Azoramide ameliorates cadmium-induced cytotoxicity by inhibiting endoplasmic reticulum stress and suppressing oxidative stress.

Background: Cadmium (Cd) is hazardous to human health because of its cytotoxicity and long biological half-life. Azoramide is a small molecular agent that targets the endoplasmic reticulum (ER) and moderates the unfolded protein response. However, its role in Cd-induced cytotoxicity remains unclear. This study was performed to investigate the protective effect of azoramide against Cd-induced cytotoxicity and elucidate its underlying mechanisms. Methods: Inductively coupled plasma‒mass spectrometry was used to measure Cd concentrations in each tissue of ICR male mice. The human proximal tubule epithelial cell line HK-2 and the human retinal pigment epithelial cell line ARPE-19 were used in the in vitro study. Cell apoptosis was determined by DAPI staining, JC-1 staining, and annexin V/propidium iodide double staining. Intracellular oxidative stress was detected by MitoSOX red staining, western blot, and quantitative real-time PCR. Moreover, ER stress signaling, MAPK cascades, and autophagy signaling were analyzed by western blot. Results: The present data showed that Cd accumulated in various organs of ICR mice, and the concentrations of Cd in the studied organs, from high to low, were as follows: liver > kidney > testis > lung > spleen > eye. Our study demonstrated that azoramide inhibited ER stress by promoting BiP expression and suppressing the PERK-eIF2α-CHOP pathway. Additionally, we also found that azoramide significantly decreased ER stress-associated radical oxidative species production, attenuated p38 MAPK and JNK signaling, and inhibited autophagy, thus suppressing apoptosis in HK-2 and ARPE-19 cells. Conclusion: Our study investigated the effect of azoramide on Cd-induced cytotoxicity and revealed that azoramide may be a therapeutic drug for Cd poisoning.

Ovatodiolide induces autophagy-mediated cell death through the p62-Keap1-Nrf2 signaling pathway in chronic myeloid leukemia cells.

Ovatodiolide is a macrocyclic diterpenoid compound with various biological activities that displays considerable anticancer potential in different tumor models. However, the underlying mechanism for this antineoplastic activity remains unclear. The aim of the present study was to investigate the anticancer effect and possible molecular mechanism of ovatodiolide in human chronic myeloid leukemia (CML). Ovatodiolide suppressed cell colony formation and induced apoptosis in the K562 and KU812 cells. We also observed that ovatodiolide enhanced the production of reactive oxygen species (ROS), activated Nrf2 signaling, and inhibited mTOR phosphorylation. Autophagic flux was shown to be enhanced after treatment with ovatodiolide in K562 cells. Furthermore, autophagy inhibition alleviated ovatodiolide-induced cell apoptosis, whereas autophagy promotion aggravated apoptosis in CML cells. These results demonstrated that ovatodiolide activates autophagy-mediated cell death in CML cells. Additionally, ovatodiolide transcriptionally activated the expression of p62, and the p62 levels were negatively regulated by autophagy. Moreover, p62-Keap1-Nrf2 signaling was confirmed to be involved in ovatodiolide-induced cell death. Accordingly, LC3B knockdown augmented the ovatodiolide-induced p62 expression, increased the p62-Keap1 interaction, and enhanced the translocation of Nrf2 into the nucleus. In contrast, p62 inhibition abolished the effects that were induced through ovatodiolide treatment. Nrf2 inhibition with ML385 diminished the protective effect of autophagy inhibition in CML cells. Collectively, our results indicate that ovatodiolide induces oxidative stress and provokes autophagy, which effectively decreases the expression of p62 and weakens the protective effect of Nrf2 signaling activation, thus contributing to apoptosis in CML cells.

Polystyrene nanoplastics exposure alters muscle amino acid composition and nutritional quality of Pacific whiteleg shrimp (Litopenaeus vannamei).

