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The oral delivery of macromolecular therapeutics to the intestinal tract requires novel, robust, and controlled formulations. Here, we report on fabrication by molding of composite hydrogel cylinders made of cellulose nanocrystals (CNCs) and chitosan (Cht) and their performance as delivery vehicles. CNCs provide excellent mechanical and chemical stress resistance, whereas Cht allows scaffold degradation by enzyme digestion. The release of a representative medium size protein (bovine serum albumin) dispersed in the hydrogel is slow and shows a sigmoidal profile; meanwhile, the hydrogel scaffold degrades according to a preferred route, that is the cylinder is eroded along the vertical axis. The cup-like, scarcely interconnected porous network, with a gradient of hardness along the cylinder axis, and the compact skin-like layer covering the lateral wall which stayed in contact with the mold during gelification, explain the preferred erosion direction and the long-term protein release. The possible effect of the molding process on hydrogel structure suggests that molding could be a simple and cheap way to favor surface compaction and directional scaffold degradation.

With the purpose of designing active patches for photodynamic therapy of melanoma, transparent and soft hydrogel membranes (HMs) have been fabricated by cation-induced gelation of rod-like cellulose nanocrystals (CNCs) bearing negatively charged carboxylic groups. Na+ , Ca2+ , Mg2+ have been used as cross-linkers of cellulose nanocrystal (CNC). The biosafety of this material and of its precursors has been evaluated in vitro in cell cultures. Morphological changes, cell organelles integrity, and cell survival with the tetrazolium salt reduction (MTT) assay were utilized as tests of cytotoxicity. Preliminary investigation was performed by addition of the hydrogel components to the cell culture medium and by incubations of the CNC-HM in direct and indirect contact with a confluent monolayer of A375 melanoma cells. Direct contact assays suffered from interference of physical stress. Careful evaluation of cytotoxicity was obtained considering the overall picture provided by microscopy and biochemical tests performed with the CNC-HM in indirect contact with two melanoma cell lines (A375, M14) and human fibroblasts. CNCs have been demonstrated to be a safe precursor material and CNC-HMs have a good biocompatibility provided that the excess of cations, in particular of Ca2+ is removed. These results indicate that CNC and can be safely used to fabricate biomedical devices such as transparent hydrogel patches, although attention must be paid to the fabrication procedure.

The great potential of graphene can be enhanced thanks to the functionalization of its surface. For this aim, different thicknesses of TiO2 were grown on graphene films by atomic layer deposition (ALD) at 200 °C using H2O and TiCl4 as precursors. The changes in electronic structure of graphene after the deposition of TiO2 and the influence of graphene in TiO2 photocatalytic activity under UV–Visible irradiation were studied. Results indicated the presence of inhomogeneity and intrinsic strain effects within the same sample. Undecorated graphene showed pre-existent strain due to the mismatch between graphene film and the underlying substrate, while non-intentional self-doping is caused by the presence of charged impurities. The deposition of TiO2 films with thickness ≤10 nm led graphene to be p-doped, while strain became the dominant effect increasing film thickness. Oxygen vacancies in the film decreased exponentially by increasing the film thickness leading to a stoichiometric O/Ti atomic ratio of TiO2 above 10 nm thickness. The combination of TiO2 and graphene enhanced the efficiency of electron-hole separation of TiO2 under UV–Visible light, leading to a higher photocatalytic activity tested for methyl red molecule degradation, making TiO2/Graphene hybrid material a promising candidate for the photodegradation of pollutants and water purification.

The influence of the physical, rheological, and process parameters on the cellulose nanocrystal (CNC) drops before and after external gelation in a CaCl2 solution was investigated. The dominant role of the CNC’s colloidal suspension properties, such as the viscous force, inertial, and surface tension forces in the fluid dynamics was quantitatively evaluated in the formation of drops and jellified beads. The similarity and difference between the behavior of carbohydrate polymers and rod-like crystallites such as CNC were enlightened. Pump-driven and centrifugally-driven external gelation approaches were followed to obtain CNC hydrogel beads with tunable size and regular shape. A superior morphological control—that is, a more regular shape and smaller dimension of the beads—were obtained by centrifugal force-driven gelation. These results suggest that even by using a simple set-up and a low-speed centrifuge device, the extrusion of a colloidal solution through a small nozzle under a centrifugal field is an efficient approach for the production of CNC hydrogel beads with good reproducibility, control over the bead morphology and size monodispersion.

