Vitamin D3 attenuates SARS-CoV-2 nucleocapsid protein-caused hyperinflammation by inactivating the NLRP3 inflammasome through the VDR-BRCC3 signaling pathway in vitro and in vivo.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection-caused coronavirus disease 2019 (COVID-19) is a global crisis with no satisfactory therapies. Vitamin D3 (VD3) is considered a potential candidate for COVID-19 treatment; however, little information is available regarding the exact effects of VD3 on SARS-CoV-2 infection and the underlying mechanism. Herein, we confirmed that VD3 reduced SARS-CoV-2 nucleocapsid (N) protein-caused hyperinflammation in human bronchial epithelial (HBE) cells. Meanwhile, VD3 inhibited the NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome activation in N protein-overexpressed HBE (HBE-N) cells. Notably, the inhibitors of caspase-1, NLRP3, and NLRP3 or caspase-1 small interference RNA (siRNA) enhanced VD3-induced NLRP3 inflammasome inactivation, with subsequent suppression of interleukin-6 (IL6) and IL1ß release in HBE-N cells, which were abolished by the NLRP3 agonist. Moreover, VD3 increased NLRP3 ubiquitination (Ub-NLRP3) expression and the binding of the VDR with NLRP3, with decreased BRCA1/BRCA2-containing complex subunit 3 (BRCC3) expression and NLRP3-BRCC3 association. VD3-induced Ub-NLRP3 expression, NLRP3 inflammasome inactivation, and hyperinflammation inhibition were improved by the BRCC3 inhibitor or BRCC3 siRNA, which were attenuated by the vitamin D receptor (VDR) antagonist or VDR siRNA in HBE-N cells. Finally, the results of the in vivo study in AAV-Lung-enhanced green fluorescent protein-N-infected lungs were consistent with the findings of the in vitro experiment. In conclusion, VD3 attenuated N protein-caused hyperinflammation by inactivating the NLRP3 inflammasome partially through the VDR-BRCC3 signaling pathway.

Protective effect of a dietary flavonoid-rich antioxidant from bamboo leaves against internal exposure to acrylamide and glycidamide in humans.

Polyphenolic antioxidants may effectively reduce acrylamide contents in processed foods. However, few studies focused on their detoxification effects via estimating the profile change of internal exposure biomarkers. Here we showed the protective effect of a water-soluble flavone-C-glycoside-rich antioxidant from bamboo leaves (AOB-w) against acrylamide-induced toxicity in college students. The participants were randomly assigned to either the AOB-w or control group and served potato chips, corresponding to 12.6 µg per kg·bw of dietary exposure to acrylamide, followed by capsules containing 350 mg AOB-w or equivalent placebo. The kinetics of acrylamide, glycidamide, and mercapturic acid metabolites was profiled, and their hemoglobin adducts were measured. The toxicokinetic study showed that AOB-w promoted the excretion of acrylamide and shortened the distribution but prolonged the excretion of N-acetyl-S-(2-carbamoylethyl)-l-cysteine (AAMA) and N-acetyl-S-(2-carbamoyl-2-hydroxyethyl)-l-cysteine. The intervention with AOB-w reduced the peak concentration and area under curve of AAMA by 42.1% and 49.8%, respectively. Besides, AOB-w gender-dependently altered the toxicokinetic profile and reduced the amount of a human-specific urinary biomarker, N-acetyl-S-(2-carbamoylethyl)-l-cysteine-sulfoxide in women. AOB-w accelerated the metabolism of hemoglobin adducts of acrylamide and glycidamide in blood of women. Compared with the baseline levels on the beginning day, we observed a significant enhancement of hemoglobin adducts on the 10th day after serving them potato chips, showing 54.5% and 20.9% higher levels of the hemoglobin adducts of acrylamide and glycidamide, respectively, which thus indicated a lower level of glycidamide-to-acrylamide ratio in blood of participants. Overall AOB-w could effectively reduce the internal exposure to acrylamide in college students, which provides advanced insights into protective functions of natural antioxidants against in vivo toxicity of chemical contaminants from diet.

Influences of personality characteristics and coping modes on anxiety in primary glaucoma patients.

AIM: To examine the influences of personality characteristics and coping modes on the anxiety of primary glaucoma patients. METHODS: A total of 200 individuals, including 50 with primary angle-closure glaucoma, 60 with primary open angle glaucoma and 90 control participants, filled out the State-Trait Anxiety Inventory, NEO Five-Factor Inventory, and Medical Coping Modes Questionnaire. Sociodemographic information was also collected. Data were analyzed via the Spearman rank correlation test and stepwise regression. RESULTS: The personality and coping variables are predictive and jointly account for a significant amount (45.3%-54.2%) of variance across the two subscales of anxiety measures. Notably, neuroticism seems to be most closely related to anxiety disturbances in glaucoma patients. The level of resignation is positively linked to anxiety scores. CONCLUSION: Some personality factors and coping modes help to predict the process of anxiety disorders in primary glaucoma patients. Recognizing the predictive role of these variables in the patients may further enrich clinical research in glaucoma and help to design more effective interventions involving both ophthalmology and psychiatry.

