Vitamin B6 Competition in the Tumor Microenvironment Hampers Antitumor Functions of NK Cells.

Nutritional factors play crucial roles in immune responses. The tumor-caused nutritional deficiencies are known to affect antitumor immunity. Here, we demonstrate that pancreatic ductal adenocarcinoma (PDAC) cells can suppress NK-cell cytotoxicity by restricting the accessibility of vitamin B6 (VB6). PDAC cells actively consume VB6 to support one-carbon metabolism, and thus tumor cell growth, causing VB6 deprivation in the tumor microenvironment. In comparison, NK cells require VB6 for intracellular glycogen breakdown, which serves as a critical energy source for NK-cell activation. VB6 supplementation in combination with one-carbon metabolism blockage effectively diminishes tumor burden in vivo. Our results expand the understanding of the critical role of micronutrients in regulating cancer progression and antitumor immunity, and open new avenues for developing novel therapeutic strategies against PDAC. SIGNIFICANCE: The nutrient competition among the different tumor microenvironment components drives tumor growth, immune tolerance, and therapeutic resistance. PDAC cells demand a high amount of VB6, thus competitively causing NK-cell dysfunction. Supplying VB6 with blocking VB6-dependent one-carbon metabolism amplifies the NK-cell antitumor immunity and inhibits tumor growth in PDAC models. This article is featured in Selected Articles from This Issue, p. 5.

Recombinant tissue-type plasminogen activator (rt-PA) effectively restores neurological function and improves prognosis in acute ischemic stroke.

OBJECTIVE: To evaluate the clinical efficacy of intravenous thrombolysis with recombinant tissue-type plasminogen activator (rt-PA) for acute ischemic stroke. METHODS: A total of 76 patients with acute ischemic stroke admitted to the Encephalopathy Dept. of Zhecheng Hospital of Traditional Chinese Medicine between February 2021 and June 2022 were recruited in this prospective trial (ClinicalTrials.gov, NCT03884410) and patients were randomized 1:1 to receive either aspirin plus clopidogrel (control group) or aspirin plus clopidogrel and rt-PA intravenous thrombolytic therapy (experimental group), with 38 cases in each group. The treatment efficiency, National Institute of Health stroke scale (NIHSS) scores, daily living ability, coagulation function, serum Lipoprotein-associated phospholipaseA2 (Lp-PLA2), homocysteine (HCY), hypersensitive C-reactive protein (hsCRP) levels, adverse events, and prognosis were evaluated and compared between the two groups. RESULTS: Intravenous thrombolysis with rt-PA resulted in a better treatment outcome of patients versus aspirin plus clopidogrel (P<0.05). Patients with rt-PA exhibited better improvement in neurological function than those with aspirin plus clopidogrel, as shown by the lower NIHSS scores (P<0.05). Intravenous thrombolysis with rt-PA resulted in a better quality of life of patients than aspirin plus clopidogrel, indicated by the higher Barthel Index (BI) levels (P<0.05). The lower von Willebrand factor (vWF) and Factor VIII (F) levels indicated better coagulation function of patients with rt-PA versus those with aspirin plus clopidogrel (P<0.05). The lower serum concentrations of Lp-PLA2, HCY, and hsCRP suggested patients with rt-PA had milder inflammatory responses versus those without rt-PA (P<0.05). There was no significant difference in the incidence of adverse events in the two groups (P>0.05). Intravenous thrombolytic therapy with rt-PA better enhanced the prognosis of patients than with aspirin plus clopidogrel (P<0.05). CONCLUSION: Compared with conventional pharmacological regimens, additional rt-PA intravenous thrombolytic therapy improves the clinical outcome of patients with acute ischemic stroke, promotes neurological recovery, and enhances patient prognosis without increasing the risk of patient-related adverse events.

Biological Selenite Reduction, Characterization and Bioactivities of Selenium Nanoparticles Biosynthesised by Pediococcus acidilactici DSM20284.

