as to a greater efficiency of the cell repair system to recognize the drug-target adducts and to repair them. Together with the previously demonstrated antineoplastic effectiveness in vitro, the findings here reported suggest PtAcacDMS as a potential alternative to cisplatin indicating, at the same time, that the choice of platinum compounds with new subcellular targets could be a strategy to prevent neurotoxicity induced by cisplatin and overcome drug resistance induced by mutations in the intrinsic apoptotic pathway.

In recent years seaweeds increasingly attracted interest in the search for new drugs and have been shown to be a primary source of bioactive natural products including antibiotics. In the present investigation the antimicrobial activity of Gracilariopsis longissima lipidic extract was assayed and its chemical characterization was carried out by means of advanced analytical techniques such as gas-chromatography and multinuclear and multidimensional NMR spectroscopy. G. longissima lipidic extract showed an antibacterial activity against several Vibrio species. These results are interesting considering both the resistance against antibiotics developed by vibrios and the need to control fish and shellfish diseases due to vibriosis. Analysis of fatty acid methyl esters performed by gas-chromatography showed that palmitic acid methyl ester (16:0) was the predominant saturated fatty acid (42%), while, among monounsaturated fatty acids, oleic acid methyl ester (18:1) prevailed (8.5%). Because the palmitic acid represents the main component of fatty acids we hypothesized its involvement in the antibacterial activity observed. However, a pure sample of palmitic acid did not show an antibacterial activity. The fatty acid profile of G. longissima revealed also an interesting composition in polyunsaturated fatty acids and in particular the ratio of omega-3 to omega-6 fatty acids was >1 thus suggesting that this macroalga may be used as a natural source of omega 3. Moreover, the H-1 NMR spectrum in CDCl3 of algal lipid fraction shows the characteristic signals of saturated and unsaturated fatty acids as well as other metabolites. Interestingly, in the lipid extract the presence of polyhydroxybutyrate, a linear biodegradable and biocompatible polyester, was clearly identified by NMR spectroscopy. In conclusion, the lipidic extract of G. longissima on account of its antimicrobial activity, nutritional value and content in biodegradable and biocompatible polyester represents an interesting potential biotechnological resource.

It was previously demonstrated that [Pt(O,O0-acac)(g-acac)(DMS)] exerted toxic effects at high doses, whilst sub-cytotoxic concentrations induced anoikis and decreased cell migration. Aim of this study was to investigate the hypothesis that [Pt(O,O0-acac)(g-acac)(DMS)] alters the [Ca2+]i and that this is linked to its ability to trigger rapid apoptosis in MCF-7 cells. Thus, cells were treated with [Pt(O,O0-acac)(gacac)( DMS)] and its effects on some of the systems regulating Ca2+ homeostasis were studied, also in cells dealing with the complex changes occurring during the Ca2+ signalling evoked by extracellular stimuli. [Pt(O,O0-acac)(g-acac)(DMS)] caused the decrease of PMCA activity (but not SERCA or SPCA) and Ca2+ membrane permeability. These two opposite effects on [Ca2+]i resulted in its overall increase from 102 12 nM to 250 24 nM after 15 min incubation. The effects of [Pt(O,O0-acac)(g-acac)(DMS)] were also evident when cells were stimulated with ATP: the changes in Ca2+ levels caused by purinergic stimulation resulted altered due to decreased PMCA activity and to the closure of Ca2+ channels opened by purinergic receptor. Conversely, [Pt(O,O0-acac)(g-acac)(DMS)] did not affect the store-operated Ca2+ channels opened by thapsigargin or by ATP. [Pt(O,O0-acac)(g-acac)(DMS)] provoked the activation of PKC-a and the production of ROS that were responsible for the Ca2+ permeability and PMCA activity decrease, respectively. The overall effect of [Pt(O,O0-acac)(g-acac)(DMS)] is to increase the [Ca2+]i, an effect that is likely to be linked to its ability to trigger rapid apoptosis in MCF-7 cells. These data reinforce the notion that [Pt(O,O0-acac)(gacac)( DMS)] would be a promising drug in cancer treatment

