Application of the novel Droplet digital PCR technology for identification of meat species

The authenticity and traceability of meat products are issues of primary importance to ensure food safety. Unfortunately, food adulteration (e.g. the addition of inexpensive cuts to minced meat products) and mislabelling (e.g. the inclusion of meat from species other than those declared) happens frequently worldwide. The aim of this study was to apply a droplet digital PCR assay for the detection and quantification (copies μL⁻¹) of the beef, pork, horse, sheep, chicken and turkey in meat products. The analysis conducted on commercial meat showed the presence of traces of DNA from other animal species than those declared. We show that the method is highly sensitive, specific and accurate (accuracy = 100%). This method could be adopted by competent food safety authorities to verify compliance with the labelling of meat products and to ensure quality and safety throughout the meat supply chain, from primary production to consumption.     

Correlation between leukotriene D4-induced contraction and cytosolic calcium elevation: a quantitative and simultaneous evaluation in smooth muscle

The role of Ca++ as an intracellular messenger in leukotriene (LT)D4-induced muscle contraction was investigated by measuring force development and elevation in cytosolic Ca++ concentration simultaneously in strips of guinea pig ileal longitudinal muscle loaded with the fluorescent calcium indicator Fura 2. Upon addition of LTD4, a simultaneous increase in tension and cytosolic calcium concentration, [Ca++]i, was observed. Cumulative applications of LTD4 induced concentration-dependent increases in both muscle tension and [Ca++]i, being the half-maximal effect reached at approximately 6 to 9 nM. A statistically significant positive correlation (r = 0.993, P < .001) exists between the two parameters examined. Removal of calcium in the bathing solution, accompanied by addition of 7.5 mM EGTA, completely prevented any increase in either calcium levels or force development, thus indicating a role for Ca++ influx, rather than a release from intracellular stores. All of the LTD4 antagonists tested were able to counteract the effect of the leukotriene on both [Ca++]i and tension increase. However, although LY171883 shifted both of the LTD4 curves to the right in a parallel fashion, FPL 55712 and ICI 198,615 behaved as non-competitive antagonists in reversing the effect of LTD4 on [Ca++]i and tension. Thus, these results strongly suggest that changes in muscle tension induced by LTD4 are attributable to changes in cytosolic free Ca++ concentrations in guinea pig ileum.     

The AFLATOX® Project: Approaching the Development of New Generation,Natural-Based Compounds for the Containment of the Mycotoxigenic Phytopathogen Aspergillus flavus and Aflatoxin Contamination

The control of the fungal contamination on crops is considered a priority by the sanitary authorities of an increasing number of countries, and this is also due to the fact that the geographic areas interested in mycotoxin outbreaks are widening. Among the different pre- and post-harvest strategies that may be applied to prevent fungal and/or aflatoxin contamination, fungicides still play a prominent role; however, despite of countless efforts, to date the problem of food and feed contamination remains unsolved, since the essential factors that affect aflatoxins production are various and hardly to handle as a whole. In this scenario, the exploitation of bioactive natural sources to obtain new agents presenting novel mechanisms of action may represent a successful strategy to minimize, at the same time, aflatoxin contamination and the use of toxic pesticides. The Aflatox^® Project was aimed at the development of new-generation inhibitors of aflatoxigenic Aspergillus spp. proliferation and toxin production, through the modification of naturally occurring molecules: a panel of 177 compounds, belonging to the thiosemicarbazones class, have been synthesized and screened for their antifungal and anti-aflatoxigenic potential. The most effective compounds, selected as the best candidates as aflatoxin containment agents, were also evaluated in terms of cytotoxicity, genotoxicity and epi-genotoxicity to exclude potential harmful effect on the human health, the plants on which fungi grow and the whole ecosystem.     

An adaptive overlay network inspired by social behaviour

Nature is a great source of inspiration for scientists, because natural systems seem to be able to find the best way to solve a given problem by using simple and robust mechanisms. Studying complex natural systems, scientists usually find that simple local dynamics lead to sophisticated macroscopic structures and behaviour. It seems that some kind of local interaction rules naturally allow the system to auto-organize itself as an efficient and robust structure, which can easily solve different tasks. Examples of such complex systems are social networks, where a small set of basic interaction rules leads to a relatively robust and efficient communication structure. In this paper, we present PROSA, a semantic peer-to-peer (P2P) overlay network inspired by social dynamics. The way queries are forwarded and links among peers are established in PROSA resemble the way people ask other people for collaboration, help or information. Behaving as a social network of peers, PROSA naturally evolves to a small world, where all peers can be reached in a fast and efficient way. The underlying algorithm used for query forwarding, based only on local choices, is both reliable and effective: peers sharing similar resources are eventually connected with each other, allowing queries to be successfully answered in a really small amount of time. The resulting emergent structure can guarantee fast responses and good query recall.     

