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We discuss upsets in erased floating gate cells, due to large threshold voltage shifts, using statistical distributions collected on a large number of memory cells. The spread in the neutral threshold voltage appears to be too low to quantitatively explain the experimental observations in terms of simple charge loss, at least in SLC devices. The possibility that memories exposed to high energy protons and heavy ions exhibit negative charge transfer between programmed and erased cells is investigated, although the analysis does not provide conclusive support to this hypothesis.

Seventy-one (71) food samples were analyzed for the mycotoxin fumonisin by a monoclonal antibody based competitive enzyme-linked immunosorbent assay (ELISA). Fumonisins were detected primarily in corn-based products with 7/12, 2/2 and 1/3 and 1/7 yellow cornmeal, blue cornmeal, corn muffin mix, and mixed grain cereal samples yielding positive results, respectively. When the positive samples and randomly selected negative samples were assessed by other methods, correlations (r values) between ELISA and gas chromatography-mass spectrometry (GC-MS), ELISA and high-pressure liquid chromatography (HPLC) and GC-MS and HPLC were 0.478 (p0.05), 0.512 (p0.05), and 0.946 (p0.01), respectively. The results suggested that although the immunoassay could be used for screening of fumonisin in food samples, higher estimates were attained by ELISA than by the other two methods particularly in the more contaminated samples. These observations may result from differences in sample preparation among the methods or because of the presence of structurally related compounds in extracts that are detectable by ELISA but not the other two methods.

Starting from all the experimental evidence we presented in Part I of this work (Resnati et al. , 2016), in this paper, we propose a new microscopic picture for cycling-induced charge trapping/detrapping in Flash memories. This picture involves structural relaxation of the charged tunnel-oxide defects as limiting step in the detrapping process, triggering the possibility for defects to exchange carriers with the substrate. This view is implemented in a statistical model able to reproduce the threshold-voltage instability coming from the phenomenon, including its dependence on cycling dose, temperature, and cell state during idle/bake periods. The model is then used to address some elaborated accelerated testing schemes commonly used to monitor charge trapping/detrapping and its related instability in Flash memories, explaining their results in the context of the new proposed picture of the phenomenon.

This paper exploits the possibility of monitoring the floating-gate (FG) charge of state-of-the-art NAND Flash arrays with a single-electron resolution to investigate in detail the program operation and some previously inaccessible technological parameters. In particular, the analysis leads to the assessment of the statistical distribution of the FG to control-gate capacitance and of the leakage current from the FG through the intergate dielectric during the programming pulses. Results highlight that resolving the FG charge at the single-electron level opens new prospects for the investigation of the operation and the parameters of NAND Flash arrays.

Rationale Several Bacillus strains, typically isolated from different food sources, represent renowned producers of a multitude of low and high molecular weight compounds, including lipopeptides and macrolactones, with an importance for their antimicrobial activity. The high homology shared by many of these compounds also occurring as closely related isoforms poses a challenge in their prompt detection. Methods Identification and structural elucidation is generally achieved by matrix-assisted laser desorption/ionization (MALDI) or liquid chromatography (LC) coupled to mass spectrometry (MS) after a pre-fractionation and/or purification step of the extract. In this paper we report the application of a method based on LC separation and high-resolution Orbitrap™-based MS for the rapid screening of raw filtrate of the strain Bacillus subtilis TR50 endowed with antimicrobial activity, without requiring any sample pre-treatment. Results Upon direct analysis of the cell-free filtrate of Bacillus subtilis TR50 by high-resolution mass spectrometry (HRMS), different compounds families, that proved to exert a remarked antimicrobial activity against several foodborne pathogens, can be readily displayed along the chromatographic run. Among them, three different classes were identified and characterized belonging to the iturin, fengycin and surfactin groups. The high resolving power and accurate mass accuracy provided by the HRMS system in use ensured an enhanced selectivity compared to other mass spectrometers. In addition, after activation of the HCD cell, the HR-MS/MS spectra can provide insights in the structural elucidation of several compounds. Conclusions The acquisition of HRMS spectra of raw filtrates of subtilis strains allows untargeted analysis of the major classes of compounds produced to be performed, thus facilitating identification of other unknown bioactive molecules after retrospective analysis. These features make this approach a fast tool applicable to the rapid screening and further identification of antimicrobial compounds released by Bacillus strains in raw filtrates. Copyright © 2015 John Wiley & Sons, Ltd.

The aim of this work was to select a yeast able to hydrolyse phytate during bread-making or under simulated gastro-intestinal (GI) conditions. High levels of cell-bound and intracellular phytate-degrading enzyme activities were found in Saccharomyces cerevisiae LC3, selected among 16 yeast strains. Phytase enzyme of LC3, purified by three chromatographic steps, had a molecular weight of ca. 48 kDa; its optimal activity, displayed on sodium phytate, was at pH 5.0 and at 40 °C. S. cerevisiae LC3 was used together with negative phytase producing Lactobacillus plantarum LP41 to make wholemeal bread that showed a marked decrease ( ca. 79%) in phytate content in comparison with bread produced with commercial baker's yeast and the same Lactobacillus strain. In addition, S. cerevisiae LC3 survived and hydrolysed phytate under simulated GI conditions. The results of this study indicate that the selected LC3 could be a promising candidate to be used as starter for the reduction of phytate content of wholemeal flour during bread-making or as a probiotic for human or animal applications.

