A rapid liquid chromatography electrospray ionization mass spectrometry(n) method for evaluation of synephrine in Citrus aurantium L. samples

Immature bitter orange fruit and its extracts have been introduced into the market as an alternative to Ephedra in weight loss products. However, the safety of the immature bitter orange fruit and its extracts is a debated argument due to the presence of synephrine, a constituent known as a sympathomimetic agent. In this paper, we describe the development of a new, rapid, and simple liquid chromatography-electrospray ionization-tandem mass spectrometry method devoted to the quantitative determination of synephrine in bitter orange samples, containing a high quantity of synephrine, and sweet orange samples, known to contain a low level of synephrine but at the same time being one of the main synephrine sources in a normal human diet. Two bitter orange dry extracts containing 5 and 6% sSynephrine and 10 sweet orange samples have been analyzed. Between the sweet orange samples, six were fresh oranges and four were fresh-squeezed juices; in these samples, the synephrine levels ranged from 0.00128 to 0.00349%.     

Protective effect of the phenolic fraction from virgin olive oils against oxidative stress in human cells

This paper reports the protective effect of the phenolic fraction extracted from extra virgin olive oils (OOPEs) against the cytotoxic effects of reactive oxygen species in human erythrocytes and Caco-2 cells, employed as model systems. Pretreatment of cells with various OOPEs, indeed, provides a remarkable protection against oxidative damages: this effect was strictly dependent on the o-diphenolic content of the extracts. Moreover, the protective effects observable in cellular systems were compared with in vitro antioxidant properties, measured by using the FRAP (ferric reducing/antioxidant power) assay; the reducing ability of OOPEs strictly parallels their o-phenolic content. The linear relationship demonstrated between biological effects and antioxidant capacity measured by the FRAP assay allows us to propose the use of this rapid colorimetric method in assessing and certifying the antioxidant power of extra virgin olive oil.     

Fusarium genetic traceability: Role for mycotoxin control in small grain cereals agro-food chains

Risks associated with mycotoxin contamination of cereals, that are included in the ten major staple foods and greatly contribute to the dietary energy intake, are of worldwide relevance. In small grain cereals, mycotoxins are produced by fungi such as Aspergillus, Penicillium, Alternaria and Fusarium that colonize the plant in the field and can grow during the post-harvest period, producing several classes of mycotoxins. The identification of mycotoxigenic fungal species and strains is essential for developing effective strategies for control. For this purpose, genetic traceability has proved to be a valuable tool that can be applied along the whole production chain, starting in the field for early diagnosis of FHB (Fusarium Head Blight) disease to the final processing steps, such as malting or pasta making. In this paper, DNA-based analytical tools that are currently available for the identification and quantification of mycotoxigenic fungal species and strains are reviewed, with particular emphasis on Fusarium, and their possible applications in mycotoxin control in small grain cereal chains are discussed.     

Rhamnose-binding lectins induce respiratory burst activity in macrophage cells from rainbow trout

Rhamnose-binding lectins (RBLs) have been isolated and characterized from various kinds of fish. RBLs recognize the structures of lipopolysaccharides and lipoteichoic acid on microbes, and have an opsonic effect, suggesting that RBLs play an important role as pattern recognition proteins in innate immunity. However, the function of RBLs in the removal of pathogens from the body has not yet been elucidated. In this study, we investigated the effect of three RBLs—named CSL1, 2, and 3—that were isolated from chum salmon Oncorhynchus keta eggs on the respiratory burst activity in a macrophage cell line. The CSLs induced reactive oxygen species in the peritoneal macrophage cell line from rainbow trout Oncorhynchus mykiss (RTM5). However, this activity was not affected by l-rhamnose or dl-threo-1-phenyl-2-palmitoylamino-3-morpholino-1-propanol, an inhibitor for glucosylceramide synthesis, indicating that the interaction between CSLs and Gb3 on cell surfaces was not involved in the induction of respiratory burst activity.     

Phytoestrogen content of foods of animal origin: dairy products, eggs, meat, fish, and seafood