Litopenaeus vannamei were exposed to 80-nm polystyrene nanoplastics (NPs) at different concentrations (0, 0.1, 1, 5, and 10 mg/L) for 28 days to study the effects on muscle nutritional quality. Our results showed that with increasing NPs concentrations, the survival rate, specific gain rate, and protein efficiency ratio decreased but the feed conversion ratio increased. There was no significant difference in moisture, ash, and crude lipid content in the muscle, and a general decrease in crude protein content was observed. However, the total amino acid and semi-essential amino acid contents decreased. The spacing between muscle fibers and the melting morphology of muscle increased. The hardness of muscle flesh texture increased, but springiness, cohesiveness, and chewiness decreased. Regarding antioxidant enzyme activity, the activity of catalase decreased, but the total antioxidant capacity, superoxide dismutase activity, and reduced glutathione first increased and then decreased. The expression level of the growth-related genes retinoid X receptor (RXR), chitin synthase (CHS), and calmodulin A (CaM) first increased then decreased, but calcium/calmodulin-dependent protein kinase I (CaMKI), ecdysteroid receptor (EcR), chitinase 5 (CHT5), cell division cycle 2 (Cdc2), and cyclin-dependent kinase 2 (CDK2) decreased. Our results suggest that exposure to NPs can inhibit growth by inducing oxidative stress, which leads to muscle tissue damage and changes in amino acid composition. These results will provide a theoretical reference for the risk assessment of NPs and the ecological health aquaculture of shrimp.

Comparative metabolomics analysis investigating the impact of melatonin-enriched diet on energy metabolism in the crayfish, Cherax destructor.

Melatonin is a multifunctional bioactive molecule present in almost all organisms and has been gradually used in the aquaculture industry in recent years. Energy metabolism is an essential process for individuals to maintain their life activities; however, the process through which melatonin regulates energy metabolism in aquatic animals remains unclear. The present study aimed to conduct a comprehensive analysis of the regulatory mechanism of melatonin for energy metabolism in Cherax destructor by combining metabolomics analysis with the detection of the key substance content, enzymatic activity, and gene expression levels in the energy metabolism process after culturing with dietary melatonin supplementation for 8 weeks. Our results showed that dietary melatonin increased the content of glycogen, triglycerides, and free fatty acids; decreased lactate levels; and promoted the enzymatic activity of pyruvate kinase (PK), malate dehydrogenase (MDH), and acetyl-CoA carboxylase. The results of gene expression analysis showed that dietary melatonin also increased the expression levels of hexokinase, PK, MDH, lactate dehydrogenase, lipase, and fatty acid synthase genes. The results of metabolomics analysis showed that differentially expressed metabolites were significantly enriched in lysine degradation and glycerophospholipid metabolism. In conclusion, our study demonstrates that dietary melatonin increased oxidative phosphorylation, improved glucose utilization, and promoted storage of glycogen and lipids in C. destructor. These lipids are used not only for energy storage but also to maintain the structure and function of cell membranes. Our results further add to the understanding of the mechanisms of energy regulation by melatonin in crustaceans.

Effects of ß-1,3-glucan on growth, immune responses, and intestinal microflora of the river prawn (Macrobrachium nipponense) and its resistance against Vibrio parahaemolyticus.

In this study, we investigated the impact of ß-1,3-glucan on the immune responses and gut microbiota of the river prawn (Macrobrachium nipponense) in the presence of Vibrio parahaemolyticus stress. Shrimps were fed one of the following diets: control (G1), 0.2% curdlan (G2), 0.1% ß-1,3-glucan (G3), 0.2% ß-1,3-glucan (G4), or 1.0% ß-1,3-glucan (G5) for 6 weeks and then challenged with V. parahaemolyticus for 96 h. Under Vibrio stress, shrimps in G4 exhibited the highest length gain rate, weight gain rate, and survival rate. They also showed increased intestinal muscle thickness and villus thickness compared to the control and 0.2% curdlan groups. The apoptosis rate was lower in G4 than in the control group, and the digestive enzyme activities (pepsin, trypsin, amylase, and lipase), immune enzyme activities (acid phosphatase, alkaline phosphatase, lysozyme, and phenoxidase), and energy metabolism (triglyceride, cholesterol, glycogen, and lactate dehydrogenase) were enhanced. Expression levels of growth-related genes (ecdysone receptor, calmodulin-dependent protein kinase I, chitin synthase, and retinoid X receptor) and immune-related genes (toll-like receptor 3, myeloid differentiation primary response 88, mitogen-activated protein kinase 7, and mitogen-activated protein kinase 14) were higher in G4 than in the control. Microbiota analysis indicated higher bacterial abundance in shrimps fed ß-1,3-glucan, as evidenced by Sob, Chao1, and ACE indices. Moreover, 0.2% ß-1,3-glucan increased the relative abundances of Bacteroidota and Firmicutes while reducing those of Corynebacteriales and Lactobacillales. In summary, ß-1,3-glucan enhances immune enzyme activities, alters immune-related gene expression, and impacts gut microbial diversity in shrimp. These findings provide valuable insights into the mechanisms underlying ß-1,3 glucan's immune-enhancing effects.

