Construction of a BiOCl/Bi2O2CO3 S-Scheme Heterojunction Photocatalyst via Sharing [Bi2O2]2+ Slabs with Enhanced Photocatalytic H2O2 Production Performance.

The construction of a close contact interface is key to enhancing the photocatalytic activity in heterojunctions. In the work, the BiOCl/Bi2O2CO3 of sharing [Bi2O2]2+ slabs S-scheme heterojunction was prepared by a HCl in situ etching method. The optimal composite photocatalyst could accomplish sizable productivity of H2O2 to 2562.95 µmol g-1 h-1 under simulated solar irradiation, higher than that of primitive Bi2O2CO3 and BiOCl. Moreover, the synthesized catalysts showed good stability. The band structures of BiOCl and Bi2O2CO3 were determined, confirming the formation of BiOCl/Bi2O2CO3 S-scheme heterojunction The BiOCl/Bi2O2CO3, which obviously improved the separation efficiency of photoinduced carriers and effectively enhanced the redox ability of the photocatalyst. In addition, density functional theory (DFT) calculations were utilized to analyze the electron transfer properties and the constitution of the built-in electric field at the interface of BiOCl and Bi2O2CO3. The photocatalytic reaction process was further researched by electron paramagnetic resonance (EPR), indicating the active species in the photocatalytic production of hydrogen peroxide. Eventually, a feasible S-scheme electron transfer mechanism on the BiOCl/Bi2O2CO3 heterojunction during the photocatalytic H2O2 production process was proposed and discussed. This work provides a reliable strategy for the fine design of the S-scheme heterojunction.

Aquaporins alteration revealed kidney damages in cerebral ischemia/reperfusion rats.

Background: Restoration of blood supply is a desired goal for the treatment of acute ischemic stroke. However, the restoration often leads to cerebral ischemia-reperfusion injury (CIR/I), which greatly increases the risk of non-neural organ damage. In particular, the acute kidney injury might be one of the most common complications. Aims: The study aimed to understand the damage occurred and the potential molecular mechanisms. Methods: The study was explored on the CIR/I rats generated by performing middle cerebral artery occlusion/reperfusion (MCAO/Reperfusion). The rats were evaluated with injury on the brains, followed by the non-neural organs including kidneys, livers, colons and stomachs. They were examined further with histopathological changes, and gene expression alterations by using RT-qPCR of ten aquaporins (Aqps) subtypes including Aqp1～Aqp9 and Aqp11. Furthermore, the Aqps expression profiles were constructed for each organ and analyzed by performing Principle Component Analysis. In addition, immunohistochemistry was explored to look at the protein expression of Aqp1, Aqp2, Aqp3 and Aqp4 in the rat kidneys. Results: There was a prominent down-regulation profile in the MCAO/Reperfusion rat kidneys. The protein expression of Aqp1, Aqp2, Aqp3 and Aqp4 was decreased in the kidneys of the MCAO/Reperfusion rats. We suggested that the kidney was in the highest risk to be damaged following the CIR/I. Down-regulation of Aqp2, Aqp3 and Aqp4 was involved in the acute kidney injury induced by the CIR/I.

Dual function of magnetic nanocomposites-based SERS lateral flow strip for simultaneous detection of aflatoxin B1 and zearalenone.

Aflatoxin B1 (AFB1) and zearalenone (ZEN) are two mycotoxins that often co-occur in corn. A surface-enhanced Raman scattering-based lateral flow immunoassay (SERS-LFIA) that can simultaneously detect AFB1 and ZEN in corn samples was developed employing the core-interlayer-satellite magnetic nanocomposites (Fe3O4@PEI/AuMBA@AgMBA) as dual-functional SERS tags. Under the optimal conditions, the detection ranges of AFB1 and ZEN in corn samples were 0.1-10 µg/kg and 4-400 µg/kg, respectively. Moreover, the test results for two mycotoxins in contaminated corn samples employing the suggested SERS-LFIA was in line with those of the HPLC technique. In view of its satisfactory sensitivity, accuracy, precision and short testing time (20 min), the developed system has a promising application prospect in the on-site simultaneous detection of AFB1 and ZEN.

Assessment of the Adverse Health Effects of Aflatoxin Exposure from Unpackaged Peanut Oil in Guangdong, China.

