Bacterial DNAemia in Alzheimer’s Disease and Mild Cognitive Impairment: Association with Cognitive Decline,Plasma BDNF Levels,and Inflammatory Response

Microbial dysbiosis (MD) provokes gut barrier alterations and bacterial translocation in the bloodstream. The increased blood bacterial DNA (BB-DNA) may promote peripheral- and neuro-inflammation, contributing to cognitive impairment. MD also influences brain-derived neurotrophic factor (BDNF) production, whose alterations contribute to the etiopathogenesis of Alzheimer’s disease (AD). The purpose of this study is to measure BB-DNA in healthy elderly controls (EC), and in patients with mild cognitive impairment (MCI) and AD to explore the effect on plasma BDNF levels (pBDNF), the inflammatory response, and the association with cognitive decline during a two-year follow-up. Baseline BB-DNA and pBDNF were significantly higher in MCI and AD than in EC. BB-DNA was positively correlated with pBDNF in AD, plasma Tumor necrosis factor-alpha (TNF-α), and Interleukin-10 (IL-10) levels in MCI. AD patients with BB-DNA values above the 50th percentile had lower baseline Mini-Mental State Examination (MMSE). After a two-year follow-up, AD patients with the highest BB-DNA tertile had a worse cognitive decline, while higher BB-DNA levels were associated with higher TNF-α and lower IL-10 in MCI. Our study demonstrates that, in early AD, the higher the BB-DNA levels, the higher the pBDNF levels, suggesting a defensive attempt; BB-DNA seems to play a role in the AD severity/progression; in MCI, higher BB-DNA may trigger an increased inflammatory response.     

IL-33 and IL-37: A Possible Axis in Skin and Allergic Diseases

Interleukin (IL)-37 and IL-33 are among the latest cytokines identified, playing a role in several inflammatory conditions, spanning from systemic conditions to tumors to localized diseases. As newly discovered interleukins, their role is still scarcely understood, but their potential role as therapeutic targets or disease activity markers suggests the need to reorganize the current data for a better interpretation. The aim of this review is to collect and organize data produced by several studies to create a complete picture. The research was conducted on the PubMed database, and the resulting articles were sorted by title, abstract, English language, and content. Several studies have been assessed, mostly related to atopic dermatitis and immunologic pathways. Collective data demonstrates a pro-inflammatory role of IL-33 and an anti-inflammatory one for IL-37, possibly related to each other in an IL-33/IL-37 axis. Although further studies are needed to assess the safety and plausibility of targeting these two interleukins for patients affected by skin conditions, the early results indicate that both IL-33 and IL-37 represent markers of disease activity.     

Downexpression of miR-200c-3p Contributes to Achalasia Disease by Targeting the PRKG1 Gene

Achalasia is an esophageal smooth muscle motility disorder with unknown pathogenesis. Taking into account our previous results on the downexpression of miR-200c-3p in tissues of patients with achalasia correlated with an increased expression of PRKG1, SULF1, and SYDE1 genes, our aim was to explore the unknown biological interaction between these genes and human miR-200c-3p and if this relation could unravel their functional role in the etiology of achalasia. To search for putative miR-200c-3p binding sites in the 3′-UTR of PRKG1, SULF1 and SYDE1, a bioinformatics tool was used. To test whether PRKG1, SULF1, and SYDE1 are targeted by miR-200c-3p, a dual-luciferase reporter assay and quantitative PCR on HEK293 and fibroblast cell lines were performed. To explore the biological correlation between PRKG1 and miR-200c-3p, an immunoblot analysis was carried out. The overexpression of miR-200c-3p reduced the luciferase activity in cells transfected with a luciferase reporter containing a fragment of the 3′-UTR regions of PRKG1, SULF1, and SYDE1 which included the miR-200c-3p seed sequence. The deletion of the miR-200c-3p seed sequence from the 3′-UTR fragments abrogated this reduction. A negative correlation between miR-200c-3p and PRKG1, SULF1, and SYDE1 expression levels was observed. Finally, a reduction of the endogenous level of PRKG1 in cells overexpressing miR-200c-3p was detected. Our study provides, for the first time, functional evidence about the PRKG1 gene as a direct target and SULF1 and SYDE1 as potential indirect substrates of miR-200c-3p and suggests the involvement of NO/cGMP/PKG signaling in the pathogenesis of achalasia.     

