The Inactivation Mechanism of Human Group IIA Phospholipase A2 by Scalaradial

Secretory phospholipases A₂ (sPLA₂s) are implicated in the pathogenesis of several inflammation diseases, such as rheumatoid arthritis, septic shock, psoriasis, and asthma. Thus, an understanding of their inactivation mechanisms could be useful for the development of new classes of chemical selective inhibitors. In the marine environment, several bioactive terpenoids possess interesting anti‐inflammatory activity, often through covalent and/or noncovalent inactivation of sPLA₂. Herein, we report the molecular mechanism of human group IIA phospholipase A₂ (sPLA₂‐IIA) inactivation by Scalaradial (SLD), a marine 1,4‐dialdehyde terpenoid isolated from the sponge Cacospongia mollior and endowed with a significant anti‐inflammatory profile. Our results have been collected by a combination of biochemical approaches, advanced mass spectrometry, surface plasmon resonance, and molecular modeling. These suggest that SLD acts as a competitive inhibitor. Indeed, the sPLA₂‐IIA inactivation process seems to be driven by the noncovalent recognition process of SLD in the enzyme active site and by chelation of the catalytic calcium ion. In contrast, covalent modification of the enzyme by the SLD dialdehyde moiety emerges as only a minor side event in the ligand–enzyme interaction. These results could be helpful for the rational design of new PLA₂ inhibitors that would be able to selectively target the enzyme active site.     

From wheat sourdough to gluten‐free sourdough: a non‐conventional process for producing gluten‐free bread

Gluten‐free (GF) sourdough was prepared from wheat sourdough and analysed both in fresh (GFS) and dried forms (DGFS). The gluten content in each GF sourdough sample was <20 mg kg^?1. The dough leavening capacity and the properties of the bread samples were investigated and compared to those of bread prepared using bakery yeast (Saccharomyces cerevisiae). Two commercial rice‐based mixtures (different for the presence/absence of buckwheat flour) were used to prepare bread samples. In GFS, lactic acid bacteria (LAB) and yeasts were found in amounts corresponding to 10⁸ and 10⁷ CFU g^?1, respectively, whereas both LAB and yeasts were detected in lower amounts (about 10⁶ CFU g^?1) in DGFS. When used in bread‐making, both GFS types produced significant dough acidification and exhibited good dough development during proofing, resulting in loaves with specific volume values between 3.00 and 4.12 mL g^?1, values similar to those obtained for reference bread (3.05÷4.15 mL g^?1). The use of GFS was effective in lowering the bread staling rate during storage for up to 7 days.     

Floating modular drug delivery systems with buoyancy independent of release mechanisms to sustain amoxicillin and clarithromycin intra-gastric concentrations

Release modules of amoxicillin and clarithromycin combined in a single dosage form designed to float in the gastric content and to sustain the intra-gastric concentrations of these two antibiotics used for the eradication of Helicobacter pylori have been studied. The modules having a disc shape with curved bases were formulated as hydrophilic matrices. Two modules of clarithromycin were assembled by sticking the concave base of one module to the concave base of the other, creating an internal void chamber. The final dosage form was a floating assembly of three modules of clarithromycin and two of amoxicillin in which the drug release mechanism did not interfere with the floatation mechanism. The assembled system showed immediate in vitro floatation at pH 1.2, lasting 5?h. The in vitro antibiotics release profiles from individual modules and assembled systems exhibited linear release rate during buoyancy for at least 8?h. The predicted antibiotic concentrations in the stomach maintained for long time levels significantly higher than the respective minimum inhibitory concentrations (MIC). In addition, an in vivo absorption study performed on beagle dogs confirmed the slow release of clarithromycin and amoxicillin from the assembled system during the assembly’s permanence in the stomach for at least 4?h.     

Isolation and Characterization of Squamous Cell Carcinoma-Derived Stem-like Cells: Role in Tumor Formation

In human epidermis, keratinocyte stem cells (KSC) are characterized by high levels of β₁-integrin, resulting in the rapid adhesion to type IV collagen. Since epithelial tumors originate from KSC, we evaluated the features of rapidly adhering (RAD) keratinocytes derived from primary human squamous cell carcinoma of the skin (cSCC). RAD cells expressed higher levels of survivin, a KSC marker, as compared to non-rapidly adhering (NRAD) cells. Moreover, RAD cells proliferated to a greater extent and were more efficient in forming colonies than NRAD cells. RAD cells also migrated significantly better than NRAD cells. When seeded in a silicone chamber and grafted onto the back skin of NOD SCID mice, RAD cells formed tumors 2–4 fold bigger than those derived from NRAD cells. In tumors derived from RAD cells, the mitotic index was significantly higher than in those derived from NRAD cells, while Ki-67 and survivin expression were more pronounced in RAD tumors. This study suggests that SCC RAD stem cells play a critical role in the formation and development of epithelial tumors.     

