Effects of Lipidation on a Proline-Rich Antibacterial Peptide

The emergence of multidrug-resistant bacteria is a worldwide health problem. Antimicrobial peptides have been recognized as potential alternatives to conventional antibiotics, but still require optimization. The proline-rich antimicrobial peptide Bac7(1-16) is active against only a limited number of Gram-negative bacteria. It kills bacteria by inhibiting protein synthesis after its internalization, which is mainly supported by the bacterial transporter SbmA. In this study, we tested two different lipidated forms of Bac7(1-16) with the aim of extending its activity against those bacterial species that lack SbmA. We linked a C12-alkyl chain or an ultrashort cationic lipopeptide Lp-I to the C-terminus of Bac7(1-16). Both the lipidated Bac-C12 and Bac-Lp-I forms acquired activity at low micromolar MIC values against several Gram-positive and Gram-negative bacteria. Moreover, unlike Bac7(1-16), Bac-C12, and Bac-Lp-I did not select resistant mutants in E. coli after 14 times of exposure to sub-MIC concentrations of the respective peptide. We demonstrated that the extended spectrum of activity and absence of de novo resistance are likely related to the acquired capability of the peptides to permeabilize cell membranes. These results indicate that C-terminal lipidation of a short proline-rich peptide profoundly alters its function and mode of action and provides useful insights into the design of novel broad-spectrum antibacterial agents.     

Graphene Grown by Chemical Vapour Deposition on Steel Substrates: Friction Behaviour

Graphene was grown by low-pressure chemical vapour deposition on polished steel samples using ethanol as a precursor at different pressures: 200 and 400 mTorr. The samples were characterized by Raman spectroscopy, and the Raman maps clearly show the formation of an inhomogeneous graphene coating with surface regions covered with single-layer, bilayer, and multilayer graphene, the last one covering most of the surface area of the sample when the sample was prepared at 400 mTorr. Scanning electron microscopy revealed the partial coverage of the sample surface when the graphene coating was prepared at a pressure of 200 mTorr. Friction measurements were taken using a micro-tribometer in a ball-on-flat contact geometry with a stainless steel ball as the counterbody. An important reduction in the coefficient of friction was verified even with partial coverage of the steel surface, and it was attributed to the formation of nanographitic structures in the wear track region and partially transferred to the counterbody.     

Steam reforming and oxidative steam reforming for hydrogen production from bioethanol over Mg2AlNiXHZOY nano-oxyhydride catalysts

Mg₂AlNi_XH_ZO_Y nano-oxyhydrides formation is evidenced during pre-treatment in H₂ at 450 °C of Mg₂AlNi_XO_Y nano-compounds leading to highly performant catalysts in ethanol conversion and H₂ formation, particularly at low temperature, through catalytic steam reforming (SRE) and oxidative steam reforming (OSRE). Total conversion of ethanol is obtained in SRE and OSRE with high stability. A higher production of H₂ (60 L h^?1 g_cat^?1) can be achieved at a reaction temperature of 300 °C in OSRE conditions compared to SRE (10 L h^?1 g_cat^?1) mainly because of a beneficial use of a high concentration of ethanol (14 mol%) in presence of O₂. Moreover, carbon formation is decreased and a much lower input of energy of 50 °C is used to get a temperature of 300 °C when O₂ is added. Different physico-chemical characterizations and in particular in H₂ (TPR, H₂-XRD, INS) and after tests allow to conclude that the presence of Ni²⁺ cations in strong interaction with other cations, anionic vacancies and hydride species on and inside the solid play an important role in the catalytic performance (conversion and selectivity) and stability.     

ANP and BNP Exert Anti-Inflammatory Action via NPR-1/cGMP Axis by Interfering with Canonical,Non-Canonical,and Alternative Routes of Inflammasome Activation in Human THP1 Cells

Dysregulated inflammasome activation and interleukin (IL)-1β production are associated with several inflammatory disorders. Three different routes can lead to inflammasome activation: a canonical two-step, a non-canonical Caspase-4/5- and Gasdermin D-dependent, and an alternative Caspase-8-mediated pathway. Natriuretic Peptides (NPs), Atrial Natriuretic Peptide (ANP) and B-type Natriuretic Peptide (BNP), binding to Natriuretic Peptide Receptor-1 (NPR-1), signal by increasing cGMP (cyclic guanosine monophosphate) levels that, in turn, stimulate cGMP-dependent protein kinase-I (PKG-I). We previously demonstrated that, by counteracting inflammasome activation, NPs inhibit IL-1β secretion. Here we aimed to decipher the molecular mechanism underlying NPs effects on THP-1 cells stimulated with lipopolysaccharide (LPS) + ATP. Involvement of cGMP and PKG-I were assessed pre-treating THP-1 cells with the membrane-permeable analogue, 8-Br-cGMP, and the specific inhibitor KT-5823, respectively. We found that NPs, by activating NPR-1/cGMP/PKG-I axis, lead to phosphorylation of NLRP3 at Ser295 and to inflammasome platform disassembly. Moreover, by increasing intracellular cGMP levels and activating phosphodiesterases, NPs interfere with both Gasdermin D and Caspase-8 cleavage, indicating that they disturb non-canonical and alternative routes of inflammasome activation. These results showed that ANP and BNP anti-inflammatory and immunomodulatory actions may involve the inhibition of all the known routes of inflammasome activation. Thus, NPs might be proposed for the treatment of the plethora of diseases caused by a dysregulated inflammasome activation.     

