Detectable Levels Research Articles

Enhancing corporate accountability through covert situational integrity testing (CSIT)

ABSTRACT This article explores the potential of ‘covert situational integrity testing’ as a mechanism for assessing corporate/organisational compliance with legal rules and standards, the goal being to enhance corporate/organisational accountability. There are major challenges to holding corporations/organisations to account for non-compliance: low detection levels, incomplete understanding of the inner workings of organisations, and the modest power of current social scientific research methodologies. This article argues there is scope for methodological innovation through the use of covert situational integrity testing, a variation of mystery shopping methodologies, to address some of these gaps, with a focus not on service quality or customer satisfaction but on compliance with regulatory and legal requirements, and as a data gathering tool on secretive and difficult to access areas of business operation. We focus on two examples: AML compliance by challenger banks, and the promotion of aggressive tax avoidance schemes. Our core argument is that covert situational integrity testing provides a research mechanism through which robust and systematic observational data can be collected and scrutinised by independent, third-party assessors to understand levels of organisational/corporate compliance with legal rules and standards, and by doing so, identify critical vulnerabilities and strengths in the compliance responses of organisations and industries.

Health risk assessment of dietary aflatoxin B1 exposure in common food matrices in Vietnam

AbstractAflatoxins, potent carcinogenic mycotoxins produced by Aspergillus species, persist in various foods, posing significant health risks upon consumption. This study assesses dietary aflatoxin B1 (AFB1) exposure in Vietnam using Margin of Exposure (MOE) and Hazard Index (HI) to evaluate risk levels in peanuts, chili powder, corn, and raisins. AFB1 levels were quantified by UPLC-FLD. AFB1 was detected in 71.5% of chili powder, 29.3% of peanuts, and 26.0% of corn samples, with no detectable levels in raisins. Peanuts and corn kernels were classified as high-risk, with MOE values below 10,000 (peanuts: 25.8–56.6; corn: 42.3–92.6) and HI values exceeding 1, potentially contributing to 20.4–44.7 and 12.5–27.3 liver cancer cases per 100,000 adults, respectively. In contrast, chili powder had MOE values between 3,208 and 7,021 and HI below 1, indicating a low public health risk. Raisins were deemed safe with MOE over 200,000 and HI at 0.01. Results also indicated higher cancer risk for women and lean individuals consuming the same AFB1 levels. These findings underscore the need for focused risk management strategies to mitigate AFB1 exposure and protect public health in Vietnam.

Association Between Dynamic Viral Rebound and Longitudinal Measures of Viral Load/CD4 Counts Among People with HIV in South Carolina.

Monitoring HIV viral rebound (VR) is crucial, as it indicates an increased risk of infection, transmission, disease progression, and drug resistance. This study aims to identify the association between dynamic VR and historical viral load (VL)/CD4 count measures. A 15-year South Carolina population-based electronic health record data were used for the study. VR was defined as the return of detectable levels of VL (>200 copies/mL) after stable viral suppression (VS) (two consecutive VS, i.e., VL ≤200 copies/mL). A generalized linear mixed model was used to evaluate the association between dynamic VR and historical time-dependent predictors, such as nadir CD4 count and comorbidities, within a year prior to each VR. Subgroup analysis for men who have sex with men (MSM) was also conducted. Among 8,185 people with HIV (PWH), 1,173 (14.3%) had a history of VR. Lower nadir CD4 count (≥500 vs. <200 cells/μL; adjusted odds ratio [aOR]: 0.51, 95% confidence interval [CI]: [0.43, 0.60]), younger age (>60 years old vs. 18-30 years old; aOR: 0.43, 95% CI: [0.29, 0.63]), and being Black (Black vs. White; aOR: 1.58, 95% CI: [1.34, 1.85]) were associated with a higher risk of VR, while MSM (MSM vs. heterosexual; aOR: 0.81, 95% CI: [0.67, 0.96]) were associated with decreased VR risk. The rate of VR among PWH in South Carolina is significant. Within-1-year VL/CD4 test is critical for identifying PWH at risk for VR. Tailored interventions are needed for PWH at risk for VR to achieve sustained suppression and better health outcomes.

