Advances in Metabolism and Metabolic Toxicology of Quinoxaline 1,4-Di-N-oxides.

Quinoxaline 1,4-di-N-Oxides (QdNOs) have been used as synthetic antimicrobial agents in animal husbandry and aquaculture. The metabolism and potential toxicity have been also concerns in recently years. The metabolism investigations showed that there were 8 metabolites of Carbadox (CBX), 34 metabolites of Cyadox (CYA), 33 metabolites of Mequindox (MEQ), 35 metabolites of Olaquindox (OLA), and 56 metabolites of Quinocetone (QCT) in different animals. Among them, Cb3 and Cb8, M6, and O9 are metabolic residual markers of CBX, MEQ and OLA, which are associated with N → O reduction. Toxicity studies revealed that QdNOs exhibited severe tumorigenicity, cytotoxicity, and adrenal toxicity. Metabolic toxicology showed that toxicity of QdNOs metabolites might be related to the N → O group reduction, and some metabolites exhibited higher toxic effects than the precursor, which could provide guidance for further research on the metabolic toxicology of QdNOs and provide a wealth of information for food safety evaluation.

Foodborne Disease Outbreaks Caused by Biotoxins in Yantai City: A 10-Year Spatiotemporal Monitoring Study.

Unsafe food causes 600 million cases of foodborne diseases and 420,000 deaths every year. Meanwhile, biological toxins such as poisonous mushrooms, saponins, and aflatoxin can cause significant damage to humans. Therefore, it is particularly important to study foodborne disease outbreaks caused by biotoxins (FDOB). We collected FDOB in Yantai City from 2013 to 2022 and further established a corresponding database. Statistical analysis was carried out according to time, place, pathogen, and contamination of pathogenic factors. There were 128 FDOB, resulting in 417 patients and 6 deaths. The third quarter was a high season for foodborne disease outbreaks, the number of events, patients and deaths accounted for 65.63% (84/128), 55.88% (233/417), and 100% (6/6) of the total number, respectively. The highest number of outbreaks per 10,000 persons was Qixia (0.41), followed by Zhifu (0.36) and Laiyang (0.33). The top three causes of outbreaks were poisonous mushroom toxin, saponins and hemagglutinin, and Lagenaria siceraria (Molina) Standl. Sixty-five (50.78%) outbreaks were attributed to poisonous mushroom toxin, 18 (14.06%) outbreaks to saponin and hemagglutinin, and 12 (9.38%) outbreaks to L. siceraria (Molina) Standl. The largest number of outbreaks, patients and deaths all occurred in families, accounting for 82.81% (106/128) outbreaks, 66.19% (276/417) patients, and 100% (6/6) deaths, respectively. Followed by catering service establishments, accounting for 14.84% (19/128), 30.22% (126/417), and 0% (0/6), respectively. The main poisoning link of outbreaks was ingestion and misuse, accounting for 72.66% (93/128), followed by improper processing, accounting for 20.31% (26/128). It is necessary to carry out targeted family publicity and education, strengthen the integration of medical and prevention, explore innovative monitoring and early warning mechanisms for foodborne diseases, and reduce the occurrence of underreporting of foodborne disease outbreaks.

Research progress on the application of different preservation methods for controlling fungi and toxins in fruit and vegetable.

Fruits and vegetables are susceptible to fungal infections during picking, transportation, storage and processing, which have a high potential to produce toxins. Fungi and toxins can cause acute or chronic poisoning after entering the body. In the field of fruit and vegetable preservation, technologies such as temperature control, modified atmosphere, irradiation, application of natural or chemical preservatives, and edible films are commonly used. In practical applications, according to the types, physiological differences and actual needs of fruits and vegetables, suitable preservation methods can be selected to achieve the effect of preservation and control of fungi and toxins. The starting point of fresh-keeping technology is to delay post-harvest senescence of fruits and vegetables, inhibit the respiratory intensity, and control the reproduction of microorganisms, which is important to control the reproduction of fungi and the production of toxins. From the three directions of physical, chemical and biological means, the article analyses and explores the effects of different external factors on the production of toxins and the effects of different preservation techniques on fungal growth and toxin production in fruits and vegetables, in order to provide new ideas for the preservation of fruits and vegetables and the control of harmful substances in food.