Quantifying the Detrimental Effects of Multiple Freeze/Thaw Cycles on Primary Human Lymphocyte Survival and Function

The use of cryopreserved peripheral blood mononuclear cells is common in biological research. It is widely accepted that primary cells are rendered unusable by several freezing cycles, although this practice might be very helpful when the biological material is valuable and its re-collection is impractical. To determine the extent to which primary cells undergoing repeated freezing cycles are comparable to one another and to fresh samples, we evaluated overall lymphocyte viability, their proliferation and cytokine production capabilities, as well as the levels of 27 cell subtypes in ten human peripheral blood mononuclear cells frozen for five years and repeatedly thawed. As expected, we observed a progressive increase in cell death percentages on three rounds of thawing, but the frequency of the main lymphocyte subsets was stable across the three thawings. Nevertheless, we observed a significant reduction of B cell frequency in frozen samples compared to fresh ones. On repeated thawings and subsequent conventional stimulation, lymphocyte proliferation significantly decreased, and IL-10, IL-6, GM-CSF, IFN-gamma, and IL-8 showed a trend to lower values.     

The P2X7R-NLRP3 and AIM2 Inflammasome Platforms Mark the Complexity/Severity of Viral or Metabolic Liver Damage

P2X7R-NLRP3 and AIM2 inflammasomes activate caspase-1 and the release of cytokines involved in viral-related liver disease. Little is known about their role in non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steato-hepatitis (NASH). We characterized the role of inflammasomes in NAFLD, NASH, and HCV. Gene expression and subcellular localization of P2X7R/P2X4R-NLRP3 and AIM2 inflammasome components were examined in histopathological preparations of 46 patients with biopsy-proven viral and metabolic liver disease using real-time PCR and immunofluorescence. P2X7R, P2X4R, and Caspase-1 are two- to five-fold more expressed in patients with NAFLD/NASH associated with chronic HCV infection than those with metabolic damage only (p ≤ 0.01 for all comparisons). The AIM2 inflammasome is 4.4 times more expressed in patients with chronic HCV infection, regardless of coexistent metabolic abnormalities (p = 0.0006). IL-2, a cytokine playing a pivotal role during chronic HCV infection, showed a similar expression in HCV and NASH patients (p = 0.77) but was virtually absent in NAFLD. The P2X7R-NLRP3 complex prevailed in infiltrating macrophages, while AIM2 was localized in Kupffer cells. Caspase-1 expression correlated with elastography-based liver fibrosis (r = 0.35, p = 0.02), whereas P2X7R, P2X4R, NRLP3, Caspase-1, and IL-2 expression correlated with circulating markers of disease severity. P2X7R and P2X4R play a major role in liver inflammation accompanying chronic HCV infection, especially when combined with metabolic damage, while AIM2 is specifically expressed in chronic viral hepatitis. We describe for the first time the hepatic expression of IL-2 in NASH, so far considered a peculiarity of HCV-related liver damage.     

Modelling and predicting the effect of social distancing and travel restrictions on COVID-19 spreading

To date, the only effective means to respond to the spreading of the COVID-19 pandemic are non-pharmaceutical interventions (NPIs), which entail policies to reduce social activity and mobility restrictions. Quantifying their effect is difficult, but it is key to reducing their social and economic consequences. Here, we introduce a meta-population model based on temporal networks, calibrated on the COVID-19 outbreak data in Italy and applied to evaluate the outcomes of these two types of NPIs. Our approach combines the advantages of granular spatial modelling of meta-population models with the ability to realistically describe social contacts via activity-driven networks. We focus on disentangling the impact of these two different types of NPIs: those aiming at reducing individuals’ social activity, for instance through lockdowns, and those that enforce mobility restrictions. We provide a valuable framework to assess the effectiveness of different NPIs, varying with respect to their timing and severity. Results suggest that the effects of mobility restrictions largely depend on the possibility of implementing timely NPIs in the early phases of the outbreak, whereas activity reduction policies should be prioritized afterwards.     