Porous silicon micro-particles (micro-pSi) with size in the range of 1–10 μm are obtained by etching of silicon wafers followed by sonication. The derivatization of the micro-pSi surface by wet chemistry (silylation and coupling with a diamine) yields an interface, which exposes negative (carboxylic) or positive (amine) groups at pH 7.4. The surface modification, beyond the introduction of groups for the drug loading by covalent or electrostatic interactions, stabilizes the intense orange luminescence characteristic of the silicon nano-crystallites. Derivatization by amines introduces also a second emission in the blue region, which follows a different excitation pathway and can be attributed to the interface defects. The micro-pSi are efficiently internalized by human dendritic cells and do not show any toxic effect even at a concentration of 1 mg mL−1. The intrinsic luminescence of the differently functionalized micro-pSi is preserved inside the cells and permits the selective and efficient tracking of the microparticles without using molecular tags and thus leaving the organic coating available for the interaction with the drug. The results obtained suggest that the functionalized micro-pSi are an efficient platform for simultaneous imaging and delivery of therapeutic agents to the disease site.

Porous silicon (pSi) microparticles obtained by porosification of crystalline silicon wafers have unique optical properties that, together with biodegradability, biocompatibility and absence of immunogenicity, are fundamental characteristics to candidate them as tracers in optical imaging techniques and as drug carriers. In this work, we focus on the possibility to track down the pSi microparticles also by MRI (magnetic resonance imaging), thus realizing a comprehensive tool for theranostic applications, i.e., the combination of therapy and diagnostics. We have developed and tested an easy, quick and low-cost protocol to infiltrate the COOH-functionalized pSi microparticles pores (tens of nanometers about) with magnetic nanospheres (SPIONs&mdash, Super Paramagnetic Iron Oxide Nanoparticles, about 5&ndash, 7 nm) and allow an electrostatic interaction. The structural properties and the elemental composition were investigated by electron microscopy techniques coupled to elemental analysis to demonstrate the effective attachment of the SPIONs along the pores&rsquo, surface of the pSi microparticles. The magnetic properties were investigated under an external magnetic field to determine the relaxivity properties of the material and resulting in an alteration of the relaxivity of water due to the SPIONs presence, clearly demonstrating the effectiveness of the easy functionalization protocol proposed.

The European food sector generates about 250 million tons/yr of by-products and waste, of which around 10% from fruit and vegetable processing, with a heavy environmental burden. The agri-food residues (AFR) contain a significant fraction of cellulose and bioactive compounds (mainly antioxidants), which, if recovered, are high added-value material components. The reduction of cellulose down to nano-sized crystalline structures (nanocellulose, NC) provides versatile building blocks, which self-assemble into new materials with superior performances. Despite wood-derived NC is generally considered a green material, its production process is environmentally unfriendly and its large scale utilization would contribute to deforestation. Therefore, more sustainable sources, such as AFR, are desired. The PANACEA project, within the frame of PRIN 2017 call supported by the Italian Ministry of University and Research, proposes an approach based on the recovery of cellulose and bioactive compounds from AFR, with high yield, at various degrees of hierarchical organization, by cascading different physical and chemical processes of increasing complexity. More specifically, physical processes and microbial digestion are exploited to obtain micro-sized cellulose structures while preserving their bioactivity. Chemical and enzymatic processes are used to isolate, purify and functionalize NC at different levels of hierarchical organization, and to design advanced functional materials such as food ingredients, edible coatings, functional colloids, biocides and flame retardants. The sustainable integrated valorization of AFR is addressed through (i) ad hoc pre-treatment of different AFR (Tomato peels, Wheat straw, Coffee residues, Rice bran, Grape marcs, Orange peels), available all year round and with different composition, (ii) application of proprietary high-pressure homogenization (HPH) techniques to micronize the residues in water and completely disintegrate the vegetable cells, with integrated physical fractionation to recover insoluble fibers, (iii) combination with chemical fractionation or high P/T autohydrolysis, or enzymatic treatments. The fabrication of NC structures with tunable size, crystallinity, and surface properties is important to bridge the gap between the production of cellulose and cellulose hybrids and the final applications and is pursued through (i) the size-reduction of NC, or enzymatic lysis, (ii) functionalization via amination or phosphorylation, or (iii) via physical immobilization or covalent bonding with limonene. The characterization of NC in terms of (i) yields, purity, physicochemical, structural, and functional properties, using a multi-technique approach, is associated to the evaluation of (ii) bio-accessibility (through the simulated digestive process) of bioactive compounds and fibers, (iii) film-forming capacity, (iv) gas-barrier properties, (v) rheological behavior, as well as (vi) energy, water, and reagents consumptions. Finally, the PANACEA project also addresses the exploitation of NC-based colloids for novel materials and applications, such as (i) edible coatings, antimicrobial varnishes, and oil structuring materials, (ii) packaging films with gas-barrier properties, and (iii) fabrics and foams with flame-retardant properties (iv) advanced nanocomposite films, (v) Pickering emulsions, and (vi) antimicrobial films and coatings. The main contribution of the PANACEA project to the advancement of the knowledge is expected in (i) the deployment of greener and sustainable, cascading processes exploiting raw AFR functionalities, (ii) the tailored valorization of different AFR by developing physical, chemical, and biological procedures for sculpting the nanostructures, and (iii) the development of sustainable, high performing advanced materials such as edible coatings and gas barrier packages for the food industry, foams, and textiles with flame retardant properties, biocides for organic pest control in agriculture.

