Combined Experimental and Multivariate Model Approaches for Glycoalkaloid Quantification in Tomatoes.

The intake of tomato glycoalkaloids can exert beneficial effects on human health. For this reason, methods for a rapid quantification of these compounds are required. Most of the methods for α-tomatine and dehydrotomatine quantification are based on chromatographic techniques. However, these techniques require complex and time-consuming sample pre-treatments. In this work, HPLC-ESI-QqQ-MS/MS was used as reference method. Subsequently, multiple linear regression (MLR) and partial least squares regression (PLSR) were employed to create two calibration models for the prediction of the tomatine content from thermogravimetric (TGA) and attenuated total reflectance (ATR) infrared spectroscopy (IR) analyses. These two fast techniques were proven to be suitable and effective in alkaloid quantification (R2 = 0.998 and 0.840, respectively), achieving low errors (0.11 and 0.27%, respectively) with the reference technique.

Kinetics of glucosinolate hydrolysis by myrosinase in Brassicaceae tissues: A high-performance liquid chromatography approach.

Glucosinolates are a group of secondary metabolites occurring in all the vegetables belonging to the Brassicaceae family. Upon tissue damage, glucosinolates are hydrolyzed by myrosinase to a series of degradation products, including isothiocyanates, which are important for their health-promoting effects in humans. The glucosinolate-myrosinase system has been characterized in several Brassica species, of which white mustard (Sinapis alba) has been studied the most. In this study, a new HPLC-UV assay to evaluate the activities and kinetics of myrosinases in aqueous extracts, which closely represent the physiological conditions of plant tissues, was developed. This method was tested on myrosinases extracted from broccoli and cauliflower inflorescences, employing sinigrin and glucoraphanin as substrates. The results showed a strong inhibition of both enzymes at high substrate concentrations. The main issues related to kinetic analysis on the glucosinolate-myrosinase system were also elucidated.

Effects of Aqueous Extract of Lycopersicum esculentum L. var. "Camone" Tomato on Blood Pressure, Behavior and Brain Susceptibility to Oxidative Stress in Spontaneously Hypertensive Rats.

Behavioral disorders affect millions of people worldwide. Hypertension contributes to both the development and progression of brain damage and cognitive dysfunction and could represent the most powerful modifiable risk factor for cerebral vessel dysfunction and consequent behavioral impairment. Tomato contains antioxidants and bioactive molecules that might play an important role in the prevention of cardiovascular and brain diseases. The effects of the combined gel and serum from Lycopersicum esculentum L. var. "Camone" tomatoes and those of purified tomato glycoalkaloids (tomatine) and an antihypertensive drug (captopril) were investigated in male spontaneously hypertensive rats (SHRs) and compared with normotensive Wistar Kyoto (WKY) rats. Body weight, systolic blood pressure, behavioral parameters, as well as brain susceptibility to oxidative stress and brain cytokine contents, were assessed. Treating hypertensive rats with tomato gel/serum or captopril for four weeks caused a significant reduction in blood pressure, decreased locomotor activity and increased grooming behavior; the last two parameters were also significantly affected by tomatine treatment. Brain slices obtained from hypertensive rats treated with tomato gel/serum were more resistant to oxidative stress and contained lower levels of inflammatory cytokines than vehicle-treated ones. In contrast, tomatine treatment had no effect. In conclusion, the tomato-derived gel/serum can be considered a dietary supplement able to drive in vivo blood pressure towards healthier values and also control some central effects such as behavior and brain oxidative stress.

Chemical Characterisation and Antihypertensive Effects of Locular Gel and Serum of Lycopersicum esculentum L. var. "Camone" Tomato in Spontaneously Hypertensive Rats.

