Strictosidine synthase: mechanism of a Pictet-Spengler catalyzing enzyme

The Pictet-Spengler reaction, which yields either a beta-carboline or a tetrahydroquinoline product from an aromatic amine and an aldehyde, is widely utilized in plant alkaloid biosynthesis. Here we deconvolute the role that the biosynthetic enzyme strictosidine synthase plays in catalyzing the stereoselective synthesis of a beta-carboline product. Notably, the rate-controlling step of the enzyme mechanism, as identified by the appearance of a primary kinetic isotope effect (KIE), is the rearomatization of a positively charged intermediate. The KIE of a nonenzymatic Pictet-Spengler reaction indicates that rearomatization is also rate-controlling in solution, suggesting that the enzyme does not significantly change the mechanism of the reaction. Additionally, the pH dependence of the solution and enzymatic reactions provides evidence for a sequence of acid-base catalysis steps that catalyze the Pictet-Spengler reaction. An additional acid-catalyzed step, most likely protonation of a carbinolamine intermediate, is also significantly rate controlling. We propose that this step is efficiently catalyzed by the enzyme. Structural analysis of a bisubstrate inhibitor bound to the enzyme suggests that the active site is exquisitely tuned to correctly orient the iminium intermediate for productive cyclization to form the diastereoselective product. Furthermore, ab initio calculations suggest the structures of possible productive transition states involved in the mechanism. Importantly, these calculations suggest that a spiroindolenine intermediate, often invoked in the Pictet-Spengler mechanism, does not occur. A detailed mechanism for enzymatic catalysis of the beta-carboline product is proposed from these data.     

Chemical Composition,Antioxidant and Anti-Inflammatory Activities of Clary Sage and Coriander Essential Oils Produced on Polluted and Amended Soils-Phytomanagement Approach

The potential of essential oils (EO), distilled from two aromatic plants—clary sage (Salvia sclarea L.) and coriander (Coriandrum sativum L.)—in view of applications as natural therapeutic agents was evaluated in vitro. These two were cultivated on a trace element (TE)-polluted soil, as part of a phytomanagement approach, with the addition of a mycorrhizal inoculant, evaluated for its contribution regarding plant establishment, growth, and biomass production. The evaluation of EO as an antioxidant and anti-inflammatory, with considerations regarding the potential influence of the TE-pollution and of the mycorrhizal inoculation on the EO chemical compositions, were the key focuses. Besides, to overcome EO bioavailability and target accession issues, the encapsulation of EO in β-cyclodextrin (β-CD) was also assessed. Firstly, clary sage EO was characterized by high proportions of linalyl acetate (51–63%) and linalool (10–17%), coriander seeds EO by a high proportion of linalool (75–83%) and lesser relative amounts of γ-terpinene (6–9%) and α-pinene (3–5%) and coriander aerial parts EO by 2-decenal (38–51%) and linalool (22–39%). EO chemical compositions were unaffected by both soil pollution and mycorrhizal inoculation. Of the three tested EO, the one from aerial parts of coriander displayed the most significant biological effects, especially regarding anti-inflammatory potential. Furthermore, all tested EO exerted promising antioxidant effects (IC₅₀ values ranging from 9 to 38 g L⁻¹). However, EO encapsulation in β-CD did not show a significant improvement of EO biological properties in these experimental conditions. These findings suggest that marginal lands polluted by TE could be used for the production of EO displaying faithful chemical compositions and valuable biological activities, with a non-food perspective.     

A study on the interactions between coenzyme Q 0 and superoxide anion. Could ubiquinones mimic superoxide dismutase (SOD)?

An electrochemical study was carried out on 1,4-benzoquinone, duroquinone, coenzymes Q ₀ and Q ₁₀ in the absence and in the presence of molecular oxygen in aprotic (DMF) and protic (DMF/H₂O 95:5 (v/v)) media. Water was added because the investigated reactions are deeply influenced by the presence of protons. Q ₀ and Q ₁₀ exhibited a similar electrochemical behaviour. Since Q ₀ is more soluble in protic medium than the biologically more important analogue Q ₁₀ , it was chosen as a model for a more detailed investigation. Voltammetric studies of Q ₀ carried out in aprotic and protic media in the presence of oxygen showed that, besides simple O₂ ^·− dismutation, the Q ₀ promoted dismutation of O₂ ^·− should also be considered. Spectroelectrochemical experiments with the same experimental conditions support the electrochemical results, showing that in the presence of superoxide and in aprotic medium semiquinone Q ₀ ^·− gives rise to a disproportionation equilibrium, while in the presence of water it tends to be reoxidized to the starting Q ₀ by OOH^·. EPR measurements are also in agreement with these results.     