Bilayer membranes for food packaging applications were prepared by depositing on polylactic acid (PLA) substrates pure cellulose nanofibers' (CNF) coatings and nanocomposite coatings made of CNF with dispersed TiO2 nanoparticles. The gas barrier properties of the bilayer membranes were studied by gas phase permeation technique using CO2, N2, D2, He and ambient air as test gases. Few micrometer thick pure and nanocomposite CNF coatings act as effective gas permeation barriers impeding the CO2, N2 and O2 transport and reducing the He and D2 transport rates by 3 orders of magnitude when compared with the PLA substrate. Gas transport through the coatings obeys the solution-diffusion mechanism: D2 permeation process shows activation energy values of 44 ± 6 kJ/mol and 40 ± 6 kJ/mol in the pure and nanocomposite CNF coating, respectively. The addition of the TiO2 nanoparticles reduces the penetrant diffusivity by increasing the tortuosity of the penetrant migration path but does not affect the gas barrier performances. UV–Vis spectroscopy analyses show that the optical transparency of the PLA support is slightly decreased by the CNF coating: at 600 nm the optical transmittance decreases, in fact, from 95% to 91%. TiO2 nanoparticles strongly absorb UV radiation and slightly reduce the PLA transparency in the optical range due to light scattering effects.

We have studied the kinetics of gas transport through films made of self-assembled cellulose nanofibrils (CNF) by a time-resolved mass spectroscopy technique. Few micrometers thick films deposited on polylactic acid (PLA) substrates act as impermeable barriers for CO2, O2, and N2 and reduce the 2H2 (deuterium) and He permeation flux by a factor of ∼103 with respect to the uncoated substrate. Penetrant transport is controlled by the solution-diffusion mechanism and the coating acts as a diffusive barrier. 2H2 and He diffusivity values are in the 10–10 and 10–9 cm2 s–1 range, respectively, and their migration occurs by thermally activated process with 39 ± 1 and 33 ± 2 kJ mol–1 activation energy. Positron annihilation lifetime spectroscopy analysis indicates that the diffusive path between the packed nanofibrils consists of elongated cavities with cross-sectional size ∼ 0.31 nm. Results evidence that the selective transport of the small size penetrants is due to sieving effects and that small penetrant tran...

Selective functionalization of the external/internal pore surface of porous silicon is of interest for the numerous potential applications of this material, in particular in pharmacology. With the aim of obtaining porous silicon platforms compatible with the aqueous environment and providing hydrophobic pores to load poorly water soluble molecules, we set-up a three step functionalization procedure consisting in two hydrosilylation reactions separated by the selective etching of the external surface. This procedure was applied both, to porous layers and porous microparticles. The characterization of the functionalized material by conventional techniques such as contact angle and FTIR showed a change of the properties of porous structures in line with the expected surface modifications. However, these techniques do not permit to clearly distinguish between internally and externally grafted functional groups. For this reason, an innovative procedure based on angle-resolved XPS was set-up and applied to differently functionalized pSi layers. By this technique, we obtained indications of prevalent grafting of hydrophilic moieties on the external surface and hydrophobic ones inside the pores.

A functionalization strategy, consisting of a silylation reaction by acrylic acid followed by diamine coupling, preserves and stabilizes the photoluminescence (PL) of porous silicon (pSi) microparticles suspended in ethanol. We found that under the condition of efficient amine coupling, besides the orange emission typical of the native pSi, an emission band in the blue region appears. The investigation of the interaction between pSi and diamine shows that diamine quenches and shifts the orange band meanwhile it induces an increase of the intensity of the blue one. PL lifetimes of the orange and blue bands are in the micro and nano second range, respectively. These values and their wavelength dependence clearly prove that the two bands have different origin: quantum confinement and nitrogen impurities introduced at silicon/silicon oxide interface, respectively. Thus, they can be used to discriminate between the pSi microparticles obtained by silylation, which expose carboxylic groups and the pSi microparticles after the diamine coupling, which bear amine functionalities at the surface. The increase in the stability of the PL emission of pSi in aqueous solution after functionalization, with quantum yields of the order of 1–2%, supports the use in biological systems of these brightly emitting, largely porous microparticles, bearing positive or negative surface charge.