Selenium (Se) is in great demand as a health supplement due to its superior reactivity and excellent bioavailability, despite selenium nanoparticles (SeNPs) having signs of minor toxicity. At present, the efficiency of preparing SeNPs using lactic acid bacteria is unsatisfactory. Therefore, a probiotic bacterial strain that is highly efficient at converting selenite to elemental selenium is needed. In our work, four selenite-reducing bacteria were isolated from soil samples. Strain LAB-Se2, identified as Pediococcus acidilactici DSM20284, had a reduction rate of up to 98% at ambient temperature. This strain could reduce 100 mg L-1 of selenite to elemental Se within 48 h at pH 4.5-6.0, a temperature of 30-40 °C, and a salinity of 1.0-6.5%. The produced SeNPs were purified, freeze-dried, and subsequently systematically characterised using FTIR, DSL, SEM-EDS, and TEM techniques. SEM-EDS analysis proved the presence of selenium as the foremost constituent of SeNPs. The strain was able to form spherical SeNPs, as determined by TEM. In addition, DLS analysis confirmed that SeNPs were negatively charged (-26.9 mV) with an average particle size of 239.6 nm. FTIR analysis of the SeNPs indicated proteins and polysaccharides as capping agents on the SeNPs. The SeNPs synthesised by P. acidilactici showed remarkable antibacterial activity against E. coli, B. subtilis, S. aureus, and K. pneumoniae with inhibition zones of 17.5 mm, 13.4 mm, 27.9 mm, and 16.2 mm, respectively; they also showed varied MIC values in the range of 15-120 µg mL-1. The DPPH, ABTS, and hydroxyl, and superoxide scavenging activities of the SeNPs were 70.3%, 72.8%, 95.2%, and 85.7%, respectively. The SeNPs synthesised by the probiotic Lactococcus lactis have the potential for safe use in biomedical and nutritional applications.

Integrated eco-physiological, biochemical, and molecular biological analyses of selenium fortification mechanism in alfalfa.

MAIN CONCLUSION: Foliar Se (IV) application at 100 mg/kg can act as a positive bio-stimulator of redox, photosynthesis, and nutrient metabolism in alfalfa via phenotypes, nutritional compositions, biochemistry, combined with transcriptome analysis. Selenium (Se) is an essential element for mammals, and plants are the primary source of dietary Se. However, Se usually has dual (beneficial/toxic) effects on the plant itself. Alfalfa (Medicago sativa L.) is one of the most important forage resources in the world due to its high nutritive value. In this study, we have investigated the effects of sodium selenite (Se (IV)) (0, 100, 200, 300, and 500 mg/kg) on eco-physiological, biochemical, and transcriptional mechanisms in alfalfa. The phenotypic and nutritional composition alterations revealed that lower Se (IV) (100 mg/kg) levels positively affected alfalfa; it enhanced the antioxidant activity, which may contribute to redox homeostasis and chloroplast function. At 100 mg/kg Se (IV) concentration, the H2O2, and malondialdehyde (MDA) contents decreased by 36.72% and 22.62%, respectively, whereas the activity of glutathione peroxidase (GPX) increased by 31.10%. Se supplementation at 100 mg/kg increased the plant pigments contents, the light-harvesting capacity of PSII (Fv/Fm) and PSI (ΔP700max), and the carbon fixation efficiency, which was demonstrated by enhanced photosynthesis (37.6%). Furthermore, alfalfa shifted carbon flux to protein synthesis to improve quality at 100 mg/kg of Se (IV) by upregulating carbohydrate and amino acid metabolic genes. On the contrary, at 500 mg/kg, Se (IV) became toxic. Higher Se (IV) disordered the plant antioxidant system, increasing H2O2 and MDA by 14.2 and 4.3%, respectively. Moreover, photosynthesis was inhibited by 20.2%, and more structural substances, such as lignin, were synthesized. These results strongly suggest that Se (IV) at a concentration of 100 mg/kg act as the positive bio-stimulator of redox metabolism, photosynthesis, and nutrient in alfalfa.

