In vitro and in vivo metabolic study of three new psychoactive ß-keto-arylcyclohexylamines.

Since the 2000s, an increasing number of new psychoactive substances have appeared on the illicit drug market. ß-keto-arylcyclohexylamine compounds play important pharmacological roles in anesthesia; however, because these new psychoactive substances have rapidly increasing illicit recreational use, the lack of detailed toxicity data are of particular concern. Therefore, analysis of their metabolites can help forensic personnel provide references and suggestions on whether a suspect has taken an illicit new psychoactive ß-keto-arylcyclohexylamine. The present study investigated the in vitro and in vivo metabolism and metabolites of three ß-keto-arylcyclohexylamines: deschloro-N-ethyl-ketamine, fluoro-N-ethyl-ketamine and bromoketamine. In vitro and in vivo models were established using zebrafish and human liver microsomes for analysis of Phase I and Phase II metabolites by liquid chromatography-high-resolution mass spectrometry. Altogether, 49 metabolites were identified. The results were applied for the subject urine samples of known fluoro-N-ethyl-ketamine consumer screen analysis in forensic cases. Hydroxy-deschloro-N-ethyl-ketamine, hydroxy-fluoro-N-ethyl-ketamine and hydroxy-bromoketamine were recommended as potential biomarkers for documenting intake in clinical and forensic cases.

Metabolism of ADB-FUBIATA in zebrafish and pooled human liver microsomes investigated by liquid chromatography-high-resolution mass spectrometry.

RATIONALE: ADB-FUBIATA is one of the most recently identified new psychoactive substance (NPS) of synthetic cannabinoids. The co-use of in vitro (human liver microsomes) and in vivo (zebrafish) models offers abundant metabolites and may give a deep insight into the metabolism of NPS. METHODS: In vivo and in vitro metabolic studies of new synthetic cannabinoid ADB-FUBIATA were carried out using zebrafish and pooled human liver microsome models. Metabilites were structurally characterized by liquid chromatography-high-resolution mass spectrometry. RESULTS: In total, 18 metabolites were discovered and identified in the pooled human liver microsomes and zebrafish, including seventeen phase I metabolites and one phase II metabolite. The main metabolic pathways of ADB-FUBIATA were hydroxylation, dehydrogenation, N-dealkylation, amide hydrolysis, glucuronidation, and combination thereof. CONCLUSION: Hydroxylated metabolites can be recommended as metabolic markers for ADB-FUBIATA because of the structural characteristics and high intensity. These metabolism characteristics of ADB-FUBIATA were useful for its further forensic or clinical related investigations.

[Expression of ß-xylosidase An-xyl from Aspergillus niger and characterization of its xylose tolerance].

The hydrolysis of xylo-oligosaccharides catalyzed by ß-xylosidase plays an important role in the degradation of lignocellulose. However, the enzyme is easily inhibited by its catalytic product xylose, which severely limits its application. Based on molecular docking, this paper studied the xylose affinity of Aspergillus niger ß-xylosidase An-xyl, which was significantly differentially expressed in the fermentation medium of tea stalks, through cloning, expression and characterization. The synergistic degradation effect of this enzyme and cellulase on lignocellulose in tea stems was investigated. Molecular docking showed that the affinity of An-xyl to xylose was lower than that of Aspergillus oryzae ß-xylosidase with poor xylose tolerance. The Ki value of xylose inhibition constant of recombinant-expressed An-xyl was 433.2 mmol/L, higher than that of most ß-xylosidases of the GH3 family. The Km and Vmax towards pNPX were 3.6 mmol/L and 10 000 µmol/(min·mL), respectively. The optimum temperature of An-xyl was 65 ℃, the optimum pH was 4.0, 61% of the An-xyl activity could be retained upon treatment at 65 ℃ for 300 min, and 80% of the An-xyl activity could be retained upon treatment at pH 2.0-8.0 for 24 h. The hydrolysis of tea stem by An-xyl and cellulase produced 19.3% and 38.6% higher reducing sugar content at 2 h and 4 h, respectively, than that of using cellulase alone. This study showed that the An-xyl mined from differential expression exhibited high xylose tolerance and higher catalytic activity and stability, and could hydrolyze tea stem lignocellulose synergistically, which enriched the resource of ß-xylosidase with high xylose tolerance, thus may facilitate the advanced experimental research and its application.