In this study we investigated the fabrication of small hydrogel objects by the coordination-driven assembly of supramolecular rod-like crystallites of nanocellulose, using ionotropic gelation as a methodological approach and Ca2+ as a gelling agent. We proved that the gelation process is diffusion-mediated and fitting the equations modelling this process to the profile of the Ca2+ front, a Ca2+ diffusion coefficient in the incipient hydrogel of (4.5 ± 1.1) × 10−6 cm2 s−1 was calculated. At the steady-state a spatially homogeneous distribution of Ca2+-crosslinked sites in the hydrogel network was observed. External ionotropic gelation produced beads, wires or disks, while core–shell capsules were obtained by inverse ionotropic gelation. We demonstrated that equilibrium and dynamics of the distribution of Ca2+ offer the opportunity to design precisely the size and shape of these small hydrogel objects. In particular, the core size and the shell thickness of the capsules can be tailored under kinetic controlled conditions. The proposed approach, with supramolecular structures of the natural source as assembling components and the water-in-water fabrication process, is fast, simple, and requires only sustainable chemistry and is easily implementable in automatic microfluidic platforms.

In recent decades, biocompatible and light emitting porous silicon (pSi) showed the possibility for use in biomedical applications. Unfortunately, until now the biological applications of light-emitting pSi have been severely limited due the immediate surface oxidation of this nanostructured material, followed by loss of optical properties and degradation in aqueous solutions. In view of drug delivery, the red-ox activity of pSi damages the carried drug. Herein, we have investigated the stabilization of pSi microparticles in aqueous solution by covalent attachment of polymers to a short-chain organic coating grafted onto inorganic silicon. The morphological, chemical and optical stability obtained using polyethylene glycol confirm this biocompatible polymer as a superior shell matrix nanostructure for use in biological environments.

Silicon is the most common semiconductor in technology. Its main limit is the lacking of efficient light emission properties. Here quantum effects are exploited to fabricate optically active silicon photonics elements. The manuscript reports recent results obtained in different research fields spanning from bio- to integrated-photonics and nonlinear optics.

Mesoporous silicon (pSi) has several interesting features that makes it suitable for various biomedical applications. In particular, the large surface area make it very sensitive to environment changes. Among other approaches, thermal oxidation is an effective way to passivate its surface. Herein, we present experimental and analytical results concerning kinetics of thermal oxidation reaction of pSi. The experiments were conducted on pSi powders produced from silicon wafer by anodization and converted to particles by sonication. Oxidation experiments were carried out at different heating rates. Structure and morphology of the samples have been investigated by XRD and SEM before and after thermal oxidation. The model-free kinetics proposed by Ozawa–Flynn–Wall (OFW) was used to determine the Arrhenius relationship for the pSi thermal oxidation. The obtained apparent activation energy by OFW was confirmed by Starink method. At low temperature, the oxidation of surface dangling bonds obeys the Avrami–Erofeev mechanism. At high temperature, oxidation is followed by classical bulk oxidation according to diffusion mechanism controlled by the diffusion of oxygen through the silicon dioxide layer on the surface of the pSi. The reaction mechanism was checked by the model fitting kinetics, which confirmed the reaction is a kind of sequential two-stage process, Avrami–Erofeev and 3D diffusion. Finally, differential thermal analysis suggests that the second oxidation step is also possibly affected by phase transformation of the silicon dioxide.

Porous silicon (pSi) is a sponge-like material obtained by electrochemical etching of a crystalline silicon wafer. Due to quantum confinement effects, this material is photoluminescent and this is a fundamental property from the perspective of bioimaging applications. Limitations in nanomedicine to the use of photoluminescent pSi structures are mainly due to optical quenching in an aqueous environment and to the adverse effects of reactive groups introduced by etching procedures. In this work, we exploited an inorganic TiO2 coating of pSi microparticles by Atomic Layer Deposition (ALD) that resulted in optical stability of pSi particles in a biological buffer (e.g. PBS). The use of a rotary reactor allows deposition of a uniform coating on the particles and enables a fine tuning of its thickness. The ALD parameters were optimized and the photoluminescence (PL) of pSi–TiO2 microparticles was stabilized for more than three months without any significant effect on their morphology. The biocompatibility of the coated microparticles was evaluated by analyzing the release of cytokines and superoxide anion (O2−) by human dendritic cells, which play an essential role in the regulation of inflammatory and immune responses. We demonstrated that the microparticles per se are unable to significantly damage or stimulate human dendritic cells and therefore are suitable candidates for nanomedicine applications. However, a synergistic effect of the microparticles with bacterial products, which are known to stimulate immune-response, was observed, indicating that a condition unfavorable to the use of inorganic nanomaterials in biological systems is the presence of infection diseases. These results, combined with the proved PL stability in biological buffers, open the way for the use of pSi–TiO2 microparticles as promising materials in nanomedicine, but their ability to increase immune cell activation by other agonists should be considered and even exploited.