Ingestion of food is considered a major route of exposure to many contaminants including mycotoxins. The amount of mycotoxin resisting to the digestion process and potentially absorbable by the systemic circulation is only a smaller part of that ingested. In vitro digestion models turn useful for evaluating mycotoxins bioaccessibility during the intestinal transit and can be intended as a valuable tool for the assessment of mycotoxin bioavailability in food. In this paper we describe a study aimed at investigating toxicity of in vitro gastro-duodenal digests of mycotoxin contaminated bread collected along the digestion time-course. Toxicity tests were carried out on a sensitive RPMI lymphoid B cell line chosen as the most suitable lineage to assess toxicity retained by gastro-duodenal digests. In parallel, a chemical quantification of T-2 and HT-2 toxins contaminating the bread digests was accomplished during the gastric and duodenal transit. The digestive fluids undergoing chemical and toxicological analysis were collected at the beginning and end of gastric phase, and after completion of the duodenal phase. Results proved that a correlation between HT-2 content and toxicity did exist although a more persistent toxic activity was displayed in the later stage of the duodenal phase. This persistent toxicity might be explained by the co-occurrence of unknown HT-2-related conjugates or metabolites formed during digestion.

This paper reviews the recent historical trends of the NAND Flash technology, highlighting the evolution of its main parameters and explaining what allowed it to become not only the most important integrated solution for nonvolatile storage of high volumes of data but also a strong rival eroding the market share of hard-disk drives. The scaling trend followed by planar arrays will be discussed with close attention, along with the major physical constraints impacting the performance and the reliability of modern deca-nanometer technologies. This will make clear why the development of further planar nodes with feature size below ~15 nm, representing today’s state of the art, can be considered less favorable than turning all the efforts toward the integration of 3-D arrays. The most promising 3-D architectures will then be reviewed, discussing their benefits and issues and addressing the impact of the change of the integration paradigm from the standpoint of the major NAND applications.

The variability in the total ionizing dose response of 25-nm single level cell NAND Flash memories from two different lots is studied. More than 1 Terabit of floating gate cells were irradiated with gamma rays and the number of errors was statistically analyzed. The behavior of the two lots is remarkably different in terms of floating gate errors. The statistical parameters such as mean, standard deviation, and shapes of the error distributions were studied. The sample-to-sample statistical distribution of TID errors is not Gaussian and it is attributed to variability in neutral cells, due to phenomena such as random discrete dopant fluctuations. Finally, the impact of scaling on variability is discussed.

Food allergen research has made giant steps in the last years thanks to the features offered by the latest technology of mass analyzers placed on the market allowing multiplex sensitive detection of proteins. Potentials and features of two mass analyzers namely a linear ion trap capable of performing a data dependent or selected reaction monitoring analysis and an Orbitrap(TM) stand-alone MS enabling a broadband fragmentation without mass selection at highest mass resolving power are herein described and applied to the multiplex screening of allergens in a type of wine chosen as a reference matrix. Quantitative and confirmative capabilities of both platforms were assessed on the specific case study, the multiple detection of egg and milk -related proteins, typically employed in white wines as fining agents. Commercial bioinformatic tools used for a quick allergen identification will be also discussed.

An experimental protocol for evaluating performances of qualitative methods through interlaboratory validation has been designed and applied to a multiplex dipstick kit for the determination of major Fusarium toxins, namely zearalenone, T-2 and HT-2 toxins, deoxynivalenol, and fumonisins B1 and B2 in wheat and maize. The test is intended for screening of cereals for the presence/absence of these mycotoxins at maximum permitted levels established by European legislation. The response of the measurement is determined with a reader device. Decision on whether a sample is classified as positive or negative is taken by comparing the measured response with a cut off value which has been established based on the results from the method validation. The collaborative study involved 12 laboratories and included a training phase enabling the participants to familiarise with the protocol. The interlaboratory validation design consisted of three steps, namely (1) estimating the precision under reproducibility conditio...

There is a raising demand for sensitive and high throughput MS based methods for screening purposes especially tailored to the detection of allergen contaminants in different food commodities. A challenging issue is represented by complex food matrices where the antibody-based kits commercially available might encounter objective limitations consequently to epitope masking phenomena due to a multitude of interfering compounds arising from the matrix. The performance of a method duly optimized for the extraction and simultaneous detection of soy, egg and milk allergens in a cookie food matrix by microHPLC-ESI-MS/MS, is herein reported. Thanks to the innovative configuration and the versatility shown by the dual cell linear ion trap MS used, the most intense and reliable peptide markers were first identified by untargeted survey experiment, and subsequently employed to design an ad hoc multitarget SRM method, based on the most intense transitions recorded for each selected precursor peptide. A sample extraction and purification protocol was optimized also including an additional step based on sonication, which resulted in a considerable improvement in the detection of milk allergen peptides. Data Dependent TM Acquisition scheme allowed to fill out a tentative list of potential peptide markers, which were further filtered upon fulfilling specific requirements. A total of eleven peptides were monitored simultaneously for confirmation purposes of each allergenic contaminant and the two most sensitive peptide markers/protein were selected in order to retrieve quantitative information. Relevant LODs were found to range from 0.1 g/g for milk to 0.3 g/g for egg and 2 g/g for soy.