Blood pressure control in hypertensive subjects calls for changes in lifestyle, especially diet. Tomato is widely consumed and rich in healthy components (i.e., carotenoids, vitamins and polyphenols). The aim of this study was to evaluate the chemical composition and antihypertensive effects of locular gel reconstituted in serum of green tomatoes of "Camone" variety. Tomato serum and locular gel were chemically characterised. The antihypertensive effects of the locular gel in serum, pure tomatine, and captopril, administered by oral gavage, were investigated for 4 weeks in male spontaneously hypertensive and normotensive rats. Systolic blood pressure and heart rate were monitored using the tail cuff method. Body and heart weight, serum glucose, triglycerides and inflammatory cytokines, aorta thickness and liver metabolising activity were also assessed. Locular gel and serum showed good tomatine and polyphenols content. Significant reductions in blood pressure and heart rate, as well as in inflammatory blood cytokines and aorta thickness, were observed in spontaneously hypertensive rats treated both with locular gel in serum and captopril. No significant effects were observed in normotensive rats. Green tomatoes locular gel and serum, usually discarded during tomato industrial processing, are rich in bioactive compounds (i.e., chlorogenic acid, caffeic acid and rutin, as well as the glycoalkaloids, α-tomatine and dehydrotomatine) that can lower in vivo blood pressure towards healthier values, as observed in spontaneously hypertensive rats.

Enriched Gellan Gum hydrogel as visco-supplement.

Viscosupplementation, i.e. intra-articular injection of hyaluronic acid derivatives, is considered as the most effective treatment for patients with mild to moderate osteoarthritis. Even if hyaluronic acid is still considered as the gold standard, research is now focusing on the development of new products with enhanced injectability and yet reasonable viscoelastic behavior for OA treatment. A Gellan Gum (GG) hydrogel was synthesized and coated with crosslinked polyvinyl alcohol (PVA) to protect the polysaccharide from degradation during sterilization and improve its performance for the foreseen application. Thermal analyses indicated that mixed hydrogel showed a higher degree of structuring than the bare polysaccharide core without losing its swelling properties, thanks to the hydrophylicity of both coating and cross-linking agent. The PVA coating increased elastic and viscous moduli of the polysaccharide core conferring it a higher resistance to shear and compression and better thixotropic properties. Despite the double crosslinking, hydrogel was injectable. Cytocompatibility towards chondrocytes was verified.

Chemical characterization and antioxidant properties of products and by-products from Olea europaea L.

The products and by-products of Olea europaea L.: olive fruits (primary agricultural product), oils (primary agro-industrial product), pomaces (agro-industrial processing by-product), and leaves (agricultural practices by-product), are promising sources of bioactive compounds. In the present study, qualitative and quantitative analyses of selected bioactive components in olive fruits, oils, and pomaces were performed. Total polyphenol content and antioxidant activity were analyzed in all samples (humid pomaces 2015: TPP, 26.0 ± 1.5-43.7 ± 3.0 g(GAEq)/kg DW; TEAC/ABTS, 189.5 ± 3.7-388.1 ± 12.0 mmol(Trx)kg DW). Radical (DPPH) quenching potential was analyzed via photometric and EPR methods, obtaining Vis/EPR signal ratio by 1.05 ± 0.45 and 1.66 ± 0.39 for fruits and pomaces, respectively. Through HPLC-UV and HPLC-MS/MS techniques, oleuropein and hydroxytyrosol, as well as selected hydroxycinnamic acids and flavonoids, were identified and quantified in olive fruits and pomaces. The main components were rutin, luteolin, and chlorogenic acid. Cytotoxic assay on fibroblast cells revealed toxic effects for selected extracts at highest tested concentrations (5%).

Ordering effect of protein surfaces on water dynamics: NMR relaxation study.

Proteins in solution affect the structural and dynamic properties of the bulk water at the protein-water interface, resulting in a contribution to the order of the hydration water. Theoretical and experimental NMR relaxation methods were developed to study the dynamic properties of water molecules in the protein hydration shell. Water non-selective and selective relaxation rates, were shown to be sensitive to contributions from ordered solvent molecules at protein surface. The average rotational correlation time of water molecules in the protein hydration shell was determined for three protein systems of different size: ribonuclease A, human serum albumin and fibrinogen. The knowledge of these properties is an important step toward the determination of the size of the water ordering contributions originate in proteins systems.