Recently, there is a growing interest towards the development of strategies for invasive seaweed control and exploitation as source of secondary metabolites. Here, we investigated the potential of exploitation in biotechnology and recycling options in eradication programs of the lipidic extract of the Mediterranean invasive seaweed Caulerpa cylindracea (Chlorophyta). The chemical characterization was carried out by means of multinuclear and multidimensional NMR spectroscopy. The fatty acid profile of C. cylindracea assessed the presence of several types of molecules known for antioxidant activity such as carotenoids, chlorophylls, pheophytins, and sterols. The NMR spectroscopy showed also the characteristic signals of saturated, unsaturated, and free fatty acids as well as other metabolites including the biopolymer polyhydroxybutyrate. The lipidic extract exerted an antioxidant activity corresponding to 552.14 ± 69.13 mmol Trolox equivalent/g (ORAC) and to 70.3 ± 2.67 mmol Trolox equivalent/g (TEAC). The extract showed an antibacterial activity against several Vibrio species, suggesting its potential use in the control of diseases in mariculture. Our results show that C. cylindracea, representing a critical hazard in coastal areas, could be transformed into a gain supporting specific management actions to reduce the effects of human pressures.

It was previously shown that [Pt(O,O'-acac)(γ-acac)(DMS)] induces apoptosis in various cancer cells and exerts antimetastatic responses in vitro. In rats, [Pt(O,O'-acac)(γ-acac)(DMS)] reaches the central nervous system in quantities higher than cisplatin causing less excitotoxicity. The aim of the present paper was to investigate whether [Pt(O,O'-acac)(γ-acac)(DMS)] is able to exert cytotoxic effects on SH-SY5Y human neuroblastoma cell line, and to study the intracellular transduction mechanisms underlying these effects. Here we have demonstrated that [Pt(O,O'-acac)(γ-acac)(DMS)] was more effective than cisplatin in provoking apoptosis characterized by: (a) mitochondria depolarization, (b) decrease of Bcl-2 expression and increase of BAX expressions with cytosol-to-mitochondria translocation, (c) activation of caspase-7 and -9 and (d) generation of reactive oxygen species (ROS). [Pt(O,O'-acac)(γ-acac)(DMS)] provoked the activation of the following signalling kinases that were interacting with each other: PKC-δ and -ɛ, ERK1/2, p38MAPK, JNK1/2, NF-κB, c-src and FAK. We found that ROS generated by NADPH oxidase was responsible for the [Pt(O,O'-acac)(γ-acac)(DMS)]-mediated PKC-δ and -ɛ activation and consequential phosphorylation of all MAPKs. [Pt(O,O'-acac)(γ-acac)(DMS)]-induced mitochondrial apoptosis was blocked when p38MAPK and JNK1/2 were inhibited, whilst the effects on Bax/Bcl-2 mRNA and protein levels were blocked inhibiting NF-κB. NF-κB nuclear translocation was blocked inhibiting MEK1/2 activity. In addition to the induction of apoptosis [Pt(O,O'-acac)(γ-acac)(DMS)] downregulated pro-survival pathway. Survival inhibition started from mitochondrial ROS generation which induced c-src, FAK and Akt activation. In conclusion, our results suggest that [Pt(O,O'-acac)(γ-acac)(DMS)] may be considered a promising compound for the treatment of neuroblastoma. Further studies are warranted to explore in detail the therapeutic potential of this compound.

Cisplatin, cis-diamminedichloroplatinum(II), is an effective antitumor drug known to exert its action by binding to the N7 of purine bases in nuclear DNA, consequently impairing replication, transcription and repair processes. Recently we have demonstrated, by the model complex [Pt(dien)(N7-5'-dGTP)] (dien = diethylenetriamine) the possible existence of an alternative mechanism for cisplatin antitumor activity, in which the metal is bonded to free purine bases, before incorporation in the newly synthesized DNA. This evidence suggests a new possible approach for the development of N7-platinated purines, as new antitumor drugs, for the treatment of cancer diseases. However, it is known that the efficacy of a new drug it is strictly related upon the transport across the cells plasma membrane barrier which strongly condition the drug pharmacokinetic and the targeted cells bioavailability. In particular in this study, we tested the possibility that N7-platinated purines could be transported into cells. At this scope we synthesized model Pt(II) coordination compounds, of the type [Pt(dien)(N7-G)] (G = Guanine derivative) and analysed the uptake of each single platinated nucleoside/nucleotide into Hela cells, by measuring Pt accumulation, by Inductively Coupled Plasma-Atomic Emission Spectrometry (ICP-AES). The quantitative analysis of Pt has provided a strong evidence for transport of the N7 platinated nucleoside [Pt(dien)(N7-dGuo)] into Hela cells (see figure). Moreover, the observed intracellular Pt accumulation was strictly sodiumdependent suggesting a carrier-mediated uptake for the [Pt(dien)(N7-dGuo)]. Overall, these results suggest that N7-platination does not compromise cellular uptake of nucleosides and may have important implications in the development of new potential nucleoside analogues in cancer treatment.