Система контроля качества и безопасности оказания медицинской помощи как один из ключевых элементов достижения целей расширенного неонатального скрининга в Российской Федерации

After analyzing the Russia-wide Extended neonatal screening (ENS) system for hereditary and/or congenital diseases results it became necessary to change the ENS tasks ranking: the highest priority among them as yet is an increase in the congenital and/or hereditary diseases detection level and a decrease in the therapy initiation age. The results of an on-site audit conducted in 20 Russia regions in 2024 showed significant differences from the target indicators that in its turn had made it possible to determine the control points and roadmap operational procedures as necessary elements for the ENS system improvement with the purpose of medical care quality and safety monitoring in order to increase the congenital and/or hereditary pathology detection level, to reduce the time span from birth to the therapy initiation and to increase the effectiveness of patient treatment.

The use of a medical information system in the detection of occupational oncological pathology in the Krasnoyarsk Territory

Introduction. The article tells about the consistently low levels of detection of occupational malignant neoplasms (OMN) for many years in the Russian Federation. This creates a picture of imaginary well-being, which does not allow us to objectively characterize the real risks to public health associated with exposure to carcinogens in the workplace. The study aims to develop and practically test an algorithm for detecting occupational malignant neoplasms using a medical information system. Materials and methods. On the basis of the medical information system used in the Krasnoyarsk Regional Center of Occupational Pathology ("Regional Clinical Hospital"), specialists have developed and put into practice a system for detecting occupational malignant diseases. The combination of information flows from the main institutions providing medical care in the field of Oncology, the selection of patient data using information about the place of work, probable professional contact with carcinogens (according to data provided by the Office of Rospotrebnadzor in the Krasnoyarsk Territory), allowed us to analyze the information received about patients. Results. In case of a positive decision of the commission on suspicion of the professional nature of the disease, taking into account the professional route, the localization of the OMN and the current list of occupational diseases, the specialists initiated a procedure for examining the connection of the disease with the profession. During the three quarters of 2024, the researchers analyzed 437 cases of diseases with subsequent registration of sanitary and hygienic characteristics in 25 cases. Of the selected cases, the link between the disease and the profession has currently been established in 9 cases, 10 cases are in the works at the time of writing. In other cases, specialists recognized the diseases as common. Limitations. The study was limited by the time frame and the number of identified cases of neoplasms submitted for detailed consideration. Conclusion. The implemented system made it possible in less than a year to identify the number of cases of occupational cancer in only one subject of the Federation (Krasnoyarsk Territory), corresponding to almost half of the number of occupational cancers detected in the whole country. The use of a medical information system expands the possibilities of diagnosing occupational diseases. Ethics. The conducted research did not require the conclusion of the Ethics Committee.

Radiation-induced nasopharyngeal necrosis combined with local recurrence in nasopharyngeal carcinoma: diagnosis and treatment strategies

ABSTRACT Background To identify the diagnosis and treatment strategies by analyzing the clinical characteristics and treatment methods of RNNCLR. Methods A total of 210 patients pathologically diagnosed with RNNCLR were retrospectively included. Clinical characteristics, MRI features, treatment methods, and survival outcomes were analyzed. Propensity score matching (PSM) analysis was performed to adjust the surgical benefit. Results Ninety-one patients (43.3%) took a single biopsy, 67 patients (31.9%) underwent repeated biopsies and 52 patients (24.8%) received endoscopic surgery to obtain pathological positive tissues. RNNCLR had characteristic imaging features distinguished from pure radiation necrosis. The interval from the previous radiotherapy was 13.2 (7.0, 23.3) months. The 1-year, 3-year, and 5-year overall survival rates were 59.6%, 32.3%, and 14.6%. Patients with reirradiation, detectable EBV-DNA level, or ICA exposure had a worse prognosis. Overall survival was significantly higher in the endoscopic surgery group than in nonsurgery group after PSM (3-year OS rates, 44.5% vs. 23.9%, p = 0.011). Conclusions Histopathological diagnosis of RNNCLR needs repeated biopsies or even surgery. Careful analysis of MRI images, correlation with interval time from last radiation, and short-term follow-up may solve the diagnostic dilemmas. Endoscopic surgery results in a survival benefit by completely resecting lesions or removing necrotic tissue to reduce necrosis-related complications.

Tailoring Infrared Light-Molecule Coupling for Highly Sensitive Cortisol Detection Employing Aptamer-Conjugated Gold Nanonails.