On the detection of card-sharing traffic through wavelet analysis and Support Vector Machines

In the last years, the interest in methods and techniques for circumventing the security of the available digital video broadcasting systems is continuously increasing. Digital TV providers are struggling to restrict access to their video contents only to authorized users, by deploying more and more sophisticated conditional access systems. At the state-of-the-art, the most significant menace is the card-sharing activity which exploits a known weakness allowing an authorized subscriber to provide access to digital contents to a potentially large group of unauthorized ones connected over a communication network. This is usually realized by using ad hoc customized devices. Detecting the presence of these illegal systems on a network, by recognizing their related traffic is an issue of primary importance. Unfortunately, to avoid the identification of such traffic, payload obfuscation strategies based on encryption are often used, hindering packet inspection techniques.This paper presents a strategy for the detection of card-sharing traffic, empowered by machine-learning-driven traffic classification techniques and based on the natural capability of wavelet analysis to decompose a traffic time series into several component series associated with particular time and frequency scales and hence allowing its observation at different frequency component levels and with different resolutions. These ideas have been used for the proof-of-concept implementation of an SVM-based binary classification scheme that relies only on time regularities of the traffic and not on the packet contents and hence is immune to payload obfuscation techniques.     

Determination of pyrethroids in chicken egg samples: development and validation of a confirmatory analytical method by gas chromatography/mass spectrometry

The aim of this study was to develop and validate an efficient analytical method based on gas chromatography/tandem mass spectrometry (GC/MS/MS) for detection and quantification of six pyrethroids residues (Phenothrin, Permethrin, Cyfluthrin, Cypermethrin, Deltamethrin and Fenvalerate) in chicken eggs. The method was based on a preliminary liquid–liquid extraction of albumen‐free yolk samples, followed by a clean‐up by solid‐phase extraction. GC/MS/MS analyses were carried out in the selected reaction monitoring mode. Validation parameters such as specificity, detection capability, decision limit, precision, recovery, stability and ruggedness were determined, resulting in compliance with Decision 2002/657/EC. No complicated apparatus are required; moreover, low volumes of organic solvents and a nonintensive manual labour are required. These low costs and simple procedure, based on rapid and safe operations, may represent a useful tool in the routine analysis of pyrethroids pesticides, in the place of the currently used conventional techniques.     

Chemical composition,sensory and cooking quality evaluation of durum wheat spaghetti enriched with pea flour

The goal of the study was to obtain spaghetti enriched with pea flour with sensory properties close to conventional pasta. To this aim, the study has been organised in two subsequent trials. In the first, the pea flour amount added to the spaghetti was continuously increased until the overall sensory quality reached its threshold value (pea flour concentration = 15%). In fact, the spaghetti samples supplemented with 15% pea flour (15%S‐P) showed less elasticity, unpleasant colour and higher firmness compared to the control sample (CTRL). The second step was aimed to improve the overall sensory quality of the 15%S‐P spaghetti by means of guar gum (GUAR). The sample with GUAR (15%S‐P/GUAR) showed a pleasant colour, odour and taste. Furthermore, the 15%S‐P/GUAR sample recorded a low starch digestibility value (i.e. 54) and a higher soluble fibre content with respect to the 15%S‐P sample.     

An LMI approach for H ∞ analysis and control of discrete-time piecewise affine systems

In this paper we investigate some analysis and control problems for discrete-time hybrid systems in the piece-wise affine form. By using arguments from the dissipativity theory for non-linear systems, we show that H X analysis and synthesis problems can be formulated and solved via linear matrix inequalities by taking into account the switching structure of the considered system. In this paper we address the generalized problem of controlling hybrid systems whose switching structure does not depend only on the state but also on the control input.