Aflatoxins are liver carcinogens and are common contaminants in unpackaged peanut (UPP) oil. However, the health risks associated with consuming aflatoxins in UPP oil remain unclear. In this study, aflatoxin contamination in 143 UPP oil samples from Guangdong Province were assessed via liquid chromatography-tandem mass spectrometry (LC-MS). We also recruited 168 human subjects, who consumed this oil, to measure their liver functions and lipid metabolism status. Aflatoxin B1 (AFB1) was detected in 79.72% of the UPP oil samples, with levels ranging from 0.02 to 174.13 µg/kg. The average daily human intake of AFB1 from UPP oil was 3.14 ng/kg·bw/day; therefore, the incidence of liver cancer, caused by intake of 1 ng/kg·bw/day AFB1, was estimated to be 5.32 cases out of every 100,000 persons per year. Meanwhile, Hepatitis B virus (HBV) infection and AFB1 exposure exerted a synergistic effect to cause liver dysfunction. In addition, the triglycerides (TG) abnormal rate was statistically significant when using AFB1 to estimate daily intake (EDI) quartile spacing grouping (p = 0.011). In conclusion, high aflatoxin exposure may exacerbate the harmful effects of HBV infection on liver function. Contamination of UPP oil with aflatoxins in Guangdong urgently requires more attention, and public health management of the consumer population is urgently required.

Entrapment of probiotic (Bifidobacterium longum) in bilayer emulsion film with enhanced barrier property for improving viability.

The gelatin/gellan gum based-bilayer emulsion film was developed in this work to improve the survivability of Bifidobacterium longum during the storage process. The baobab seed oil (BO) was added to the gelatin (GE) matrix to develop emulsion film as the barrier outer layer. The blueberry anthocyanin extract (BE) was incorporated into the gellan gum (GG)-based inner layer to enhance the viability of B. longum. The SEM and FTIR results revealed that the probiotics were successfully entrapped in BO/BE-loaded bilayer film. The greatest survivability and viable cell numbers of the B. longum during the storage period were observed in the BO/BE loaded bilayer film. Furthermore, the stability of the colorful patterns by electrochemical writing was also evaluated in this work. Finally, the GE/BO-GG/BE/BM maintain satisfactory probiotic viability in steamed bread coating application. Hence, the GE/BO-GG/BE/BM bilayer film could be considered a novel material to deliver and protect the probiotics in food applications.

Fabrication and characterization of polyvinyl alcohol/sodium alginate/zein/ chitosan bilayer film for dynamic visualization of pork quality.

The development of real-time and convenient meat freshness indication technology is crucial to ensure food safety. A novel antibacterial visualized intelligent film was designed based on polyvinyl alcohol (PA), sodium alginate (SA), zein (ZN), chitosan (CS), alizarin (AL) and vanillin (VA) using layer-by-layer assembly (LBL) method for real-time and in situ monitoring of pork freshness. The fabricated film had various advantageous properties, including an excellent hydrophobicity with a water contact angle (WCA) of 91.59°, improved color stability, excellent water barrier properties and increased mechanical performance (TS = 42.86 MPa). The fabricated film also demonstrated effective antibacterial properties with a bacteriostatic circle diameter of 13.6 mm for Escherichia coli. Moreover, the film can perceive and visualize the antibacterial effect through color changes, enabling dynamic visual monitoring of the antibacterial effect. A good correlation (R2 = 0.9188) between the color changes (ΔE) and total viable count (TVC) of pork was documented. Conclusively, fabricated multifunctional film improves the accuracy and versatility of freshness indication and had great potential for food preservation and freshness monitoring. The outcomes of this research provides a new perspective for the design and development of multifunctional intelligent films.

Novel gellan gum-based probiotic film with enhanced biological activity and probiotic viability: Application for fresh-cut apples and potatoes.

A novel probiotic film based on gellan gum (GN), cranberry extract (CE), and Lactococcus lactis (LA) was developed in the present study. The fluorescence and SEM image results showed that GN/CE film containing LA was successfully fabricated. The incorporation of LA significantly enhanced the antibacterial activity of the film. The presence of CE strengthened the antioxidant activity and LA survivability in the film. The combination of LA (0-1.0 %) and CE (0.5-1.0 %) improved the mechanical property of the film through the formation of density structure. The best comprehensive properties were obtained with the film containing 2.0 %LA and 0.5 %CE. The GN/2.0 %LA/0.5 %CE film also showed the optimal preservation effect on fresh-cut potatoes and apples. Hence, GN/2.0 %LA/0.5 %CE probiotic film has proved to be suitable for fruit and vegetable preservation.