Chemically synthesized gold nanoantennas possess easy processability, low cost, and suitability for large-area fabrication, making them advantageous for surface-enhanced infrared (SEIRA) biosensing. Nevertheless, current gold nanoantennas face challenges with limited enhancement of biomolecular signals that hinder their practical applications. Here, we demonstrate that the coupling rate between antennas and molecules critically impacts the enhancement of molecular signals based on temporal coupled mode theory. To improve this coupling rate, we synthesized gold nanonails with sharp tips, significantly amplifying the localized electric fields of antenna resonance modes. Modulating the nanonail aspect ratio allows us to tailor antenna resonance frequencies to match molecular vibrational frequencies. Additionally, we introduced specific aptamers on antenna surfaces through solution exchange methods to control the antenna-molecule distances. These combined strategies enabled noninvasive, label-free detection with high sensitivity for the biomarker cortisol. Experiments revealed 3 orders of magnitude enhancement in cortisol detection levels upon increasing coupling efficiency, achieving a detection limit of 0.1 ng/mL, notably lower than the normal cortisol concentration in human saliva (0.398 ng/mL). In addition to demonstrating a novel strategy for cortisol detection, this study provides a viable approach to biomarker detection for future applications in disease diagnosis and human health monitoring.

Bisphenol A in Indian Take-Out Soups: Compliance, Implications and Sustainable Solutions

This research investigates the migration of Bisphenol A (BPA) from packaging containers into take-out vegetable soups and premixed tomato soups through three replicate studies. The samples underwent extraction using solid-phase extraction (SPE) cartridges, followed by separation on a C18 column. BPA concentrations in the soups were assessed at 15, 30, and 45-minute intervals, consistently revealing undetectable levels (&lt;LOQ). Plastic packaging samples, known for BPA utilization in production, remained below the Specific Migration Limit (SML) set at 0.5 mg.kg-1, irrespective of material type or contact conditions. These results, conforming to EC regulations, suggest that food-contact materials (FCMs) in the Indian market pose no apparent health hazards during initial use. The absence of detectable BPA levels is attributed to the limited time-temperature relationship during the study. However, caution is warranted as BPA migration can occur with repeated use, emphasizing the importance of considering material quality and intended use of FCMs. The study underscores the significance of understanding BPA leaching under varied conditions, necessitating further research to explore long-term implications. Overall, the findings provide valuable insights for regulators, manufacturers, and consumers, contributing to the ongoing discourse on food safety and using plastic materials in food packaging.

Halobenzoquinone-induced potential carcinogenicity associated with p53-mediated cell cycle pathway

2,6-Dibromo-1,4-benzoquinone (2,6-DBBQ) and 2,6-dichloro-1,4-benzoquinone (2,6-DCBQ), two emerging halobenzoquinones (HBQs), have the highest detection frequencies and levels in drinking water among all HBQs. They are more toxic than the regulated disinfection byproducts. Quantitative structure toxicity relationship analysis predicted that HBQs are a class of potential bladder carcinogens. However, direct experimental evidence for the carcinogenicity of 2,6-DBBQ and 2,6-DCBQ is lacking and the associated toxicity mechanisms remain unclear. In this study, we confirmed the potential carcinogenicity of 2,6-DBBQ and 2,6-DCBQ using an in vitro malignant transformation assay, evaluated their cytotoxicity and genotoxicity, and investigated their toxicity mechanisms. The results showed that 2,6-DBBQ and 2,6-DCBQ significantly decreased the viability of human uroepithelial SV-HUC-1 cells and induced DNA damage in SV-HUC-1 cells, and chromosomal damage in HepG2 cells, and malignant transformation of SV-HUC-1 cells. Moreover, transcriptome sequencing revealed that 2,6-DBBQ and 2,6-DCBQ activated the p53-mediated cell cycle pathway in bladder cancer. In the p53-mediated cell cycle pathway, 2,6-DBBQ and 2,6-DCBQ induced cell cycle arrest at the S phase by downregulating p53 and upregulating p21. Additionally, 2,6-DBBQ and 2,6-DCBQ may have produced excessive reactive oxygen species, damaging DNA and chromosomes. These results not only first confirm the potential carcinogenicity of 2,6-DBBQ and 2,6-DCBQ but also provide an important reference for exploring the cytotoxicity and genotoxicity mechanisms of these HBQs.

Evaluation of the analytic performance and macroprolactin sensitivity of a new prolactin immunoassay.