T-2 and HT-2 toxins are Fusarium mycotoxins frequently occurring in cereal-based products with a toxic effect ascertained on different biological systems. Despite other mycotoxins, to date bioaccessibility of T-2 and HT-2 toxins has never been investigated. In order to provide insights on T-2 and HT-2 stability and bioaccessibility along digestion in the upper intestinal tract, differently contaminated bread samples produced at laboratory scale were submitted to in vitro digestion experiments. Two different contaminated bread models were prepared with this aim. One employed naturally contaminated wheat flour for production of naturally contaminated bread, whereas in the other model dough was fortified with both mycotoxins during bread preparation before undergoing the leavening and baking process. Gastro-duodenal fluids were collected at different time-points along simulated digestion and mycotoxin content determined by LC-High Resolution–Mass Spectrometry. Our data report that HT-2 content in the beginning of the gastric phase was not significantly different from what was recorded at the end of the duodenal phase, although an apparent decrease of the signal was displayed in the early duodenal phase in both bread models tested. By contrast, the consistent drop of approximately 50% observed for T-2 when the duodenal digestion started remained constant until the digestion reached completion. In conclusion both approaches provided similar results thus considering incurred food models valid alternatives to undertake such investigations. Finally, bioaccessibility of T2 and HT2 was calculated in both food models tested with a higher bioaccessibility found in artificially contaminated bread samples. Industrial Relevance: Food is very complex both in composition and structure; therefore, generic realistic models that can mimic this complexity are required. Such models would greatly facilitate evaluation of the impact of changing composition or processing conditions on nutrition and safety. The primary scope of this work (part of the EU funded DREAM project) is to integrate experimental and mathematical approaches elsewhere optimised to develop a cereal-based food model realistic enough to be used by the industry and sufficiently versatile to be used as predictive tool of food behaviour. This is would be also applicable to study the release of contaminants from food such as mycotoxins. In general, food parameters may affect mycotoxins bioavailability from foods. A number of mycotoxins are currently regulated in foodstuffs by the European Commission and as a consequence there is growing interest and concern of the public health authorities for the presence of mycotoxins in human food. Due to the different chemical structure and stability, the fate of mycotoxins during food processing and the relevant bioaccessibility upon digestion process vary considerably depending on the different food preparations. Limited data are currently available on the fate of trichothecenes during food processing and human digestion. Therefore, the fate of these mycotoxins during processing of some specific food models is of paramount importance to be assessed. The bioaccessibility of trichothecenes (deoxynivalenol, T2 and HT-2 toxins) from a selected cereal based food model evaluated by an in vitro digestion model is also important to understand the safety of the final food.

The synthesis of partially hydrolyzed fumonisins (PHFB1 and PHFB2) and hydrolyzed fumonisins (HFB1 and HFB2) by chemical hydrolysis of pure fumonisins (FB1 and FB2) is reported together with the isolation and characterization by liquid chromatography–high-resolution mass spectrometry (LC–HRMS). Two structural isomers of partially hydrolyzed forms of FB1 and FB2 were identified, namely PHFB1a and PHFB1b and PHFB2a and PHFB2b. Reaction yields were 21% for PHFB1 (sum of the two isomers), 52% for HFB1, 31% for PHFB2 (sum of the two isomers) and 30% for HFB2. Purity of each isolated compound was >98%. An LC–HRMS method for the simultaneous determination of fumonisins and their partially and totally hydrolyzed derivatives was applied to 24 naturally contaminated samples of maize and maize-based products. The majority of samples (18 out of 24) were contaminated with fumonisins B1 and B2. Fumonisins co-occurred with both partially hydrolyzed and hydrolyzed fumonisins in four nixtamalized samples (three masa flours and one tortilla chips). Co-occurrence of fumonisins with partially hydrolyzed fumonisins was also recorded in one sample of maize kernels and four samples of maize-based products (i.e. maize meal, cous-cous, corn-cakes and cornflakes). Mycotoxins levels ranged from 60 to 5700 µg/kg for fumonisins (sum of FB1 and FB2), from 10 to 210 µg/kg for partially hydrolyzed fumonisins (sum of PHFB1 and PHFB2) and from 30 to 200 µg/kg for hydrolyzed fumonisins (sum of HFB1 and HFB2). This is the first report of the isolation of PHFB2 and the co-occurrence of FB1, FB2, PHFB1, PHFB2, HFB1 and HFB2 in maize products. Considering the growing use of nixtamalized and maize-based products, the monitoring of fumonisins and their partially and totally hydrolyzed forms in these products may represent an important contributing factor in evaluating the relevant human risk exposure. Copyright © 2014 John Wiley & Sons, Ltd.

This letter highlights a random telegraph noise-induced sensitivity of the data retention threshold-voltage transient of nand Flash memory cells to their position in the programmed array distribution. This sensitivity appears with a reduction of the threshold-voltage loss of the cells in the lower part of the programmed distribution of the memory array and an increase of that of the cells in the upper part of the distribution. The experimental evidence is explained considering the impact of random telegraph noise on the programmed array distribution in the stretch of time in-between the program-and-verify operation and the first read operation used as reference for data retention assessment.