Macrophage-Mediated Melanoma Reduction after HP-NAP Treatment in a Zebrafish Xenograft Model

The Helicobacter pylori Neutrophil Activating Protein (HP-NAP) is endowed with immunomodulatory properties that make it a potential candidate for anticancer therapeutic applications. By activating cytotoxic Th1 responses, HP-NAP inhibits the growth of bladder cancer and enhances the anti-tumor activity of oncolytic viruses in the treatment of metastatic breast cancer and neuroendocrine tumors. The possibility that HP-NAP exerts its anti-tumor effect also by modulating the activity of innate immune cells has not yet been explored. Taking advantage of the zebrafish model, we examined the therapeutic efficacy of HP-NAP against metastatic human melanoma, limiting the observational window to 9 days post-fertilization, well before the maturation of the adaptive immunity. Human melanoma cells were xenotransplanted into zebrafish embryos and tracked in the presence or absence of HP-NAP. The behavior and phenotype of macrophages and the impact of their drug-induced depletion were analyzed exploiting macrophage-expressed transgenes. HP-NAP administration efficiently inhibited tumor growth and metastasis and this was accompanied by strong recruitment of macrophages with a pro-inflammatory profile at the tumor site. The depletion of macrophages almost completely abrogated the ability of HP-NAP to counteract tumor growth. Our findings highlight the pivotal role of activated macrophages in counteracting melanoma growth and support the notion that HP-NAP might become a new biological therapeutic agent for the treatment of metastatic melanomas.     

Chemical deactivation of V2O5/WO3–TiO2 SCR catalysts by additives and impurities from fuels, lubrication oils and urea solution: Part II. Characterization study of the effect of alkali and alkaline earth metals

A characterization study on a practice-oriented V₂O₅/WO₃–TiO₂ SCR catalyst deactivated by Ca and K, respectively, was carried out using NH₃-TPD, DRIFT spectroscopy, and XPS as well as theoretical DFT calculations. It was found from NH₃-TPD experiments that strongly basic elements like K or Ca drastically affect the acidity of the catalysts. Detailed DRIFT spectroscopy experiments revealed that these poisoning agents mostly interact with the Brønsted acid sites of the V₂O₅ active phase, thus affecting the NH₃ adsorption. Moreover, these experiments also indicated that the V⁵⁺ = O sites are much less reactive on the poisoned catalysts. XPS investigations of the O 1s binding energies showed that the oxygen atoms of the V⁵⁺ = O sites are affected by the presence of the poisoning agents. Based on these results and on DFT calculations with model clusters of the vanadia surface, the poisoning mechanism is explained by the stabilization of the non atomic holes of the (0 1 0) V₂O₅ phase as a result of the deactivation element. Consequently, V–OH Brønsted acid sites and V⁵⁺ = O sites are inhibited, which are both of crucial importance in the SCR process. The deactivation model also gives an explanation to the very low concentrations of potassium needed to deactivate the SCR catalyst, since one metal atom sitting on such a non-atomic hole site deactivates up to four active vanadium centers.     

Molecular Mechanisms and Physiological Changes behind Benign Tracheal and Subglottic Stenosis in Adults

Laryngotracheal stenosis (LTS) is a complex and heterogeneous disease whose pathogenesis remains unclear. LTS is considered to be the result of aberrant wound-healing process that leads to fibrotic scarring, originating from different aetiology. Although iatrogenic aetiology is the main cause of subglottic or tracheal stenosis, also autoimmune and infectious diseases may be involved in causing LTS. Furthermore, fibrotic obstruction in the anatomic region under the glottis can also be diagnosed without apparent aetiology after a comprehensive workup; in this case, the pathological process is called idiopathic subglottic stenosis (iSGS). So far, the laryngotracheal scar resulting from airway injury due to different diseases was considered as inert tissue requiring surgical removal to restore airway patency. However, this assumption has recently been revised by regarding the tracheal scarring process as a fibroinflammatory event due to immunological alteration, similar to other fibrotic diseases. Recent acquisitions suggest that different factors, such as growth factors, cytokines, altered fibroblast function and genetic susceptibility, can all interact in a complex way leading to aberrant and fibrotic wound healing after an insult that acts as a trigger. However, also physiological derangement due to LTS could play a role in promoting dysregulated response to laryngo-tracheal mucosal injury, through biomechanical stress and mechanotransduction activation. The aim of this narrative review is to present the state-of-the-art knowledge regarding molecular mechanisms, as well as mechanical and physio-pathological features behind LTS.     

Hamiltonian-type Lyapunov functions

A flexible and systematic way for the construction of Lyapunov functions which lends itself to the use of nonlinear programming techniques is presented. The class of Lyapunov functions obtained is shown to be energy functions (Hamiltonians) for an equivalent system.