In a biological cell, cellular functions and the genetic regulatory apparatus are implemented and controlled by complex networks of chemical reactions involving genes, proteins, and enzymes. Accurate computational models are indispensable means for understanding the mechanisms behind the evolution of a complex system, not always explored with wet lab experiments. To serve their purpose, computational models, however, should be able to describe and simulate the complexity of a biological system in many of its aspects. Moreover, it should be implemented by efficient algorithms requiring the shortest possible execution time, to avoid enlarging excessively the time elapsing between data analysis and any subsequent experiment. Besides the features of their topological structure, the complexity of biological networks also refers to their dynamics, that is often non-linear and stiff. The stiffness is due to the presence of molecular species whose abundance fluctuates by many orders of magnitude. A fully stochastic simulation of a stiff system is computationally time-expensive. On the other hand, continuous models are less costly, but they fail to capture the stochastic behaviour of small populations of molecular species. We introduce a new efficient hybrid stochastic-deterministic computational model and the software tool MoBioS (MOlecular Biology Simulator) implementing it. The mathematical model of MoBioS uses continuous differential equations to describe the deterministic reactions and a Gillespie-like algorithm to describe the stochastic ones. Unlike the majority of current hybrid methods, the MoBioS algorithm divides the reactions' set into fast reactions, moderate reactions, and slow reactions and implements a hysteresis switching between the stochastic model and the deterministic model. Fast reactions are approximated as continuous-deterministic processes and modelled by deterministic rate equations. Moderate reactions are those whose reaction waiting time is greater than the fast reaction waiting time but smaller than the slow reaction waiting time. A moderate reaction is approximated as a stochastic (deterministic) process if it was classified as a stochastic (deterministic) process at the time at which it crosses the threshold of low (high) waiting time. A Gillespie First Reaction Method is implemented to select and execute the slow reactions. The performances of MoBios were tested on a typical example of hybrid dynamics: that is the DNA transcription regulation. The simulated dynamic profile of the reagents' abundance and the estimate of the error introduced by the fully deterministic approach were used to evaluate the consistency of the computational model and that of the software tool.

Sol-gel transition of carboxylated cellulose nanocrystals has been investigated using rheology, SAXS, NMR and optical spectroscopies to unveil the distinctive roles of ultrasound treatments and addition of various cations. Besides cellulose fiber fragmentation, sonication treatment induces fast gelling of the solution. The gelation is independent of the addition of cations, while the final rheological properties are highly influenced by the type, concentration and sequence of the operations since the cations must be added prior to sonication to produce stiff gels. The gel elastic modulus was found to increase proportionally to the ionic charge rather than the cationic size. In cases where ions were added after sonication, SAXS analysis of the Na+ hydrogel and Ca2+ hydrogel indicated the presence of structurally ordered domains in which water is confined, and 1H-NMR investigation showed the dynamics of water exchange within the hydrogels. Conversely, separated phases containing essentially free water were characteristic of the hydrogels obtained by sonication after Ca2+ addition, confirming that this ion induces irreversible fiber aggregation. The rheological properties of the hydrogels depend on the duration of the ultrasound treatments, enabling the design of programmed materials with tailored energy dissipation response.

α-Tocopherol, the main component of vitamin E, traps highly reactive radicals which otherwise might react with lipids present in plasmatic lipoproteins or in cell membranes. The α-tocopheroxyl radicals generated by this process have also a pro-oxidant action which is contrasted by their reaction with ascorbate or by bimolecular self-reaction (dismutation). The kinetics of this bimolecular self-reaction were explored in solution such as ethanol, and in heterogeneous systems such as deoxycholic acid micelles and in human plasma. According to ESR measurements, the kinetic rate constant (2k(d)) of the bimolecular self-reaction of α-tocopheroxyl radicals in micelles and in human plasma was calculated to be of the order of 10(5) M(-1) s(-1) at 37 °C. This value was obtained considering that the reactive radicals are confined into the micellar pseudophase and is one to two orders of magnitude higher than the value we found in homogeneous phase. The physiological significance of this high value is discussed considering the competition between bimolecular self-reaction and the α-tocopheroxyl radical recycling by ascorbate.

The structures of copper amine oxidases from various sources show good similarity, suggesting similar catalytic mechanisms for all members of this enzyme family. However, the optimal substrates for each member differ, depending on the source of the enzyme and its location. The structural factors underlying substrate selectivity still remain to be discovered. With this in view, we examined the kinetic behaviour of pea seedling amine oxidase with cadaverine and hexylamine, the first bearing two, and the second only one, positively charged amino group. The dependence of Km and catalytic constant (kc) values on pH, ionic strength and temperature indicates that binding of the monoamine is driven by hydrophobic interactions. Instead, binding of the diamine is strongly facilitated by electrostatic factors, controlled by polar side-chains and two titratable residues present in the active site. The position of the docked substrate is also essential for the participation of titratable amino acid residues in the following catalytic steps. A new mechanistic model explaining the substrate-dependent kinetics of the reaction is discussed.