Marangoni Instability of a Drop in a Stably Stratified Liquid.

Marangoni instabilities can emerge when a liquid interface is subjected to a concentration or temperature gradient. It is generally believed that for these instabilities bulk effects like buoyancy are negligible compared to interfacial forces, especially on small scales. Consequently, the effect of a stable stratification on the Marangoni instability has hitherto been ignored. Here, however, we show that they can matter. We report, for an immiscible drop immersed in a stably stratified ethanol-water mixture, a new type of oscillatory solutal Marangoni instability that is triggered once the stratification has reached a critical value. We experimentally explore the parameter space spanned by the stratification strength and the drop size and theoretically explain the observed crossover from levitating to bouncing by balancing the advection and diffusion around the drop. Finally, the effect of the stable stratification on the Marangoni instability is surprisingly strongly amplified in confined geometries, leading to an earlier onset.

Toxicological evaluation of S. involucrata culture: Acute, 90-day subchronic and genotoxicity studies.

Saussurea involucrata is an endangered plant that is used in traditional Chinese medicine. Through the use of plant cell culture techniques, preparations of Saussurea involucrata (S. involucrata) cell cultures have been developed and used to generate medicinal preparations. There have been few evidence-based analyses of the toxicological effects of S. involucrata culture conducted to date. Here, we conducted the experiments designed to assess the acute, subchronic, and genotoxic toxicological effects of S. involucrata culture. The genotoxic study was assessed through Ames, marrow micronucleus, and sperm malformation assays. The acute toxicity was assessed by orally administering in rats and mice at dose of 7500 mg/kg. Subchronic toxicity studies were then conducted by administering rats at doses of 500, 1000, or 1500 mg/kg for 90 days. No genotoxicity was observed at any tested dose levels, nor was any evidence of acute toxicity detected in treated mice or rats. Similarly, subchronic study of S. involucrata culture administration was not associated with any changes in rat food intake, weight, hematological parameters, organ weight, or organ histology. Then, we determined that the no observed adverse effect level of S. involucrata culture was greater than 1500 mg/kg in our 90-day toxicity study.

Monostable superrepellent materials.

Superrepellency is an extreme situation where liquids stay at the tops of rough surfaces, in the so-called Cassie state. Owing to the dramatic reduction of solid/liquid contact, such states lead to many applications, such as antifouling, droplet manipulation, hydrodynamic slip, and self-cleaning. However, superrepellency is often destroyed by impalement transitions triggered by environmental disturbances whereas inverse transitions are not observed without energy input. Here we show through controlled experiments the existence of a "monostable" region in the phase space of surface chemistry and roughness, where transitions from Cassie to (impaled) Wenzel states become spontaneously reversible. We establish the condition for observing monostability, which might guide further design and engineering of robust superrepellent materials.

Bouncing Oil Droplet in a Stratified Liquid and its Sudden Death.

Droplets can self-propel when immersed in another liquid in which a concentration gradient is present. Here we report the experimental and numerical study of a self-propelling oil droplet in a vertically stratified ethanol-water mixture: At first, the droplet sinks slowly due to gravity, but then, before having reached its density matched position, jumps up suddenly. More remarkably, the droplet bounces repeatedly with an ever increasing jumping distance, until all of a sudden it stops after about 30 min. We identify the Marangoni stress at the droplet-liquid interface as responsible for the jumping: its strength grows exponentially because it pulls down ethanol-rich liquid, which in turn increases its strength even more. The jumping process can repeat because gravity restores the system. Finally, the sudden death of the jumping droplet is also explained. Our findings have demonstrated a type of prominent droplet bouncing inside a continuous medium with no wall or sharp interface.

Metabolic Profile, Bioavailability and Toxicokinetics of Zearalenone-14-Glucoside in Rats after Oral and Intravenous Administration by Liquid Chromatography High-Resolution Mass Spectrometry and Tandem Mass Spectrometry.

