[Determination of amanita peptide and tryptamine toxins in wild mushrooms by high performance liquid chromatography-tandem mass spectrometry].

The discovery and identification of mushroom toxins has long been an important area in the fields of toxicology and food safety. Mushrooms are widely favored for their culinary and medicinal value; however, the presence of potentially lethal toxins in some species poses a substantial challenge in ensuring their safe consumption. Therefore, the development of a robust and sensitive analytical method is necessary for accurately identifying the risks associated with mushroom consumption. The study of mushroom toxins, which are characterized by their diversity and substantial variations in chemical structures, present a considerable challenge for achieving precise and high-throughput analysis. To address this issue, the present study employed a robust approach combining a solid-phase extraction (SPE) purification technique with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) to establish an analytical method for the detection and quantification of five amatoxins and two tryptamines (psilocybin and bufotenine) present in some mushrooms. Several optimization procedures were undertaken, including optimizing the chromatographic conditions, mass spectrometric parameters, and sample extraction and purification. The procedure involved the extraction of dry mushroom powder with methanol containing 0.3% formic acid, followed by purification using a strong cation exchange cartridge (SCX). The analytes were separated on a T3 chromatographic column (100 mm×2.1 mm, 1.8 µm) using mobile phases of acetonitrile and 5 mmol/L ammonium acetate solution containing 0.1% formic acid. The multiple reaction monitoring (MRM) mode was employed for data acquisition. Amatoxins were quantified using matrix-matched standard calibration curves, whereas isotopic internal standards were used to quantify tryptamine. The results showed that all seven toxins exhibited good linearities (r2>0.99) within the optimized concentration range. The limits of detection (LODs) for bufotenine, psilocybin, and amatoxins were determined as 2.0, 5.0, and 10 µg/kg, respectively, while the limits of quantification (LOQs) were determined as 5.0, 10, and 20 µg/kg, respectively. The LOD and LOQ values further underscore the ability of the method to detect minute quantities of toxins, making it particularly well suited for screening food samples for potential contamination. Using dried shiitake mushroom powder as the matrix, the recoveries of the two tryptamines ranged from 80.6% to 117%, with relative standard deviations (RSDs) ranging from 1.73% to 5.98%, while the recoveries of amatoxins ranged from 71.8% to 115%, with RSDs varying from 2.14% to 9.92% at the three concentration levels. The consistent and satisfactory recoveries of amatoxins and tryptamines demonstrated the ability of this method to accurately quantify the target analytes even in a complex matrix. Comparison with the results of supplementary test method recognized by State Administration for Market Regulation for food (BJS 202008) demonstrated comparable results, indicating no significant differences (p>0.05) in amatoxin contents. The newly developed method is rapid, accurate, precise, meets the required standards, and is suitable for the detection of seven toxins in wild mushrooms. As part of the application of this method, a comprehensive investigation of the distribution of toxins in wild mushrooms from Fujian Province was undertaken. In this study, 59 wild mushroom samples from nine cities were collected in the Fujian province. Species identification was conducted using rDNA-internal transcribed space (rDNA-ITS) molecular barcode technology, which revealed the presence of toxins in the two samples. Notably, one specimen named Amanita fuligineoides contained α-amanitin, ß-amanitin, and phalloidin in quantities of 607, 377, and 69.0 mg/kg, respectively. Additionally, another sample, identified as Tricholomataceae, had a psilocybin concentration of 12.6 mg/kg.

Comparative Analysis of Clinical and CT Findings in Patients with SARS-CoV-2 Original Strain, Delta and Omicron Variants.

OBJECTIVES: To compare the clinical characteristics and chest CT findings of patients infected with Omicron and Delta variants and the original strain of COVID-19. METHODS: A total of 503 patients infected with the original strain (245 cases), Delta variant (90 cases), and Omicron variant (168 cases) were retrospectively analyzed. The differences in clinical severity and chest CT findings were analyzed. We also compared the infection severity of patients with different vaccination statuses and quantified pneumonia by a deep-learning approach. RESULTS: The rate of severe disease decreased significantly from the original strain to the Delta variant and Omicron variant (27% vs. 10% vs. 4.8%, p < 0.001). In the Omicron group, 44% (73/168) of CT scans were categorized as abnormal compared with 81% (73/90) in the Delta group and 96% (235/245, p < 0.05) in the original group. Trends of a gradual decrease in total CT score, lesion volume, and lesion CT value of AI evaluation were observed across the groups (p < 0.001 for all). Omicron patients who received the booster vaccine had less clinical severity (p = 0.015) and lower lung involvement rate than those without the booster vaccine (36% vs. 57%, p = 0.009). CONCLUSIONS: Compared with the original strain and Delta variant, the Omicron variant had less clinical severity and less lung injury on CT scans.