For the first time a variety of foods, characterized by high antioxidant activity, have been tested for the reactivity by which the system of antioxidants present in these foods competes for peroxyl radicals with poly-unsaturated fatty acids. The oxygraphic method we have used, on the basis of a rigorous kinetic model, permits to obtain the reactivity that is the Peroxyl Radical Trapping Efficiency (PRTE), beyond the Peroxyl Radical Trapping Capacity (PRTC) and to assign to each food characteristic values of these parameters, so facilitating their inter-comparison. In the analyzed foods the PRTE/PRTC ratio spans more than one order of magnitude, so reflecting the quality of antioxidants present in foods. According to the PRTE values and on the basis of the serving size the ranking of antioxidant food efficiency in trapping peroxyl radicals was blueberry > red chicory > coffee > pineapple ≈ red wine ≥ orange ≈ dark chocolate ≈ apple ≥ tea > pomegranate.

Porous silicon solid supports with pore diameter 0.5–1 μm, infiltrated with Ag nanostructures for surface enhanced Raman scattering (SERS) were prepared according to two procedures: spontaneous Ag+ reduction on the surface of freshly etched porous silicon immersed in Ag+ aqueous solutions, or anchoring colloidal Ag nanoparticles on the surface previously functionalized by aminosilane. Using Rhodamine 6G (RH6G) as analyte the detection limits were of the order of 20 μM and 20 nM with porous silicon metalized by the first and second procedure, respectively. This large increase of sensitivity notwithstanding a reduced surface density of Rhodamine 6G obtained on porous silicon metalized by the second procedure is discussed in terms of better hot spot efficiency and reduced aspecific binding out of the hot regions obtained depositing the colloids on the aminosilane functionalized surface.

Silver nanocrystallites are obtained through immersion of porous silicon samples in AgNO 3 solutions and a successive thermal annealing. The efficiency of nanostructures as surface enhanced Raman scattering (SERS) substrates is checked on cyanine-based dyes and horseradish peroxidase, evidencing detectable concentrations as low as 10 −7 to 10 −8 M. The substrate efficiency is strictly related to the Ag particle morphology, which could yield to either local surface plasmons (LSP) coupled to individual particles or to inter-particle short-range interaction.

In this paper, we report on a method to evaluate the activity of water soluble and H-atom donor antioxidants as peroxyl radical scavengers in a micelle system reproducing the conditions occurring in the upper small intestine in humans, during digestion and absorption of lipids. This method, which overcomes some of the problems of the total radical trapping antioxidant parameter (TRAP) assays, measures the peroxyl radical trapping capacity (n) and the peroxyl radical trapping efficiency IC 50 − 1 of antioxidants, that is the number “n” of peroxyl radicals trapped by one molecule of the studied antioxidant and the reciprocal of the antioxidant concentration that halves the steady-state concentration of peroxyl radicals, respectively. These two fundamental parameters characterizing the radical chain breaking of many water soluble antioxidants, among which dietary polyphenols, can be obtained with relatively good precision from a single experiment, on the basis of a rigorous treatment of the kinetic data.

The inhibition by anthocyanins of the free radical-mediated peroxidation of linoleic acid in a SDS micelle system was studied at pH 7.4 and at 37 degrees C, by oxygraphic and ESR tecniques. The number of peroxyl radicals trapped by anthocyanins and the efficiency of these molecules in the trapping reaction, which are two fundamental aspects of the antioxidant action, were measured and discussed in the light of the molecular structure. In particular the contribution of the substituents to the efficiency is -OH-OCH(3)-H. By ESR we found that the free radicals of anthocyanins are generated in the inhibition of the peroxidation of linoleic acid. The life time of these radical intermediates, the concentration of which ranges from 7 to 59 nM under our experimental conditions, is strictly correlated with the anthocyanin efficiency and with the heat of formation of the radical, as calculated by a semiempirical molecular orbital approach.

Nanoporous materials are ideal candidates to fabricate sensors with high sensitivities. Using nanoporous materials both the strength and the type of interactions between analytes and host matrix can be tuned at will, and their large surface area permit to achieve ultralow detection limit and to keep high loading capacity and, thus, large dynamic range. Despite these intriguing possibilities their use in sensing technology is still limited because several fundamental aspects are not fully understood. By comparing supported and free standing Porous Silicon membranes, we demonstrate that sensor’s sensitivity is clearly overestimated if the sample is not properly washed. Moreover we underline the fact that, despite the reduced Q-factor of the resonant cavities, the strong interaction between the analyte and the optical modes in porous materials produces a large net sensitivity of the sensor. Finally, we show that the non-specific signal may have a linear response with the analyte concentration and it thus extremely difficult to separate it from the specific one. The work presented here is of importance in the design of nanoporous materials for sensing application. In fact it demonstrates that the final sensitivity of the device is not defined by its optical quality alone and that several other effects have to be considered to fabricate a reliable sensor.

Aqueous solutions of silicon nanocrystals have been obtained, by sonication, from porous Si (p-Si) aged in air for various times. The photoluminescence of these solutions changes with the aging time of p-Si. These changes correlate with nanocrystal core dimensions, i.e. with the oxidation of the nanocrystals. Infrared spectra show that the reaction with water depends on the age of the starting p-Si sample, since the native superficial oxide layer on p-Si inhibits these reactions.