Metabolism of dipentylone in zebrafish and human liver microsomes determined by liquid chromatography-high resolution mass spectrometry.

The consumption of novel psychoactive substances (NPS) is exceedingly prevalent in society, as these substances are sold and distributed as "legal highs." One novel synthetic cathinone emerging in the market is 1-(1,3-benzodioxol-5-yl)-2-(dimethylamino) pentan-1-one (dipentylone). The goal of this work was to study the in vivo and in vitro metabolism of dipentylone in zebrafish and human liver microsomes (HLMs) by liquid chromatography-high resolution mass spectrometry (LC-HRMS). The zebrafish and HLM samples contained 14 dipentylone metabolites, specifically 12 phase Ⅰ metabolites and 2 phase Ⅱ metabolites. The main metabolic pathways included monohydroxylation (M1 and M2), N-dealkylation (M3), hydroxylation of the aromatic ring and dealkoxylation of M3 (M4), O-dealkylation (M5), N-dealkylation of M5 (M6), reduction of carboxide (M7), monohydroxylation of M5 (M8), dehydrogenation (M9), dealkoxylation (M10), N-dealkylation of M10 (M11), dealkoxylation of M9 (M12), glucuronidation of M5 (M13), and sulfation (M14). The monohydroxylated metabolite (M2) can be recommended as metabolic markers for dipentylone. This study is the first to identify a target compound for monitoring the abuse of dipentylone and to determine the essential chemical structure of the metabolites for further toxicological research.

Development of an LC-MS/MS method for the determination of multiple mycotoxins in human urine.

A sensitive liquid chromatography-tandem mass spectrometry method was developed for the determination of 17 mycotoxins in human urine. The method incorporates a two-step liquid-liquid extraction with ethyl acetate:acetonitrile (7:1), which had good extraction recovery. The LOQs of all mycotoxins ranged from 0.1 to 1 ng/mL. Intra-day accuracy ranged from 94 to 106%, and intra-day precision ranged from 1 to 12% for all mycotoxins. Inter-day accuracy and precision were 95-105% and 2-8%, respectively. The method was successfully applied to investigate the urine levels of 17 mycotoxins from 42 volunteers. Deoxynivalenol (DON, 0.97-9.88 ng/mL) was detected in 10 (24%) urine samples and zearalenone (ZEN, 0.13-1.11 ng/mL) in 2 (5%) urine samples.

Physical Literacy for Communities (PL4C): physical literacy, physical activity and associations with wellbeing.

BACKGROUND: Physical literacy (PL) is considered an important determinant of children's physical activity through which health benefits may be derived. The purpose of this study is to describe a sample of Canadian children's baseline levels of PL and movement behaviors, and explore whether the associations between PL and their mental wellbeing, if any, are mediated by moderate-to-vigorous physical activity (MVPA). METHODS: All grade two children in 14 elementary schools in the West Vancouver School District, Canada were invited to participate in a two-year longitudinal project. PL was assessed through PLAYfun and PLAYself tools. Physical activity was measured by wrist-worn accelerometers (GT3X + BT) for seven days. Children's mental well-being was assessed using the Strengths and Difficulties Questionnaire (SDQ). A score of total difficulties was aggregated for internalizing and externalizing problems. RESULTS: A total of 355 children aged 7-9 (183 boys, 166 girls, 6 non-binary) participated with 258 children providing valid accelerometer data. Children exhibited an average of 111.1 min of MVPA per day, with 97.3% meeting the physical activity guidelines. Approximately 43% (108/250) of participants were meeting the Canadian 24-h movement guidelines. Children were at an 'emerging' level of overall physical competence (45.8 ± 5.6) and reported a mean score of 68.9 (SD = 12.3) for self-perceived PL, with no significant differences between boys and girls. PL was significantly associated with MVPA (r = .27) and all SDQ variables (rs = -.26-.13) except for externalizing problems. Mediation analyses showed PL was negatively associated with internalizing problems and total difficulties when the association with MVPA was considered. However, the mediating role of MVPA was found only between PL and internalizing problems, ß = -.06, 95%CI [-.12, -.01]. CONCLUSIONS: Although most of our sample was physically active and showed higher adherence to 24-H movement guidelines than comparable population data, the motor competence and self-perceived PL of our sample were similar to those of previous studies. PL has an independent association with children's internalizing problems and total difficulties. Ongoing assessment will investigate the relationships between PL and children's mental health from a longitudinal perspective.