Dietary melatonin positively impacts the immune system of crayfish, Cherax destructor, as revealed by comparative proteomics analysis.

Melatonin, an indoleamine with various biological activities, is being used increasingly in the aquaculture industry for its broad immune effects. Cherax destructor is an emerging economically cultured crayfish that faces many problems in the breeding process. Previous work found that dietary melatonin has positive effects on the growth and immunity of C. destructor, but the specific mechanism involved remained unclear. In this study, proteomics was used to determine the mechanism of action of melatonin in C. destructor. Results showed that dietary melatonin resulted in decreased levels of hydrogen peroxide, alanine aminotransferase, and aspartate aminotransferase, but increased levels of glutathione peroxidase, acid phosphatase, and glutathione S-transferases. In total, 608 proteins were differentially expressed (418 upregulated and 190 downregulated), and were enriched in three main categories: innate immunity (B cell receptor signaling pathway and natural killer cell-mediated cytotoxicity), glucose metabolism (pentose phosphate pathway, pentose and glucuronate interconversions, and propionate metabolism), and amino acid metabolism (valine, leucine, and isoleucine degradation, and cysteine and methionine metabolism). In addition, dietary melatonin was also involved in the regulation of the mTOR signaling pathway, and upregulated the expression of genes encoding key factors, such as Ras-related GTP-binding protein A/B, eukaryotic initiation factor 4E, eukaryotic initiation factor 4E-binding protein, and p70 ribosomal S6 kinase. Overall, this study demonstrates the role of melatonin in the physiological regulation of C. destructor, laying the foundation for the development of melatonin as a feed additive in the aquaculture of this species.

Comparison of Growth Performance and Biochemical Components between Low-Salinity-Tolerant Hybrid and Normal Variety of Pacific White Shrimp (Penaeus vannamei).

Penaeus vannamei, a high-yield economical shrimp, is confronting germplasm degradation in the culture environments found in China, which results in a sharp drop in production. Genetic improvement by hybridization is an effective way to solve this problem. In this study, we selected the hybrid species adapted to low-salinity culture obtained by intraspecific crossing as the experimental group. The control group consisted of normal variety from the Hainan Lutai Company. The two groups of shrimps were cultured for three months under salinities of 1 PSU, 5 PSU, and 15 PSU. Growth-performance-related indicators, biochemical composition, and molting-related gene expression were examined. The results showed that at salinities of 1 PSU and 5 PSU, the survival rate and growth performance of the low-salt breeding group were better than those of the normal variety population. The digestive enzyme activity in the low-salt breeding group was higher, which was consistent with its better growth performance, and was also associated with higher triglyceride, total cholesterol, and glycogen content. Lower levels of lactic acid indicated less anaerobic metabolism and better adaptability to the environment. The amino acid and fatty acids analysis showed that levels of essential amino acids and high unsaturated fatty acids were both higher in the low-salt breeding group than in the normal variety shrimp cultured in a low-salinity environment. The expression levels of genes associated with molting (CHS, CaMKI, RXR, EcR, HSP60, and HSP70) were also higher in the low-salt breeding group than in the control group. The results indicated that the hybrid shrimp showed better growth performance and nutritional advantages compared with the normal shrimp under salinities of 1 PSU and 5 PSU. This research provides a valuable reference for subsequent genetic breeding and shrimp culture.

Effects of dietary soluble ß-1,3-glucan on the growth performance, antioxidant status, and immune response of the river prawn (Macrobrachium nipponense).