Metal-Ligand Recognition Index Determination by NMR Proton Relaxation Study.

In this study, we developed and validated a new proposed parameter quantifying the interaction strength between natural and/or synthetic molecules with paramagnetic metal ions. The Metal ion Recognition Index, Miri, is a quantitative parameter to describe the proton environment and to define their involvement in the inner and/or outer sphere of the paramagnetic metal ion. The method is based on the analysis of NMR proton spin-lattice relaxation rates of a specific ligand in both the diamagnetic and paramagnetic conditions. The proposed procedure is also useful to calculate the ligand proton spin-lattice relaxation rate in the paramagnetic bound conditions, which is typically very difficult to determine experimentally. Miri was used to compare the ligand proton involvement toward different paramagnetic species, in particular the Copper(II)-Piroxicam system. Copper(II)-Piroxicam complex is one of the most active anti-inflammatory and anti-arthritic species. Miri provides an opportunity to improve our knowledge of metal-ligand complexes that play a fundamental role in bioinorganic interactions.

Chemical characterization of liposomes containing nutraceutical compounds: Tyrosol, hydroxytyrosol and oleuropein.

Tyrosol, hydroxytyrosol and oleuropein are among the major phenolic compounds in fruits, leaves and oils from Olea europaea L. These natural antioxidants molecules revealed several beneficial effects on human health, but a low bioavailability and accessibility to targeted site. Liposomes are drug/nutraceutical delivery carriers, used for driving bioactive molecules to desired target tissues, decreasing potential side effects and protecting the encapsulated molecule from enzymatic metabolic processes. In this study, zwitterionic liposomes containing tyrosol, hydroxytyrosol and oleuropein were synthesized and characterized for their size and surface charge. Particular attention was devoted to the determination of encapsulation efficiency (EE%), quantifying the loaded Tyr, HTyr and Ole amount, by using three different techniques: direct UV spectrophotometry, High Performance Liquid Chromatography and Trolox Equivalent Antioxidant Capacity assay. The results revealed higher EE% for oleuropein. Cyto-toxicity and cyto-compatibility of liposomes were also tested on human chondrocyte cells.

Ionic Exchange Resins and Hydrogels for Capturing Metal Ions in Selected Sweet Dessert Wines.

Samples of sweet and dessert wines, Vin Santo (VSR) from Malvasia grapes, and Granello (GR) from Sauvignon grapes were collected and analyzed for the content of selected macro- and micro-nutrients (Na, K, Mg, Ca, Mn, Fe, Cu and Zn) and of Pb. GR wines had low levels for Fe, Cu and Zn, when compared to VSR and in particular Zn was two orders of magnitude lower. Methods to decrease the content of Zn and Cu in VSR, as well as those for reducing, at the same time, the concentrations of Ca, Mg and K in both VSR and GR, to avoid the formation of opalescence and depots of metal tartrates, were studied. Synthetic hydrogels containing l-histidine residue were tested. The overall relative lowering effects were by ca 4, 23, and 12% for K, Mg and Ca contents, and ca 6, 27 and 10%, for Mn, Cu and Zn contents, in GR wine samples. Commercial ion exchange resin Lanxess Lewatit L-207 and L-208 were then assayed, being legally allowed in the agro-food industry. The L-207 resin revealed great lowering effects on the concentrations of Mn, Cu and Zn, being 75, 91 and 97%, respectively, in VSR wines and 77, 76 and 92%, respectively, in GR wines. The content of Zn was reduced from 49.3 ± 1.2 mg/L in the original wine, down to 1.1 ± 0.1 mg/L, within 48 h soaking. The effects on the character of the dessert wines by the resin L-207 was also taken under control, measuring pH and color index. The color index changed by ca 15% and pH by ca 6% upon treatment of VSR wine with L-207 resins (48 h).