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AimsPlant cover and microclimatic conditions can profoundly alter the balance between productivity and decay, with relevant effects on soil C and N pools. In this contest, the aim of the present study was to assess how, in Mediterranean areas, soil properties and C and N sequestration differs between afforested pine forests and natural shrublands in different microclimatic conditions at low and high elevations.MethodsThe study was performed along the soil profile distinguishing between the organic layers, fermentation and humic layers, and surface mineral soils. The comparison between plant covers and elevations were carried out for C and N pools and soluble fractions, molecular characterization by solid state 13C NMR of organic layers and by 1HNMR of soil soluble fractions, potential mineralization rates and microbial and fungal amounts.ResultsOur data confirm that coniferous tree species sequester C faster than shrubs and herbaceous species especially at low elevation under favourable microclimatic conditions. Soil C and N pools reflect changes in the chemical composition of the upper organic layers and of soil soluble organic matter. In pine forests, the higher concentration of N in the upper organic layer speeds up the N loss in the fermentative layer and stimulates humus formation and C accumulation at low elevations.ConclusionsPlant cover and microclimatic conditions drive the C sequestration rate and the soil organic matter stability. Chemical changes highlighted by nuclear magnetic resonance spectroscopy can clarify patterns of decay processes and help to make predictions in a climate change scenario.

Neglected and underutilized species (NUS) offer largely unexplored opportunities for providing nutritious plant food, while making agro-ecosystems more diverse and resilient to climate change. The aim of this work was to explore the potential of two typical Mediterranean underutilized species, purslane and borage, as novel vegetable product (microgreens). Micro-scale production of edible plants is spreading due to the simplicity of their management, rapid cycle, harvest index, and phytochemical value of the edible product. Microgreens, therefore, represent an opportunity to link NUS, nutrition, and agricultural and dietary diversification. By analyzing yield, antioxidants activities, mineral composition, and main phenolic acids and flavonoids, our work indicated that the two species provide interesting results when compared with those reported for crops and horticultural species. Specifically, purslane should be considered highly nutritional due to the amount of phenolic compounds and ascorbic acid, and to potential good β-carotene bioavailability. Borage microgreens have a very high fresh yield and a more composite and balanced phenolic profile. In conclusion, our work provided evidence for implementing new ways to expand the NUS market-chains and for developing added-value food products.

Curcumin is a bioactive natural compound with a wide range of pharmacological properties, including antitumor activity; however, its clinical application has been limited because of its low solubility, stability, and bioavailability. In this study, a solid phase approach was proposed for the combinatorial synthesis of a mini library of the mimics of curcumin in good purity and yield. The non-effective findings in pancreatic cancer cells switched to strong growth inhibition and cell death efficacy for PC3 prostate cancer cells, and mimic 9, in which tyrosol (TYR) and homovanillyl alcohol (HVA) units were linked by a phosphodiester bond, was quite effective not only in cell growth inhibition but also in causing strong cell death under the study conditions and treatments that were not effective in PANC1 cells. The results got more exciting when we also consider the findings in SW480 human colorectal carcinoma cell line, where the growth inhibitor effects were more in line with that of the PC3 cells, but the lack of cell death effect was more in line with the PANC1 cells.

The global world olive oil industry produces large amounts of wastes and wastewaters. However, the disposal of these polluting by-products is a significant environmental problem owing to their high polyphenolic compounds. In order to learn possible ways of exploiting this waste, we investigated the phytochemical profiles, the phytotoxicity and the antioxidant activities of olive wastes (leaves, mill wastewater (OMWW) and mill solid waste (OMSW)). Leaf extracts exhibited the highest phytotoxic and antioxidant effects, which are mainly due to its phenolic content. The chromatographic profiles depicted eleven phenolic compounds belonging to secoiridoids, phenyl alcohols, phenolic acids, phenylethanoid glycoside, and flavonoids. Hydroxytyrosol and oleuropein were the main components in leaf methanol and OMWW extracts, respectively, exhibiting an amount of 13.05 and 4.39 mg/g DW. These results will help to demonstrate the possibility of utilizing olive wastes, particularly the leaf, as an inexpensive natural product, in food and agricultural applications. Graphic Abstract: [Figure not available: see fulltext.].

Afforestation of new unconsolidated volcanic deposits is a practice used to stabilize barren areas and enhance the accumulation of organic matter in the developing soil. Changes in soil carbon (C) and nitrogen (N) pools, including the soluble and microbial fractions, within the first decades since afforestation have been poorly investigated. Therefore the objective of the present study was to investigate how key C and N pools vary in litter and soil of four forests planted on barren volcanic deposits from recent Mount Vesuvius eruptions. We examined three forest stands (40, 70 and 100 years old) afforested with Stone pine (Pinus pinea L.) and a 40-year old forest of Black pine (Pinus nigra Arn.). As a baseline of C and N pools prior to afforestation, data from treeless sites were included in the study. Both the inputs with litter fall and soil C and N stocks increased with forest age in the Stone pine stands. In the mineral soil, C concentration per gram soil dry weight and C:N ratio increased with age from treeless sites to the oldest forest. Microbial biomass C and fungal biomass as a fraction of organic carbon (OC) and respiration per unit OC (an index of organic matter mineralization potential) decreased significantly with stand age. The results suggest that a main driver of C accumulation in the mineral soil is the decline with increasing stand age of the microbial fraction of organic matter and its activity. The comparison between the two pine species revealed that litter production was more abundant in the Black pine than in the even-aged, 40-year-old, Stone pine stand; moreover Black pine litter was more acidic and had a higher stable residue than Stone pine litter. Therefore a different pattern of C sequestration occurs with a higher C stock in the organic layers and a lower C stock in the mineral soil of Black pine compared to Stone pine.

Polyphenols are active compounds from medicinal plants widely distributed in nature. Many reports suggest that the mechanisms by which plant polyphenols exert their protective actions against various diseases are in part due to their redox properties [1]. Silibinin, belonging to flavonolignan class, is the major component of Silymarin complex extracted from the fruits of milk thistle (Silybum marianum) [2]. Silibinin is a diastereoisomeric mixture of two flavonolignans, Silybin A and Silybin B, in a ratio of approximately 1:1; this metabolite has long been studied for its large variety of pharmacological properties ranging from the antioxidant to neuroprotective [3] and antiviral activities. Unfortunately, many drug candidates including Silibinin, even if have been reported to possess a strong therapeutic potential in vitro, fail in vivo because of their poor bioavailability, often connected to a low water solubility. The design and synthesis of water-soluble pro-drugs are a powerful approach in order to overcome these limits and to improve the pharmacological potentialities of these compounds [4]. In this regard, here, we report the synthesis and characterization of new phosphodiester Silibinin conjugates by the insertion of a phosphate group at the 9” position. This, besides improving the Silibinin water solubility, allows conjugation of different molecules (Figure) able to recognize cell targets and to improve Silibinin pharmaceutical properties [5-8]. The new derivatives have been tested in different radical scavenging assays. A antiviral and neuroprotective activities have also been explored.