The effects of dietary ß-1,3-glucan on the growth performance, body composition, hepatopancreas tissue structure, antioxidant activities, and immune response of the river prawn (Macrobrachium nipponense) were investigated. In total, 900 juvenile prawns were fed one of five diets with different contents of ß-1,3-glucan (0%, 0.1%, 0.2%, and 1.0%) or 0.2% curdlan for 6 weeks. The growth rate, weight gain rate, specific growth rate, specific weight gain rate, condition factor, and hepatosomatic index of juvenile prawns fed 0.2% ß-1,3-glucan were significantly higher than those fed 0% ß-1,3-glucan and 0.2% curdlan (p < 0.05). The whole-body crude lipid content of prawns supplemented with curdlan and ß-1,3-glucan was significantly higher than that of the control group (p < 0.05). The antioxidant and immune enzyme activities of superoxide dismutase (SOD), total antioxidant capacity (T-AOC), catalase (CAT), lysozyme (LZM), phenoloxidase (PO), acid phosphatase (ACP), and alkaline phosphatase (AKP) in the hepatopancreas of juvenile prawns fed 0.2% ß-1,3-glucan were significantly higher than those of the control and 0.2% curdlan groups (p < 0.05), and tended to increase and then decrease with increasing dietary ß-1,3-glucan. The highest malondialdehyde (MDA) content was observed in juvenile prawns without ß-1,3-glucan supplementation. The results of real-time quantitative PCR indicated that dietary ß-1,3-glucan promoted expression of antioxidant and immune-related genes. Binomial fit analysis of weight gain rate and specific weight gain rate showed that the optimum ß-1,3-glucan requirement of juvenile prawns was 0.550%-0.553%. We found that suitable dietary ß-1,3-glucan improved juvenile prawns growth performance, antioxidant capacity, and non-specific immunity, which provide reference for shrimp healthy culture.

CSF TNF-α Levels Were Associated with Longitudinal Change in Brain Glucose Metabolism Among Non-Demented Older People.

PURPOSE: Emerging studies have suggested that tumor necrosis factor-alpha (TNF-α) is implicated in the pathogenesis of Alzheimer's disease (AD), and that cerebral glucose hypometabolism is a key feature of AD. However, the association of CSF TNF-α levels with changes in cerebral glucose metabolism has not been studied among non-demented older people. PATIENTS AND METHODS: At baseline, there were a total of 214 non-demented older people from Alzheimer's Disease Neuroimaging Initiative (ADNI) study. We examined the cross-sectional and longitudinal associations of CSF TNF-α with global cognition (as assessed by mini-mental state examination), verbal memory (as assessed by Rey Auditory Verbal Learning Test-total learning score), and cerebral glucose metabolism (as measured by FDF-PET). Linear mixed-effects models were used to examine the longitudinal association of CSF TNF- α with change in each outcome over time with adjustment of age, educational level, gender, and APOE4 status. RESULTS: In the cross-sectional study, CSF TNF-α was negatively associated with MMSE scores, but not verbal memory or FDG-PET. In the longitudinal study, higher CSF TNF- α at baseline was associated with a faster decline in cerebral glucose metabolism, but not MMSE scores or RAVLT total learning scores. CONCLUSION: Higher CSF TNF-α levels were associated with a steeper decline in cerebral glucose metabolism among non-demented older people.

CSF VEGF Was Positively Associated with Neurogranin Independent of ß-Amyloid Pathology.

PURPOSE: Increasing evidence suggests that both vascular endothelial growth factor (VEGF) and synaptic failure have been involved in the pathogenesis of Alzheimer's disease (AD). However, it is not clear whether CSF VEGF levels are associated with synaptic function in living human. PATIENTS AND METHODS: In the present study, we included a total of 291 older individuals, including 83 individuals with normal cognition (NC), 143 individuals with mild cognitive impairment (MCI) and 65 patients with AD. Several linear regression models were conducted to examine the associations of CSF VEGF with CSF neurogranin levels (NG, reflecting synaptic degeneration) when controlling for other potential confounding factors, including age, gender, years of education, clinical diagnosis, APOE4 genotype and CSF ß-amyloid 42 (Aß 42) levels. RESULTS: There was no significant difference in VEGF levels between the three diagnostic groups. In the pooled sample, females had significantly lower levels of VEGF than males. Aß-positive (CSF Aß 42 < 192 pg/mL) individuals had lower levels of VEGF than Aß-negative individuals. However, the relationships between VEGF and NG levels were not modified by disease stage. Finally, we found that CSF VEGF levels were associated with NG levels with adjustment of age, gender, years of education, clinical diagnosis, APOE4 genotype and CSF Aß 42 levels. CONCLUSION: CSF VEGF levels were associated with NG independent of AD pathology and disease stage.