The results of the present study suggest that DmTpc1 is actively implicated in the specific uptake of free cytoplasmic Pt bonded nucleotides, and therefore could be linked to the mechanism of action of some platinum-based antitumor drugs. Although DmTpc1 has a low affinity for model [Pt(dien)(N7-5′-dGTP)] and cis-[Pt(NH3)2(py)(N7-5′-dGTP)] compared to dATP it's well known that DNA platination level of few metal atoms per double-stranded molecule may account for the pharmacological activity of platinum based antitumor drugs. This is the first investigation where it has been demonstrated that a mitochondrial carrier is directly involved in the transport of metalated purines related with the cisplatin mechanism of action. Moreover it is shown as a lower hindrance of nucleotide bonded platinum complexes could strongly enhance mitochondrial uptake. Furthermore, a new application of ICP-AES addressed to measure the transport of metalated nucleobases, by using a recombinant protein reconstituted into liposomes, has been here, for the first time, developed and compared with a standard technique such as the liquid scintillation counting.

Abstract: Volatile metabolites from mold contamination have been proposed for the early identification of toxigenic fungi to prevent toxicological risks, but there are no such data available for Fusarium poae. F. poae is one of the species complexes involved in Fusarium head blight, a cereal disease that results in significant yield losses and quality reductions. The identification of volatile organic compounds associated with F. poae metabolism could provide good markers to indicate early fungal contamination. To this aim, we evaluated the volatile profile of healthy and F. poae-infected durum wheat kernels by SPME-GC/MS analysis. The production of volatile metabolites was monitored for seven days, and the time course analysis of key volatiles was determined. A total of 29 volatile markers were selected among the detected compounds, and multivariate analysis was applied to establish the relationship between potential volatile markers and fungal contamination. A range of volatile compounds, including alcohols, ketones, esters, furans and aromatics, were identified, both in contaminated and in healthy kernels. However, the overall volatile profile of infected samples and controls differed, indicating that the whole volatile profile, rather than individual volatile compounds, could be used to identify F. poae contamination of durum wheat grains.

Pt prodrugs: We synthesized new cationic complexes [PtCl(η2-C2H4)(R,R-chxn)]Cl (1) and [PtCl(η2-C2H4) (S,S-chxn)]Cl (2), which are organometallic analogues of the drug oxaliplatin. Complexes 1 and 2 can be considered antitumor prodrugs, as we demonstrate that they can decompose to give the same metabolites as those of oxaliplatin.

Abstract The single crystal X-ray structure of the pentacoordinate complex [PtBr2(η2-CH2double bond; length as m-dashCH2)(Me2phen)], Me2phen = 2,9-dimethyl-1,10-phenanthroline, is here reported for the first time. Comparison of the complete series of [PtX2(η2-CH2double bond; length as m-dashCH2)(Me2phen)] (X = Cl, Br, I) X-ray structures shows a very low variability of the bond lengths and angles, in the trigonal equatorial plane (where η2-olefin and Me2phen are bound), on varying the coordinated axial halogens. In first approximation, this suggests describing as independents and not interacting the two subsystems constituted by the metal bonds with axial (X–Pt–X) and equatorial ligands (Me2phen–Pt–η2-ethene). This means that the electric charge donated to the metal, by the axial ligands, cannot substantially modify the bonds of the metal with the ligands in the trigonal equatorial plane. The 1H, 13C, 15N and 195Pt NMR chemical shifts variations, studied as a function of the ionic radius of the axial halides are here discussed. The NMR data strongly suggest the existence of electric pseudo-ring currents circulating around the Pt–X axes and modulated by the ionic radius of the coordinated halides.