Validation of a 1-Day Analytical Diagnostic Real-Time PCR for the Detection of Salmonella in Different Food Meat Categories

Foodborne disease caused by Salmonella represents a worldwide public health problem. In Europe, salmonellosis is still the second most commonly recorded zoonosis. Since the standard culture method for detecting Salmonella (ISO 6579:2002) requires up to 5 days to produce results, the need to develop rapid methods represents an important issue for the authorities and the producers. The aim of the present study was the in-house validation, according to ISO 16140, of an open-formula diagnostic real-time PCR for the detection of Salmonella in all the different meat categories reported in the EU Regulations relative to microbiological criteria for food safety. The assay employed specific primers and a probe target within the ttrRSBCA locus, which allows the tetrathionate respiration in Salmonella. Selectivity, relative accuracy, relative sensitivity and relative specificity were established by testing 110 bacterial strains and 175 various edible meat samples. Results showed 100 % selectivity, 100 % relative accuracy, 100 % relative sensitivity and 100 % relative specificity of the real-time PCR when compared to the standard culture method used as reference. In addition, in order to minimize the effect of the competitive micro-flora naturally present on meat samples, a highly nutritious and selective commercial medium (ONE Broth Salmonella, Oxoid) was evaluated in comparison with the classical non-selective pre-enrichment broth (buffered peptone water). Results demonstrated that the ONE Broth Salmonella medium increases the growth of Salmonella in the presence of competitive micro-flora.     

Identification of polyphenolic compounds in red and white grape varieties grown in R. Macedonia and changes of their content during ripening

The composition of polyphenols in the skin, seed and pulp extracts of the grapes of Vranec and Smederevka varieties, and Merlot and Chardonnay as well, was analyzed by liquid chromatography-diode array detection-mass spectrometry (HPLC-DAD-MS) and MS/MS techniques. Thirty-one phenolic compounds, including anthocyanins, flavonols and flavan-3-ols, as well as phenolic acids derivatives, have been identified in the extracts prepared from the grapes at physiological maturity, by mass spectrometry applying electrospray ionization operated in alternating ion mode and by performing tandem MS experiments in the ion trap mass analyzer. Grapes were analyzed at three different phases: (i) veraison, (ii) physiological ripeness and (iii) late harvest, in order to follow the evolution of polyphenolic content during berry development, applying spectrophotometric methods. Vranec had a higher polyphenolic content compared to Merlot due to the higher levels of total phenolics and flavonoids in the skins and seeds as well as a higher content of flavan-3-ols and anthocyanins in the skins, allowing discriminating the varieties. HPLC-DAD-MS analysis presented significantly higher relative amounts of anthocyanin monoglucosides and p-coumaroylglucosides in Vranec grapes. Smederevka seeds had a higher amount of flavan-3-ols than Chardonnay, while a higher content of total phenolics and flavonoids was found in the skins of the Chardonnay variety. Anthocyanin content in both red varieties increased during the berry ripening, while flavan-3-ols in seeds were mainly accumulated in the veraison phase followed by decrease of the content with ripening.     

Identification of novel pyranoanthocyanins in berry juices

In most fruit and berry products the attractive red color is unstable and easily susceptible to degradation. The colors of strawberry and raspberry juices can be enhanced and stabilized by the addition of cinnamic acids. The aim of this study was to identify the novel anthocyanin products responsible for the improved juice color. The pyranoanthocyanins formed were detected using high-performance liquid chromatography, and the fractions from this were analyzed using a nano-ESI tandem MS technique. The pyranoanthocyanins identified were formed from reactions of the main anthocyanins of strawberry and raspberry juices with ferulic acid or sinapic acid. The new anthocyanin derivatives were the 4-vinylguaiacol and 4-vinylsyringol adducts of pelargonidin and cyanidin. This is the first time that pelargonidin 3-glucoside-based vinylphenol pyranoanthocyanins and pyranoanthocyanins with more complex sugar residues have been found. This is also the first time pyranoanthocyanins have been detected in strawberry and raspberry juices.     

The effect of cocoa fermentation and weak organic acids on growth and ochratoxin A production by Aspergillus species

The acidic characteristics of cocoa beans have influence on flavor development in chocolate. Cocoa cotyledons are not naturally acidic, the acidity comes from organic acids produced by the fermentative microorganisms which grow during the processing of cocoa. Different concentrations of these metabolites can be produced according to the fermentation practices adopted in the farms, which could affect the growth and ochratoxin A production by fungi. This work presents two independent experiments carried out to investigate the effect of some fermentation practices on ochratoxin A production by Aspergillus carbonarius in cocoa, and the effect of weak organic acids such as acetic, lactic and citric at different pH values on growth and ochratoxin A production by A. carbonarius and Aspergillus niger in culture media. A statistical difference (ρ<0.05) in the ochratoxin A level in the cured cocoa beans was observed in some fermentation practices adopted. The laboratorial studies demonstrate the influence of organic acids on fungal growth and ochratoxin A production, with differences according to the media pH and the organic acid present. Acetic acid was the most inhibitory acid against A. carbonarius and A. niger. From the point of view of food safety, considering the amount of ochratoxin A produced, fermentation practices should be conducted towards the enhancement of acetic acid, although lactic and citric acids also have an important role in lowering the pH to improve the toxicity of acetic acid.     