Eight varieties of Cichorium genus vegetables (five heavily red colored, one red spotted, and two fully green) were investigated for their phenolic content (by HPLC and UV-vis spectrophotometry) and for their antioxidant activity. In particular, the capacity (that is, the amount of trapped peroxyl radicals) and the efficiency (that is, the amount of antioxidant necessary to halve the steady-state concentration of peroxyl radicals) were measured. All of the studied chicories are characterized by the presence of a large amount of hydroxybenzoic and hydroxycinnamic acids, whereas the red color is due to cyanidin glycosides. The presence of these phenolics in red chicories confers to them an exceptionally high peroxyl radical scavenging activity in terms of both capacity and efficiency, particularly in their early stage of growth, and makes this popular and low-cost foods comparable or superior to many foods having well-known antioxidant properties such as red wine, blueberry, and tomato.

TPP+ (tetraphenylphosphonium ion) and its analogues were found to act as powerful competitive inhibitors of BSAO (bovine serum amine oxidase). The binding of this new class of inhibitors to BSAO was characterized by kinetic measurements. TPP+ can bind to the BSAO active site by hydrophobic and by coulombian interactions. The binding probably occurs in the region of the ‘cation-binding site’[Di Paolo, Scarpa, Corazza, Stevanato and Rigo (2002) Biophys. J. 83, 2231–2239]. Under physiological conditions, the association constant of TPP+ for this site is higher than 106 M−1, the change of enthalpy being the main free-energy term controlling binding. Analysis of the relationships between substrate structure and extent of inhibition by TPP+ reveals some new molecular features of the BSAO active site.

Thirty-two experimental red wines, obtained from eight cultivars and aged in bottles for 2 and 7 years, were examined for the presence of stable free radicals (SFR), for the peroxyl radical trapping capacity (PRTC), and for the concentrations of some important polyphenol families. Aging significantly increases SFR, polyphenol polymers with n ≥ 5 (HMWP), and PRTC and is accompanied by a strong decrease of free anthocyanins. Multivariate regression analyses show that HMWP and SFR are independently associated with PRTC while HMWP and anthocyanins are independently associated with the formation of SFR. These results indicate that polymeric polyphenols generated from anthocyanins and proanthocyanidins during wine aging are able to convert highly reactive free radicals into nonreactive radicals through electron delocalization. The strict correlation between SFR and antioxidant activity that we found suggests that these characteristics are related to the functional properties of food. Keywords: Free radical; ant...

The assessment of kidney viability before transplantation (with a view of discarding nonviable organs) remains an obstacle to confidently extending organ harvesting to marginal donors. In the present study phosphorus magnetic resonance spectroscopy was used to monitor metabolic changes in (31)P-containing metabolites in isolated porcine kidneys. After various warm ischemia times, the organs were stored at 0 degrees C. Time-dependent changes in the phosphomonoester/inorganic-phosphate ratio were recorded at 0 degrees C were shown to follow a biexponential decay. The first-order kinetic rate constant of the short-time decay was strongly dependent on the warm ischemia time, a result that was discreted in terms of the underlying biochemistry. The metabolic events responsible for the dramatic decrease in phosphomonoester/inorganic phosphate ratio that occur immediately after organ perfusion and storage, suggest that any procedure to minimize organ damage must occur immediately after harvesting.

A steady-state kinetic study of bovine serum amine oxidase activity was performed, over a wide range of pH values (5.4-10.2) and ionic strength (10-200 mM), using various (physiological and analogue) substrates as specific probes of the active-site binding region. Relatively small changes in k (cat) values (approx. one order of magnitude) accompanied by marked changes in K(m) and k(cat)/K(m) values (approx. six orders of magnitude) were observed. This behaviour was correlated with the presence of positively charged groups or apolar chains in the substrates. In particular, it was found that the docking of the physiological polyamines, i.e. spermidine and spermine, appears to be modulated by three amino acid residues of the active site, which we have named L(-)H(+), G(-)H(+) and IH(+), characterized by p K (a) values of 6.2+/-0.2 [Di Paolo, Scarpa, Corazza, Stevanato and Rigo (2002) Biophys. J. 83, 2231-2239], 8.2+/-0.3 and 7.8+/-0.4 respectively. The electrostatic interaction between the protonated substrates and the enzyme containing the residues L(-)H(+), G(-)H(+) and IH(+) in the deprotonated form, the on/off role of the IH(+) residue and the role of hydrophobic interactions with substrates characterized by apolar chains are discussed.