The abnormal deposition of Aβ amyloid deposits in the brain is a hallmark of Alzheimer's disease (AD). Based on this evidence, many current therapeutic approaches focus on the development of small molecules halting Aβ aggregation. However, due to the temporary and elusive structures of amyloid assemblies, the rational design of aggregation inhibitors remains a challenging task. Here we combine ThT assays and MD simulations to study Aβ aggregation in the presence of the natural compounds tyrosol (TY), 3-hydroxytyrosol (HDT), and 3-methoxytyrosol (homovanillyl alcohol - HVA). We show that albeit HDT is a potent inhibitor of amyloid growth, TY and HVA catalyze fibril formation. An inspection of MD simulations trajectories revealed that the different effects of these three molecules on Aβ1-40 aggregation are ascribable to their capacity to arrange H-bonds network between the ligand (position C-3) and the peptide (Glu22). We believe that our results may contribute to the design of more effective and safe small molecules able to contrast pathogenic amyloid aggregation.

Recently, many studies highlighted the consistent finding of irbesartan in effluents from wastewater treatment plants (WWTPs) and in some rivers and lakes in both Europe and North America, suggesting that no N80% can be removed by specific treatments. The present investigation attempts to study the chemical fate of irbesartan in a simulated chlorination step, mimicking the conditions of a WWTP. A total of six disinfection by-products were identified, five were completely new, and separated on a C-18 column by employing a gradient HPLC method. Initially, a complete mass fragmentation pathway of the drug was established with the help of MS/ TOF, and subsequently, the disinfection by-products were subjected to MS/TOF mass studies to obtain their mass and fragment pattern. The MS results helped to assign tentative structures to the disinfection products, which were verified through 1D and 2D NMR experiments. The chemical structures of the new compounds have been justified by a proposed mechanism of formation. A preliminary ecotoxicity assessment with the crus- tacean Daphnia magna showed that some of the identified by-products were up to 12-times more toxic than irbesartan.

Beer is one of the most consumed alcoholic beverages in the world, rich in chemical compounds of natural origin with high nutritional and biological value. It is made up of water, barley malt, hops, and yeast. The main nutrients are carbohydrates, amino acids, minerals, vitamins, and other compounds such as polyphenols which are responsible for the many health benefits associated with this consumption of drinks. Hops and malt are one of the raw materials for beer and are a source of phenolic compounds. In fact, about 30% of the polyphenols in beer comes from hops and 70%-80% from malt. Natural compounds of foods or plants exert an important antioxidant activity, counteracting the formation of harmful free radicals. In the presence of an intense stressing event, cells activate specific responses to counteract cell death or senescence which is known to act as a key-task in the onset of age-related pathologies and in the loss of tissue homeostasis. Many studies have shown positive effects of natural compounds as beer polyphenols on biological systems. The main aims of our research were to determine the polyphenolic profile of three fractions, coming from stages of beer production, the mashing process (must), the filtration process (prehopping solution), and the boiling process with the addition of hops (posthopping solution), and to evaluate the effects of these fractions on Dental-derived Stem Cells (D-dSCs) and human intestinal epithelial lines (Caco-2 cells). Furthermore, we underline the bioavailability of beer fraction polyphenols by carrying out the in vitro intestinal absorption using the Caco-2 cell model. We found an antioxidant, proliferating, and antisenescent effects of the fractions deriving from the brewing process on D-dSCs and Caco-2 cells. Finally, our results demonstrated that the bioavailability of polyphenols is greater in beer than in the control standards used, supporting the future clinical application of these compounds as potential therapeutic tools in precision and translational medicine.

Diclofenac (DCF) is the most widely prescribed non-steroidal anti-inflammatory drug in the world and it has been detected in drinking and surface waters. In this paper, the effect of chlorination process on DCF in aqueous solutions was investigated and the structures of 14 isolated degradation by-products (DPs), of which nine are new, have been determined from combining mass spectrometry and nuclear magnetic resonance data and justified by a proposed mechanism of formation beginning from the parent drug. Some degradation by-products show only one phenyl, others are dimers or trimers of the parental compound, which has undergone oxidative decarboxylation of the side chain and/or chlorination of this or one or both aromatic rings. Ecotoxicological bioassays evidenced the following sensitivities D. magna < R. subcapitata < A. fischeri. The isolated DPs (DP1−8, except for DP9) exhibited effects ≥ 50 % in the exposed microalgae and crustaceans showing toxicities mainly ranked from slight to acute.

Plant cover and microclimatic conditions can profoundly alter the balance between productivity and decay, with relevant effects on soil carbon (C) and nitrogen (N) pools. Despite their importance, the combined effects of plant cover and microclimatic conditions on soil C and N pools have been poorly assessed, especially in the Mediterranean environment. Therefore, the aims of the present study were to assess how, in Mediterranean areas, C and N pools differ between afforested pine forests and natural shrublands and to highlight the different impacts of plant cover on C and N sequestration at low and high elevations Along a forest floor-mineral soil continuum, plant cover and elevation effects were evaluated through measurements of C and N pools and water-soluble fractions, molecular characterization by 13C and 1H NMR, and microbial and fungal amounts and activities. Our data show that C accumulated more in afforested pine forest soils than in shrubland soils, especially at low elevations. In pine forests, the higher content of aromatic and O alkyl compounds in the upper organic layers and the abundance of aromatic and carboxylic components in the soluble fraction suggest a greater stability of soil organic matter than in shrublands. Additionally, the high concentration of N in the upper organic layer and its reduction in the fermentative layer stimulate soil C accumulation mainly in pine forests at low elevations. The abundant organic mass at high elevations in pine forests improves microbial growth, whereas the greater recalcitrance of organic residues at low elevations in pine forests leads to a decrease in the bacterial component compared with the fungal component. In conclusion, the plant cover effect appears to be strongly conditioned by elevation, and afforestation with pine at low elevations could favour long-term soil C storage.

In the last decade, a large number of evidences points to the misfolding, aggregation and accumulation of structurally abnormal proteins, termed amyloid, as a common pathogenic mechanism of a range of increasingly common human disorders, including Alzheimer`s Disease (AD) and type II diabetes mellitus (T2DM). In particular, AD and T2DM are characterized by the accumulation of the amyloid-β (Aβ) peptide in neuronal tissues and islet amyloid polypeptide (IAPP) in pancreatic cells, respectively (1), implicitly suggesting that targeting protein misfolding and self-assembly would cure the diseases. However, the failure of all clinical trials focusing on anti-aggregating drugs has clearly demonstrated that a deeper understanding of the phenomena involved in proteome maintenance is needed. It is widely known that cells express an integrated array of proteolytic machineries that control protein homeostasis (proteostasis). As a matter of fact, the latest studies suggest that pathological conditions occur when the equilibrium between the production and the clearance of the involved protein results unbalanced. The ubiquitin–proteasome system (UPS) is the major quality control pathway responsible for cellular homeostasis; it is the primary proteolytic route for misfolded proteins and ubiquitination is utilized as a degradation signal (2). Many researchers have screened a number of molecules using the whole UPS as target (proteasome activators, inhibitors of deubiquitinating enzymes DUBs or ubistatins); however, very few molecules, if any, selected by using this strategy have shown to have the potential to be pipelined to clinical trials. But, due to the multifactorial nature of protein diseases, it may be difficult to find an effective drug by screening molecules on a single target. This study evaluates the ability of silybins A and B, components of the natural product silymarin, to rescue the proteostatic balance of the amyloidogenic peptides Aβ and IAPP against several targets: proteasome, polyubiquitination, membrane protection, amyloid aggregation and oxidative stress. The obtained results evidence the important role of the stereochemistry, demonstrating the high potential of these natural compounds in anti-AD therapeutics. Our multi-target strategy to rescue proteostasis opens the door to a new approach to face these important pathologies.