Cross-sectional imaging of individual layers and buried interfaces of graphene-based heterostructures and superlattices

By stacking various two-dimensional (2D) atomic crystals on top of each other, it is possible to create multilayer heterostructures and devices with designed electronic properties. However, various adsorbates become trapped between layers during their assembly, and this not only affects the resulting quality but also prevents the formation of a true artificial layered crystal upheld by van der Waals interaction, creating instead a laminate glued together by contamination. Transmission electron microscopy (TEM) has shown that graphene and boron nitride monolayers, the two best characterized 2D crystals, are densely covered with hydrocarbons (even after thermal annealing in high vacuum) and exhibit only small clean patches suitable for atomic resolution imaging. This observation seems detrimental for any realistic prospect of creating van der Waals materials and heterostructures with atomically sharp interfaces. Here we employ cross sectional TEM to take a side view of several graphene-boron nitride heterostructures. We find that the trapped hydrocarbons segregate into isolated pockets, leaving the interfaces atomically clean. Moreover, we observe a clear correlation between interface roughness and the electronic quality of encapsulated graphene. This work proves the concept of heterostructures assembled with atomic layer precision and provides their first TEM images.     

Saliva improves Streptococcus mitis protective effect on human gingival fibroblasts in presence of 2-hydroxyethyl-methacrylate

This study aimed to investigate the effect of saliva on Streptococcus mitis free cells and on S. mitis/human gingival fibroblasts (HGFs) co-culture model, in presence of 2-hydroxyethyl-methacrylate (HEMA). The bacterial aggregation both in the planktonic phase and on HGFs, as well as the apoptotic and necrotic eukaryotic cells amount were analyzed, in presence of saliva and/or HEMA. The aggregation test revealed a significant saliva aggregation effect on S. mitis strains compared to the untreated sample. No significant differences were recorded in the amount of culturable bacteria in all studied conditions; however, from microscopy images, the saliva/HEMA combining effect induced a significant bacterial aggregation and adhesion on HGFs. HEMA treatment decreased viable eukaryotic cell number with a parallel increment of necrotic cells, but when saliva was added to the co-culture, the viable cells percentage increased to a value comparable to the control sample.     

The Influence of Different Processing Stages on Particle Size,Microstructure, and Appearance of Dark Chocolate

The effect of different process stages on microstructural and visual properties of dark chocolate was studied. Samples were obtained at each phase of the manufacture process: mixing, prerefining, refining, conching, and tempering. A laser light diffraction technique and environmental scanning electron microscopy (ESEM) were used to study the particle size distribution (PSD) and to analyze modifications in the network structure. Moreover, colorimetric analyses (L*, h°, and C*) were performed on all samples. Each stage influenced in stronger way the microstructural characteristic of products and above all the PSD. Sauter diameter (D [3.2]) decreased from 5.44 μm of mixed chocolate sample to 3.83 μm, of the refined one. ESEM analysis also revealed wide variations in the network structure of samples during the process, with an increase of the aggregation and contact point between particles from mixing to refining stage. Samples obtained from the conching and tempering were characterized by small PS, and a less dense aggregate structure. From color results, samples with the finest particles, having larger specific surface area and the smallest diameter, appeared lighter and more saturated than those with coarse particles.     

Analysis of extra virgin olive oils from stoned olives

This paper focuses on comparing the main chemical characteristics of 16 fresh commercial samples of extra virgin olive oil obtained from four harvest years (1999–2002) and derived from both stoned and whole fruits. The qualitative and quantitative contents of minor polar compounds (MPCs) together with other reference analytical parameters (acidity, peroxides, UV absorption values and Rancimat test) were evaluated. An investigation of the MPCs and oil composition obtained from only stoned olives was also carried out. The acidity values of the oils from stoned fruits were always similar to or lower than those of the corresponding oils from whole fruits. For almost all the samples from stoned olives a better resistance to oxidation was revealed in comparison with the corresponding traditional oils. Five pairs obtained from the 2000 and 2001 harvests showed higher concentrations of both MPCs and hydroxytyrosol derivatives in the oils from stoned fruits, in agreement with their higher Rancimat values. Overall, our findings with regard to acidity values, % hydrolysis, the Rancimat test and the qualitative and quantitative distribution of MPCs suggest a higher antioxidant capacity of the oils from stoned olives. Copyright © 2004 Society of Chemical Industry     

Design of Sonotrode Ultrasound-Assisted Extraction of Phenolic Compounds from <Emphasis Type="Italic">Psidium guajava</Emphasis> L. Leaves

Psidium guajava L. has gained a special attention as health plant due to the presence of phenolic compounds. Box-Behnken design (BBD) has been applied for the extraction of target compounds from guava leaves via sonotrode ultrasound-assisted extraction (UAE). Different extraction times (5, 30, and 55 min), ratios of ethanol/water (50, 75, and 100% (v/v)), and ultrasound (US) power (80, 240, and 400 W) were tested to find their effect on the sum of phenolic compound (SPC), flavonols and flavan-3-ols via HPLC-ESI-QqQ-MS, and antioxidant activity (DPPH and TEAC assays). The best process conditions were as follows: 40 min, 60% ethanol/water (v/v), and 200 W. Established method has been used to extract phenolic compounds in two guava leaves varieties (pyrifera and pomifera). Pyrifera var. showed greater values of the SPC via HPLC-ESI-QqQ-MS (49.7 mg/g leaf dry weight (d.w.)), flavonols (12.51 mg/g d.w.), flavan-3-ols (7.20 mg/g d.w.), individual phenolic compounds, and antioxidant activity (8970 ± 5 and 465 ± 6 μmol Trolox/g leaf d.w, respectively) than pomifera var. Conventional extraction showed lower amounts of phenolic compounds (7.81 ± 0.03 and 4.64 ± 0.01 mg/g leaf d.w. for flavonols and flavan-3ols, respectively) in comparison to the ultrasound-assisted ones.