This letter provides the first direct experimental detection of single-electron charging of the floating gate of a mainstream Flash memory cell. The detection is shown to be easily achievable through conventional and very simple measurement techniques on state-of-the-art technologies. Results represent a milestone for the investigation of the physics of Flash memory operation, opening the possibility for direct analyses of the piling up of single electrons in the floating gate during cell programming.

Subsistence farmers are exposed to a range of mycotoxins. This study applied novel urinary multi-mycotoxin LC-MS/MS methods to determine multiple exposure biomarkers in the high oesophageal cancer region, Transkei, South Africa. Fifty-three female participants donated part of their maize-based evening meal and first void morning urine, which was analysed both with sample clean-up (single and multi-biomarker) and by a 'dilute-and-shoot' multi-biomarker method. Results were corrected for recovery with LOD for not detected. A single biomarker method detected fumonisin B1 (FB1) (87% incidence; mean ± standard deviation 0.342 ± 0.466 ng/mg creatinine) and deoxynivalenol (100%; mean 20.4 ± 49.4 ng/mg creatinine) after hydrolysis with b-glucuronidase. The multi-biomarker 'dilute-and-shoot' method indicated deoxynivalenol-15-glucuronide was predominantly present. A multi-biomarker method with b-glucuronidase and immunoaffinity clean-up determined zearalenone (100%; 0.529 ± 1.60 ng/mg creatinine), FB1 (96%; 1.52 ± 2.17 ng/mg creatinine), a-zearalenol (92%; 0.614 ± 1.91 ng/mg creatinine), deoxynivalenol (87%; 11.3 ± 27.1 ng/mg creatinine), b-zearalenol (75%; 0.702 ± 2.95 ng/mg creatinine) and ochratoxin A (98%; 0.041 ± 0.086 ng/mg creatinine). These demonstrate the value of multi-biomarker methods in measuring exposures in populations exposed to multiple mycotoxins. This is the first finding of urinary deoxynivalenol, zearalenone, their conjugates, ochratoxin A and zearalenols in Transkei.

Fusarium langsethiae, formally described as a new species over a decade ago, has been identified as the main producer of HT-2 (HT2) and T-2 (T2) toxins in Europe in small cereal grains. Mycotoxin contamination caused by this Fusarium species can represent a food safety hazard that deserves further attention. In the present work, the mycotoxin profile in wheat cultures of F. langsethiae is presented with particular reference to the production of major type-A trichothecenes and their glucosyl derivatives. F. langsethiae isolates, representative of the major Italian wheat cultivation areas, were tested for the production of T2, HT2, diacetoxyscirpenol (DAS) and neosolaniol (NEO), and relevant glucosyl derivatives. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used for the identification and chemical characterization of these metabolites. F. langsethiae isolates under investigation resulted to be potent producers of T2, HT2 and NEO. Furthermore, a well-defined set of isolates, all originating from Central Italy, produced also DAS. All isolates were found to be able to produce HT2 glucosyl derivatives, whereas only traces of T2 glucoside were detected in one sample. Furthermore, two mono-glucosyl derivatives of NEO and one mono-glucoside derivative of DAS were identified and characterized. The screening for the presence/absence of glucosylated trichothecenes in analyzed fungal extracts revealed a general co-occurrence of these derivatives with the parent toxin at levels that could be roughly estimated to account up to 37% of the relevant unconjugated toxin. This is the first report of the production of glucosylated trichothecenes by F. langsethiae cultured on small grains.

The results of a proficiency test for the LC-MS/(MS) determination of up to 11 mycotoxins (aflatoxins B1, B2, G1 and G2, fumonisins B1 and B2, ochratoxin A, deoxynivalenol, T-2 and HT-2 toxins and zearalenone) in maize were evaluated to identify possible strengths and weaknesses of various methodologies used by the 41 participating laboratories. The majority of laboratories (56%) used mixtures of acetonitrile:water for extraction. Other laboratories used methanol:water mixtures (17%) or performed two consecutive extractions with phosphate buffer solution (PBS) followed by methanol (15%). Few laboratories used mixtures of acetonitrile:water:methanol (7%), water:ethyl acetate (2.5%) or PBS alone (2.5%). The majority of laboratories (58%) used a clean-up step prior to chromatography. The remaining laboratories analysed crude extracts (37%) or used a mixed approach (5%). The amount of sample equivalent injected into LC-MS/(MS) ranged between 0.1-303 mg for purified extracts and 0.08-20 mg for directly analysed crude extracts. External (54%), matrix-matched (22%) or stable isotope-labelled internal standards calibration (24%) were used for toxin quantification. In general, extraction mixtures of water with acetonitrile, methanol or both provided good results for quantitative extraction of mycotoxins from maize. Laboratories using sample extract clean-up reported acceptable results for the majority of mycotoxins. Good results were also obtained by laboratories that analysed crude extracts although a high variability of results was observed for all tested mycotoxins. Matrix-matched calibration or isotope-labelled internal standards efficiently compensated matrix effects whereas external calibration gave reliable results by injecting ≤10 mg of matrix equivalent amounts. Unacceptable high recovery and high variability of fumonisin results were obtained by the majority of laboratories, which could not be explained and thus require further investigation. These findings provide the basis for the optimization and selection of methods to be used in future interlaboratory validation studies to derive their performance characteristics for simultaneous determination of mycotoxins in maize.