During the last years, the widespread occurrence of personal care products, pesticides and stimulating drugs in water has gained much attention, especially for the evaluation of the environmental risk based on the calculation of the Predicted Environmental Concentrations (PEC) obtained from consumption data [1]. Data on the occurrence of these emerging pollutants were reported by Xing et al. (2018) in sewage treatment plant influents, effluents and sludge in many countries of the world, where they were found until to hundreds ng/L concentrations [2]. Often the products obtained by disinfection treatment could be more toxic than parent compounds especially using the chlorination process [3]. Moreover, the potential risk posed by these pollutants could be increased by their susceptibility to produce photo-transformation products [4]. Tramadol is a synthetic, centrally acting analgesic agent used for the relief of acute and chronic pain, which has been used in both human and veterinary medicine. Recent papers have reported a study on the increasing tramadol utilization associated to the mortality over the past 15 years in the UK, highlighting the high toxicity of this drug [5]. Our research is focused on the evaluation of the environmental effects of tramadol and its transformation products, mimicking the most common experimental conditions in wastewater treatment plants. After disinfection treatment, the main transformation products have been isolated and characterized by NMR and ESI-MS analyses. Furthermore, in order to evaluate the environmental eects of products, aquatic acute and chronic toxicity have been tested on Brachionus calyciorus and Ceriodaphnia dubia as well as their mutagenesis and genotoxicity tested on bacterial strains.

In the last years, G-rich oligonucleotides (GROs) able to form G-quadruplexes, have attracted considerable interest in biological and therapeutic fields. G-quadruplexes are among the most studied DNA structures because of the large variety of their biological properties ranging from anticancer to antiviral activities.(1) Recently, new findings on the anti-HIV-1 activity of ODNs forming G-quadruplex have been reported.(2) In this frame, here it is reported the synthesis of a mini-library of 5' and 3'-modified oligonucleotides with the Riboflavin (Vitamin B). The choice to insert the Riboflavin into lead sequence, has been inspired by the similarity with the hydrogen bond pattern of guanine, in addition to its excellent fluorescence properties that may be an useful tools for in vitro and in vivo studies. In order to allow the Riboflavin moiety similar to a nucleoside unit, the ribityl chain is rigidified by 2',4'-O-benzylidene group. The synthetic strategy is focused on the synthesis of two key intermidiates, the Riboflavin-Nucleoside phosphoramidite (I), useful for the synthesis of the 5'-end modified sequences and the CPG (Control Pore Glass) support anchoring the 5'-O-DMT-2',4'-O-benzyldene-riboflavin unit (II), fruitful for the synthesis of 3'-modified sequences. Using the phosphoramidite coupling automated protocol, the building block I was incorporated into oligonucleotides with a good yield. The new modified ODNs were assembled by standard automated DNA synthesis and after RP-HPLC purifications, they were analyzed by MALDI-TOF. Preliminary biophysical studies (CD, DSC and fluorescence studies) were carried out in order to evaluate the ability of modified ODNs to fold into G-quadruplex structures. Finally, their anti-HIV activity were evaluated.

Agrocybe aegerita (V. Brig.) Singer, commonly known as Pioppino, is a popular edible mushroom, known in the Campania Region (Italy). Despite its habitual consumption, little nutritional and biochemical information is available. Thus, nutritional values, anti-radical properties and chemical composition of the wild Pioppino were compared to those of the cultivated Agaricus bisporus (J.E. Lange) Imbach (known as Champignon), equally analysed.; Results: Macronutrient components (proteins, carbohydrates and lipids), free and protein amino acids and fatty acid content of poplar mushroom were achieved. Total phenol content of a defatted Pioppino alcoholic extract (PM) was determined, whereas DPPH and ABTS methods were applied to determine the radical scavenging capabilities of the extract. Ferricyanide and ORAC-fluorescein methods were also performed. Finally, LC-HRMS was used to identify and quantify the main metabolites in the extract. PM was mainly constituted of disaccharides, hexitol derivatives and malic acid. Coumaric acid isomers and C6 C1 compounds were also detected.; Conclusion: All data revealed that wild Pioppino is an excellent functional food, by far exceeding that of the Champignon. Therefore, these data are useful to promote the consumption of this mushroom encouraging thus its biological cultivation, due to wild availability is strongly compromised by the extensive use of fungicides. © 2017 Society of Chemical Industry.; © 2017 Society of Chemical Industry.

The aim of this work is to compare new and traditional extraction methods to obtain silymarin from

Advanced analytical data on microgreens' response to different light spectra constitutes a valuable resource for designing future crop-specific spectral management systems. The current study defined variation in productivity, nutritive and functional quality (mineral-carotenoid-polyphenolic profiles and antioxidant capacity) of novel microgreens (amaranth, cress, mizuna, purslane) in response to select spectral bandwidths (red, blue, blue-red), and appraised clustering patterns configured by the genotype-light-spectrum nexus. Growth parameters dependent on primary metabolism were most favored by blue-red light's efficiency in activating the photosynthetic apparatus. Nitrate accumulation was higher under monochromatic light owing to the dependency of nitrite reductase on the light-driven activity of PSI, most efficiently promoted by blue-red light. Although mineral composition was mostly genotype-dependent, monochromatic red and blue lights tended to increase K and Na and decrease Ca and Mg concentrations. Lutein, β-carotene, and lipophilic antioxidant capacity were generally increased by blue-red light putatively due to the coupling of heightened photosynthetic activity to increased demand for protection against oxidative stress; the disparate response however of purslane highlights the importance of genotype specificity in these responses and calls for additional investigation. Analysis of polyphenols by Orbitrap LC-MS/MS revealed substantial genotypic differences. Most abundant phenolics were chlorogenic acid ( x ¯ = 5503 µg g-1 dw), feruloylquinic acid ( x ¯ = 974.1 µg g-1 dw), and caffeoyl feruloyl tartaric acid ( x ¯ = 993 µg g-1 dw). Hydroxycinnamic acids accounted for 79.0% of the mean total phenolic content across species, flavonol glycosides for 20.7%, and flavone glycosides for 0.3%. The general response across species was a decrease in individual polyphenolic constituents, particularly flavonol glycosides, and total polyphenols under blue-red light. The pronounced effectiveness of monochromatic blue light in eliciting synthesis of flavonoids could be linked to their capacity for absorbing shorter wavelengths thereby quenching generated photo-oxidation potential. The light-induced stimulation of the phenylpropanoid pathway by monochromatic blue light through epigenetic mechanisms or redox signaling in the photosynthetic apparatus warrants further investigation. The current work highlights how optimized genetic background combined with effective light management might facilitate the production of superior functional quality microgreens.