3rd Ceramic Leadership Summit

The Ceramic Leadership Summit is designed to explore business opportunities, emerging technologies, and critical issues that challenge the ceramic and glass materials community. In 2012, the 3rd Ceramic Leadership Summit was held in conjunction with the 4th International Congress on Ceramics (ICC4). The one‐day program focused on technology transfer, entrepreneurship, and product innovation. Important topics discussed included strategies for creating and sustaining a small business, intellectual property protection, technology transfer, and mechanisms for government–university–industry partnerships. International speakers in the case study session addressed these topics from the perspective of their local and national policies and practices. Several of the case study presentations highlighted the use of “waste materials” in the creation and application of new technologies and products.     

Influence of Processing Parameters on the Sintering and Electrical Properties of Pb(Zn1/3Nb2/3)O3-Based Ceramics

Pb(Zn_1/3Nb_2/3)O₃-based ceramics have been prepared by two different processing methods: conventional (PZN-C) and reaction-sintering (PZN-RS). The conventionally prepared PZN-based ceramics densified at lower temperatures (950°C) than the reaction-sintered samples (1100°C), but the perovskite/pyrochlore ratio was always higher in PZN-RS. The presence of a substantial amount of pyrochlore phase in PZN-C ceramics caused a decrease in the electrical properties. The maximum dielectric constant values in PZN-C ceramics were 10%–15% lower than those of PZN-RS, despite a similar average grain size, 7 ± 0.2 μm. The temperature of the maximum of the dielectric constant ( T _max) was lower than that expected from the mixing rule because of the possible formation of Ba–Nb clusters. The higher chemical homogeneity in PZN-RS ceramics is the main reason for the higher dielectric constant, T _max and electromechanical response, as well as for the lower difference between T _max and the depolarization temperature ( T _d) and the lower diffusiveness parameter (δ).     

Cytoskeletal Protein 4.1G Is Essential for the Primary Ciliogenesis and Osteoblast Differentiation in Bone Formation

The primary cilium is a hair-like immotile organelle with specific membrane receptors, including the receptor of Hedgehog signaling, smoothened. The cilium organized in preosteoblasts promotes differentiation of the cells into osteoblasts (osteoblast differentiation) by mediating Hedgehog signaling to achieve bone formation. Notably, 4.1G is a plasma membrane-associated cytoskeletal protein that plays essential roles in various tissues, including the peripheral nervous system, testis, and retina. However, its function in the bone remains unexplored. In this study, we identified 4.1G expression in the bone. We found that, in the 4.1G-knockout mice, calcium deposits and primary cilium formation were suppressed in the trabecular bone, which is preosteoblast-rich region of the newborn tibia, indicating that 4.1G is a prerequisite for osteoblast differentiation by organizing the primary cilia in preosteoblasts. Next, we found that the primary cilium was elongated in the differentiating mouse preosteoblast cell line MC3T3-E1, whereas the knockdown of 4.1G suppressed its elongation. Moreover, 4.1G-knockdown suppressed the induction of the cilia-mediated Hedgehog signaling and subsequent osteoblast differentiation. These results demonstrate a new regulatory mechanism of 4.1G in bone formation that promotes the primary ciliogenesis in the differentiating preosteoblasts and induction of cilia-mediated osteoblast differentiation, resulting in bone formation at the newborn stage.     

Hybrid Nanoparticles and Composite Hydrogel Systems for Delivery of Peptide Antibiotics

The growing number of drug-resistant pathogenic bacteria poses a global threat to human health. For this reason, the search for ways to enhance the antibacterial activity of existing antibiotics is now an urgent medical task. The aim of this study was to develop novel delivery systems for polymyxins to improve their antimicrobial properties against various infections. For this, hybrid core–shell nanoparticles, consisting of silver core and a poly(glutamic acid) shell capable of polymyxin binding, were developed and carefully investigated. Characterization of the hybrid nanoparticles revealed a hydrodynamic diameter of approximately 100 nm and a negative electrokinetic potential. The nanoparticles demonstrated a lack of cytotoxicity, a low uptake by macrophages, and their own antimicrobial activity. Drug loading and loading efficacy were determined for both polymyxin B and E, and the maximal loaded value with an appropriate size of the delivery systems was 450 µg/mg of nanoparticles. Composite materials based on agarose hydrogel were prepared, containing both the loaded hybrid systems and free antibiotics. The features of polymyxin release from the hybrid nanoparticles and the composite materials were studied, and the mechanisms of release were analyzed using different theoretical models. The antibacterial activity against Pseudomonas aeruginosa was evaluated for both the polymyxin hybrid and the composite delivery systems. All tested samples inhibited bacterial growth. The minimal inhibitory concentrations of the polymyxin B hybrid delivery system demonstrated a synergistic effect when compared with either the antibiotic or the silver nanoparticles alone.     

Impact of Endocrine Disruptors upon Non-Genetic Inheritance

Similar to environmental factors, EDCs (endocrine-disrupting chemicals) can influence gene expression without modifying the DNA sequence. It is commonly accepted that the transgenerational inheritance of parentally acquired traits is conveyed by epigenetic alterations also known as “epimutations”. DNA methylation, acetylation, histone modification, RNA-mediated effects and extracellular vesicle effects are the mechanisms that have been described so far to be responsible for these epimutations. They may lead to the transgenerational inheritance of diverse phenotypes in the progeny when they occur in the germ cells of an affected individual. While EDC-induced health effects have dramatically increased over the past decade, limited effects on sperm epigenetics have been described. However, there has been a gain of interest in this issue in recent years. The gametes (sperm and oocyte) represent targets for EDCs and thus a route for environmentally induced changes over several generations. This review aims at providing an overview of the epigenetic mechanisms that might be implicated in this transgenerational inheritance.