Prolactin measurement is essential in endocrine diagnostics. Challenges such as the hook effect and reactivity to macroprolactin, which varies according to the reagent, complicate accurate measurement. The present study evaluated a newly marketed reagent to detect prolactin, IDS Prolactin, comparing it to an established reagent, Roche Elecsys Prolactin, assessing its behavior toward macroprolactin and polyethylene glycol (PEG) treatment, and establishing reference intervals. The IDS Prolactin and Roche Elecsys Prolactin assays were compared using 44 samples containing macroprolactin confirmed on gel filtration chromatography (macroprolactin: BBPRL) and 104 samples for which the diagnosis of macroprolactin was excluded (monomeric prolactin: MNPRL). Analytic performance of the IDS Prolactin assay was also assessed. The new reagent showed satisfactory analytic performance, meeting EFLM standards for repeatability and intermediate imprecision. Comparison between the two methods found robust correlation for monomeric samples (y=1.060x-18.28; r2=0.993). Compared to the Roche assay, which is particularly low in its reaction to macroprolactin, the IDS assay displayed a higher level of detection. PEG precipitation effectively separated monomeric and macroprolactin samples when a cut-off of 65% recovery was used or at the threshold of 444 mIU/L (20.9 µg/L) for post-PEG monomeric prolactin upper limit of normal. Reference intervals were established for women, with ROC curve analysis demonstrating high sensitivity and specificity. The IDS Prolactin assay showed excellent analytic performance and satisfactory characteristics on macroprolactinemic samples.

Ship detection and water level measurement system based on 3D point cloud

ABSTRACT With the continuous development of the water transportation and shipping industries, the number of ships in rivers has steadily multiplied, followed by the increasing complexity of the ship routes. These changes have highlighted the growing importance of ship detection and water level measurement systems. Such systems not only enhance the management efficiency of waterborne traffic, ensure navigation safety, and reduce congestion and collision accidents, but also effectively safeguard the integrity of riverside and bridge structures. Ship detection and river elevation measurement based on 3D point clouds can directly acquire depth information without being affected by lighting conditions. So it has a good research prospect. Therefore, this paper proposes a novel ship detection algorithm based on improved PointRCNN and a novel method for riverbank line extraction and water level measurement based on 3D point clouds, respectively. For ship detection, the improved PointRCNN algorithm can increase the performance of data processing and keypoint extraction techniques, and make the network to keep more foreground point clouds and learn more effective features. This improves the recognition capability of distant ships. Compared to the original PointRCNN algorithm, the improved PointRCNN algorithm has achieved a 3.84% increase in detection precision in practical scenarios. Regarding riverbank extraction and water level measurement, the proposed method based on 3D point clouds can directly extract riverbank lines with depth information, obtaining water level height without direct contact with the river surface. Within a distance range between 15 and 45 m from the LiDAR, the average absolute error using this measurement method is less than 5 cm, demonstrating the good detection accuracy of this method.

Development of dihydropyrimidine based chemosensor for Hg2+ ions

The detection and monitoring of heavy metal ions in water media is critical due to their toxic effects on human health and the environment. In this study, we propose a green approach for synthesizing a dihydropyrimidine-based ligand by utilizing the concept of the Biginelli reaction and incorporating Ag nanoparticles as activators. This synthesized ligand demonstrates exceptional potential as a chemosensor for detecting poisonous Hg2+ ions using a simple spectrofluorometer tool. The ligand exhibits high selectivity and sensitivity towards Hg2+ over other transition metal ions in aqueous solution. This innovative method aligns with green chemistry principles, emphasizing the importance of using a straightforward and efficient sensing method of fluorescence. This is the first report where a dihydropyrimidine-based moiety detects toxic heavy metal ions, Hg+2(minimum detection level is 5X10−7 M).