RATIONALE A method based on High-Resolution Mass Spectrometry was developed for the simultaneous determination of fining agents containing potentially allergenic milk (casein) and egg-white (lysozyme and ovalbumin) proteins, added to commercial white wines at sub-ppm levels. Selected tryptic peptides were used as quantitative markers. An evaluation of protein digestion yields was also performed by implementing the 15 N-valine-labelled analogues of the best peptide markers identified for αS1-casein and ovalbumin. METHODS The method was based on the combination of ultrafiltration (UF) of protein-containing wines, tryptic digestion of the dialyzed wine extracts and liquid chromatography/high resolution mass spectrometry (LC/HRMS) analysis of tryptic digests. Peptides providing the most intense electrospray ionization (ESI)-MS response were chosen as quantitative markers of the proteins under investigation. RESULTS Six-point calibrations were performed by adding caseinate and egg-white powder in the concentration range between 0.25 and 10 µg/mL, to an allergen-free white wine. The following three peptide markers, LTEWTSSNVMEER, GGLEPINFQTAADQAR and ELINSWVESQTNGIIR, were highlighted as best markers for ovalbumin, while GTDVQAWIR and NTDGSTDYGILQINSR for lysozyme and YLGYLEQLLR, GPFPIIV and FFVAPFPEVFGK for caseinate. Limits of detection (LODs) ranged from 0.4 to 1.1 µg/mL. CONCLUSIONS The developed method is suited for assessing the contemporary presence of allergenic milk and egg proteins characterizing egg white and caseinate, fining agents typically employed for wine clarification. The LODs of the method enable the detection of sub-ppm concentrations of residual fining agents, that could represent a potential risk for allergic consumers. Copyright © 2013 John Wiley & Sons, Ltd.

Maize harvested in the Centane region of the former Transkei, Eastern Cape Province, South Africa, by subsistence farmers has been shown over many seasons to be contaminated with fumonisin mycotoxins. However, there are limited data on the presence of other mycotoxins. Two multimycotoxin LC-MS/MS methods were applied to good and moldy maize samples, as separated by the farmers themselves from the 2011 harvest. One method involved extract cleanup on multitoxin immunoaffinity columns before LC-MS/MS analysis for aflatoxins, fumonisins, deoxynivalenol (DON), zearalenone (ZEN), and T-2 and HT-2 toxins. The other method was based on a "dilute-and-shoot" approach for the above mycotoxins and a wide range of other fungal secondary metabolites. Both methods showed high incidences of fumonisins B1 and B2 (FB1 and FB2) in good maize (100% for both by the first method, means were 2083 and 927 μg/kg for the two analogues; 93% for both by the second method, positive means of 2764 and 1050 μg/kg, respectively). All samples of moldy maize were contaminated (mean FB1 of 27.64 and 35.98 mg/kg, respectively; mean FB2 of 10.58 and 14.14 mg/kg, respectively). Comparison of the two methods for FB1 and FB2 over the entire range of samples gave R(2) values 0.9144 and 0.8859, respectively. Low levels of DON were found by both methods (positive means of 12 and 4.7 μg/kg in good maize, respectively, and of 14 and 5.8 μg/kg in moldy maize, respectively). ZEN was determined with positive means of 108 and 25 μg/kg in good maize, respectively, and of 111 and 135 μg/kg in moldy maize, respectively. No aflatoxins, OTA, or T-2 or HT-2 toxins were detected. A wide range of other Fusarium , Aspergillus , Alternaria , and Penicillium mycotoxins and secondary metabolites were determined.

The multi-biomarker approach was used to validate urinary biomarkers in piglets administered boluses contaminated with mixtures of deoxynivalenol (DON), aflatoxin B1 (AFB1), fumonisin B1 (FB1), zearalenone (ZEA) and ochratoxin A (OTA) at different concentrations. Boluses contaminated with mycotoxins were prepared by slurrying and freezedrying feed material fortified with culture extracts of selected toxigenic fungi. Piglets were individually placed in metabolic cages to collect urine before gavage and 24 h post dose. Urine samples were hydrolysed with β-glucuronidase and analysed by a multi-biomarker LC-MS/MS method developed and validated to identify and measure biomarkers of FB1, OTA, DON, ZEA and AFB1. Urinary levels of FB1, OTA, DON + de-epoxy-deoxynivalenol, ZEA + alphazearalenol and aflatoxin M1 were selected as biomarkers of FB1, OTA, DON, ZEA and AFB1, respectively. Mean percentages of dietary mycotoxins excreted as biomarkers in 24 h post dose urine were 36.8% for ZEA, 28.5% for DON, 2.6% FB1, 2.6% for OTA and 2.5% for AFB1. A good correlation was observed between the amount of mycotoxins ingested and the amount of relevant biomarkers excreted in 24 h post dose urine. Linear dose-response correlation coefficients ranged between 0.68 and 0.78 for the tested couples of mycotoxin/biomarker. The good sensitivity of the LC-MS/MS method and the good dose-response correlations observed in this study permitted to validate the selected mycotoxin biomarkers in piglets at dietary levels close to the maximum permitted levels reported in Commission Directive 2003/100/EC for AFB1 and the guidance values reported in Commission Recommendation 2006/576/EC for DON, ZEA, OTA and FB1.