The aim of this work is to compare new and traditional extraction methods to obtain silymarin from Silybum marianum, a biennial herbaceous plant of the Asteraceae family, present throughout the Mediterranean basin and used to treat several diseases. Silymarin primarily contains flavonolignans and flavonoids and is used in some pharmaceutical preparations to improve of liver function and as a protective against some hepatotoxins. In six extracts obtained by new and traditional extraction methods, the total contents of silymarin and its main flavonolignans, total phenols and condensed tannins were evaluated in addition to their respective antioxidant capacities. By the Naviglio extractor, that is a rapid solid-liquid dynamic extraction method, it is possible to obtain a fraction quantitatively more abundant than other methods, and with a lower content of tannins and phenolic compounds but with a higher content of flavonolignans, rare and expensive, and therefore easier to separate and purify.

Phenylpropanoids are secondary metabolites widely distributed in plants. They are believed to protect human cells against oxidative stress and in turn, to prevent various diseases associated with it, such as cancer, cardiovascular, and neurodegenerative disorders as well [2,3]. Recently, many natural phenylpropanoids have emerged as a particularly promising class of small molecules able to inhibit β-amyloid aggregation and disrupt preformed amyloid fibrils that represent important targets in the development of pharmacological treatments of Alzheimer’s disease (AD) [4]. Unfortunately, natural phenylpropanoids often have poor pharmacokinetic properties and low bioavailability. In this frame an interesting approach in the design of new bioactive compounds could be the combination of two or more polyphenolic "fragments". This "natural-fragment-based drug-discovery" approach would allow the assembly, also in a combinatorial manner, of libraries based on complex polyphenols in a few steps. This strategy provides us with the possibility of easily modifying both scaffold and decorations and modulating pharmacodynamic and pharmacokinetic properties [5]. Therefore, we planned to join two "tyrosol-fragments" by a phosphodiester bridge to obtain a new class of phosphate-linked compounds [6], which are potentially more bioactive than the corresponding precursors. We report here the general synthetic strategy to obtain new phosphate-linked tyrosol dimers in good yield and their full characterization. Since oxidative stress, as well as metal ion dyshomeostasis, are known to play important roles in AD pathogenesis, preliminary studies are focused on the evaluation of their antioxidant activity, metal chelating ability and their ability to inhibit β-amyloid aggregation.

Outer membrane vesicles (OMVs) are nanostructures of 20– 200 nm diameter deriving from the surface of several Gram- negative bacteria. OMVs are emerging as shuttles involved in several mechanisms of communication and environmental adaptation. In this work, OMVs were isolated and charac- terized from Novosphingobium sp. PP1Y, a Gram-negative non-pathogenic microorganism lacking LPS on the outer membrane surface and whose genome was sequenced and annotated. Scanning electron microscopy performed on sam- ples obtained from a culture in minimal medium highlighted the presence of PP1Y cells embedded in an extracellular ma- trix rich in vesicular structures. OMVs were collected from the exhausted growth medium during the mid-exponential phase, and purified by ultracentrifugation on a sucrose gra- dient. Atomic force microscopy, dynamic light scattering and nanoparticle tracking analysis showed that purified PP1Y OMVs had a spherical morphology with a diameter of ca. 150 nm and were homogenous in size and shape. Moreover, pro- teomic and fatty acid analysis of purified OMVs revealed a specific biochemical “fingerprint”, suggesting interesting de- tails concerning their biogenesis and physiological role. More- over, these extracellular nanostructures do not appear to be cytotoxic on HaCaT cell line, thus paving the way to their future use as novel drug delivery systems.

Silibinin is the major component of an extract, known as Silymarin, obtained from the seeds of the milk thistle (Silybum marianum). Structurally, Silibinin is diastereoisomeric equimolar mixture of two flavonolignans (Silybin A and Silybin B). As extensively reported in literature, Silibinin displays multiple biological activities, most of them related to its radical scavenging activity. In the past two decades, in addition to hepatoprotective effects, Silibinin has demonstrated remarkable anti-cancer as well as cancer chemopreventive efficacy in pre-clinical cell culture and animal models of several epithelial cancers, including skin, bladder, colon, prostate, and lung [2]. Oral administration of chemotherapeutic agents is the mainstay for the treatment of disease. Sustained release formulations have been crucial for the safe and effective dosing of orally administered drugs [3]. In this frame, the synthesis of phosphodiester derivatives can improve the oral bioavailability of poorly water-soluble metabolites maintaining their pharmacologic activity [4,5]. Here, we reported the synthesis of new Silibinin conjugates with 3'-ribonucleotide units. The phosphodiester junction is typically used in pro-drug strategies, in which the phosphate group is susceptible to hydrolysis by endogenous phosphatases, allowing the release of the active compound. The new conjugates were prepared in few steps and in good yields, starting from the suitable Silibinin or Silybin building blocks and nucleotide phosphoramidites. All compounds were full characterized by NMR and Maldi-TOF and their serum stability was evaluated by HPLC analyses.

Microgreens constitute novel gastronomic ingredients that combine visual, kinesthetic and bioactive qualities. The definition of the optimal developmental stage for harvesting microgreens remains fluid. Their superior phytochemical content against mature leaves underpins the current hypothesis of significant changes in compositional profile during the brief interval of ontogeny from the appearance of the first (S1) to the second true leaf (S2). Microgreens of four brassicaceous genotypes (Komatsuna, Mibuna, Mizuna and Pak Choi) grown under controlled conditions and harvested at S1 and S2 were appraised for fresh and dry yield traits. They were further analyzed for macro- and micromineral content using inductively coupled plasma optical emission spectrometry (ICP-OES), carotenoid content using high-performance liquid chromatography with a diode-array detector (HPLC-DAD), volatile organic compounds using solid-phase microextraction followed by gas chromatography-mass spectrometry (SPME-GC/MS), anthocyanins and polyphenols using liquid chromatography-high resolution-tandem mass spectrometry (LC-MS/MS) with Orbitrap technology and for chlorophyll and ascorbate concentrations, well as antioxidant capacity by spectrophotometry. Analysis of compositional profiles revealed genotype as the principal source of variation for all constituents. The response of mineral and phytochemical composition and of antioxidant capacity to the growth stage was limited and largely genotype-dependent. It is, therefore, questionable whether delaying harvest from S1 to S2 would significantly improve the bioactive value of microgreens while the cost-benefit analysis for this decision must be genotype-specific. Finally, the lower-yielding genotypes (Mizuna and Pak Choi) registered higher relative increase in fresh yield between S1 and S2, compared to the faster-growing and higher-yielding genotypes. Although the optimal harvest stage for specific genotypes must be determined considering the increase in yield against reduction in crop turnover, harvesting at S2 seems advisable for the lower-yielding genotypes.

