Engineering of Janus-Like Dendrimers with Peptides Derived from Glycoproteins of Herpes Simplex Virus Type 1: Toward a Versatile and Novel Antiviral Platform

Novel antiviral nanotherapeutics, which may inactivate the virus and block it from entering host cells, represent an important challenge to face viral global health emergencies around the world. Using a combination of bioorthogonal copper-catalyzed 1,3-dipolar alkyne/azide cycloaddition (CuAAC) and photoinitiated thiol–ene coupling, monofunctional and bifunctional peptidodendrimer conjugates were obtained. The conjugates are biocompatible and demonstrate no toxicity to cells at biologically relevant concentrations. Furthermore, the orthogonal addition of multiple copies of two different antiviral peptides on the surface of a single dendrimer allowed the resulting bioconjugates to inhibit Herpes simplex virus type 1 at both the early and the late stages of the infection process. The presented work builds on further improving this attractive design to obtain a new class of therapeutics.     

High Glucose Exposure Impairs L-Cell Differentiation in Intestinal Organoids: Molecular Mechanisms and Clinical Implications

Intestinal organoids are used to analyze the differentiation of enteroendocrine cells (EECs) and to manipulate their density for treating type 2 diabetes. EEC differentiation is a continuous process tightly regulated in the gut by a complex regulatory network. However, the effect of chronic hyperglycemia, in the modulation of regulatory networks controlling identity and differentiation of EECs, has not been analyzed. This study aimed to investigate the effect of glucotoxicity on EEC differentiation in small intestinal organoid platforms. Mouse intestinal organoids were cultured in the presence/absence of high glucose concentrations (35 mM) for 48 h to mimic glucotoxicity. Chronic hyperglycemia impaired the expression of markers related to the differentiation of EEC progenitors (Ngn3) and L-cells (NeuroD1), and it also reduced the expression of Gcg and GLP-1 positive cell number. In addition, the expression of intestinal stem cell markers was reduced in organoids exposed to high glucose concentrations. Our data indicate that glucotoxicity impairs L-cell differentiation, which could be associated with decreased intestinal stem cell proliferative capacity. This study provides the identification of new targets involved in new molecular signaling mechanisms impaired by glucotoxicity that could be a useful tool for the treatment of type 2 diabetes.     

An efficient self-timed adder realized using conventional CMOS standard cells

Usually, efficient self-timed adders are realized using the dynamic differential cascode voltage switch logic. This allows the end-completion to be easily detected, but it makes circuit design and testing very complex, compelling the production of full-custom layouts and leading to a very long time before marketing. This paper presents a new 56-bit high-speed self-timed adder realized with conventional AMS 0.35 μm CMOS standard cells. The proposed circuit uses overlapped execution circuits, which exploit the initialization time that always elapses between two consecutive addition operations. Compared to several self-timed adders existing in the literature, the addition circuit proposed here shows brilliant advantages in terms of speed-performance, silicon area occupancy and power dissipation.     

Identification of new epidemiological molecular markers by comparative proteomics of serogroup A meningococcal isolates from three pandemic waves

We previously described the first reference map for the proteome of one strain of serogroup A Neisseria meningitidis (MenA), a major cause of epidemic meningitis in humans. As a preliminary finding, in that work we noted that 2‐DE protein maps of closely related MenA isolates from different epidemics spreads could be easily compared to detect minor differences and that 2‐DE phenotypes attributable to the well‐known epidemiological marker tbpB agreed with the genoclouds model of MenA epidemiological variation during pandemic waves. We explored here the possibility that an extended comparative study of 2‐DE maps of isolates representative of the nine genoclouds described by Achtman and collaborators could be used to discriminate between strains otherwise undistinguishable. We showed the example of 14 proteins with different 2‐DE spot patterns in different genoclouds that could be considered as putative tracers for alike‐strains discrimination. We introduce the novel concept that comparative proteomics can be useful in identifying new epidemiological markers for N. meningitidis.     

A 3D moisture-stress FEM analysis for time dependent problems in timber structures

This paper presents a 3D moisture-stress numerical analysis for timber structures under variable humidity and load conditions. An orthotropic viscoelastic-mechanosorptive material model is specialized on the basis of previous models. Both the constitutive model and the equations needed to describe the moisture flow across the structure are implemented into user subroutines of the Abaqus finite element code and a coupled moisture-stress analysis is performed for several types of mechanical loads and moisture changes. The presented computational approach is validated by analyzing some wood tests described in the literature and comparing the computational results with the reported experimental data.     

Molecular characterization of Italian rice cultivars

Random amplified polymorphic DNA (RAPD) is an efficient tool to detect DNA polymorphisms and to distinguish different clones and cultivars. We have applied the RAPD technique to distinguish, starting from a single rice seed, among some Italian rice cultivars from market and from National Institute for research on food and nutrition, Italy. We also estimated the genomic similarity among the rice cultivars object of this study taking advantage of the amplification patterns. Despite the low genomic variability among Italian rice cultivars, RAPD technique allowed us to distinguish among them. This indicates that RAPD can be used routinely to verify the identity of Italian rice cultivars in order to prevent fraudulent commercial activity.     