In this work, we present clear experimental results pointing to a new microscopic picture for cycling-induced charge trapping/detrapping in Flash memories. In particular, the evidence gathered from experiments designed to investigate the dependence of charge detrapping on cell threshold-voltage reveals that the simple and widely used model based only on carrier exchange between oxide defects and substrate is not enough to explain the main features of the phenomenon. We then propose a new microscopic description of the detrapping phenomenology, including structural relaxation of oxide defects as a limiting step enabling carrier exchange. This new microscopic picture for the oxide defects is, finally, implemented in a statistical model able to reproduce the charge trapping/detrapping dynamics and the consequent threshold-voltage instabilities along the memory array lifetime.

By using our 16-nm NAND Flash technology as a test vehicle, in this paper, we summarize all the experimental evidence that we have gathered so far on the phenomenology of cycling-induced charge trapping/detrapping in Flash memories. In particular, results from experiments conceived to explore the dependence of charge detrapping on cell memory state reveal that the phenomenon involves more than the simple capture and emission of charge carriers in tunnel-oxide defects. Besides, the thermal activation of charge trapping is highlighted for the first time, along with the detailed dynamics leading to the increase of the trapped charge as cell cycling proceeds. These results allow to lay some cornerstones for the microscopic description of cyclinginduced charge trapping/detrapping in Flash memories. From these cornerstones, in the companion paper (Resnati et al. , 2016), a new physical picture for the trapping/detrapping processes will be proposed and implemented in a statistical model able to reproduce their corresponding threshold-voltage instabilities over array lifetime.

A new analytical method for the determination of ochratoxin A (OTA) in red wine has been developed by using a double-extract cleanup and a fluorometric measurement after spectral deconvolution. Wine samples were diluted with a solution containing 1% polyethylene glycol and 5% sodium hydrogencarbonate, filtered, and purified by immunoaffinity and aminopropyl solid-phase column. OTA contents in the purified extract were determined by a spectrofluorometer (excitation wavelength, 330 nm; emission wavelength, 470 nm) after deconvolution of fluorescence spectra. Average recoveries from wine samples spiked with OTA at levels ranging from 0.5 to 3.0 ng/mL were 94.5-105.4% with relative standard deviations (RSD) of15% (n = 4). The limit of detection (LOD) was 0.2 ng/mL, and the total time of analysis was 30 min. The developed method was tested on 18 red wine samples (naturally contaminated and spiked with OTA at levels ranging from 0.4 to 3.0 ng/mL) and compared with AOAC Official Method 2001.01, based on immunoaffinity column cleanup and HPLC with fluorescence detector. A good correlation (r(2) = 0.9765) was observed between OTA levels obtained with the two methods, highlighting the reliability of the proposed method, the main advantage of which is the simple OTA determination by a benchtop fluorometer with evident reductions of cost and time of analysis.

The analytical performances of a novel DNA-ligand system using the time-resolved fluorescence (TRF) response of ochratoxin A (OTA)-terbium-DNA aptamer interaction were tested for the quantitative determination of OTA in wheat. Wheat was extracted with acetonitrile/water (60:40, v/v) followed by clean-up through affinity columns containing a DNA-aptamer-based oligosorbent. Then, OTA was detected by TRF spectroscopy after reaction with a terbium fluorescent solution containing the DNA-aptamer probe. The entire procedure was performed in less than 30 min, including sample preparation, and allowed analysis of several samples simultaneously with a 96-well microplate reader. The average recovery from samples spiked with 2.5-25 ?g kg(-1) OTA was 77%, with a relative standard deviation lower than 6% and a quantification limit of 0.5 ?g kg(-1). Comparative analyses of 29 naturally contaminated (up to 14 ?g kg(-1)) wheat samples using the aptamer-affinity column/TRF method or the immunoaffinity column/high-performance liquid chromatography method showed good correlation (r = 0.985) in the range tested. The trueness of the aptamer-based method was additionally assessed by analysis of two quality control wheat materials for OTA. The DNA-ligand system is innovative, simple and rapid, and can be used to screen large quantities of samples for OTA contamination at levels below the EU regulatory limit with analytical performances satisfying EU criteria for method acceptability.

The presence of glucoside derivatives of T-2 and HT-2 toxins (type A trichothecene mycotoxins) in naturally contaminated wheat and oats is reported for the first time. The use of advanced high-resolution mass spectrometry based on Orbitrap technology allowed to obtain molecular structure details by measuring exact masses of main characteristic fragments, with mass accuracy lower than 2.8 ppm (absolute value). A monoglucoside derivative of T-2 toxin and two monoglucoside derivatives of HT-2 toxin were identified and characterized. The analysis of their fragmentation patterns provided evidence for glucosylation at C-3 position for T-2 toxin and at C-3 or C-4 position for HT-2 toxin. A screening for the presence of these new masked forms of mycotoxins was carried out on a set of naturally contaminated wheat and oats samples. On the basis of peak area ratio between glucoside derivatives and free T-2 and HT-2 toxins, the presence of glucoside derivatives was more likely in wheat than in oats samples. The present work confirms the widespread occurrence of trichothecene glucosides in cereal grains naturally contaminated with the relevant unconjugated toxins, thus suggesting the importance of developing suitable analytical methods for their detection. Besides toxicity studies, tracking down these new masked forms of trichothecenes along the food/feed chain would enable to collect information on their relevance in human/animal exposure to mycotoxin risk. Copyright © 2012 John Wiley & Sons, Ltd.