The inhibitory effect of anthocyanins has been investigated in the peroxidation of linoleic acid in micelles in the presence and in the absence of (+)-catechin. The peroxidation was initiated by thermal decomposition of 2,2(')-azobis[2-(2-imidazolin-2-yl)propane], and the kinetics of peroxidation were followed by measuring the rate of oxygen consumption and the rate of disappearance of the antioxidant. The analysis of the antioxidant effect of various anthocyanins, alone or in the presence of catechin, demonstrates that catechin, which is relatively inefficient at inhibiting linoleic acid oxidation, regenerates the highly efficient antioxidant malvidin 3-glucoside and, at a lower extent, peonidin 3-glucoside. The malvidin 3-glucoside recycling by catechin strongly increases the antioxidant efficiency of these two antioxidants. This protective mechanism appears specific for malvidin and peonidin 3-glucosides. The high unpaired spin density of the phenolic O atoms in the radicals generated by these anthocyanins, calculated by the semiempirical quantum chemical AM1 method, may explain the observed behavior.

In this paper, we report on the presence of cation binding areas on bovine serum amine oxidase, where metal ions of the groups IA and IIA, such as Na(+), K(+), Cs(+), Mg(2+), and Ca(2+), bind with various affinities. We found a cation-binding area that influences the enzyme activity if occupied, so that the catalytic reaction may be altered by some physiologically relevant cations, such as Ca(2+) and K(+). This binding area appears to be localized inside the enzyme active site, because some of these cations act as competitive inhibitors when highly charged amines, such as spermine and spermidine, are used as substrates. In particular, dissociation constant values (K(d)) of 23 and 27 mM were measured for Cs(+) and Ca(2+), respectively, using, as substrate, spermine, a polyamine of plasma. An additional cation-binding area, where metal ions such as Cs(+) (K(d) congruent with 0.1 mM) and Na(+) (K(d) congruent with 54 mM) bind without affecting the enzyme activity, was found by NMR.

In this article we describe the fabrication of free standing n-type porous silicon microcavity (MC) and their properties as liquid sensors. We have optimized the etching recipe to keep both large pore size and high quality factor (Q-factor). Thus the fabricated porous layers have pore size in the range of 40 to 110 nm and are thus compatible with mass transport across the porous layer. We found that MC with a Q-factor of 60 can measure down to 1.1*10 -5 refractive index variations. Furthermore we analyze the role of non specific binding by comparing flow through versus flow over geometries. We compare these two approaches using different techniques and we show that flow over assay systematically overestimates the sensitivity of the device because of an inefficient rinse of the sample. Our work clearly indicates a limit in the reliability of measurements performed in flow over geometry unless specific controls are taken into account. Keywords: Porous silicon, Free standing membrane, Microcavity, Non specific signal, flow through, mass transport, electrochemical etching

A patient with a severe amnesic syndrome following a glioma of the splenium of the corpus callosum is reported. The long-term memory deficit involved anterograde as well as retrograde events dating back to 40 years and causing topographical disorientation. Short-term memory test performance was in the normal range, with the exception of tactile memory which was severely impaired. The patient also showed disconnection symptoms, due to severing of occipito-parietal and parieto-temporal connections, while parieto-parietal connections were undamaged.

Porous materials are ideal hosts to fabricate high sensitivity devices. Their large specific area and the possibility to modify the type and the strength of the matrix–analyte interactions allow the realization of sensors with finely tailored characteristics. In this article, we investigate how mass transport across the nanoporous structure influences the response due to the non-specific signal by comparing flow-through versus flow-over geometries. We observed a systematic overestimation of the sensitivity for porous substrate devices made of closed-ended pores compared with open-ended pore ones. Our analysis shows that such an effect is due to (unbound) analytes or contaminants that remain trapped within the pores and are not removed by rinsing of the sample. This result was verified by measuring similar samples in both flow through and flow over configurations, as well as their residual response after blockage of all their active sites. We also notice that sensors based on free-standing membranes show similar results independent of the fact that mass transport is induced by either an external pressure source or simply by Brownian motions.

The isoenzyme I of cytosolic Cu,Zn-superoxide dismutase (SOD) from Nicotiana plumbaginifolia (tobacco) leaves has been purified to apparent homogeneity. The relative molecular mass of the native isoenzyme, determined by gel filtration chromatography, is about 33.2 kDa. SDS-polyacrylamide gel electrophoresis shows that the enzyme is composed of two equal subunits of 16.6 kDa The isolectric point, assayed by isoelectric focusing, in the pH range of 3.5–6.5, is 4.3. The enzyme stability was tested at different temperatures, pH, and concentration of inhibitors (KCN and H 2 O 2 ). The catalytic constant ( k cat ) was 1.17 ± 0.14 × 10 9 M −1 s −1 at pH 9.9 and 0.1 M ionic strength. The activation energy of the thermal denaturation process is 263 kJ mol −1 . The electrostatic surface potential of the modeled tobacco Cu,Zn-SOD I was calculated showing that the functional spatial network of charges on the protein surface has been maintained, independently of the amino acid substitution around the active sites.