SERS determination of hydroxy-α-sanshool in spicy hotpot seasoning: The strategy to restrain the interference of capsaicin and its mechanism.

Hydroxy-α-sanshool (α-SOH) is the principal ingredient responsible for the numbing sensation in spicy hotpot. However, utilizing surface-enhanced Raman scattering (SERS) to analyze the α-SOH in hotpot seasoning is challenging due to the significant interference of capsaicin (CAP). Therefore, two schemes were proposed to address CAP interference in hotpot seasoning, namely laccase-catalyzed conversion and metal-organic framework (MOF) interaction. Among them, Fe-BTC MOF exhibited significant anti-interference effect and the underlying mechanism is elucidated. The motion of CAP aromatic ring was constrained by steric hindrance and electrostatic interactions of Fe-BTC. Additionally, the interaction between CAP aromatic ring and conjugated triene group in α-SOH was quenched, enhancing the α-SOH SERS signal. The proposed method had a significant anti-interference effect on α-SOH quantification in the presence of CAP, significantly enhancing the α-SOH SERS signal in a range of 0.85 to 4.00 × 107. The linearity and reproducibility of the proposed hotpot seasoning testing method were also validated.

Saccharomyces cerevisiae-incorporated and sucrose-rich sodium alginate film: An effective antioxidant packaging film for longan preservation.

A sodium alginate (SA) film incorporated with Saccharomyces cerevisiae (SE) and sucrose (SU) was fabricated to control the quality and pericarp browning of longan. The SE with satisfactory glutathione production was selected as the antioxidant agent. The scanning electron microscopy (SEM) results revealed that the SU-rich SA film could be used as an effective carrier to protect the cell integrity of SE. The FTIR and mechanical property results indicated that the SA-SE film with the incorporation of SU has good flexibility due to the existence of hydrogen bonds. Notably, the cell viability of the SE was significantly improved with the addition of SU, which positively affects the antioxidant property of the film during the storage period. Finally, the SA-SE-3.0%SU films obviously improved the quality and pericarp browning of longan. The SA-based film incorporated with SU and SE may be established as a novel antioxidant fruit packaging material.

A Novel Gas Sensor for Detecting Pork Freshness Based on PANI/AgNWs/Silk.

A novel, operational, reliable, flexible gas sensor based on silk fibroin fibers (SFFs) as a substrate was proposed for detecting the freshness of pork. Silk is one of the earliest animal fibers utilized by humans, and SFFs exposed many biological micromolecules on the surface. Thus, the gas sensor was fabricated through polyaniline (PANI) and silver nanowires (AgNWs) and deposited on SFFs by in-suit polymerization. With trimethylamine (TMA) as a model gas, the sensing properties of the PANI/AgNWs/silk composites were examined at room temperature, and the linear correlativity was very prominent between these sensing measures and the TMA measures in the range of 3.33 µg/L-1200 µg/L. When the pork sample is detected by the sensor, it can be classified into fresh or stale pork with the total volatile basic nitrogen (TVB-N) as an index. The result indicated that the gas sensor was effective and showed great potential for applications to detect the freshness of pork.

High- sensitivity bilayer nanofiber film based on polyvinyl alcohol/sodium alginate/polyvinylidene fluoride for pork spoilage visual monitoring and preservation.

A colorimetric bilayer film for pork freshness detection and preservation was developed using electrospinning technique. The bilayer film consisted of a layer with polyvinyl alcohol - sodium alginate - alizarin as sensor layer and a layer with polyvinylidene fluoride - vanillin as antibacterial layer. The water contact angle of bilayer film was larger than the single colorimetric layer. The color sensitivity to the ammonia of the bilayer film was higher, with an ΔE value of 47.99. The film could display color shifts from yellow to purple with the naked eye is critical for checking pork freshness. In addition, the bilayer film exhibited sensitive antibacterial activity, with an inhibition zone against S. aureus (8.3 mm) and E. coli (14.7 mm), respectively. Finally, the bilayer film was applied to freshness monitoring of pork. The film displayed significant color changes and prolonged the pork shelf life by 24 h at 25 °C.

A cell-based electrochemical sensor for assessing immunomodulatory effects by atrazine and its metabolites.