Comparison of Antibacterial and Antioxidant Properties of Red (cv. Negramaro) and White (cv. Fiano) Skin Pomace Extracts

Wine pomace has attracted the attention of the food industry, due to its high content in bioactive compounds, and its multiple healthy activities. In this work, whole and separated skin pomaces from fermented (red) and un-fermented (white) grape by-products were characterized for their antioxidant and antimicrobial activities in order to exploit them as functional food ingredient. Antioxidant activity, measured by both ORAC and TEAC assays, was higher in whole than in skin pomace extracts. The characterization of phenolic composition in whole and skin pomace extracts confirmed the peculiarity of some compounds such as anthocyanins (107.84 + 10.3 mg/g TP) in red skin pomace and a great amount of flavanols (80.73 + 4.04 mg/g TP) in white skin pomace. Whole and skin pomace extracts displayed the same antibacterial activity at 250 µg gallic acid equivalents (GAE)/mL. Red and white skin pomace extracts showed a Minimum Inhibitory Concentration (MIC) of 31.25–62.5 GAE/mL against Staphylococcus aureus and Enterococcus faecalis. Pseudomonas spp. were more sensitive to red skin pomace extracts rather than white skin pomace extracts. Given these results, both red and white pomace extracts could be exploited for future application in food, pharmaceutical and cosmetic industry.     

An extendable poroelastic plate formulation in dynamics

A strategy is presented, which allows deriving poroelastic plate formulations of any desired level of approximation. Starting point are the governing three-dimensional (3D) equations of poroelasticity in frequency domain developed by Biot. In order to reduce the dimension of the problem from 3D to 2D, all unknown quantities are approximated by series expansions in thickness direction. This avoids the need for any engineering assumptions. The reduction in the dimension can then be achieved by an integration over the thickness. After truncating the series, a special plate formulation is retrieved. Results are presented for a square, clamped plate which show excellent agreement with the solution of the 3D equations and a considerable saving in computation time.     

Synthesis and chiroptical properties of optically active photochromic methacrylic polymers bearing in the side chain the (S)-3-hydroxypyrrolidinyl group conjugated with the trans-azoaromatic chromophore

Novel optically active polymethacrylates, namely poly[(S)-3-methacryloyloxy-1-(4-azobenzene)pyrrolidine] and poly[(S)-3-methacryloyloxy-1-(4′-nitro-4-azobenzene)pyrrolidine], have been synthesized by radical polymerization of the corresponding monomers, prepared in turn through a synthetic route preserving the asymmetric center by any racemization reaction. These homopolymers are characterized by the presence in the side chain of an optically active pyrrolidinyl ring linked to the trans-azoaromatic system through the nitrogen atom. The optical activity of the polymers in solution appears much higher than that observed with the low molecular weight models, purposely synthesized for comparison. Circular dichroism spectra of the synthesized products demonstrated that, in solution, the macromolecules assume highly homogeneous conformations with a prevailing chirality to a larger extent with respect to analogous systems previously investigated. © 1999 John Wiley & Sons, Inc. J Polym Sci A: Polym Chem 37: 3257–3268, 1999     

On starting and stopping criteria for nested primal-dual iterations

Numerical Algorithms - The importance of an adequate inner loop starting point (as opposed to a sufficient inner loop stopping rule) is discussed in the context of a numerical optimization...     

Microimaging FT-IR spectroscopy on pathological breast tissues

Microimaging Fourier transform infrared spectroscopy is able to monitor differentiation between normal and malignant tissues. All the specimens, previously submitted to histological analysis, displayed abnormal spectra compared with the corresponding normal tissues with changes in many diagnostic bands like those arising from phosphate, C–O and CH stretching vibrational modes. The comparison between cancer (K) and connective (C) spectra evidenced the following differences: in the vCH region 3000–2800 cm⁻¹ no hypomethylation effect was evident in K; the convolution of the bands of connective indicated an expected higher membrane fluidity; in the neoplastic zone, Amide I and II modes showed convoluted bands with maxima at 1651 and 1547 cm⁻¹, respectively, indicating an α-helix conformation of proteins due to changes in the secondary structure proteins upon carcinogenesis. Other signature bands, such as the deformation O–P–O phosphate band at 965 cm⁻¹, suggested DNA conformational changes in solid cancer, infiltrating cancer and neoplasia in the region 1350–800 cm⁻¹. These characteristic bands have been monitored as a function of the degree of cancer progression. Chemometric methods, such as principal component analysis (PCA) and hierarchical clustering analysis (HCA) have been used in order to distinguish spectra of neoplastic and normal zones.     

Innovative Eco-Friendly Hydrogel Film for Berberine Delivery in Skin Applications

Hydrogel formulations (masks or patches, without tissue support) represent the new frontier for customizable skin beauty and health. The employment of these materials is becoming popular in wound dressing, to speed up the healing process while protecting the affected area, as well as to provide a moisturizing reservoir, control the inflammatory process and the onset of bacterial development. Most of these hydrogels are acrylic-based at present, not biodegradable and potentially toxic, due to acrylic monomers residues. In this work, we selected a new class of cellulose-derived and biodegradable hydrogel films to incorporate and convey an active compound for dermatological issues. Films were obtained from a combination of different polysaccharides and clays, and berberine hydrochloride, a polyphenolic molecule showing anti-inflammatory, immunomodulatory, antibacterial and antioxidant properties, was chosen and then embedded in the hydrogel films. These innovative hydrogel-based systems were characterized in terms of water uptake profile, in vitro cytocompatibility and skin permeation kinetics by Franz diffusion cell. Berberine permeation fitted well to Korsmeyer–Peppas kinetic model and achieved a release higher than 100 µg/cm² within 24 h. The latter study, exploiting a reliable skin model membrane, together with the biological assessment, gained insights into the most promising formulation for future investigations.