Zearalenone-14-glucoside (ZEN-14G), a key modified mycotoxin, has attracted a great deal of attention due to the possible conversion to its free form of zearalenone (ZEN) exerting toxicity. In this study, the toxicokinetics of ZEN-14G were investigated in rats after oral and intravenous administration. The plasma concentrations of ZEN-14G and its major five metabolites were quantified using a validated liquid chromatography tandem mass spectrometry (LC-MS/MS) method. The data were analyzed via non-compartmental analysis using software WinNonlin 6.3. The results indicated that ZEN-14G was rapidly hydrolyzed into ZEN in vivo. In addition, the major parameters of ZEN-14G following intravenous administration were: area under the plasma concentration-time curve (AUC), 1.80 h·ng/mL; the apparent volume of distribution (VZ), 7.25 L/kg; and total body clearance (CL), 5.02 mL/h/kg, respectively. After oral administration, the typical parameters were: AUC, 0.16 h·ng/mL; VZ, 6.24 mL/kg; and CL, 4.50 mL/h/kg, respectively. The absolute oral bioavailability of ZEN-14G in rats was about 9%, since low levels of ZEN-14G were detected in plasma, which might be attributed to its extensive metabolism. Therefore, liquid chromatography high-resolution mass spectrometry (LC-HRMS) was adopted to clarify the metabolic profile of ZEN-14G in rats' plasma. As a result, eight metabolites were identified in which ZEN-14-glucuronic acid (ZEN-14GlcA) had a large yield from the first time-point and continued accumulating after oral administration, indicating that ZEN-14-glucuronic acid could serve a potential biomarker of ZEN-14G. The obtained outcomes would prompt the accurate safety evaluation of ZEN-14G.

Tracing major metabolites of quinoxaline-1,4-dioxides in abalone with high-performance liquid chromatography tandem positive-mode electrospray ionization mass spectrometry.

BACKGROUND: Owing to the comprehensive application of quinoxaline-1,4-dioxides (QdNOs) in aquaculture, QdNOs and metabolites are often detected in marine food, including abalone. QdNOs are reported to exhibit cytotoxicity, photoallergy, mutagenicity, and carcinogenicity activities. To monitor for contamination of QdNOS in abalone and assess dietary exposure, a simple and reliable analytical method for the detection of QdNOs and their major metabolites was developed. RESULTS: This work is the first to present a simple and fast pretreatment procedure coupled with high-performance liquid chromatography tandem positive-mode electrospray ionization mass spectrometry (LC-MS/MS) for tracing of major metabolites of QdNOs in abalone. Extraction steps were simplified by the use of methanol and ethyl acetate containing 0.1% formic acid instead of more complicated acidolysis and enzymolysis pretreatment procedures. High-sensitive characters were obtained with limits of detection ranged from 0.16 to 2.1 µg kg-1 for QdNOs and their major metabolites. CONCLUSION: These results indicate that the LC-MS/MS method developed could be applied for QdNOs and major metabolites detection in actual samples. Considering the large production and consumption of abalone in Shandong Province, China, this work will also contribute to the further understanding of the often-ignored exposure pathway of QdNOs. © 2019 Society of Chemical Industry.

Cysticercosis in Shandong Province, Eastern China.

We analyzed demographic and clinical data and estimated the incidence of cysticercosis in Shandong Province, China, during 1975-2014. Our analyses showed that a cysticercosis-endemic area is present in Shandong Province, especially in its western regions. Improved surveillance and control are needed to address the elevated risk for cysticercosis in this region.

Unraveling the Metabolic Routes of Retapamulin: Insights into Drug Development of Pleuromutilins.