Implementation of response surface methodology to optimise extraction of onion (Allium cepa) solid waste phenolics

An experimental setup based on a 2³-full factorial, central composite design was implemented with the aim to optimising recovery of polyphenols from onion solid wastes (OSW). In order to allow for the establishment of a sustainable process, reusable and non-toxic solutions composed of water/ethanol/citric acid were employed as extracting media. The factors considered were (i) the pH of the medium, (ii) the extraction time and (iii) the ethanol concentration. The model obtained produced a satisfactory fitting of the experimental data with regard to total polyphenol extraction (R² = 0.97, p = 0.0025) and the reducing power of the extracts (R² = 0.97, p = 0.0033), but not with the antiradical activity (R² = 0.89, p = 0.0592). The 2nd order polynomial equations obtained after elaboration of the experimental data indicated that all parameters considered were significant in respect with the efficiency of total polyphenol recovery. The highest total polyphenol yield was theoretically predicted to be 9342 ± 1435 mg gallic acid equivalents per 100 g dry weight, under optimal conditions (60% EtOH, pH 2 and 4.2 h). Liquid chromatography-electrospray ionisation mass spectrometry of the optimally obtained extract revealed that the principal phytochemicals recovered were quercetin 3,4′-diglucoside, quercetin 4′-glucoside and quercetin. Simple linear regression analysis between the total polyphenol and the antiradical activity of the OSW extracts showed that there was no correlation in a statistically significant manner, as opposed to reducing power.Industrial relevanceThe recovery of value-added substances from agri-food industrial wastes is an issue with importance pertaining to both the reduction of the waste load released to the environment, and the development of novel, natural food additives with functional properties. Up to date, the examinations pertaining to the clarification of factors that can affect extractability were based on rather unilateral assessment, while it is generally accepted that the retrieval depends on several parameters, which render the phenomenon a particular complexity. In this view, the implementation of factorial design with respect to investigating in parallel several factors pertaining to efficient polyphenol recovery becomes imminent. Thus the establishment of models on such a sound experimental basis is expected to provide a reliable background for more costand resource-effective processes, with a potential direct industrial applicability.     

Influence of mediator immobilization on the electrochemically assisted microbial dechlorination of trichloroethene (TCE) and cis‐dichloroethene (cis‐DCE)

BACKGROUND: A bioelectrochemical process for trichloroethene (TCE) dechlorination was developed. In this new process, a solid‐state electrode polarized to ?450 mV versus the standard hydrogen electrode (SHE), in combination with a redox mediator (i.e., methyl viologen, MV) is employed as an electron donor for the microbial reductive dechlorination of TCE. In this study we compared the performance of the process with the redox mediator immobilized at the surface of electrodes or dissolved in the bulk liquid, using both a culture highly enriched in Desulfitobacterium spp., capable of dechlorinating TCE to cis‐dichloroethene (cis‐DCE), and a culture highly enriched in Dehalococcoides spp. capable of dechlorinating cis‐DCE to ethene. RESULTS: Short‐term potentiostatic (?450 mV versus SHE) experiments showed that TCE or cis‐DCE was dechlorinated both in the presence of soluble (500 µmol L^?1) and immobilized MV. However, TCE or cis‐DCE dechlorination rates with MV‐modified electrodes were remarkably lower than with soluble MV. Both cultures produced significant amounts of H₂ in the presence of electrically reduced, soluble MV, whereas no H₂ was produced when the mediator was immobilized at the electrode surface, regardless of the potential applied to the electrode, in the range ?425 to ?500 mV versus SHE. CONCLUSIONS: The process, operated with modified electrodes, supports the microbial dechlorination of TCE to ethene. Immobilization not only allows retention of the mediator within the system, but also increases process efficiency by preventing bioelectrochemical H₂ formation. On the other hand, strategies to increase dechlorination rates with modified electrodes need to be developed. Copyright © 2009 Society of Chemical Industry     

Content of phytoestrogens in soy-based dietary supplements

The isoflavone content of 14 soy-based dietary supplements intended to help alleviate perimenopausal and menopausal symptoms on sale in Italy were analysed using HPLC with UV detection. The aim was to quantify soy isoflavones after hydrolysis as aglycones, which are the bioactive part of isoflavone molecules. In the examined products, the amounts of isoflavones were frequently expressed ambiguously, and none of the products stated whether the isoflavone content of the product was expressed as aglycones or as conjugates. Each product revealed a different aglycone concentration profile. These supplements have different “fingerprints”, probably due to different sources of raw materials and to methods used in processing and preparation of extracts. In more than half the supplements tested, the actual values contained were below those stated and below those expected to help alleviate perimenopausal and menopausal symptoms.     

Variations of iridoid oleuropein in Italian olive varieties during growth and maturation

This research paper reports on the concentrations of oleuropein in fruits of seven Italian olive cultivars. These oleuropein levels, during fruit growth and maturation, were monitored by taking olive samples from the same olive trees grown in Middle Italy. A novel high‐resolution gas chromatography (HRGC) oleuropein quantification method and a new high‐performance liquid chromatography (HPLC) method were developed. The olive fruits showed high contents of oleuropein, which were greatly influenced by the olive cultivar (genetic store) factor. The stage of fruit growth and maturation was another factor strongly affecting the contents of iridoid oleuropein in olive drupes, whatever the olive cultivar considered. Green olives had much higher concentrations of this biofunctional glucoside, which decreased regularly during fruit growth and ripeness. No differences were observed between the two developed oleuropein quantification methods, which gave, for all the olive cultivars, average values of fruit oleuropein contents that were not statistically different. These two reliable repetitive methods, noticeably the HPLC one (which also showed a lower running time), could be used for routine determination of oleuropein in olive drupes. We observed that, in general, olive cultivars having high oleuropein concentrations in the fruits were also characterized by marked oleuropein contents in the leaves.