Retrofitting of R/C shear walls by means of high performance jackets

A new technique for the strengthening of existing R/C shear walls based on the application of thin high performance jackets is presented in this paper. The strengthening jacket is made of high performance concrete, having a compression resistance higher than 150 MPa, and reinforced by means of an high strength steel mesh. The experimental study is carried out on a 1:3 scale R/C wall, proportioned to resist vertical loads only, and reinforced by means of a 15 mm thick high performance jacket. Cyclic loads of increasing magnitude are applied to the experimental shear wall up to collapse. The effectiveness of the technique is also verified numerically. The results show the efficiency of the proposed solution in significantly increasing the structure resistance, deformation capacity and ductility.     

Blue fish burgers: nutritional characterisation and sensory optimisation

In order to reduce waste of marine resources, valorise fishing discards and move towards fish consumption, a new ready-to-cook seafood product was developed. In particular, a chemical investigation on chub mackerel and hake was conducted to the aim of mixing the two fish species to produce nutritionally balanced fish burgers. In addition, to evaluate consumer opinion about these new fresh products and best acceptability of different proportions of mackerel and hake, focus group interviews and consumer test were also performed. The results of the chemical analyses achieved on both selected fish species gave rise to minimally processed fish products with a high amount of chub mackerel, therefore providing the desired nutritional properties and low level of hake in order to obtain firmer and wetter burgers. By means of focus group interviews, the novelty and high level of convenience of the developed fresh commodity was highlighted. Consumer test declared that the most successful sample was that with 70% chub mackerel and 30% hake.     

Effectiveness of active and modified atmosphere packaging on shelf life extension of a cheese tart

This study evaluated the shelf life extension of a cheese cake subjected to modified atmosphere (MAP) and active packaging (AP). Cheese cakes were packaged under different N₂/CO₂ ratios (70/30 and 20/80) (MAP batches), by placing a sachet of an iron oxide-based oxygen absorber inside trays (AP batch) and with air (air batch). Changes in microbial growth, in-package gas composition, chemical–physical parameters including texture and sensory attributes were monitored for 48 days at 20 °C. AP allowed a mould-free cheese cake shelf life of 48 days, MAP extended the shelf life of samples packaged under 30% and 80% CO₂ up to 14 and 34 days, respectively, whereas tarts stored with air spoiled after 7 days. MAP resulted in a significant increase in hardness after 14 days of storage, whereas AP tarts recovered the initial texture after 48 days. Panellists judged AP tarts over the acceptability threshold during the whole shelf life of cheese tarts.     

Phase Evolution in Synthesis of Manganese Ferrite Nanoparticles

Physico-chemical equilibria that influence oxide powders' precipitation from an aqueous solution can be substantially altered when the process is carried out in a microemulsion system. To obtain nanosized MnFe₂O₄ and gain information about the physico-chemical characteristics of products, Mn²⁺ and Fe³⁺ metal ions were induced to precipitate in a toluene/water/sodium dodecylbenzenesulfonate microemulsion system. Portions of the precipitated powder were differently treated, both in solution and in the solid state, and the role of restricted aqueous domains in the obtained materials was investigated. X-ray diffraction profile-fitting methods and chemical analysis were applied to characterize the powder particles. Samples obtained from the selected microemulsion were identified as nanosized mixed hydroxide compounds. A low metal content and a limited matter exchange among aqueous nanodroplets appear to inhibit hydroxide to oxide transformation inside the selected micellar system. A calcination process of precipitated powder was required to obtain a manganese ferrite compound.     

Technological properties and non-enzymatic browning of white lupin protein enriched spaghetti

Spaghetti was prepared by replacing semolina with different amounts of lupin protein, in order to increase the protein content. A detailed investigation of the rheological properties of the dough and the cooking quality of pasta was performed in comparison to standard semolina spaghetti. Moreover, the effect of the addition of lupin protein on non-enzymatic browning was evaluated by measuring ε-furoylmethyllysine (furosine) and 5-hydroxymethyl-2-furancarboxaldehyde (HMF), which are considered useful indices of semolina quality and pasta processing conditions. Dried spaghetti fortified with 5% of lupin protein isolate has a colour and rheological features comparable with the semolina sample and also the behaviour during cooking results to be satisfactory. As far as the thermal damage is concerned, the furosine values of fortified spaghetti differ only marginally from standard pasta and the percentage lysine loss is quite small (ranging from 12.1% to 15.7%).