A three-dimensional (3D) cell-based electrochemical sensor was developed to evaluate the immunomodulatory effects of atrazine and its metabolites by monitoring nitric oxide (NO) release. Nafion/Fe(III) meso-tetra (4-carboxyphenyl) porphyrin/reduced graphene oxide (Nafion/FeTCP@RGO) was functionalized on the screen-printed carbon electrode (SPCE) as a working electrode to enhance the NO selectivity and sensitivity. RAW264.7 cells encapsulated in a gelatin methacrylate (GelMA) hydrogel, forming a 3D cell culture system, were immobilized on the Nafion/FeTCP@RGO/SPCE to serve as biorecognition elements. The proposed sensor presented the induced NO expression as a distinct single differential pulse voltammetric (DPV) anodic peak in the selected physiological lipopolysaccharide (LPS) concentration range (0.01-2000 ng/mL). Furthermore, the peak current intensity of the NO was linearly correlated with the LPS concentration logarithm, which was further validated using a Griess reagent kit. The proposed sensor was utilized in optimized LPS-induction conditions to compare the immunosuppression effect of atrazine with its main metabolites (i.e. desethylatrazine (DEA), deisopropylatrazine (DIA), and diaminochlorotriazine (DACT)). The results demonstrated that the order of 30% inhibitory concentrations was atrazine (25.71 ± 1.08 µg/mL) < DEA (48.63 ± 2.17 µg/mL) ≤ DACT (49.11 ± 1.98 µg/mL) ≤ DIA (52.36 ± 2.34 µg/mL). Overall, this work provided a potential in vitro approach to evaluate the immunotoxicity of pesticides and their metabolites.

Organ Specific Differences in Alteration of Aquaporin Expression in Rats Treated with Sennoside A, Senna Anthraquinones and Rhubarb Anthraquinones.

Senna and rhubarb are often used as routine laxatives, but there are differences in mechanism of action and potential side effects. Here, we studied metabolites of senna anthraquinones (SAQ), rhubarb anthraquinones (RAQ) and their chemical marker, sennoside A (SA), in a rat diarrhea model. In in vitro biotransformation experiments, SAQ, RAQ and SA were incubated with rat fecal flora solution and the metabolites produced were analyzed using HPLC. In in vivo studies, the same compounds were investigated for purgation induction, with measurement of histopathology and Aqps gene expression in six organs. The results indicated that SAQ and RAQ had similar principal constituents but could be degraded into different metabolites. A similar profile of Aqps down-regulation for all compounds was seen in the colon, suggesting a similar mechanism of action for purgation. However, in the kidneys and livers of the diarrhea-rats, down-regulation of Aqps was found in the RAQ-rats whereas up-regulation of Aqps was seen in the SAQ-rats. Furthermore, the RAQ-rats showed lower Aqp2 protein expression in the kidneys, whilst the SA-rats and SAQ-rats had higher Aqp2 protein expression in the kidneys. This may have implications for side effects of SAQ or RAQ in patients with chronic kidney or liver diseases.

Novel roles of an intragenic G-quadruplex in controlling microRNA expression and cardiac function.

Simultaneous dysregulation of multiple microRNAs (miRs) affects various pathological pathways related to cardiac failure. In addition to being potential cardiac disease-specific markers, miR-23b/27b/24-1 were reported to be responsible for conferring cardiac pathophysiological processes. In this study, we identified a conserved guanine-rich RNA motif within the miR-23b/27b/24-1 cluster that can form an RNA G-quadruplex (rG4) in vitro and in cells. Disruption of this intragenic rG4 significantly increased the production of all three miRs. Conversely, a G4-binding ligand tetrandrine (TET) stabilized the rG4 and suppressed miRs production in human and rodent cardiomyocytes. Our further study showed that the rG4 prevented Drosha-DGCR8 binding and processing of the pri-miR, suppressing the biogenesis of all three miRs. Moreover, CRISPR/Cas9-mediated G4 deletion in the rat genome aberrantly elevated all three miRs in the heart in vivo, leading to cardiac contractile dysfunction. Importantly, loss of the G4 resulted in reduced targets for the aforementioned miRs critical for normal heart function and defects in the L-type Ca2+ channel-ryanodine receptor (LCC-RyR) coupling in cardiomyocytes. Our results reveal a novel mechanism for G4-dependent regulation of miR biogenesis, which is essential for maintaining normal heart function.

Bilayer pH-sensitive colorimetric films with light-blocking ability and electrochemical writing property: Application in monitoring crucian spoilage in smart packaging.