Toxicokinetics and internal exposure of acrylamide: new insight into comprehensively profiling mercapturic acid metabolites as short-term biomarkers in rats and Chinese adolescents.

Acrylamide is classified as a probable carcinogen to humans and generated from Maillard reaction. Currently, the short-term exposure to acrylamide was evaluated via external diet sources in vitro or two main mercapturic acid metabolites: N-acetyl-S-(2-carbamoylethyl)-L-cysteine (AAMA) and N-acetyl-S-(2-carbamoyl-2-hydroxyethyl)-L-cysteine (GAMA) in vivo. In the present work, we comprehensively profiled four mercapturic acid metabolites and evaluated their internal exposure in rats and Chinese adolescents. The cumulative excretion of mercapturic acid metabolites contributes 38.4-73.0 and 43.8-63.6 % of total in vivo metabolites of acrylamide in male and female rats, respectively, when 1, 10, and 50 mg/kg bw of acrylamide were orally administered. Toxicokinetic study revealed that the conversion of acrylamide into glycidamide and glutathione coupling process is highly related to the gender and oral gavage dose via evaluating kinetic parameters, accumulative excretion percentages, and molar ratios of oxidative to reductive metabolism. In human study, a total of 101 Chinese adolescents (41 men and 60 women) were enrolled and served with a meal of potato chips, corresponding to a single-dose (12.6 µg/kg bw) exposure to acrylamide. Toxicokinetic work showed that AAMA is an early and predominant metabolite appearing as a biomarker in urine. N-acetyl-S-(2-carbamoylethyl)-L-cysteine-sulfoxide (AAMA-sul), an oxidative product from AAMA, exhibits a higher peak concentration than GAMA and N-acetyl-S-(1-carbamoyl-2-hydroxyethyl)-L-cysteine (iso-GAMA) during the whole 48-h toxicokinetic period. The internal exposure via four mercapturic acid metabolites is associated with the gender and body mass index characteristics. Thus, current study aims at mercapturic acid metabolites as urinary biomarkers and provides comprehensive insights into the short-term internal exposure to acrylamide.

Decreased expression of sestrin 2 predicts unfavorable outcome in colorectal cancer.

Sestrin 2 is a conserved antioxidant protein that is involved in p53­dependent antioxidant defenses and protects cells against oxidative stresses. The present study was conducted to examine the expression of sestrin 2 in colorectal cancer (CRC) and investigate a possible relationship between sestrin 2 expression and prognosis in CRC. The expression of sestrin 2 in human CRC tissues and cell lines was evaluated by immunohistochemical or immunofluorescent staining and western blot analysis. The correlations between sestrin 2 expression in human CRC tissues and clinicopathological variables, including overall survival (OS) and disease­free survival (DFS), were analyzed. Both human CRC tissues and cell lines showed a decreased expression of sestrin 2. Furthermore, a low expression of sestrin 2 was significantly correlated with advanced tumor stage, lymphatic invasion, lymph node metastasis, vascular invasion and liver metastasis. Survival analysis showed that patients with low sestrin 2 staining had a significantly worse DFS and OS. Additionally, early or advanced stage CRC patients with a low expression of sestrin 2 had a shorter survival. In univariate analysis, the patients with low sestrin 2 expression, advanced tumor stage, lymphatic invasion, lymphatic node metastasis, vascular invasion, liver metastasis and peritoneal metastasis had shorter OS and DFS. In multivariate analysis, only low sestrin 2 expression, advanced tumor stage, lymphatic node metastasis, vascular invasion and liver metastasis remained as independent prognostic factors of poor OS and DFS. The findings suggested that a decreased expression of sestrin 2 is associated with an unfavorable prognosis, which suggests that it is a novel and crucial predictor for CRC metastasis.

High expression of CASK correlates with progression and poor prognosis of colorectal cancer.

Calcium/calmodulin-dependent serine protein kinase (CASK), which localizes at cell-cell adhesion sites and binds to the heparan sulfate proteoglycan syndecan-2, is involved in cell proliferation, cytoskeletal remodeling, and cell migration. To demonstrate the role of CASK in colorectal cancer (CRC) carcinogenesis, we examined the expression of CASK and its binding protein syndecan-2 in human CRC tissues. The expression of CASK was measured in CRC specimens and the controls from adenomas and normal mucosae by immunohistochemical staining and Western blot analysis. Syndecan-2 protein level was tested in CRC samples and the controls by Western blot analysis. The correlations between CASK expression and clinicopathological variables, including disease-free survival (DFS) and overall survival (OS), were analyzed. Compared to the controls, both CASK and syndecan-2 expression were enhanced in CRC tissues. Furthermore, high expression of CASK and syndecan-2 was significantly correlated with advanced tumor stage, lymphatic invasion, lymph node metastasis, vascular invasion, liver metastasis, and unresectable metastatic CRC. Survival analysis showed that patients with low CASK staining had a significantly better survival compared to patients with high CASK staining. In multivariate analysis, CASK overexpression, advanced tumor stage, lymph node metastasis, vasvular invasion, and liver metastasis were independent prognostic factors of poor DFS and OS. Our present study indicates that CASK overexpression is associated with an unfavorable prognosis. CASK is an independent prognostic factor for CRC, which suggests that it is a novel and crucial predictor for CRC metastasis.