Sartans are chemical compounds with antagonistic action towards the AT1 receptors of angiotensin II, a multifunctional peptide with vasoconstrictor action and one of the main peptides responsible for regulating blood pressure. Losartan (progenitor of the group), valsartan, irbesartan, candesartan, eprosartan, telmisartan and a few other compounds belong to this class. The use of sartans, introduced in the mid-90s of the last century, has quickly spread mainly due to the alleged reduction in the incidence of certain adverse effects of varying severity (cough, angioedema) associated with the intake of ACE inhibitors. Their very strong commercial success together with their low metabolic degradation and high environmental persistence has consecrated them as emerging pollutants in just 20 years. This review presents a complete picture of sartans in relation to their chemical structure, chemical-physical properties and by-products in aqueous solutions as well as under oxidative and photodegradative conditions, their environmental presence and knowledge known to date and their fate in wastewater treatment plants.

In recent years, many studies have highlighted the consistent finding of amoxicillin in waters destined for wastewater treatment plants, in addition to superficial waters of rivers and lakes in both Europe and North America. In this paper, the amoxicillin degradation pathway was investigated by simulating the chlorination process normally used in a wastewater treatment plant to reduce similar emerging pollutants at three different pH values. The structures of 16 isolated degradation byproducts (DPs), one of which was isolated for the first time, were separated on a C-18 column via a gradient HPLC method. Then, combining mass spectrometry (MALDI-MS/TOF) and nuclear magnetic resonance, we compared commercial standards and justified a proposed formation mechanism beginning from the parent drug. Microbial growth inhibition bioassays with Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus were performed to determine the potential loss of antibacterial activity in isolated degradation byproducts. An increase of antibacterial activity in the DPs was observed compared to the parent compound.

Polyphenols are active compounds of the medicinal plants widely distributed in nature. Despite the few and common biosynthetic origins, polyphenols encompass many subclasses of structurally different entities with a variety of pharmacological properties. Many studies suggests that the mechanisms by which plant polyphenols exert their protective actions against various diseases, in part are due to their redox properties. Silibinin belongs to flavonolignan class and is a major component (approximately 30%) of the Silymarin complex extracted from milk thistle [Silybum marianum (L.) Gaertn. Carduus marianus L., Asteraceae]. Silibinin exists as a mixture of two diastereomers: Silybin A and Silybin B. Silibinin has long been studied for a large variety of pharmacological properties ranging from its antioxidant activity to neuroprotective and antiviral activities. Unfortunately, its in vivo applications are rather hampered by its very low bioavailability mainly due to low water solubility. In an attempt to improve its biological properties and facilitate the in vivo applications, we have developed an efficient strategy for the synthesis of Silibinin modified namely Phosphate-Linked Silibinin dimers (PLSd) in which the Silibinin monomer units are linked by phosphodiester bond (Figure). Exploiting the selective protection of Silibinin’s hydroxyl groups, we carried out a synthetic approach using the phosphoramidite chemistry. Water solubility of dimers, radical scavenger activity as well as their ability to react with reactive oxygen species (ROS) were determined. In particular, dimers reactivity with 1O2 and HO∙ was determined by use of Rose Bengal (RB) and hydrogen peroxide (H2O2) as respective ROS sources. For this purpose, laser flash photolysis (LFP) experiments were coupled with a kinetic competition approach. In addition to scavenging activity, the serum stability and cytoprotective (X/XO assay on HepG2 cells) behavior of dimers were evaluated.

Many drug candidates have been reported to possess a strong therapeutic potential in vitro but they have failed in vivo because of their poor pharmacokinetic behaviour, very often due to the low water solubility. There are many formulations and drug design strategies that can be used to overcome solubility issues. The design and synthesis of soluble pro-drugs is one of the most common approach used. In this frame, the phosphate group is a useful tool for the enhancement of aqueous solubility of phenolic and other metabolites, in addition it displayed excellent chemical stability and rapid bioconversion in vivo to the parent drug by phosphatases. On the other hand, also the conjugation of specific molecules recognized by a receptor on the target cell could be a successful strategy. In our studies, we have combined both aspects through the synthesis of new phosphodiester modified Silibinin with a good water solubility, as well as, attractive antioxidant properties. In the past decade, Silibinin has received more attention due to its large variety of activities ranging from anticancer and chemopreventive actions to hypocholesterolemic, cardioprotective and neuroprotective activities. Unfortunately, the bioavailability and the therapeutic efficiency of Silibinin are rather limited by its low water-solubility. In this work, we present the synthesis of a new library of modified Silibinins and related studies of their redox behaviour. Exploiting the selective protection of the hydroxyl groups of the Silibinin, we developed an efficient strategy for the synthesis of Silibinin phosphate-based flavonolignans consisting of phosphodiester glycoconjugates and dimers of Silibinin. The water solubility, the radical scavenger efficiency and the ability to scavenge different reactive oxygen species (ROS) have been evaluated for the new phosphodiester modified Silibinins in comparison to Silibinin. Moreover, the serum stability, and their cytoprotective (X/XO assay on HepG2 cells) behaviours have been studied. The remarkable antioxidant activity and the high water solubility (compared to Silibinin) make Silibinin phosphatebased flavonolignans promising molecules for future studies.

In the last years, G-rich oligonucleotides (GROs) able to form G-quadruplexes, have attracted considerable interest in biological and therapeutic fields. G-quadruplex are among the most studied DNA structures because they are thought to be involved in important biological processes such as the modulation of gene expression; in addition they present a large variety of biological properties ranging from anticancer to antiviral activities. In all cases, the G-quadruplex formation is a crucial prerequisite for the biological effects. Recently, we reported the synthesis and full characterization of a mini-library of G quadruplex ODNs carrying aryl groups at the 5’-end through a phosphodiester bond and endowed with prominent anti-HIV activity. In this frame, we report here, the synthesis of new Riboflavin-Nucleoside (RF, Vitamin B) building block in which the ribityl chain is rigidified by 2',4' benzylidene group, the 5'-OH function is protected with DMT group and the 3’ position is functionalized with phosphoramidite. The new Riboflavin building block is incorporated into different ODN sequences able to form Gquadruplex structures using the well known phosphoramidite chemistry. This research project has been encouraged by recent studies on the use of RF as useful tool for both in vivo and in vitro studies: the Riboflavin is internalized through RF transporters and is one of the most efficient natural photosensitizers. Several modified ODN sequences have been synthesized with a solid phase synthetic approach in good yields; their biophysical and biological characterization has been carried out in order to evaluate the effect of RF-nucleotide insertion into G-quadruplex arrangements and their anti-HIV activity.

The synthesis of (1'S,2'S,4'R) and (1'S,2'R,4'R) stereoisomers of 2'-fluoro-3'-thiacytidine [(2' S)-F-3TC and (2' R)-F-3TC], the earliest examples of oxathiolanyl nucleosides with a fluorine atom in the 'sugar' backbone, is herein reported. From of a variety of synthetic routes devised for their preparation, the Pummerer rearrangement of protected lamivudine sulfoxides was successfully exploited for fluorine atom introduction. Despite the presence of three potentially labile stereocenters in such a small molecule, the (2'R)-isomer of F-3TC exhibited good chemical stability after protective group cleavage. Conversely, the (2'S)-epimer suffered from weak stability, owing to the formation of an undesired cyclization product. Based on the remarkable antiviral efficacy of the parent drugs, the access to 2'-fluorinated oxathiolanyl nucleosides (and more generally, 2'-fluorinated heterocyclic nucleosides) may provide a new source of candidates with antiviral potential.