Bilayer colorimetric films were developed for monitoring fish spoilage by using gelatin (GN) incorporated with ZnO nanoparticles as the upper layer (GN-ZnO), and gellan gum (GG) incorporated with mulberry anthocyanins (MBA) as the lower layer (GG-MBA). The color stability of the bilayer colorimetric films under visible and ultraviolet light was improved with the increase of ZnO nanoparticles content. Meanwhile, the bilayer films had good NH3 sensitivity. The limit of detection of the GG-MBA/GN-2.0% ZnO film to NH3 was 0.01 mM. The electrochemical writing ability of the bilayer films was also identified, indicating the feasibility of inks-free printing on biopolymer films. Finally, the GG-MBA/GN-2.0% ZnO film with an electrochemical writing pattern was used to monitor crucian spoilage. The GG-MBA/GN-2.0% ZnO film with electrochemical writing pattern showed visible color changes with the crucian spoilage. In conclusion, the bilayer colorimetric film was expected to be a good fish spoilage indicator in smart packaging.

Biomonitoring Study of Deoxynivalenol Exposure in Chinese Inhabitants.

Objective: To investigate the levels of a deoxynivalenol (DON) biomarker in the urine of subjects living in two China provinces with different geographic locations and dietary patterns, and estimate their dietary DON exposures and health risks. Methods: First morning urine samples were collected on three consecutive days from 599 healthy subjects-301 from Henan province and 298 from Sichuan province-to analyze the total DON concentrations (tDON) after ß-glucuronidase hydrolysis using a high-performance liquid chromatography tandem mass spectrometry-based method. The consumption of cereal foods in the previous 24 h before each urine collection was recorded using a duplicate diet method. DON exposure levels were estimated based on the urinary tDON concentrations. Results: Total DON were detected in 100% and 92% of the urine samples from Henan and Sichuan, respectively. Mean urinary tDON concentrations were 52.83 ng/mL in Henan subjects and 12.99 ng/mL in Sichuan subjects, respectively. The tDON levels were significantly higher in the urine of Henan subjects than that of the Sichuan subjects (p < 0.001). Urinary tDON levels were significantly different among age groups in both areas (Henan: p < 0.001; Sichuan: p = 0.026) and were highest in adolescents aged 13-17 years, followed by children aged 7-12 years. Based on the DON biomarker and exposure conversion reported by the European Food Safety Authority (EFSA), the mean estimated dietary intakes of DON were 1.82 µg/kg bw/day in Henan subjects and 0.45 µg/kg bw/day in Sichuan subjects. A total of 56% of Henan subjects and 12% of Sichuan subjects were estimated to exceed the PMTDI of 1 µg/kg bw/day. Consistent with urinary tDON levels, the highest estimated dietary DON intakes were also in children and adolescents aged 7-17 years. For all kinds of wheat-based foods except dumplings, the consumptions were significantly higher in Henan than those in Sichuan. The mean consumption of steamed buns was 8.4-fold higher in Henan (70.67 g/d) than that in Sichuan (8.45 g/d). The mean consumption of noodles in Henan (273.91 g/d) was 3.6-fold higher than that in Sichuan (75.87 g/d). Conclusions: The levels of urinary DON biomarker and the estimated dietary DON intakes in Henan province were high and concerning, especially for children and adolescents. The overall exposure level of Sichuan inhabitants was low.

Geospatial visualisation of food contaminant distributions: Polychlorinated naphthalenes (PCNs), potentially toxic elements (PTEs) and aflatoxins.

Large volume of multidimensional data can be summarised, both in terms of tabulated statistics, and as graphic geospatial visualisations. The latter approach allows rapid interpretation and communication of complex information to stake-holders such as regulators, risk assessors and policy makers. In the main study on polychlorinated naphthalene (PCN), individual samples representing different edible fish species were analysed from around the UK. PCNs were observed in all samples with nearly all of the twelve measured congeners being detected. Summed congener concentrations ranged from 0.7 ng/kg ww (turbot) to 265 ng/kg ww (sprats). The highest contamination levels were recorded for sprats and mackerel with mean summed concentrations of 67 ng/kg ww and 68 ng/kg ww respectively. Two ancillary studies, on potentially toxic elements (PTEs) in crabs from China and aflatoxin in children's blood from Tanzania, demonstrate the wide applicability of this approach. The PTE contents in crab showed strong dependence on the tested tissues and elements, and crabs from Tai and Yangcheng Lakes showed obviously higher PTE levels than the other lakes. Geospatial distribution of the aflatoxin biomarker AF-alb in children's serum from 3 locations showed how individual anthropometric or socio-economic data reveals the relationship between family size, socio-economic score and magnitude of serum aflatoxin levels. In addition to facilitating the flow of interpreted data to stakeholders, these techniques can direct the formulation of risk mitigation activities and help with the identification of data gaps. When combined with hierarchical cluster analyses, correlations within the data can also be predicted.