Elucidation of the Active Agents in a West African Ground Herbal Medicine Formulation That Elicit Antimalarial Activities in In Vitro and In Vivo Models

Agunmu (ground herbal medicine) is a form of West African traditional medicine consisting of a cocktail of herbs. The goal of this study is to evaluate a formulation of Agunmu made from M. indica, A. repens, E. chlorantha, A. boonei, and B. ferruginea, sold in the open market and commonly used for the treatment of malaria by the locals, for its antimalarial effects and to determine the active principles that may contribute to the antimalarial effect. The ethanolic extract obtained from this formulation (Ag-Iba) was analyzed, using TLC, LC-MS, and Tandem-MS techniques, to determine its phytochemical properties. The extract was tested in vitro against representative bacteria strains, cancer and normal human cell lines, and susceptible (D6) and resistant (W2) Plasmodium falciparum. In subsequent in vivo experiments, graded doses of the extract were used to treat mice infected with chloroquine-susceptible (NK-65) and chloroquine-resistant (ANKA) strains of Plasmodium berghei. Bacteria growth was monitored with a disc diffusion assay, cancer cell viability was determined with MTS assay, and percentage parasitemia and parasite clearance were determined by microscopy. Bound heme content, host mitochondria permeability transition (mPT) pore opening, F0F1-ATPase, and lipid peroxidation were determined via spectrophotometry. Indices of oxidative stress, anti-oxidant activities, toxicity, cell death, and inflammatory responses were obtained using biochemical and ELISA techniques. The histology of the liver and spleen was performed using the standard method. We elucidated the structures of the critical active principles in the extract to be flavonoids: kaempferol, quercetin, myricetin, and their glycosides with little or no detectable levels of the toxic Aristolochic acids that are found in Aristolochia repens, one of the components of the formulation. The extract also showed anti-plasmodial activity in in vitro and in vivo models. Furthermore, the extract dose-dependently decreased mitochondrial dysfunction, cell death, and inflammatory and oxidative damage but increased antioxidant potentials. Presumably, the active principles in the extract work as a combinatorial therapy to elicit potent antimalarial activity. Overall, our study unraveled the active components from a commercial herbal formulation that could be reformulated for antimalarial therapy.

Mycotoxin concentrations in rice are affected by chalkiness, grain shape, processing type, and grain origin.

Mycotoxins such as aflatoxins (AFs), fumonisins (FBs), zearalenone (ZEN), and deoxynivalenol (DON) pose a risk to public health due to their carcinogenic potency (AFs and FBs) and anti-nutritional effects. The hazards associated with mycotoxins are accentuated where food management practices, control, and regulatory systems from farm to plate are sub-optimal. Information on the frequency of these mycotoxins in rice commercialized in markets in sub-Sahara Africa (SSA) is limited. The current study examined AF concentrations in 527 rice samples collected from 54 markets in five SSA countries. Grain quality characteristics, processing methods, and origin of samples were contrasted with toxin levels. In total, 72% of the samples had detectable AFs levels (range = 3.0 to 89.8µg/kg). Forty-seven percent (47%) of the samples had AFs above 4µg/kg, the European Union maximum level (ML), and were evaluated for cooccurrence with FBs, ZEN, and DON. Total AFs and ZEN cooccurred in 40% of the samples, and 30% of the positive ZEN samples had concentrations above the ML of 75µg/kg. Total AFs did not co-occur with FBs and DON. Multivariate analysis revealed that length-to-width ratio (p < 0.0001), mixed variety for width (p = 0.04), and chalkiness (p = 0.009) significantly influenced aflatoxin concentrations. Slender grains had higher AFs concentrations than bold and medium grains (p < 0.0001). Possible strategies to mitigate mycotoxin contamination in rice include improving grain quality traits and practicing proper drying and hermetic storage before and after milling. These findings provide valuable insights for both domestic and international actors in establishing and strengthening regulations and management systems to mitigate rice mycotoxin contamination.

Assessing Environmental Contamination through Pollen Analysis of Solitary Cavity-nesting Resin Bees Around Kolhapur City

This study examines how pollen-nectar mixtures collected by solitary cavity-nesting resin bees can serve as bioindicators of environmental pollution. Researchers utilized paper straw trap nests across various locations, including agricultural (Shirgaon to Kandalgaon road in Gokul Shirgaon, Kolhapur), industrial (MIDC Shiroli, Kolhapur), urban (Dabolkar Corner, Kolhapur City), and a control area (KIT's College of Engineering, Gokul Shirgaon, Kolhapur). The nests, installed in February 2024, contained empty straws and some partially packed with larvae from the control area. These nests were designed to mimic natural habitats, allowing mature bees to build nests and create pollen-nectar mixtures for their offspring. Analysis aimed to detect environmental contaminants in these mixtures. Only one sample from the industrial area (Trap Nest Location 5 at Quality Castings, MIDC Shiroli) was recovered, showing elevated levels of iron, manganese, chromium, and nickel, consistent with industrial emissions. In contrast, the control area produced viable samples with detectable levels of magnesium, zinc, iron, aluminum, and barium. The absence of samples from urban and agricultural areas suggests adverse conditions affecting bee populations. This underscores the sensitivity of resin bees to pollution and highlights their role as bioindicators. The study emphasizes the need for effective environmental management to enhance habitat quality and protect both bees and human health.