Sideritis species have been used in folk medicine for their antimicrobial, antiulcerogenic, digestive and anti-inflammatory properties. Over the years, the phytochemistry of the genus Sideritis has been studied, and various terpenoids, sterols, coumarins and especially flavonoid aglycones and glycosides have been identified. In particular, species from the Balkan Peninsula have been studied and were found to be rich in flavonoids, with valuable antioxidant activity. In the folk medicine of the Balkan countries, Sideritis raeseri is used as a herbal tea in the treatment of inflammation, gastrointestinal disorders and coughs, and also as a tonic, whereas extracts are used as a component of dietary supplements for anaemia. Its dried inflorescences are used to prepare a beverage called 'mountain tea'. In light of the considerable interest generated in the chemistry, pharmacological properties and commercial value of S. raeseri Boiss. & Heldr., we review and summarise the available literature on these plants. The review details the chemical composition of the essential oil, its mineral and polyphenol contents, the naming of these plants and their physicochemical characterisation, and the nuclear magnetic resonance spectral data and biological properties associated with the plant extracts, with a focus on their potential chemotherapeutic applications. © 2016 Society of Chemical Industry.

Selenium (Se) is considered essential for human nutrition as it is involved in the metabolic pathway of selenoproteins and relevant biological functions. Microgreens, defined as tender immature greens, constitute an emerging functional food characterized by overall higher levels of phytonutrients than their mature counterparts. The nutraceutical value of microgreens can be further improved through Se biofortification, delivering Se-enriched foods and potentially an enhanced content of bioactive compounds. The current study defined the effect of sodium selenate applications at three concentrations (0, 8, and 16 &mu, M Se) on the bioactive compounds and mineral content of coriander, green basil, purple basil, and tatsoi microgreens grown in soilless cultivation. Analytical emphasis was dedicated to the identification and quantification of polyphenols by UHPLC-Q-Orbitrap-HRMS, major carotenoids by HPLC-DAD, and macro micro-minerals by ICP-OES. Twenty-seven phenolic compounds were quantified, of which the most abundant were: Chlorogenic acid and rutin in coriander, caffeic acid hexoside and kaempferol-3-O(caffeoyl) sophoroside-7-O-glucoside in tatsoi, and cichoric acid and rosmarinic acid in both green and purple basil. In coriander and tatsoi microgreens, the application of 16 &mu, M Se increased the total phenols content by 21% and 95%, respectively, moreover, it improved the yield by 44% and 18%, respectively. At the same Se dose, the bioactive value of coriander and tatsoi was enhanced by a significant increase in rutin (33%) and kaempferol-3-O(feruloyl)sophoroside-7-O-glucoside (157%), respectively, compared to the control. In green and purple basil microgreens, the 8 &mu, M Se application enhanced the lutein concentration by 7% and 19%, respectively. The same application rate also increased the overall macroelements content by 35% and total polyphenols concentration by 32% but only in the green cultivar. The latter actually had a tripled chicoric acid content compared to the untreated control. All microgreen genotypes exhibited an increase in the Se content in response to the biofortification treatments, thereby satisfying the recommended daily allowance for Se (RDA-Se) from 20% to 133%. The optimal Se dose that guarantees the effectiveness of Se biofortification and improves the content of bioactive compounds was 16 &mu, M in coriander and tatsoi, and 8 &mu, M in green and purple basil.

Research Highlights: Plant cover drives the activity of the microbial decomposer community and affects carbon (C) sequestration in the soil. Despite the relationship between microbial activity and C sequestration in the soil, potential inhibition of soil microbial activity by plant cover has received little attention to date. Background and Objectives: Differences in soil microbial activity between two paired stands on soil at a very early stage of formation and a common story until afforestation, can be traced back to the plant cover. We hypothesized that in a black locust (Robinia pseudoacacia L.) stand the high-quality leaf litter of the tree, and that of the blackberry (Rubus fruticosus L.) understory had an inhibitory effect on soil microbial community resulting in lower mineralization of soil organic matter compared to the paired black pine (Pinus nigra Arn.) stand. Materials and Methods: We estimated potential mineralization rates (MR), microbial (MB), and active fungal biomass (AFB) of newly-shed litter, forest floor, and mineral soil. We tested the effects of litters&rsquo, water extracts on soil MR, MB, AFB and its catabolic response profile (CRP). Results: Newly-shed litter of black locust had higher MR than that of blackberry and black pine, MR, MB, and AFB were higher in forest floor and in mineral soil under black pine than under black locust. Water extracts of black locust and blackberry litter had a negative effect on the amount, activity of microorganisms, and CRP. Conclusions: The results demonstrate the potential for black locust and blackberry litter to have a marked inhibitory effect on decomposer microorganisms that, in turn, reduce organic matter mineralization with possible consequences at the ecosystem level, by increasing C sequestration in mineral soil.

The biological relevance of tetramolecular G-quadruplexes especially as anti-HIV agents has been extensively reported in the literature over the last years. In the light of our recent results regarding the slow G-quadruplex folding kinetics of ODNs based on d(TGGGAG) sequence, here we report a systematic anti-HIV screening to investigate the impact of the G-quadruplex folding on their anti-HIV activity. In particular, varying the single stranded concentrations of ODNs, it has been tested a pool of ODN sample solutions with different G-quadruplex concentrations. The anti-HIV assays have been designed favouring the limited kinetics involved in the tetramolecular G4-association based on the d(TGGGAG) sequence. Aiming to determine the stoichiometry of G-quadruplex structures in the same experimental conditions of the anti-HIV assays, a native gel electrophoresis was performed. The gel confirmed the G-quadruplex formation for almost all sample solutions while showing the formation of high order G4 structures for the more concentrated ODNs solutions. The most significant result is the discovery of a potent anti-HIV activity of the G-quadruplex formed by the natural d(TGGGAG) sequence (IC50 = 14 nM) that, until now, has been reported to be completely inactive against HIV infection.

We evaluated the short-term effects of a flavanol-rich cocoa (FRC) on lipid profile and selected oxidative stress biomarkers such as oxidized low-density lipoprotein (oxLDL), glutathione (GSH), and F2-isoprostane. We also assessed whether FRC modulates plasma levels of polyunsaturated fatty acids (PUFA) in healthy individuals. The subjects (n=48) were randomly assigned to a low-cocoa group (1 g/d; ~55 mg flavanols) (n=16), middle-cocoa group (2 g/d; ~110 mg flavanols) (n=16), or a high-cocoa group (4 g/d; ~220 mg flavanols) (n=16). The samples were collected at baseline, at 1, 2, and 4 h post initial consumption of FRC, and after 4 weeks of FRC supplementation. The peak plasma concentration of (−)-epicatechin metabolites reached a maximum level (578±61 nM; Pb.05) at 2 h after ingestion of FRC. After 4 weeks, total cholesterol (−12.37±6.63; Pb.0001), triglycerides (−3.81±2.45; Pb.0001), plasma LDL (−14.98±6.77; Pb.0001), and oxLDL (−95.61±41.69; Pb.0001) decreased in the high-cocoa group, compared with baseline. We also found that plasma high-density lipoprotein (HDL) (+3.37±2.06; Pb.0001) concentrations increased significantly in the same group. Total GSH significantly increased in all FRC-treated groups (+209.73±146.8; Pb.0001), while urinary F2-isoprostane levels decreased in the middle- (−0.73±0.16; Pb.0001) and high-cocoa (−1.62±0.61; Pb.0001) groups. At the end of the four-week study, a significant reduction of arachidonic acid (AA)/eicosapentaenoic acid (EPA) ratio was observed in the low- (−2.62±2.93; P=.003), middle- (−5.24±2.75; Pb.0001) and high-cocoa (−7.76±4.96; Pb.0001) groups, compared with baseline. Despite the small sample size used in this study, these data extend previous clinical and experimental studies, providing new insights into the health benefits of cocoa flavanols.

