Exposure to b-LED Light While Exerting Antimicrobial Activity on Gram-Negative and -Positive Bacteria Promotes Transient EMT-like Changes and Growth Arrest in Keratinocytes

While blue LED (b-LED) light is increasingly being studied for its cytotoxic activity towards bacteria in therapy of skin-related infections, its effects on eukaryotic cells plasticity are less well characterized. Moreover, since different protocols are often used, comparing the effect of b-LED towards both microorganisms and epithelial surfaces may be difficult. The aim of this study was to analyze, in the same experimental setting, both the bactericidal activity and the effects on human keratinocytes. Exposure to b-LED induced an intense cytocidal activity against Gram-positive (i.e, Staphylococcus aureus) and Gram-negative (i.e., Pseudomonas aeruginosa) bacteria associated with catheter-related infections. Treatment with b-LED of a human keratinocyte cell line induced a transient cell cycle arrest. At the molecular level, exposure to b-LED induced a transient downregulation of Cyclin D1 and an upregulation of p21, but not signs of apoptosis. Interestingly, a transient induction of phosphor-histone γ-H2Ax, which is associated with genotoxic damages, was observed. At the same time, keratinocytes underwent a transient epithelial to mesenchymal transition (EMT)-like phenotype, characterized by E-cadherin downregulation and SNAIL/SLUG induction. As a functional readout of EMT induction, a scratch assay was performed. Surprisingly, b-LED treatment provoked a delay in the scratch closure. In conclusion, we demonstrated that b-LED microbicidal activity is associated with complex responses in keratinocytes that certainly deserve further analysis.     

Rheological and sensory performance of a protein-based sweetener (MNEI), sucrose,and aspartame in yogurt

Sweeteners and flavors are generally added to yogurt to make them more palatable. However, the addition of these ingredients may affect the fermentation process of yogurt as well as its physical and sensory characteristics. Consumers prioritize yogurt products that are “natural.” A modified single-chain form of the natural sweet protein monellin extracted from the fruit of Dioscoreophyllum cumminsii, called MNEI, could be a useful alternative to artificial sweeteners. The aim of the present work was to evaluate new rapid sensory methods in combination with rheology to assess the viability of using MNEI to develop sweetened yogurts without the calories of sugar. We studied the gelation and cooling kinetics of 4 yogurt samples (unsweetened or sweetened with MNEI, aspartame, or sucrose) by using a rheometer. Furthermore, the 4 yogurts, with and without addition of a flavoring agent, were characterized from a sensory perspective using a combination of 2 rapid sensory methods, ultra flash profile and flash profile. Rheological results showed that, when added at typical usage levels, aspartame, sucrose, and MNEI did not generally affect the yogurt fermentation process or its rheological properties. Sensory results demonstrated that texture attributes of yogurts with aspartame and sucrose were strongly linked to sweetness and flavor perception, but this was not true for MNEI-sweetened yogurts. In contrast to results obtained from samples sweetened with sucrose and aspartame, MNEI protein did not sweeten the yogurt when added before fermentation. This study highlights the enhancing effect of flavor on sweetness perception, supporting previous reports that noted synergistic effects between sucrose or aspartame and flavors. Hence, future studies should be conducted to determine how sweet proteins behave in yogurt when added after fermentation.     

Modulation of the biological activity of microRNA-210 with peptide nucleic acids (PNAs)

Herein we describe the activity of a peptide nucleic acid (PNA) that targets microRNA‐210 (miR‐210), which is associated with hypoxia and is modulated during erythroid differentiation. PNAs directed against miR‐210 were designed to bind with high affinity to the target RNA strand and to undergo efficient uptake in target cells. A polyarginine–PNA conjugate directed against miR‐210 (Rpep‐PNA‐a210) showed both very high affinity for RNA and efficient uptake into target cells without the need for transfection reagents. An unmodified PNA of the same sequence displayed the ability to bind RNA, but cellular uptake was very poor. Consistent with this, only Rpep‐PNA‐a210 strongly inhibited miR‐210 activity, as evaluated by assays on undifferentiated K562 cells and on cells treated with mithramycin, which was found to induce erythroid differentiation and miR‐210 overexpression. Targeting miR‐210 by Rpep‐PNA‐a210 resulted in: 1) a decrease in miR‐210 levels as measured by RT‐PCR, 2) up‐regulation of raptor mRNA, 3) a decrease in γ‐globin mRNA, and 4) decreased expression of differentiated functions (i.e., proportion of benzidine‐positive cells, content of embryo‐fetal hemoglobins). The efficient delivery of anti‐miR PNAs through a suitable peptide carrier (Rpep‐PNA‐a210) leads to the inhibition of miR‐210 activity, altering the expression of miR‐210‐regulated erythroid functions.     