A multi-stage supply chain disruption mitigation strategy considering product life cycle during COVID-19.

The global pandemic of COVID-19 has caused severe damage to the supply chain such that manufacturers may face long-term supply disruptions. In this paper, a disruption recovery strategy of a supply chain system is investigated from the perspective of product change, in which the life cycle and design change time of a new product are both considered in order to minimize the losses of manufacturer after disruptions. A mixed-integer linear programming (MILP) model is presented to address the disruption recovery problem for this multi-period, multi-supplier, and multi-stage supply chain system. A two-stage heuristic algorithm is designed to solve the problem. Experimental results show that the proposed disruption mitigation strategy can effectively reduce the profit loss of manufacturer due to supply disruption, and demonstrate the impact of product life cycle in the selection of new product design planning. A sensitivity analysis is performed to ensure the applicability of the model in the actual environment, which illustrates the effect of different parameter changes on the results. This work can help manufacturers establish an optimal recovery strategy whenever the supply chain system experiences supply disruptions.

Effectiveness of peer-mediated intervention on social skills for children with autism spectrum disorder: a randomized controlled trial.

Background: Peer-mediated intervention (PMI) is an intervention that teaches normally developing peers to help children with autism spectrum disorder (ASD) actively participate in social interactions. Previous studies have shown that PMI applied to school settings is effective for children with ASD, but more multiple-baseline single-subject design. Many questions are still not clear due to the large clinical variability in children with ASD. This study investigated the effectiveness of PMI on social skills of children with ASD at varying symptom levels and analyzed the specific changes. Methods: This study used a randomized, single-blind, parallel-controlled design to analyze the effect of PMI in a hospital setting. Fifty-five children aged 4-12 years were diagnosed with ASD by clinicians using the fifth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-5) and stratified randomly allocated to either the experimental group or the control group using the envelope method. The experimental group utilized PMI, whereas the control group utilized behavioral therapy based on applied behavior analysis (ABA) [early intensive behavioral intervention (EIBI)]. This study primarily utilized the Social Responsiveness Scale (SRS) to evaluate the social performance of autistic children prior to and after the intervention. Results: Fifty-five participants were recruited and analyzed, the experimental group (n=29; mild to moderate n=18, severe n=11) and the control group (n=26; mild to moderate n=15, severe n=11). After the intervention, the experimental group's SRS score fell significantly more than the control group's (t=-3.918, P=0.000), d=-1.043; the mild to moderate subgroup experienced the same situation (H=17.811, P=0.009), d=-1.642. At the same time, the decline in social communication scores was significantly greater in the experimental group compared to the control group (t=-3.869, P=0.000), and the 95% confidence interval was -10.067 to -3.193. The social motivation of the mild-to-moderate subgroup of the experimental group (H=16.894, P=0.011), -3.000 (25th percentile, 75th percentile: -3.000, 0.000), and the behavioral patterns of autism (H=18.150, P=0.006), -3.000 (25th percentile, 75th percentile: -5.000, 0.000), the decreased value was significantly larger. Conclusions: PMI therapy can increase social motivation in children with mild to moderate ASD, minimize undesirable behavior patterns, effectively improve overall social skills and enhance effective social communication with others. Trial Registration: Chinese Clinical Trial Registry ChiCTR2100049185.

Acute mushroom poisoning of Amanita pseudosychnopyramis: A case report from Fujian, China with exact species identification and descriptive study.

Mushroom poisoning is a deeply concerning food safety problem that affects the public in China every year. Although there are statistics on the number of poisonings and incidents, there is a lack of data on the types of toxic mushrooms, clinical manifestations and toxins. A case of wild mushroom poisoning occurred in Xiamen. Descriptive epidemiological investigation, toxins detection, and morphological and phylogenetic identification were immediately performed. The patients exhibited typical neurotoxic symptoms after consuming wild mushrooms, including chills, vertigo, drowsiness, salivation and coma. The average incubation period was 30 min. Treatments that were adopted included fluid infusion, gastric lavage, catharsis, and liver protection treatment. All patients recovered within 10 days. The species was identified as Amanita pseudosychnopyramis, and its contents of muscarine, muscimol and ibotenic acid were 170.3 ± 5.9 mg/kg, 835.4 ± 43.1 mg/kg and 637.9 ± 54.8 mg/kg in dry weight, respectively, as detected by ultrahigh-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). To our knowledge, this is the first report of Amanita pseudosychnopyramis poisoning worldwide.