Association of Clusterin Levels in Cerebrospinal Fluid with Synaptic Degeneration Across the Alzheimer's Disease Continuum.

PURPOSE: Although emerging evidence has suggested that clusterin is involved in the pathogenesis of Alzheimer's disease (AD), the association of clusterin with synaptic degeneration in living human is unclear. In the present study, we aimed to examine the association of CSF clusterin levels with synaptic degeneration in individuals with different severities of cognitive impairment. PATIENTS AND METHODS: In the present study, we compared levels of clusterin in CSF among individuals with normal cognition (NC), mild cognitive impairment (MCI), and AD. Further, linear regression models were performed to examine the association of CSF clusterin with neurogranin (NG, reflecting synaptic degeneration) with adjustment of several potential confounders. RESULTS: We found that CSF clusterin levels were positively correlated with NG in the NC and MCI groups, but not the AD group. In all subjects, linear regression models suggested that clusterin levels were positively associated with NG levels independent of age, gender, apolipoprotein E4 (APOE4) genotype, clinical diagnosis, and CSF Aß42 levels. CONCLUSION: Our data indicated that clusterin was associated with CSF NG levels among older individuals with different severities of cognitive impairment.

Stress Hyperglycemia and Outcome of Non-diabetic Patients After Acute Ischemic Stroke.

Background and Purpose: Stress hyperglycemia is relative hyperglycemia after suffering an acute illness such as stroke, even without preexisting diabetes. Our study aimed to determine the relationship between stress hyperglycemia and outcome of non-diabetic patients with acute ischemic stroke. Methods: Data were derived from the ACROSS-China (Abnormal Glucose Regulation in Patients with Acute Stroke across China) registry. Patients with ischemic stroke but without a history of diabetes mellitus were included in this analysis. Stress hyperglycemia was measured by glucose-to-glycated hemoglobin (HbA1c) ratio which was calculated by fasting plasma glucose (FPG) divided by HbA1c. We investigated the association between quartiles of glucose-to-HbA1c ratio and stroke recurrence and all-cause death at 12 months after stroke onset. Results: We included a total of 999 patients, among which there were 105 (10.9%) recurrent strokes and 76 (7.6%) deaths at 12 months. Using the lowest quartile of glucose-to-HbA1c ratio as the reference, patients with the highest quartile were associated with an increased risk of stroke recurrence [16.0 vs. 8.3%; adj.hazards ratio (HR) = 2.19, 95% confidence interval (CI): 1.26-3.83] and death (13.0 vs. 4.3%; adj.HR=2.86, 95%CI: 1.38-5.90) at 12 months after adjusted for potential covariates. We also observed that a higher level of glucose-to-HbA1c ratio was associated with an elevated risk of stroke recurrence and death. Conclusion: Stress hyperglycemia, measured by glucose-to-HbA1c ratio, was related to an elevated risk of stroke recurrence and all-cause death in patients with acute ischemic stroke but without diabetes.

Elevated Neutrophil and Presence of Intracranial Artery Stenosis Increase the Risk of Recurrent Stroke.