Dynamics of PCV2 and PCV3 in the Serum and Oral Fluids of Pigs After PCV2 Vaccination in a Commercial Farm

Objectives: This study investigated the dynamics of porcine circovirus type 2 (PCV2) and PCV3 on a commercial farm following PCV2 vaccination. Methods: Serum samples from 35 pigs, starting at 3 weeks of age, were collected weekly until 21 weeks of age. Oral fluids from six pens of pigs of the same age were also analyzed. Viral DNA was assessed in pooled sera and individual oral fluid samples, while antibodies (IgG and IgA) were measured in the serum and oral fluids. Productive parameters, including weekly mortality and cumulative mortality, were evaluated. Results: The results revealed that PCV2 and PCV3 co-infection was detected in pigs at 8 weeks of age, with PCV3 being detected in oral fluids two weeks earlier. PCV3 DNA was detected in oral fluids at 4 weeks of age. PCV2 IgG antibodies in the serum increased gradually after vaccination, peaking at 7 weeks of age, then declined and stabilized until 21 weeks of age. PCV3 IgG antibodies fluctuated early but were uniformly positive after 13 weeks of age. In oral fluids, PCV2 IgG and IgA antibodies showed a strong response only at 3 and 23 weeks of age. In contrast, a strong and consistent IgG response was observed in oral fluids in the absence of PCV2 and PCV3 co-infection of pigs at 3 to 11 weeks of age. The farm’s productive parameters remained stable throughout the study. Conclusions: These findings suggest that PCV2 and PCV3 co-infection, along with high PCV3 detection levels in serum and oral fluids, may have an impact on the efficacy of PCV2 vaccination.

Comprehensive host cell proteins profiling in biopharmaceuticals by a sensitivity enhanced mass spectrometry strategy using TMT-labeling and signal boosting

BackgroundHost Cell Proteins (HCPs) are impurities expressed in host cells during the biopharmaceutical production process, whichmay compromise product quality and potentially leading to immunogenic reactions or other adverse effects. Mass spectrometry (MS)-based strategy is more and more considered as a promising method for HCPs analysis, since it is capable of simultaneously quantifying thousands of proteins in a single test. However, considering the large excess biopharmaceutical product protein present in the system and the extremely low abundance of HCPs, sensitive MS methods are urgently needed in HCPs analysis. ResultsIn this work, we introduced a novel approach that leveraged host cell lysate as a boosting channel to enhance the MS signal of the residue HCPs in biopharmaceutical products using isobaric TMT labeling, thereby elevating the low-abundant HCPs to detectable and quantifiable levels of current MS without using enrichment or depletion method to avoid disturbance of the original concentration of the HCPs. Our method surpassed previous benchmarks by identifying a significantly higher number (23844 unique peptides for 3475 proteins) compared to existing records (4541 unique peptides for 848 proteins) for HCPs analysis in RM8671 NIST monoclonal antibody (mAb), demonstrating unparalleled sensitivity and robustness. Furthermore, our workflow successfully identified 44 of 48 UPS1 proteins across a concentration range of 0.32–4.15 ppm in monoclonal antibodies (mAbs), proving its effectiveness for in-depth HCPs analysis in biopharmaceuticals. SignificancePresent even at sub-ppm levels, HCPs may compromise the stability and safety of product proteins and alter pharmacokinetics or neutralization of therapeutic effects. Our MS signal enhancing method presented an advancement in HCP analysis, combining improved sensitivity and increased scale of HCPs with a streamlined and robust workflow. This method allowed HCPs quantification at <1 ppm level without disturbance of the original HCPs concentration, which is still rare in the field.