Potatoes are a staple food for over a billion people worldwide, a primary dietary source of carbohydrates, and a vital crop to the agricultural economy of South America, Africa, East Asia and Central Asia. Potatoes occupy third place (after rice and wheat) on the list of foods on which the world depends for food security. In particular, glyco-alkaloids contents in potatoes are potentially toxic and secondary metabolites such as α-solanine and α-chaconine are reported to be dangerous to human health. This paper describes new methods for α-solanine and α-chaconine reduction in the potato cultivar known as Marabel. The potatoes were incubated at room temperature in the dark for 24 h and the optimal experimental condition was achieved with NaOH solution at pH 12. In the assayed samples, α-solanine and α-chaconine reduction was 43% and 27% respectively. The process proposed here allows to minimize the total content of glyco-alkaloids, with respect to mode of collection, storage and cleaning of Marabel potatoes.

Objective: Berries are a rich source of anthocyanins, and clinical data suggest that a polyphenol-rich diet may exert health-promoting effects by reducing oxidative stress. The aim of this study was to elucidate the effects of dietary supplementation with Delphinol (trademark owned by MNL Chile) standardized maqui berry (Aristotelia chilensis) extract on products of lipid peroxidation in healthy, overweight, and smoker subjects.Methods: In a double-blind, placebo-controlled design, 42 participants (age 45-65years) consumed in random order either a standardized extract of maqui berry (162mg anthocyanins) or a matched placebo, given 3times daily for 4weeks. The samples were collected at baseline, after the end of the supplementation, and 40days after the end of the study. Primary outcome was the measure of oxidized low-density lipoprotein (Ox-LDL) and F-2-isoprostanes in plasma and urine, respectively. Secondary outcomes included anthropometric measures, blood pressure, and lipid profile.Results: Delphinol supplementation was associated with reduced levels of Ox-LDL in the anthocyanin group compared to baseline (p < 0.05). There was also a decrease in urinary F-2-isoprostanes (8-iso-prostaglandin F2) at 4weeks versus baseline in the Delphinol-supplemented group (p < 0.05). However, no differences in primary outcomes were evident at 40days of follow-up. In the fourth week of the intervention, no significant differences were noted for anthropometric characteristics, ambulatory blood pressure, and lipid profile.Conclusions: Our observations suggest that dietary interventions with maqui berry extract may improve oxidative status (Ox-LDL and F-2-isoprostanes) in healthy adults, overweight adults, and adult smoker

BACKGROUND Epidemiological studies have proved that tomato consumption is associated with a lower risk of developing several diseases (for example, certain types of cancers, cardiovascular diseases, macular degeneration, age-related eye disease). Many micronutrients and bioactive compounds are mainly present in peel and seeds and are lost during the processing into sauce, puree, paste and juice. RESULTS The addition of lyophilized and powdered tomato pomace enhances the properties of puree. In this paper we report the chemical and physicochemical characterization of a puree enriched with 2% dry pomace. Comparison of the analytical data of starting puree with the enriched puree showed a significant increase of all micronutrients, without the taste and appearance being compromised or altered negatively. CONCLUSION The product obtained is an example of a functional food rich in health-promoting phytochemicals, with the significant aspect of recovering a waste fraction of the tomato processing that would normally be disposed of in landfill, with associated costs and environmental impact. © 2015 Society of Chemical Industry

Introduction.Plant extracts may be suitable alternative treatments for caries.Aims. To investigate thein vitroandin vivoantimicrobial effects ofPlantago lanceolataherbal tea (from flowers and leaves) on cariogenic bacteria and to identify the major constituents ofP. lanceolataplant.Materials and Methods.The MIC and MBC against cariogenic bacteria were determined forP. lanceolatatea. Subsequently, a controlled random clinical study was conducted. Group A was instructed to rinse with aP. lanceolatamouth rinse, and Group B received a placebo mouth rinse for seven days. The salivary colonisation by streptococci and lactobacilli was investigated prior to treatment and on the fourth and seventh days. Finally, theP. lanceolatatea was analysed for its polyphenolic content, and major phenolics were identified.Results and Discussion. P. lanceolatateas demonstrate goodin vitroantimicrobial activity. Thein vivotest showed that Group A subjects presented a significant decrease in streptococci compared to Group B. The phytochemical analysis revealed that flavonoids, coumarins, lipids, cinnamic acids, lignans, and phenolic compounds are present inP. lanceolatainfusions.Conclusions. P. lanceolataextract could represent a natural anticariogenic agentviaan antimicrobial effect and might be useful as an ancillary measure to control the proliferation of cariogenic flora.

In ‘90s, Hotoda et al. identified a variety of tetramolecular G-quadruplexes based on the sequence d(TGGGAG) and modified at the 5’-end with aromatic groups, targeting the glycoprotein of HIV-1, gp120. In 2011 Di Fabio et al. reported the synthesis and characterization of a new mini-library of d(TGGGAG) ODNs carrying aryl groups at the 5’-end through a phosphodiester bond and endowed with prominent anti-HIV activity[2]. In order to have more information on the G-quadruplex folding depending on the modifications at 5’-end of d(TGGGAG), we have explored the kinetic aspects involved into G-quadruplex folding using electrospray mass spectrometry (ESI-MS). The studies are conducted on more active 5’-end modified ODNs in comparison with unmodified sequence d(TGGGAG). By ESI-MS experiments, we could obtain information about the number of strands, and the inner cations for each complex, moreover it is possible to detect all possible intermediate complexes and to determine their equilibrium binding constants. We followed the kinetics of G-quadruplex formation during 14 days in the presence of an internal reference (dT6), selecting some time points after addition of different buffers (150mM NH4OAc and 150mM TMAA/1mM KCl). Overall data, have reported a rapid conversion of the single strand into dimer and trimer, and slower conversion in a tetramer and into an unexpected G4-supramolecular structure: the octamer complex. The formation of octamer is confirmed also in solution by native PAGE.