PDK1 inhibition reduces autophagy and cell senescence through the PI3K/AKT signalling pathway in a cigarette smoke mouse emphysema model.

A number of previous studies have demonstrated the pivotal role of PI3K/AKT signalling in cigarette smoke (CS)-induced emphysema, where phosphoinositide dependent protein kinase 1 (PDK1) is a critical component of this pathway. Therefore, the present study aimed to investigate the effects of a PDK1 inhibitor (GSK-2334470) on the expression levels of PI3K, AKT, cyclin-dependent kinase inhibitor 2A (p16) and LC3B in a CS + CS extract (CSE)-induced mouse emphysema model. CS exposure and intraperitoneal injections of CSE were combined for 4 weeks to establish an emphysema model. Mice (n=35) were randomly divided into the normal control, emphysema (CS), PI3K inhibitor (CS3) and PDK1 inhibitor (CS1) groups. Immunohistochemistry staining of lung tissues was used to measure the expression of the PI3K, PDK1 and AKT proteins in airway epithelial tissues. Immunofluorescence staining was also used to measure the levels of p16 and LC3BII protein expression in the airway epithelial tissues. In addition, PI3K, PDK1, AKT, p16 and LC3B protein expression was semi-quantified using western blotting. The expression of PDK1, PI3K and AKT proteins in the airway epithelial tissues was significantly increased in the CS + CSE group compared with that in the control group. The expression levels of p16 and LC3B were also increased as well in the CS + CSE group compared with those in the control group. The expression levels of PI3K, PDK1, AKT, LC3B and p16 in the airway epithelial tissues of the CS3 group were lower compared with those in the CS + CSE group. A decrease in the expression levels of PDK1, AKT, p16 and LC3B in the airway epithelial tissues of the CS1 group compared with those in the CS + CSE group was also observed. However, there were no significant differences in the expression levels of PI3K between the CS1 and the CS groups. The present study concluded that the inhibition of PDK1 can potentially reduce autophagy and cell senescence by downregulating the expression of PI3K/AKT pathway related proteins in airway epithelial cells, thereby protecting against CS + CSE-induced emphysema in mice.

The Effectiveness of Internet-Guided Self-help Interventions to Promote Physical Activity Among Individuals With Depression: Systematic Review.