The FDA-Approved Anthelmintic Pyrvinium Pamoate Inhibits Pancreatic Cancer Cells in Nutrient-Depleted Conditions by Targeting the Mitochondria.

Pancreatic ductal adenocarcinoma (PDAC) is a lethal aggressive cancer, in part due to elements of the microenvironment (hypoxia, hypoglycemia) that cause metabolic network alterations. The FDA-approved antihelminthic pyrvinium pamoate (PP) has previously been shown to cause PDAC cell death, although the mechanism has not been fully determined. We demonstrated that PP effectively inhibited PDAC cell viability with nanomolar IC50 values (9-93 nmol/L) against a panel of PDAC, patient-derived, and murine organoid cell lines. In vivo, we demonstrated that PP inhibited PDAC xenograft tumor growth with both intraperitoneal (IP; P < 0.0001) and oral administration (PO; P = 0.0023) of human-grade drug. Metabolomic and phosphoproteomic data identified that PP potently inhibited PDAC mitochondrial pathways including oxidative phosphorylation and fatty acid metabolism. As PP treatment reduced oxidative phosphorylation (P < 0.001), leading to an increase in glycolysis (P < 0.001), PP was 16.2-fold more effective in hypoglycemic conditions similar to those seen in PDAC tumors. RNA sequencing demonstrated that PP caused a decrease in mitochondrial RNA expression, an effect that was not observed with established mitochondrial inhibitors rotenone and oligomycin. Mechanistically, we determined that PP selectively bound mitochondrial G-quadruplexes and inhibited mitochondrial RNA transcription in a G-quadruplex-dependent manner. This subsequently led to a 90% reduction in mitochondrial encoded gene expression. We are preparing to evaluate the efficacy of PP in PDAC in an IRB-approved window-of-opportunity trial (IND:144822).

Effects of incremental endosulfan sulfate exposure and high fat diet on lipid metabolism, glucose homeostasis and gut microbiota in mice.

The influence of pollutants on metabolic diseases such as type 2 diabetes mellitus is an emerging field in environmental medicine. Here, we explored the effects of a low-dose endosulfan sulfate (ES), a major metabolite of the pesticide endosulfan and a bio-persistent contaminant detected in environmental and human samples, on the progress of obesity and metabolic disorders. Pregnant CD-1 mice were given ES from gestational day 6 to postnatal day 21 (short-term). After weaning, male pups of exposed dams were provided with a low-fat or a high-fat diet (LFD or HFD) and assessed after an additional 12 weeks. At the same time, one group of male pups continuously received ES (long-term). Treatment with low-dose ES, short or long-term, alleviated the development of obesity and accumulation of hepatic triglycerides induced by HFD. Analysis of gene expression, metabolic profile and gut microbiome indicates that ES treatment inhibits adipogenesis induced by HFD due to enhanced lipid catabolism, fatty acid oxidation and disturbance of gut microbiota composition. However, impaired glucose and insulin homeostasis were still conserved in HFD-fed mice exposed to ES. Furthermore, ES treatment impaired glucose tolerance, affected hepatic gene expression, fatty acids composition and serum metabolic profile, as well as disturbed gut microbiota in LFD-fed mice. In conclusion, ES treatment at levels close to the accepted daily intake during fetal development directly impact glucose homeostasis, hepatic lipid metabolism, and gut microbiome dependent on the type of diet consumed. These findings provide a better understanding of the complex interactions of environmental pollutants and diet at early life stages also in the context of metabolic disease.

The effects of hexaconazole and epoxiconazole enantiomers on metabolic profile following exposure to zebrafish (Danio rerio) as well as the histopathological changes.