Retapamulin, a semisynthetic pleuromutilin derivative, is exclusively used for the topical short-term medication of impetigo and staphylococcal infections. In the present study, we report that retapamulin is adequately and rapidly metabolized in vitro via various metabolic pathways, such as hydroxylation, including mono-, di-, and trihydroxylation, and demethylation. Like tiamulin and valnemulin, the major metabolic routes of retapamulin were hydroxylation at the 2ß and 8α positions of the mutilin moiety. Moreover, in vivo metabolism concurred with the results of the in vitro assays. Additionally, we observed significant interspecies differences in the metabolism of retapamulin. Until now, modifying the side chain was the mainstream method for new drug discovery of the pleuromutilins. This approach, however, could not resolve the low bioavailability and short efficacy of the drugs. Considering the rapid metabolism of the pleuromutilins mediated by cytochrome P450 enzymes, we propose that blocking the active metabolic site (C-2 and C-8 motif) or administering the drug in combination with cytochrome P450 enzyme inhibitors is a promising pathway in the development of novel pleuromutilin drugs with slow metabolism and long efficacy.

Entrapment and Dissolution of Microbubbles Inside Microwells.

The formation and evolution of immersed surface micro- and nanobubbles are essential in various practical applications, such as the usage of superhydrophobic materials, drug delivery, and mineral flotation. In this work, we investigate the entrapment of microbubbles on a hydrophobic surface, structured with microwells, when water flow passes along, and the subsequent microbubble dissolution. At entrapment, the microbubble is initially pinned at the edge of the microwell. At some point, the three-phase contact line detaches from one side of the edge and separates from the wall, after which it further recedes. We systematically investigate the evolution of the footprint diameter and the contact angle of the entrapped microbubbles, which reveals that the dissolution process is in the constant contact angle mode. By varying the gas undersaturation level, we quantify how a high gas undersaturation enhances the dissolution process, and compare with simplified theoretical predictions for dissolving bubbles on a plane surface. We find that geometric partial blockage effects of the diffusive flux out of the microbubble trapped in the microwell lead to reduced dissolution rates.

Deglucosylation of zearalenone-14-glucoside in animals and human liver leads to underestimation of exposure to zearalenone in humans.

Zearalenone-14-glucoside (ZEN-14G), the modified mycotoxin of zearalenone (ZEN), has attracted considerable attention due to its high potential to be hydrolyzed into ZEN, which would exert toxicity. It has been confirmed that the microflora could metabolize ZEN-14G to ZEN. However, the metabolic profile of ZEN-14G and whether it could be deglucosidated in the liver are unknown. To thoroughly investigate the metabolism of ZEN-14G, in vitro metabolism including phase I and phase II metabolism was studied using liquid chromatography coupled to high-resolution mass spectrometry. Additionally, in vivo metabolism of ZEN-14G was conducted in model animals, rats, by oral administration. As a result, 29 phase I metabolites and 6 phase II metabolites were identified and significant inter-species metabolic differences were observed as well. What is more, ZEN-14G could be considerably deglucosidated into its free form of ZEN after the incubation with animals and human liver microsomes in the absence of NADPH, which was mainly metabolized by human carboxylesterase CES-I and II. Furthermore, results showed that the major metabolic pathways of ZEN-14G were deglucosylation, hydroxylation, hydrogenation and glucuronidation. Although interspecies differences in the biotransformation of ZEN-14G were observed, ZEN, α-ZEL-14G, ß-ZEL-14G, α-ZEL, ZEN-14G-16GlcA and ZEN-14GlcA were the major metabolites of ZEN-14G. Additionally, a larger yield of 6-OH-ZEN-14G and 8-OH-ZEN-14G was also observed in human liver microsomes. The obtained data would be of great importance for the safety assessment of modified mycotoxin, ZEN-14G, and provide another perspective for risk assessment of mycotoxin.

Genotoxicity testing of sodium formononetin-3'-sulphonate (Sul-F) by assessing bacterial reverse mutation, chromosomal aberrations and micronucleus tests.