Background and Purpose- The association of neutrophil and intracranial artery stenosis (ICAS) with the prognosis of stroke is uncertain. This study evaluated the relationship between neutrophil levels with and without ICAS and the prognosis of patients with minor stroke or transient ischemic attack. Methods- Data from the CHANCE trial (Clopidogrel in High-Risk Patients With Acute Nondisabling Cerebrovascular Events) was reviewed. Patients were divided into 4 groups according to the dichotomy of neutrophil counts and status of ICAS. The primary outcome was a new stroke (ischemic or hemorrhagic), and secondary outcomes included a new composite vascular event (stroke, myocardial infarction, or cardiovascular death) and ischemic stroke. Safety outcome was any hemorrhage at 90 days. The association between neutrophil counts with and without ICAS and risk of any outcome was analyzed by Cox regression models. Results- Of 1034 patients included in this subgroup analysis, 91 had recurrent strokes. Compared with the lower neutrophil levels without ICAS, higher neutrophil levels with ICAS significantly increased the risk of stroke recurrence (adjusted hazard ratio, 2.26; 95% CI, 1.19-4.31; P=0.01) and composite outcome (adjusted hazard ratio, 1.98; 95% CI, 1.06-3.67; P=0.03). However, there was no safety issue. Conclusions- Concomitant presence of higher neutrophil levels and ICAS was associated with the increased risk of stroke recurrence, and combined adverse outcome events in patients already had minor ischemic stroke or high-risk transient ischemic attack. Clinical Trial Registration- URL: https://www.clinicaltrials.gov . Unique identifier: NCT00979589.

Neutrophil counts, neutrophil ratio, and new stroke in minor ischemic stroke or TIA.

OBJECTIVE: Evidence about whether neutrophil counts or neutrophil ratio is associated with new stroke is scant. The aim of this study is to assess the association of neutrophil counts or neutrophil ratio with a new stroke in patients with minor stroke or TIA. METHODS: We derived data from the Clopidogrel in High-Risk Patients With Acute Nondisabling Cerebrovascular Events trial. Patients with a minor stroke or TIA were categorized into 4 groups according to the quartile of neutrophil counts or neutrophil ratio. The primary outcome was a new stroke (ischemic or hemorrhagic), and secondary outcomes included a new composite vascular event (stroke, myocardial infarction, or death resulting from cardiovascular causes) and ischemic stroke during the 90-day follow-up. We assessed the association between neutrophil counts, neutrophil ratio, and risk of new stroke. RESULTS: A total of 4,854 participants were enrolled, among whom 495 had new strokes at 90 days. Compared with the first quartile, the second, third, and fourth quartiles of neutrophil counts were associated with increased risk of new stroke (adjusted hazard ratio 1.40 [95% confidence interval (CI) 1.05-1.87], 1.55 [95% CI 1.17-2.05], and 1.69 [95% CI 1.28-2.23], respectively, p for trend <0.001). Similar results were observed for the endpoint of composite events and ischemic stroke. Parallel results were found for neutrophil ratio. CONCLUSION: High levels of both neutrophil counts and neutrophil ratio were associated with an increased risk of new stroke, composite events, and ischemic stroke in patients with a minor ischemic stroke or TIA.

[Status and drug resistance surveillance of enterococcus infection in patients with severe hepatitis].

OBJECTIVE: To analyze the infection status and the drug resistance of enterococci in patients with severe hepatitis to guide future treatment. METHODS: All bacteria from infected patients with severe hepatitis were cultured with BacT/Alert120 automation instrument (Aksu) and identified with Vitek-AMS60 (Biomerieux). Drug sensitivities of the isolated enterococci were tested with 11 antibacterial agents. RESULTS: Among the 112 isolated enterococci, Enterococcus faecalis was the most preponderant bacterium, and the second was E. faecium. Their isolation rates were 79.5% and 14.3%, respectively. 57.1% of all the enterococci were found in the ascetic fluid of patients with severe hepatitis. Fifty-eight (51.8%) isolated enterococci were found to be high level aminoglycoside resistant (HLAR), 19 (17.0%) enterococci were ampicillin-resistant enterococcus (ARE) and 7 (6.3%) were both HLAR and ARE. The susceptive rates of the enterococci to vancomycin and teicoplanin were very high, namely 96.4% and 100%, respectively. No vancomycin or teicoplanin resistant enterococci were found, but 4 enterococci were mildly sensitive to vancomycin. CONCLUSION: Enterococcus faecalis is the most prevalent species isolated in severe hepatitis patients infected with enterococcal infection. From our study, vancomycin and teicoplanin are the drugs of first choice to treat those infections.