Effect of cooking on the total antioxidant capacity and phenolic profile of some whole‐meal African cereals

BACKGROUND: At present, sorghum, fonio and millet are not placed as important commodities in the North American and European food basket, but their importance as ingredients in multigrain and gluten‐free cereal products is highlighted. Therefore in this study the phenolic profile (evaluated by liquid chromatography/tandem mass spectrometry), total phenolic content (assessed by Folin–Ciocalteu assay) and total antioxidant capacity were measured in three African whole grains, i.e. sorghum (Sorghum bicolor ssp. bicolor), fonio (Digitaria exilis) and pearl millet (Pennisetum glaucum L.), before and after a cooking procedure. RESULTS: After the cooking process, soluble phenolic acids increased significantly in sorghum, whereas bound ones and anthocyanins decreased significantly. In millet the cooking process significantly enhanced soluble phenolic acids without affecting those bound, whereas in fonio a slight but significant decrease in almost all soluble phenolic acids was observed along with a significant increase in bound ones. Finally, the cooking process negatively affected both total phenolic content and total antioxidant capacity. CONCLUSION: This is one of the few reports dealing with the antioxidant compounds of these three African whole grains in which the effect of cooking was also evaluated. The data suggested that, to improve their antioxidant properties, specific cultivars should be selected and the cooking procedures carefully considered. Copyright © 2012 Society of Chemical Industry     

Dual Stimuli Responsive Gelatin‐CNT Hybrid Films as a Versatile Tool for the Delivery of Anionic Drugs

Hybrid hydrogel films able to modulate the release of anionic species, ketoprofen (Ket) and bovine serum albumin (BSA), as a function of their molecular size are prepared by UV‐induced polymerization of methacrylated gelatin in the presence of oxidized multiwalled carbon nanotubes (MWNT_COOH). The dual‐stimuli responsive composites can modulate the release in response to the variation of temperature and the application of an external voltage. Drug release profiles, analyzed by mathematical models, show that different temperatures (25 and 45 °C) and applied voltages (0 and 36 V) have a different effect on the release of the two therapeutics. Ket release is enhanced at 45 °C (77% vs 22% at 25 °C) with the maximum value recorded when 36 V is applied (94%). An increase in the amount of released BSA is recorded at 45 °C (96% vs 50% at 25 °C), while the application of the external voltage reduces this value (39%).

     

Probabilistic modelling of hybrid solar/wind power system with solar tracking system

This paper presents a probabilistic model applied to a hybrid solar-wind power system (HSWPS), which is equipped with either a one-axis or a two-axis solar tracking system.Within the framework of a case study, the potential of the developed probabilistic approach is presented, and the effect of the solar tracking systems on the annual energy gain is discussed.Specifically, the impact of a tracking system on the probability density function (PDF) of the power produced by a photovoltaic system (PVS) is evaluated through the first four moments (mean, variance, skewness and kurtosis) of a PDF.Finally, to estimate the impact of a tracking system on HSWPS energy performance, a reliability analysis is performed using the energy index of reliability (EIR), which is directly related to energy expected not supplied (EENS), given different annual load scenarios.     

Aminolysis of Styrene Oxide Over Heterogeneous Acidic Catalysts

Silica alumina and silica zirconia mixed oxides are shown to be effective and regioselective catalysts for the aminolysis of styrene oxide under very mild experimental conditions, giving the corresponding primary ??-amino-alcohol in good to excellent yield.     

Cellular uptakes, biostabilities and anti-miR-210 activities of chiral arginine-PNAs in leukaemic K562 cells

A series of 18‐mer peptide nucleic acids (PNAs) targeted against micro‐RNA miR‐210 was synthesised and tested in a cellular system. Unmodified PNAs, R₈‐conjugated PNAs and modified PNAs containing eight arginine residues on the backbone, either as C2‐modified (R) or C5‐modified (S) monomers, all with the same sequence, were compared. Two different models were used for the modified PNAs: one with alternated chiral and achiral monomers and one with a stretch of chiral monomers at the N terminus. The melting temperatures of these derivatives were found to be extremely high and 5 M urea was used to assess differences between the different structures. FACS analysis and qRT‐PCR on K562 chronic myelogenous leukaemic cells indicated that arginine‐conjugated and backbone‐modified PNAs display good cellular uptake, with best performances for the C2‐modified series. Resistance to enzymatic degradation was found to be higher for the backbone‐modified PNAs, thus enhancing the advantage of using these derivatives rather than conjugated PNAs in the cells in serum, and this effect is magnified in the presence of peptidases such as trypsin. Inhibition of miR‐210 activity led to changes in the erythroid differentiation pathway, which were more evident in mithramycin‐treated cells. Interestingly, the anti‐miR activities differed with use of different PNAs, thus suggesting a role of the substituents not only in the cellular uptake, but also in the mechanism of miR recognition and inactivation. This is the first report relating to the use of backbone‐modified PNAs as anti‐miR agents. The results clearly indicate that backbone‐modified PNAs are good candidates for the development of very efficient drugs based on anti‐miR activity, due to their enhanced bioavailabilities, and that overall anti‐miR performance is a combination of cellular uptake and RNA binding.