Dietary phytoestrogens may be involved in the occurrence of chronic diseases. Reliable information on the phytoestrogen content in foods is required to assess dietary exposure and disease risk in epidemiological studies. However, existing analyses have focused on only one class of these compounds in plant-based foods, and there is only little information on foods of animal origin, leading to an underestimation of intake. This is the first comprehensive study of phytoestrogen content in animal food. We have determined the phytoestrogen content (isoflavones: biochanin A, daidzein, formononetin, genistein, and glycitein; lignans: secoisolariciresinol and matairesinol; coumestrol; equol; enterolactone; and enterodiol) in 115 foods of animal origin (including milk and milk-products, eggs, meat, fish, and seafood) and vegetarian substitutes using liquid chromatography-mass spectrometry (LC-MS) with (13)C-labeled internal standards. Phytoestrogens were detected in all foods analyzed; the average content was 20 microg/100 g of wet weight (isoflavones, 6 microg/100 g; lignans, 6 microg/100 g; equol, 3 microg/100 g; and enterolignans, 6 microg/100 g). In infant soy formula, 19 221 microg/100 g phytoestrogens were detected (compared to 59 microg/100 g in non-soy formula). Our study shows that all foods analyzed contained phytoestrogens and most foods (except for fish, seafood, and butter) contained mammalian phytoestrogens (enterolignans and equol). This is the first comprehensive study of phytoestrogen content of foods of animal origin and will allow for a more accurate estimation of exposure to dietary phytoestrogens.     

A reliable assay for the detection of soft wheat adulteration in Italian pasta is based on the use of new DNA molecular markers capable of discriminating between Triticum aestivum and Triticum durum

Italian pasta must be prepared using exclusively durum wheat. According to current Italian rules, only a maximum of 3% Triticum aestivum is allowed to account for cross-contamination that may occur during the agricultural process. Efficient methods for the detection of accidental or intentional contamination of common wheat to durum wheat products are therefore required. This article describes a novel approach for the detection and quantification of soft wheat adulteration in whole grain durum flours and dried pasta. The assay relies on the presence of intron-specific DNA length polymorphisms in the plant β-tubulin gene family, which can be highlighted through the PCR-based TBP (Tubulin-Based Polymorphism) method. In wheat, the TBP method produces species-specific amplification products, which can be either directly used as new DNA molecular markers capable of discriminating between T. aestivum and Triticum durum or analyzed at the sequence level for the design of species-specific probes. The latter approach allowed the development of new sequence-specific targets that can be exploited in RT-PCR assays for a rapid and accurate quantification of soft wheat adulteration in durum wheat pasta.     

Anaerobic soil disinfestation is an alternative to soil fumigation for control of some soilborne pathogens in strawberry production

Alternatives to soil fumigation are needed for soilborne disease control. The aim of this study was to test anaerobic soil disinfestation (ASD) as an alternative to soil fumigation for control of critical soilborne pathogens in Californian strawberry production. Controlled environment experiments were conducted at 25 and 15 °C to test different materials as carbon sources for ASD using soil inoculated with Verticillium dahliae. Field trials were conducted in three locations comparing ASD with 20 Mg ha^?1 rice bran (RB) against fumigated and untreated controls, steam, mustard seed meal and fish emulsion. In ASD‐treated soils, temperature and extent of anaerobic conditions were critical for control of V. dahliae, but multiple carbon inputs reduced inoculum by 80–100%. In field trials, ASD with RB provided control of a number of pathogens, and in three of four trials produced marketable fruit yields equivalent to fumigation. Little weed control benefit from ASD was found. ASD with RB also induced changes in the soil microbiome that persisted through the growing season. When equivalent yields were obtained, net returns above harvest and treatment costs with ASD RB were 92–96% of those with bed fumigation based on average prices over the previous 5 years. ASD can be a viable alternative for control of some soilborne pathogens. Growers are adopting ASD in California strawberry production, but research to determine optimal soil temperatures, anaerobicity thresholds and carbon sources for effective control of specific pathogens is needed.     

CATASAN Is a New Anti-Biofilm Agent Produced by the Marine Antarctic Bacterium Psychrobacter sp. TAE2020

The development of new approaches to prevent microbial surface adhesion and biofilm formation is an emerging need following the growing understanding of the impact of biofilm-related infections on human health. Staphylococcus epidermidis, with its ability to form biofilm and colonize biomaterials, represents the most frequent causative agent involved in infections of medical devices. In the research of new anti-biofilm agents against S. epidermidis biofilm, Antarctic marine bacteria represent an untapped reservoir of biodiversity. In the present study, the attention was focused on Psychrobacter sp. TAE2020, an Antarctic marine bacterium that produces molecules able to impair the initial attachment of S. epidermidis strains to the polystyrene surface. The setup of suitable purification protocols allowed the identification by NMR spectroscopy and LC-MS/MS analysis of a protein–polysaccharide complex named CATASAN. This complex proved to be a very effective anti-biofilm agent. Indeed, it not only interferes with cell surface attachment, but also prevents biofilm formation and affects the mature biofilm matrix structure of S. epidermidis. Moreover, CATASAN is endowed with a good emulsification activity in a wide range of pH and temperature. Therefore, its use can be easily extended to different biotechnological applications.