As part of a safety evaluation, we evaluated the potential genotoxicity of sodium formononetin-3'-sulphonate (Sul-F) using bacterial reverse mutation assay, chromosomal aberrations detection, and mouse micronucleus test. In bacterial reverse mutation assay using five strains of Salmonella typhimurium (TA97, TA98, TA100, TA102 and TA1535), Sul-F (250, 500, 1000, 2000, 4000 µg/plate) did not increase the number of revertant colonies in any tester strain with or without S9 mix. In a chromosomal assay using Chinese hamster lung fibroblast (CHL) cells, there were no increases in either kind of aberration at any dose of Sul-F (400, 800, and 1600 µg/mL) treatment groups with or without S9 metabolic activation. In an in vivo bone marrow micronucleus test in ICR mice, Sul-F at up to 2000 mg/kg (intravenous injection) showed no significant increases in the incidence of micronucleated polychromatic erythrocytes, and the proportion of immature erythrocytes to total erythrocytes. The results demonstrated that Sul-F does not show mutagenic or genotoxic potential under these test conditions.

Repeated sub-chronic oral toxicity study of xylooligosaccharides (XOS) in dogs.

In this study, Beagle dogs were administered xylooligosaccharide (XOS, CAS # 87099-0) at doses of 0, 1250, 2500, and 5000 mg/kg/day by oral gavage for 26 weeks. A 4-week recovery period was added to observe delayed or reversible toxicity. Measurements included body weight, food consumption, clinical observations, temperature, electrocardiogram (ECG), urinalysis, blood chemistry, hematology, organ weight, gross necropsy, and histopathological examination. Except for transient diarrhea or vomiting, no treatment-related adverse effects were noted. In the mid-dose groups, transitional diarrhea was observed in the initial 1-2 weeks. In the high-dose groups, diarrhea and/or vomiting were observed episodically over the duration of treatment. However, they disappeared after XOS was withdrawn in the recovery period. Although there was a tendency toward less weight gain in the high-dose group animal group, this is typical in animals and humans fed non-digestible carbohydrates. This chronic toxicity study demonstrated that the no observed adverse effect level (NOAEL) of XOS is 2500 mg/kg body weight (BW)/day. Based on body surface area (conversion factor of 0.54 for dogs to human), this corresponds to daily doses of 1350 mg/kg BW or 81-108 g XOS in human adults weighing 60-80 kg.

High-sensitivity chemiluminescent immunoassay investigation and application for the detection of T-2 toxin and major metabolite HT-2 toxin.

BACKGROUND: T-2 toxin is a widely distributed mycotoxin in cereals. HT-2 toxin is the major metabolite, which is also a contaminant in cereals. T-2 toxin and HT-2 toxin have been identified as having carcinogenic, hepatotoxic, teratogenic and immunotoxic properties. To reduce the risk of contamination, a rapid, highly sensitive and inexpensive assay for the detection is required. RESULTS: In this study a high-sensitivity chemiluminescent enzyme-linked immunoassay (CL-ELISA) of T-2 toxin and HT-2 toxin was developed. With the help of the chemiluminescent substrate, this protocol showed a highly sensitive character with an IC50 as low as 33.28 ng mL-1 and 27.27 ng mL-1 for T-2 and HT-2, respectively. In addition, this method had no cross-reaction with other structurally related mycotoxins. CONCLUSION: These results indicated that the developed CL-ELISA could be applied for the detection of T-2 toxin and HT-2 toxin in actual samples without complicated steps. © 2016 Society of Chemical Industry.

Associations of proteins relevant to MAPK signaling pathway (p38MAPK-1,HIF-1 and HO-1) with coronary lesion characteristics and prognosis of peri-menopausal women.