Introduction: Recently, we reported the synthesis and characterization of a new mini-library of tetramolecular G quadruplexes based on the d(TGGGAG) sequence, which was previously shown to target the HIV-1 glycoprotein gp120. Oligodeoxynucleotides (ODNs) carrying aryl groups at the 5’-end through a phosphodiester bond were endowed with prominent anti-HIV activity. Biophysical studies on these aptamers demonstrated that there is no relationship between thermal stability of G4 structures and their anti-HIV activity. To identify the properties that make these tetramolecular G-quadruplexes good anti-HIV aptamers, we studied by ESI-MS the stoichiometry and the self-assembly kinetics of G-quadruplex structures based on the most active 5’-end modified d(TGGGAG) aptamers [5]. ESI-MS results showed that the conjugation of d(TGGGAG) at the 5’-end does not necessarily increase the folding rate of the G-quadruplex structure. Unexpectedly, the folding kinetics of the inactive G4 (unmodified sequence) was similar to that of the 5’-end modified sequences. In addition to the slow G4-kinetics revealed by ESI-MS studies, PAGE experiments on the modified d(TGGGAG) highlighted the strong effect of ODN concentration (single stranded) on the G4-folding kinetics. Results and Discussion: In order to clarify the role of G-quadruplex structures in the anti-HIV activity on the basis of the above kinetic studies, we investigated the anti-HIV activity of ODNs at different concentrations of G-quadruplex structure. All ODNs, as well as the unmodified sequence, were evaluated against a variety of HIV-1 viruses and resistant strains. SPR experiments were also carried out to evaluate their binding to the HIV envelope gp120 and gp41 proteins. Almost all compounds showed a good anti-HIV activity, suggesting the absence of a straight correlation between the amount of available G-quadruplex and anti-HIV activity of ODNs. Surprisingly, we found a strong anti-HIV activity of the unmodified sequence d(TGGGAG), in contrast to what has been extensively reported by Hotoda and others, who showed a lack of anti-HIV activity of the d(TGGGAG) sequence without 5’-end modifications. Also, by SPR experiments no straight correlation between gp120 binding and anti HIV activity was observed. Conclusions: The anti-HIV activity results highlight the absence of a straight correlation between the formation of G-quadruplex structures and the antiviral potential of ODNs, suggesting instead that the G4 is not the specie providing anti-HIV activity. In addition, anti-HIV data do not strictly correlate with the SPR results, suggesting that gp120 and gp41 is not the primary target of these aptamers.

New silibinin phosphodiester glyco-conjugates were synthesized by efficient phosphoramidite chemistry and were fully characterized by 2D-NMR. A wide-ranging study focused on the determination of their pKa and E° values as well as on their radical scavenging activities by different assays (DPPH, ABTS+ and HRSA) was conducted. The new glyco-conjugates are more water-soluble than silibinin, and their radical scavenging activities are higher than those of silibinin. The conjugation therefore improves both the water solubilities and antioxidant activities of the flavonolignan moieties. The serum stability was evaluated under physiological conditions, and the glyco-conjugates degraded with half-lives of 40-70 h, making them useful in pro-drug approaches. We started by treating androgen-dependent prostate cancer (PCa) LNCaP cells and then expanded our studies to androgen-independent PCa PC3 and DU145 cells. In most cases, the new derivatives significantly reduced both total and live cell numbers, albeit at different levels. Anti-HIV activities were evaluated and the glucosamine-phosphate silibinin derivative showed higher activity (IC50 = 73 μM) than silibinin.

International audience; By exploiting the regioselective protection of the hydroxyl groups of silibinin along with the well-known phosphoramidite chemistry, we have developed an efficient strategy for the synthesis of new silibinin-modified species, which we have named Phosphate-Linked Silibinin Dimers (PLSd), in which the monomer units are linked by phosphodiester bonds. The antioxidant abilities of the new PLSd were estimated on HepG2 cells using DPPH free radical scavenging and xanthine/xanthine oxidase assays. The new phosphate-metabolites showed a higher anti-oxidant activity than the silibinin, as well as very low toxicity. The ability to scavenge reactive oxygen species (ROS) such as singlet oxygen () and hydroxyl radical () reveals that the two dimers are able to scavenge about two times more effectively than silibinin. Finally, solubility studies have shown that the PLSd present good water solubility (more than 20 mg·L−1) under circumneutral pH values, whereas the silibinin was found to be very poorly soluble (less than 0.4 mg·L−1) and not stable under alkaline conditions. Together, the above promising results warrant further investigation of the future potential of the PLSd as anti-oxidant metabolites within the large synthetic polyphenols field.

Polyphenols are active compounds of the medicinal plants widely distributed in nature. Despite the few and common biosynthetic origins, polyphenols encompass many subclasses of structurally different entities with a variety of pharmacological properties. Many studies suggest that the mechanisms by which plant polyphenols exert their protective actions against various diseases, are in part due to their redox properties.1Silibinin belongs to flavonolignan class, it is a major component of the Silymarin complex extracted from the milk thistle (Silybum marianum). Silibinin has long been studied for its large variety of pharmacological properties ranging from the antioxidant activity to neuroprotective and antiviral activities. In order to expand the repertoire of the modified Silibinins with high molecular diversity and good water solubility, here, we present a new and efficient method for the synthesis of dimers and trimers of the Silibinin, in which the monomer units are linked by phosphodiester bond Exploiting the selective protection of the Silibinin hydroxyl groups, we carried out a new synthetic approach using the well-established phosphoramidite chemistry. Aiming at the discovery of new pharmacological activities of these molecules, the water solubility, the radical scavenger activity by DPPH test, as well as, their ability to scavenge different reactive oxygen species (1O2 and HO●) have been studied. Additionally, the serum stability and cytoprotective behaviour (X/XO assay on HepG2 cells) of dimers have been evaluated. The combination of both striking antioxidant ability and high water solubility, makes these molecules, promising synthetic metabolites for future therapeutic applications.

The interaction of the porphyrin derivative H2TCPPSpm4, having four spermines in the meso positions, with the G-quadruplex (GQ) structure formed by the DNA aptamer TGGGAG, a sequence that show anti-HIV activity, has been investigated by means of UV, CD and PAGE studies. The results demonstrate that porphyrin substituted with positively charged side chains is capable of inducing the formation of different complexes with parallel GQs, including higherordered complexes, depending on the method used to achieve their relative stoichiometry (titration vs single addition). Indeed the CD melting experiments indicated different thermal stability fort the 1:1 H2TCPPSpm4:GQ and the 2:1 H2TCPPSpm4:GQ species obtained by titration. Interestingly also the 2:1 H2TCPPSpm4:GQ complexes obtained by titration or by direct mixing show different behavior.

The interaction of the porphyrin derivative H(2)TCPPSpm4, having spermine pendants in the four meso positions, with the G-quadruplex (GQ) structure formed by the DNA aptamer TGGGAG has been investigated by means of UV, electronic circular dichroism and PAGE studies. The results reported here demonstrate that the porphyrin derivative is capable of stabilizing or destabilizing the higher-ordered structures of parallel GQs, depending on the method used to reach their relative stoichiometry (titration vs. single addition). Noteworthily, when two equivalents of H(2)TCPPSpm4 were mixed directly with one equivalent of the (TGGGAG)(4) GQ to reach a 2 : 1 H(2)TCPPSpm4: GQ ratio T-1/2 higher than 80 degrees C was also observed confirming the presence of higher-ordered GQ structures.