Creating of novel Wx allelic variations significantly altering Wx expression and rice eating and cooking quality

Granule-bound starch synthase I (GBSSI) encoding gene Waxy (Wx), which largely regulates the amylose content of rice grains, is a master module determining rice eating and cooking quality (ECQ). Fine-tuning amylose level of grains is an ideal strategy to improve rice quality. Through fine editing of Wxa promoter and 5′UTR by CRISPR/Cas9 system, we created 14 types of novel Wx allelic variations, of which MT7 and MT13 were able to alter Wx expression and amylose content of grains. MT7 showed fragment deletion and base insertions in CAAT-boxes, hardly detectable expression levels of GBSSI mRNA and protein, and generated 5.87% amylose in grains. MT13 had fragment deletions in the A-box and the TATA-box, low expression levels of GBSSI mRNA and protein, and generated 9.61% amylose in grains. Besides of the amylose content, MT7 and MT13 significantly reduced protein content and increased lipid content of grains compared with Wxa. A comparison of MT7, MT13 and other allelic lines demonstrated the importance of base insertion around the second CAAT-box and 31bp-deletion following the second TATA-box in modulating Wx expression. Thus, our study generated two novel Wx allelic variations which significantly alter Wx expression and amylose content of rice grains, providing not only new germplasms for soft rice breeding, but also insights into candidate cis elements of Wx.

Simple and rapid determination of 3-monochloropropane-1,2-diol in food contact papers based on polydopamine-polyethyleneimine copolymerization

The aim of this work was to develop a fluorescence method based on the polydopamine-polyethyleneimine (PDA-PEI) copolymerization, which was subsequently applied for the determination of 3-monochloropropane-1,2-diol (3-MCPD) in food contact papers (FCMs). PEI could provide an alkaline environment and then react with dopamine (DA) to produce copolymers by Michael addition and Schiff-base reactions. This copolymer has a strong fluorescence emission at 527 nm. We found that amino groups of DA and PEI could also react with 3-MCPD in an alkaline medium, which improved the morphology and fluorescence intensity of PDA-PEI copolymers. The fluorescence intensity of the polymers was linear but inversely proportional to the concentration of 3-MCPD in the range of 10.0–500.0 μg kg−1 and the detection limit was 2 μg kg−1. The standard addition method was used in FCMs to demonstrate the practical applicability and the spiked recoveries ranged from 99.8 to 110.3 %. Finally, the levels of 3-MCPD in different FCMs (n = 70) were determined by the proposed method. The detection frequencies ranged from 25 % to 100 % and both the highest detection frequency and levels were observed in kitchen papers. More than half of the samples did not comply with the limits recommended by the German Federal Institute for Risk Assessment, suggesting that 3-MCPD released from FCMs is a major route of human exposure.

Co-Occurrence of Mycotoxins in the Diet and in the Milk of Dairy Cows from the Southeast Region of Brazil.

Mycotoxins are toxic fungi secondary metabolites that develop on feedstuffs and can be transferred into milk, thus representing a public health risk. The objective of this study was to assess the co-occurrence of mycotoxins in the diet and in the milk of dairy cows from the southeast region of Brazil. Samples of total mixed ration (TMR, n = 70) and milk (n = 70) were collected in dairy farms and subjected to multi-mycotoxin analysis using liquid chromatography coupled to tandem mass spectrometry. The aflatoxins (AFs), ochratoxin A (OTA), and T-2 and HT-2 toxins were not detected in TMR samples. In contrast, fumonisins (FBs), zearalenone (ZEN), and deoxynivalenol (DON) were detected in 100, 93, and 24% of TMR samples at mean levels of 336.7 ± 36.98, 80.32 ± 16.06 µg/kg and 292.1 ± 85.68 µg/kg, respectively. Ninety-two percent of TMR samples exhibited co-occurring mycotoxins. In milk, 54% of samples (n = 38) had detectable levels of mycotoxin, while 43% (n = 30) contained two or more types of mycotoxins. DON, FB, and ZEN metabolites (α-zearalenol and β-zearalenol) were the most frequent mycotoxins detected in the milk samples analyzed, at mean concentrations of 0.562 ± 0.112, 2.135 ± 0.296 µg/kg, 2.472 ± 0.436 µg/kg, and 0.343 ± 0.062 µg/kg, respectively. However, none of the analyzed milk samples had levels higher than the maximum permitted limit for AFM1 in Brazil (0.5 µg/L). The results of this trial highlight the concern about the co-occurrence of multiple mycotoxins in TMR and in milk, due to the possible additive or synergistic effects of these compounds. The presence of co-occurring mycotoxins in milk underscores the need for stringent preventive practices to avoid mycotoxin contamination in the diet of dairy cows in Brazil.