This study examines the diversity of spore-forming bacteria isolated from raw materials/bread using molecular methods along with a rapid and innovative technology, the FT-NIR spectroscopy. Microbiological analysis showed that 23% of semolina and 42% of other raw materials (including grain, brewer yeast, improvers) contained more than 100 spores/g and more than 50% of each kind of sample was contaminated at a level ranging from 1 to 100 spores/g. A high bacterial diversity characterized raw materials. In total 176 isolates were collected and characterized: 13 bacterial species belonging to Bacillus (10) and Paenibacillus (3) genera were identified by sequencing of 16S rRNA, gyrA or gyrB genes. The two closely related species Bacillus amyloliquefaciens (strain N45.1) and Bacillus subtilis (strain S63) were also analyzed by the spectroscopic technique FT-NIR. This analysis gave clear discrimination between the strains in the score plot obtained by the PCA and allowed to identify the spectral region 5600–4000 cm− 1 as the information-rich region for discrimination. B. amyloliquefaciens, possibly misidentified as B. subtilis in previous studies, was recognized as the most frequent species, found also in ropy bread. Moreover, the screening test for rope production indicated that mainly B. amyloliquefaciens, together with B. subtilis and Bacillus pumilus, could cause spoilage in bread, even if the last two species were represented by a low number of isolates. The Bacillus cereus group and Bacillus megaterium showed a lower percentage (30–70%) of isolates potentially able to cause the rope, but considering the high number of B. cereus group isolates detected in this study, this bacterial group should also be considered important in rope spoilage. In conclusion, results demonstrate that raw materials used to produce bread represent a rich source of spore-forming bacteria, therefore their microbiological quality should be monitored before use. Moreover, this study highlights for the first time the importance of the species B. amyloliquefaciens in rope spoilage and indicates that other species may also cause this alteration although strains of the same species may behave differently.

A sensitive and accurate method employing a single stage high resolution mass spectrometer equipped with a high-energy collision-dissociation cell (HCD) for the simultaneous determination of deoxynivalenol (DON), T-2 toxin (T-2) and HT-2 toxin (HT-2) in a processed bread model food has been developed. Two sample pre-treatment routes for the extraction of these mycotoxins were investigated, based on Mycosep(®) column clean up or QuEChERS-like procedure, respectively. The former approach suffered less from matrix effects and allowed to achieve in bread samples LODs of 7, 12 and 17 ng/g for T-2, HT-2 and DON, respectively, with 0.5 ppm mass accuracy. Two acquisition modes, full scan MS and all ion fragmentation, exploiting the fragmentation features offered by an HCD chamber and integrated within the Orbitrap analyser, were compared for quantitative purposes. The method was applied to investigate the degradation of these mycotoxins during bread processing using a bread model food. Most T-2 hydrolyzed to HT-2 during dough preparation, and about 20-30% of HT-2 and DON was degraded during bread baking.

Single event effects are investigated in 90-nm phase change memories. The cells are shown to be insensitive to heavy-ion strikes and will likely remain so for a few more generations. Possible physical mechanisms leading to upsets in future generations are discussed. Errors and functional interrupts, in addition to single event latch-up, were observed during read, word and buffer program due to strikes in the peripheral circuitry.

We studied the angular dependence of heavy-ion induced errors and threshold voltage shifts in Flash memories. Combining experiments and Geant4-based simulations, we provide new insight about the sensitive volume in floating gate cells. At high LET, the sensitive volume for cells belonging to the heavy-ion induced secondary peak corresponds to a large part of the floating gate. The sensitive volume for upsets may be larger or smaller, depending on the relative position of the secondary peak with respect to the relevant reference voltage.

International audience; A liquid chromatography - high resolution mass spectrometry (LC-HRMS) method for the simultaneous determination of aflatoxins (B1, B2, G1, G2), ochratoxin A, deoxynivalenol, zearalenone, T-2 and HT-2 toxins in wheat flour, barley flour and crisp bread has been developed. Data presented show the process of obtaining. Mycotoxin fragmentation patterns obtained by High Energy Collision Dissociation (HCD) have been investigated to obtain quantitative and confirmatory information (two characteristic masses per mycotoxin) using OrbitrapTM based high resolution mass spectrometry. LC-HRMS (full scan) detection carried out by HCD allowed to monitor the pseudo-molecular ion and an additional characteristic fragment (for each mycotoxin) with mass accuracy in the range 0.1 - 3.9 ppm, meeting the current European regulatory requirements for LC-MS confirmatory analysis. A sample preparation procedure based on polymeric solid phase extraction cartridges was applied, allowing recoveries higher than 74% for all tested toxins, with relative standard deviation lower than 13%. Detection limits in the range 0.5 - 3.4 µg/kg were obtained for all tested matrices. A critical comparison between the proposed method and a validated method based on triple quadrupole mass spectrometry showed similar performances in terms of detection limits, recoveries and repeatability, and matrix effects. Based on an efficient sample extraction and clean up, the LC-HCD-HRMS method developed herein represents a reliable and robust alternative tool for mycotoxin analysis in food matrices as compared to well established triple quadrupole based approaches.