BACKGROUND: Depression is a prevalent and debilitating mental disorder and a leading cause of disability worldwide. Physical activity (PA) interventions have been shown to alleviate depressive symptoms. However, not all patients have access to PA programing tailored for depression. Internet-guided self-help (IGSH) interventions may be an effective option for increasing PA among people with depression who cannot or prefer not to access supervised exercise treatment. OBJECTIVE: We aimed to evaluate the effectiveness of IGSH interventions in increasing PA and alleviating depressive symptoms in people with depression. METHODS: A systematic literature search was conducted for randomized controlled trials and quasiexperimental studies using 9 electronic databases. The review was registered in PROSPERO (2020 CRD42020221713). RESULTS: A total of 4 randomized controlled trials (430 participants) met the inclusion criteria. Of these, 3 were web-based and 1 was app-based. Three studies found IGSH interventions to have medium to large effects on decreasing depressive symptoms but not on increasing PA compared with waitlist or usual care. One study showed increased self-reported PA but no significant difference in depressive symptoms in the intervention group compared with the control group. Goal setting was the most common behavior change technique used in the interventions. Dropout rates within the intervention groups were relatively low (0%-19%). CONCLUSIONS: Our findings suggested that IGSH PA interventions are feasible and have the potential to reduce depressive symptoms in people with depression. More well-designed and tailored interventions with different combinations of behavior change techniques, particularly those targeting the emotion domain, are needed to assess the overall effectiveness and feasibility of using IGSH interventions to increase PA among people with depression. TRIAL REGISTRATION: PROSPERO CRD42020221713; https://tinyurl.com/ysaua5bu.

Non-invasive brain stimulation combined with psychosocial intervention for depression: a systematic review and meta-analysis.

OBJECTIVES: This review investigates the efficacy and safety of non-invasive brain stimulation (NIBS) combined with psychosocial intervention on depressive symptoms. MATERIALS AND METHODS: We systematically searched five electronic databases from their inception to June 2021: PubMed, Embase, PsycINFO, Web of Science, and Medline. Randomized or non-randomized clinical trials in which NIBS plus psychosocial intervention was compared to control conditions in people with depressive symptoms were included. RESULTS: A total of 17 eligible studies with 660 participants were included. The meta-analysis results showed that NIBS combined with psychosocial therapy had a positive effect on moderate to severe depression ([SMD = - 0.46, 95%CI (- 0.90, - 0.02), I2 = 73%, p < .01]), but did not significantly improve minimal to mild depression ([SMD = - 0.12, 95%CI (- 0.42, 0.18), I2 = 0%, p = .63]). Compared with NIBS alone, the combination treatment had a significantly greater effect in alleviating depressive symptoms ([SMD = - 0.84, 95%CI (- 1.25, - 0.42), I2 = 0%, p = .93]). However, our results suggested that the pooled effect size of ameliorating depression of NIBS plus psychosocial intervention had no significant difference compared with the combination of sham NIBS [SMD = - 0.12, 95%CI (- 0.31, 0.07), I2 = 0%, p = .60] and psychosocial intervention alone [SMD = - 0.97, 95%CI (- 2.32, 0.38), I2 = 72%, p = .01]. CONCLUSION: NIBS when combined with psychosocial intervention has a significant positive effect in alleviating moderately to severely depressive symptoms. Further well-designed studies of NIBS combined with psychosocial intervention on depression should be carried out to consolidate the conclusions and explore the in-depth underlying mechanism.

[Degradation and bioaccumulation characteristics of ciprofloxacin in soil-vegetable system].

A pot experiment was conducted to investigate the effects of applying ciprofloxacin (CIP) on the growth of flowering Chinese cabbage (Brassica campestris), radish (Raphanus sativus), tomato (Solanum lycopersicum) and long bean (Vigna unguiculata), the bioaccumation of CIP in the vegetables, and the degradation of the antibiotics in soil. The results showed that applying 5, 30 and 75 mg x kg(-1) of CIP had no significant effects on the plant biomass of flowering Chinese cabbage, tomato and long bean, but the high concentration of CIP (> or = 30 mg x kg(-1)) inhibited the radish growth significantly. The degradation of CIP in soil was fast at beginning, but slowed down gradually since then. The DT50 and DT90 of CIP in long bean soil were 18.3-43.9 d and 260-738 d, respectively. The CIP could be absorbed by the roots of all the four vegetables and allocated to their aboveground parts, and the CIP residues in the vegetables in creased with the concentration of CIP in soil. The CIP concentration in flowering Chinese cabbage roots was higher than that in the cabbage aboveground parts, that in radish showed a higher concentration in aboveground parts than in roots, and the CIP concentration in tomato and long bean was in the same order of root > stem > leaf.