BACKGROUND: The present study was intended to explore whether three proteins within MAPK signaling pathway (i.e. p38MAPK-1, HIF-1 and HO-1) were correlated with peri-menopausal women's coronary lesion features and prognosis. METHODS: Altogether 1449 peri-menopausal women were divided into non-coronary artery disease (CAD) group (n = 860) and CAD group (n = 589), including 167 pre-menopausal CAD populations and 422 post-menopausal CAD populations. General information about CAD risk parameters were gathered, including age, family history of CAD or hypertension or diabetes mellitus, bilirubin, cholesterol, triglyceride, high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) and so on. Coronary angiography results were judged, and CAD score was calculated with application of Genisin scoring method. Besides, detection of MAPK-1 levels was implemented with Strept Avidin-Biotin Complex (SABC) method, while HIF-1 and HO-1 expressions in the serum were determined utilizing ELISA detection kit. Correlations among protein expressions, characteristics of coronary lesions and prognosis of CAD populations were finally evaluated. RESULTS: Hypertension, hyperlipoidemia, diabetes and smoking history were more prevalent among postmenopausal CAD women than premenopausal CAD women (P < 0.05). Furthermore, postmenopausal women seemed to be significantly associated with multiple (i.e. double and triple) vessel lesions and severe lesion types (type B and C), when compared with premenopausal CAD group (P < 0.05). Similarly, remarkably elevated expressions of p38MAPK-1, HIF-1 and HO-1 were found within postmenopausal CAD populations in comparison to premenopausal ones (P < 0.05). The internal CysC, hs-CRP, TG and LDL-C concentrations all accorded with the following tendency: postmenopausal CAD women > premenopausal CAD women > non-CAD women. Moreover, p38MAPK-1, HIF-1 and HO-1 expressions were up-regulated with increasing number of vessel lesions and severity of coronary lesions among peri-menopausal women. Besides, among both pre-menopausal and post-menopausal CAD groups, positive correlations could be observed between MAPK-1 and TG (r s = 0.271; r s = 0.476), between HIF-1α and LDL-C (r s = 0.077; r s = 0.470), as well as between HO-1 and CysC (r s = 0.492; r s = 0.190) or hs-CRP (r s = 0.569; r s = 0.542) (all P < 0.05). MAPK-1, HIF-1α and HO-1 were also, respectively, positively correlated with CysC (r s = 0.415), hs-CRP (r s = 0.137), and TG (r s = 0.142), regarding post-menopausal CAD women (all P < 0.05). Finally, only SBP and TG were regarded as independent risk factors for CAD prognosis (i.e. high Genisin score) among premenopausal women (OR = 1.02, 95%CI: 1.01-1.18, P = 0.043; OR = 1.82, 95%CI: 1.01-3.33, P = 0.047). CONCLUSIONS: Expressions of p38MAPK-1, HIF-1 and HO-1 could serve as predictive roles for coronary lesions among peri-menopausal women.

From coffee rings to coffee eyes.

We discuss how the stain left after evaporation of a suspension evolves with heating of the glass or plastic on which the liquid has been deposited. Upon increasing the substrate temperature, it is found that the stain gradually changes from the usually observed ring to an "eye" shape, that is, a combination of the thick central stain and the thin outer ring. Both the size and the relative volume of the central stain increase with the substrate temperature. The main mechanism for this phenomenon is proposed to be an enhanced Marangoni recirculation flow on hot substrates. These findings can be exploited to continuously tune the morphology of coffee stains, with potential applications in self-assembly and ink-jet printing.

Integrating a Multiple Isotopologue Reaction-Monitoring Technique and LC-MS/MS for Quantitation of Small Molecules: Ten Mycotoxins in Cereals as an Example.

Accurate quantification of mycotoxin in cereals is crucial for ensuring food safety and human health. However, the preparation of traditional multisample external calibration curves (MSCCs) is labor-intensive and error-prone. Here, a multiple isotopologue reaction-monitoring (MIRM)-LC-MS/MS method for accurate quantitation of ten major mycotoxins in cereals was successfully developed and validated, where a novel one-sample multipoint calibration curve (OSCC) strategy is used instead of MSCCs. The OSCC can be established by examining the correlation between the calculated theoretical isotopic abundances and the measured abundance across various MIRM channels. In comparison to the MSCC, the OSCC strategy exhibits outstanding performance including superior selectivity, accuracy (78.4-108.6%), and precision (<12.5%). Furthermore, the proposed OSCC-MIRM-LC-MS/MS method was successfully applied to investigate mycotoxin contamination in cereal samples in China. Considering the advantages of simplified workflows and improved throughput, the OSCC-MIRM-LC-MS/MS methodology holds great promise for accurately quantifying chemical contaminants in foods.