Hexaconazole and epoxiconazole are the worldwidely used fungicides. However, limited information is known about the toxicological effects of their enantiomers on aquatic organisms. In this study, zebrafish were separately exposed to 100 and 1000 µgL-1 hexaconazole and epoxiconazole enantiomers for 21 d 1H NMR-based metabolomics analysis showed that the exposure of low and high dose of hexaconazole enantiomers altered energy metabolism, lipid metabolism and amino acid metabolism of zebrafish, with the different metabolic profiles resulted from the same dose of (+)-hexaconazole and (-)-hexaconazole. Similar to hexaconazole enantiomers, the metabolic profiles, including the changes related to energy metabolism, lipid metabolism and amino acid metabolism, were demonstrated in low and high dose epoxiconazole enantiomers treatment groups. There are differences in the metabolic profiles of zebrafish between exposed to (+)-epoxiconazole and (-)-epoxiconazole of the same dose. The results of histological examination revealed that the exposure of both enantiomers for hexaconazole and epoxiconazole resulted in the similar histopathological changes. The exposure of hexaconazole and epoxiconazole enantiomers at low and high dose resulted the vacuolization and swell in the liver of the female and male zebrafish. Compared to female zebrafish, more liver damage was found in male zebrafish in the hexaconazole enantiomers exposure groups. The reduction of spermatids was observed in hexaconazole and epoxiconazole enantiomers treatment groups of both doses. Hexaconazole enantiomers exposure of low and high dose resulted the increase in the number of mature eggs, while such effect was not observed in epoxiconazole enantiomers exposure groups. Hexaconazole and epoxiconazole enantiomers exposure resulted in no changes in brains of female and male zebrafish. As a result, both triazole-based chiral bactericides, hexaconazole and epoxiconazole, have similar toxicological effects but their mechanisms of action are not exactly the same. The above results will play an important part in making the differences in toxic effects of hexaconazole and epoxiconazole enantiomers clear. What's more, it is an indispensable part for an integrated environmental risk assessment.

Ecological risk assessment of alpha-cypermethrin-treated food ingestion and reproductive toxicity in reptiles.

Pesticides are proposed as one of the many causes for the global decline in reptile population. To understand the potential impact of alpha-cypermethrin (ACP) in reptiles, in the current study, we used a tri-trophic food web (plants - herbivores - natural enemies of predators) to examine the reproductive toxicity and biomarker changes. Based on the Maximum Residue Limit (MRL) of ACP in several agricultural products, we designed three concentrations 0, 2 (MRL), and 20 mg/kg wet weight as three treatment groups for this research. Male and female lizards were fed ACP contaminated or uncontaminated diets for eight weeks during the breeding phase. The number of deaths was different among the three groups, and a dose-dependent trend was found. Decreases in food consumption of 26.6% and 28.1% were observed in the low- and high-dose group, respectively. Dietary exposure significantly induced a dose-dependent decrease in body mass index in lizards. Significant variations in glutathione-S-transferaseb activities, catalase activities, and malondialdehyde levels in gonads, suggest that lizards were under oxidative stress. In addition, ACP exposure altered sexual hormone levels in males, reduced reproductive output of females, and induced histopathological changes in testes. These negative effects highlight that ACP dietary exposure is a potential threat to lizards' reproduction.

NMR- and LC-MS/MS-based urine metabolomic investigation of the subacute effects of hexabromocyclododecane in mice.

In the present study, both untargeted and targeted metabolomics approaches were used to evaluate the subacute effects of hexabromocyclododecane (HBCD) on mice urine metabolome. Untargeted metabolomics based on (1)H NMR showed that HBCD exposure disturbed mice metabolism in both dosed groups, especially in high dosed group. The low-dose HBCD led to a decrease in alanine, malonic acid, and trimethylamine (TMA). High-dose HBCD-treated mice developed high levels of citric acid and 2-ketoglutarate, together with decreased alanine, acetate, formate, TMA, 3-hydroxybutyrate, and malonic acid. Targeted metabolomics for metabolic profiling of 20 amino acids identified alanine, lysine, and phenylalanine as significantly disturbed metabolites. These results indicated that subchronic exposure to HBCD caused a disturbance of mice metabolism, especially in TCA cycle, lipid metabolism, gut microbial metabolism, and homeostasis of amino acids, and the application of untargeted and targeted metabolomics combined with conventional toxicology approaches to evaluate the subacute effects of pollutants will provide more comprehensive information and aid in predicting health risk of these pollutants.