A method based on LC-ESI-high-resolution (HR)-MS analysis, using a single-stage Orbitrap mass spectrometer, was developed for the quantification of casein allergens potentially present in white wines as a result of fining by caseinate. The method consists of (1) extraction from the matrix by ultrafiltration, (2) digestion with trypsin and (3) detection/quantification of residual caseins, obtained by monitoring the LC-MS response of representative tryptic peptides (peak areas in extracted-ion chromatograms). Method linearity was assessed first on caseinate solutions prepared either in water or in wine matrix (the ultrafiltration residue of a protein-free white wine). Limits of detection (LOD) ranged from 0.1 to 0.3 µg ml(-1) (S/N = 3) in water, and between 0.15 and 0.7 µg ml(-1) in wine matrix, depending on the selected peptide. Method repeatability and reproducibility, measured as response variability (standard deviation) due to LC-MS analysis alone and to both enzymatic digestion and LC-MS analysis, were assessed on caseinate standard solutions in water and ranged from 5 to 12% and from 8 to 20%, respectively. A higher variability was usually observed for the peptide marker response in the case of matrix-matched samples, the only exception being peptide GPFPIIV from β-casein, the marker also providing the highest sensitivity. The method was finally applied to a casein-free white wine ('Greco di Tufo') fined with caseinate at different concentrations, after discarding the precipitate due to casein-wine components aggregation. Minimum detectable added caseinate concentrations (i.e. those corresponding to responses with S/N = 3) were estimated between 39 and 51 µg ml(-1), according to the peptide marker chosen. These limits are compatible with caseinate concentrations typically adopted for wine-fining purposes. Moreover, a cross-check with the calibration performed in wine matrix led to an estimation of the concentration of dissolved caseinate to be in the low ng ml(-1) range.

The effect of processing on mycotoxin content in milling fractions has been investigated in 10 samples of durum wheat contaminated with T-2 and HT-2 toxins at levels ranging from 97 to 5,954 ?g/kg (sum of T-2 and HT-2 toxins). Either naturally contaminated samples or samples artificially inoculated with Fusarium sporotrichioides under field conditions were used. A method based on liquid chromatography-tandem mass spectrometry coupled with immunoaffinity column cleanup was validated in-house for the simultaneous analysis of both toxins in a variety of matrices, including uncleaned wheat, cleaned wheat, screenings, bran, red dog, fine middlings, and semolina. Mean recoveries from samples spiked with T-2 and HT-2 toxins at levels of 100 ?g/kg ranged from 85 to 107%, with relative standard deviations (RSDs) lower than 14%. The milling process led to an increase of T-2 and HT-2 toxin contents up to 13-and 5-fold in screenings and bran, respectively, compared with occurrence in the uncleaned wheat; however, an overall reduction of T-2 and HT-2 toxins by 54% (RSD, 20%) and 89% (RSD, 3%) was observed in cleaned wheat and in semolina, respectively. Copyright © International Association for Food Protection.

Humans and animals can be simultaneously exposed through the diet to different mycotoxins, including aflatoxins, ochratoxin A, deoxynivalenol, zearalenone, and fumonisins, which are the most important. Evaluation of the frequency and levels of human and animal exposure to these mycotoxins can be performed by measuring the levels of the relevant biomarkers in urine. Available data on the toxicokinetics of these mycotoxins in animals suggest that aflatoxin M(1) (AFM(1)), ochratoxin A (OTA), deoxynivalenol (DON)/de-epoxydeoxynivalenol (DOM-1), alpha-zearalenol (α-ZOL)/beta-zearalenol (β-ZOL), and fumonisin B(1) (FB(1)) can be used as urinary biomarkers. A liquid chromatographic-tandem mass spectrometric method has been developed for simultaneous determination of these mycotoxin biomarkers in human or animal urine. Urine samples were purified and concentrated by a double cleanup approach, using a multitoxin immunoaffinity column and a reversed-phase SPE Oasis HLB column. Separation of the biomarkers was performed by reversed-phase chromatography using a multi-step linear methanol-water gradient containing 0.5% acetic acid as mobile phase. Detection and quantification of the biomarkers were performed by triple quadrupole mass spectrometry (LC-ESI-MS/MS). The clean-up conditions were optimised to obtain maximum analyte recovery and high sensitivity. Recovery from spiked samples was performed at four levels in the range 0.03-12 ng mL(-1), using matrix-matched calibration curves for quantification. Mean recoveries of the biomarkers tested ranged from 62 to 96% with relative standard deviations of 3-20%. Enzymatic digestion with β-glucuronidase/sulfatase resulted in increased concentrations of the biomarkers, in both human and pig urine, in most samples containing measurable concentrations of DON, DOM-1, OTA, α-ZOL, or β-ZOL. A highly variable increase was observed between individuals. Co-occurrence of OTA and DON in human urine is reported herein for the first time.