A colorimetric hydrogen sulfide sensor based on gellan gum-silver nanoparticles bionanocomposite for monitoring of meat spoilage in intelligent packaging.

A colorimetric hydrogen sulfide (H2S) sensor based on gellan gum capped silver nanoparticles was developed to real-time monitor meat spoilage. The colorimetric sensor strategy was attributed to the ultrastrong binding ability of Ag with H2S to form Ag2S. The sensor enables the analysis of H2S with a limit of detection (LOD) of 0.81 µM, and it exhibited excellent selectivity toward H2S against other volatile components generated from chicken breast and silver carp during spoilage. By these virtues, the sensor presented visible color changes from yellow to colorless by in situ and nondestructively sensing H2S generated from chicken breast and silver carp in a packaging system. This strategy provided a simple but useful, non-destructive, robust, cost-effective, and user-friendly platform to real time monitor meat spoilage for intelligent food packaging.

ß2-Adrenergic Stimulation Compartmentalizes ß1 Signaling Into Nanoscale Local Domains by Targeting the C-Terminus of ß1-Adrenoceptors.

RATIONALE: ßARs (ß-adrenergic receptors) are prototypical GPCRs (G protein-coupled receptors) that play a pivotal role in sympathetic regulation. In heart cells, ß1AR signaling mediates a global response, including both l-type Ca2+ channels in the sarcolemma/T tubules and RyRs (ryanodine receptors) in the SR (sarcoplasmic reticulum). In contrast, ß2AR mediates local signaling with little effect on the function of SR proteins. OBJECTIVE: To investigate the signaling relationship between ß1ARs and ß2ARs. METHOD AND RESULTS: Using whole-cell patch-clamp analyses combined with confocal Ca2+ imaging, we found that the activation of compartmentalized ß2AR signaling was able to convert the ß1AR signaling from global to local mode, preventing ß1ARs from phosphorylating RyRs that were only nanometers away from sarcolemma/T tubules. This offside compartmentalization was eliminated by selective inhibition of ß2AR, GRK2 (GPCR kinase-2), ßarr1 (ß-arrestin-1), and phosphodiesterase-4. A knockin rat model harboring mutations of the last 3 serine residues of the ß1AR C terminus, a component of the putative ßarr1 binding site and GRK2 phosphorylation site, eliminated the offside compartmentalization conferred by ß2AR activation. CONCLUSIONS: ß2AR stimulation compartmentalizes ß1AR signaling into nanoscale local domains in a phosphodiesterase-4-dependent manner by targeting the C terminus of ß1ARs. This finding reveals a fundamental negative feed-forward mechanism that serves to avoid the cytotoxicity of circulating catecholamine and to sharpen the transient ß1AR response of sympathetic excitation.

Dietary Deoxynivalenol Exposure Assessment in University Students from Japan.

This study was conducted to give a preliminary estimation of deoxynivalenol (DON) dietary exposure in Japanese university students (n = 30, aged 22-25 years) using a biomarker approach and to examine the correlation between wheat food intake and DON exposure levels. Spot urine samples were collected from 30 students of Azabu University, Tokyo. Urine samples were treated with enzyme digestion (for total DON measurement) and without (for unconjugated DON analysis) before clean-up using an immuno-affinity column and analysis using an LC-MS method, with a 13C15- DON internal standard used for accurate quantification. The limit of detection for this method is 0.5 ng/mL urine. The geometric mean (95% CI) of DON concentration was 2.03 (1.64 - 6.87) ng per mL urine. Ninety of the urine samples had detectable levels of urinary DON. The DON dietary intake exposure estimation suggested that one out of the 30 subjects had an intake of DON that exceeded Joint FAO/WHO Expert Committee on Food Additives (JECFA) provisional maximum tolerable daily intake (PMTDI) level. Mean ratio of free DON to total DON was determined to be 19%. Wheat intake assessed using a basic food frequent questionnaire method did not show a significant correlation with the urinary DON level.