The Hypoglycemic and Synergistic Effect of Loganin, Morroniside, and Ursolic Acid Isolated from the Fruits of Cornus officinalis.

Hypoglycemic activity-guided separation of ethanol extracts from the fruits of Cornus officinalis Sieb. et Zucc (CO) led to the isolation of loganin, morroniside, and ursolic acid. The antidiabetic capacity of CO extracts and related compounds was further investigated in diabetes mellitus mice. The results suggested that both CO extracts and pure compounds could ameliorate diabetes-associated damages and complications. Oral administration of loganin and morroniside decreased fasting blood glucose levels in diabetes mellitus mice. Ursolic acid exhibited the highest reactive oxygen species scavenging activity and α-glucosidase inhibitory activity. Notably, we noticed an interesting synergistic effect between loganin and ursolic acid. Given these favorable hypoglycemic properties, C. officinalis, a food and medicinal plant in China, may be used as a valuable food supplement for the treatment of diabetes mellitus.

[Effects of alkaloids from coptidis rhizoma on blood lipid metabolism and low-denstity lipoprotein receptor mRNA in golden hamsters].

To study the effects of alkaloids from Coptidis Rhizoma on low-density lipoprotein receptor (LDLR) mRNA expression and antihyperlipedemic levels. The LDLR mRNA expression were detected by real time fluorescence quantitative PCR, and the levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL-c) and high-density lipoprotein cholesterol (HDL-c) in serum were measured at the first and last examination. The results show that, after the drug treatment, compared with the model group, each drug group showed a lipid-lowering effect. Especially, coptisine, palmatine, jatrorrhinze were significantly reduced TC, TG, LDL-c (P < 0.05, P < 0.01), and increased HDL-c (P < 0.01). In addition, they also increased mRNA expression of the LDLR in liver and HepG2 cells. The results showed that alkaloids from Coptidis Rhizoma can regulate lipid metabolism disorder, and coptisine have the best lipid-lowering effect.

[Effect of different parts, harvesting time and processing technologies on alkaloids content of Coptis chinensis adventitious root].

OBJECTIVE: To investigate the effect of different parts, harvesting time and processing technologies on alkaloids content of Coptis chinensis adventitious root. METHODS: The content of alkaloids were analyzed by HPLC. RESULTS: The content of total alkaloids in adventitious root harvested in different time was ranged from 2.5% to 2.9%, in which that of berberine and coptisine were the highest, reaching to 1%, and that of palmatine was only 0.1%. It suggested there was no significant difference of total alkaloids at different harvesting time. Nevertheless, the difference of the alkaloids content from different parts was much significant. The content of total alkaloid of adventitious root near to rhizome was about 4%, 2 times higher than that away from rhizome (only 2%). In addition, different processing technologies would affect alkaloids content obviously. There was hardly loss of alkaloids when the fresh adventitious root was washed with water, but it would decrease alkaloids content when the dried adventitious root was washed. CONCLUSION: Medicine value of Coptis chinensis adventitious root near to rhizome is higher than that away from rhizome. And fresh Coptis chinensis adventitious root can be washed with water.

The antihypercholesterolemic effect of jatrorrhizine isolated from Rhizoma Coptidis.

Current work was conducted to evaluate the safety and antihypercholesterolemic activity of jatrorrhizine extracted from Rhizoma Coptidis (RC) and its potential mechanism on regulating cholesterol metabolism. It was found that the LD50 of jatrorrhizine in mice was more than 5,500 mg/kg and there were no influences on clinical signs, organ weight changes, urinalysis and hematological parameters, gross necropsy and histological alterations in jatrorrhizine-treated rats during the 3-month period, compared to the control group. Jatrorrhizine showed a strong lipid-lowering effect in a dose-dependent manner. Oral administration of 70.05 mg/kg of jatrorrhizine on Mesocricetus auratus (Syrian golden hamsters) exhibited significant decrease in TC, TG, and LDL-c levels by 20%, 43%, and 19%, respectively, and increase in HDL-c and total bile acids (TBA) content in feces (p<0.01), compared to high-fat and high-cholesterol (HFHC) group. Besides, jatrorrhizine dose-dependently slowed the rate of weight gain. The results of qRT-PCR, western blotting and ELISA revealed that jatrorrhizine significantly up-regulated the mRNA and protein expression of LDLR and CYP7A1, but exhibited no significant effect on mRNA and protein expression of HMGR and ASBT in hamsters. In conclusion, jatrorrhizine was a safe and potential antihypercholesterolemic agent from RC which could improve the utilization and excretion of cholesterol by up-regulating the mRNA and protein expression of LDLR and CYP7A1.

[Study on optimal processing technology of three kinds of processed coptidis rhizoma].

OBJECTIVE: To optimize the processing technology of Coptidis Rhizoma and study the effects of different adjuvants on alkaloids during processing. METHODS: The moistening time of adjuvants (A), processing temperature (B) and processing time (C) were investigated by using the single factor test method and L9 (3(4)) orthogonal experiment design with the contents of four alkaloids as indexes. RESULTS: The sequence of importance of the factors that affect the wine processed Coptidis Rhizoma (WC) was C > B > A, turmeric processed Coptidis Rhizoma (TC) and dogwood processed Coptidis Rhizoma (DC) was B > A > C. CONCLUSION: The optimal processing technology of WC, TC and DC are drying for 4 h at 130 degrees C after moistening for 90 min, drying for 3 h at 100 degrees C after moistening for 60 min and drying for 2 h at 160 degrees C after moistening for 90 min, respectively.

Safety evaluation of main alkaloids from Rhizoma Coptidis.

ETHNOPHARMACOLOGICAL RELEVANCE: Rhizoma Coptidis (RC), a widely used traditional Chinese medicine, has been used for the treatment of heat-clearing and detoxifying, but there is very little information on its safety. AIM OF THE STUDY: To provide information on the safety of RC, we evaluated the toxicity of the crude RC and RC alkaloids (berberine, coptisine, palmatine and epiberberine) including cytotoxicity, acute toxicity in mice and sub-chronic toxicity in rats. MATERIALS AND METHODS: The cytotoxicity of RC alkaloids was tested in HepG2 and 3T3-L1 cells by the MTT assay. The acute toxicity of RC alkaloids was tested in mice and the mortality was calculated at the end of experiment. For sub-chronic toxicity study, the rats were treated with the RC alkaloids at a dose of 156 mg/kg/day and RC at a dose of 521 mg/kg/day for 90 days. Mortality, clinical signs, body weight changes, organ weights, urinalysis and hematological parameters, gross necropsy and histopathology were monitored during the study period. RESULTS: The cell assay indicates that the IC(50) values of berberine, coptisine, palmatine and epiberberine in HepG2 cells were 48.17, 64.81, 112.80 and 120.58 µg/mL, which in 3T3-L1 cells were 41.76, 56.48, 84.32 and 104.18 µg/mL, respectively. In the acute toxicity assay, the LD(50) values of four alkaloids were 713.57, 852.12, 1533.68 and 1360 mg/kg, respectively. However, in the sub-chronic toxicity study, no mortality and morbidity were observed which could be related to RC alkaloids and RC treatment. Besides, there was no abnormality in clinical signs, body weights, organ weights, urinalysis, hematological parameters, gross necropsy and histopathology in any of the animals after the oral administration of RC alkaloids and RC. CONCLUSIONS: Taking these results together, we came to the conclusion that the toxicity of berberine is the maximum and palmatine is the minimal in four RC alkaloids. The currently recommended doses of RC alkaloids and RC consumed are relatively safe.