S/O/W Emulsion with CAPE Ameliorates DSS-Induced Colitis by Regulating NF-κB Pathway, Gut Microbiota and Fecal Metabolome in C57BL/6 Mice.

Inflammatory bowel disease (IBD) has attracted much attention worldwide due to its prevalence. In this study, the effect of a solid-in-oil-in-water (S/O/W) emulsion with Caffeic acid phenethyl ester (CAPE, a polyphenolic active ingredient in propolis) on dextran sulfate sodium (DSS)-induced colitis in C57BL/6 mice was evaluated. The results showed that CAPE-emulsion could significantly alleviate DSS-induced colitis through its effects on colon length, reduction in the disease activity index (DAI), and colon histopathology. The results of ELISA and Western blot analysis showed that CAPE-emulsion can down-regulate the excessive inflammatory cytokines in colon tissue and inhibit the expression of p65 in the NF-κB pathway. Furthermore, CAPE-emulsion promoted short-chain fatty acids production in DSS-induced colitis mice. High-throughput sequencing results revealed that CAPE-emulsion regulates the imbalance of gut microbiota by enhancing diversity, restoring the abundance of beneficial bacteria (such as Odoribacter), and suppressing the abundance of harmful bacteria (such as Afipia, Sphingomonas). The results of fecal metabolome showed that CAPE-emulsion restored the DSS-induced metabolic disorder by affecting metabolic pathways related to inflammation and cholesterol metabolism. These research results provide a scientific basis for the use of CPAE-emulsions for the development of functional foods for treating IBD.

Universal organophosphate pesticides detection by peptide based fluorescent probes.

In practical applications, the rapid and efficient detection of universal organophosphorus pesticides (OPs) can assist inspectors in quickly identifying the presence of OPs in samples. However, this presents a challenge for most well-established methods, typically designed to detect only a specific type of organophosphorus molecule at a time. In this proof-of-concept study, we draw inspiration from the structural similarities among OPs to develop innovative peptide-based fluorescence probes for the first time, which could efficiently detect a broad range of OPs within a mere 3 min. Analysis of fluorescence curve fitting reveals a clear linear correlation between the fluorescent intensity of the peptide probes and the concentration of OPs. Additionally, the selectivity analysis indicates that these peptide fluorescent probes exhibit an excellent response to various OPs while maintaining sufficient selectivity for detecting other pesticide types. Accurate sample analysis has also highlighted the potential of these peptide probes as practical tools for the rapid detection of OPs in actual vegetable samples. In summary, this proof-of-concept study presents an innovative approach to designing and developing ultrafast, universally peptide-based OP probes. These custom-designed peptide probes may facilitate rapid sample screening and offer initial quantification for OPs, potentially saving valuable time and effort in practical OP detection.

Enhancing the stability of natural anthocyanins against environmental stressors through encapsulation with synthetic peptide-based gels.

The instability of anthocyanin to environmental stressors severely limits its applications as a natural bioactive pigment. To overcome these limitations, this proof-of-concept study utilizes the high biocompatibility of peptide molecules and the unique gel microstructure to develop innovative peptide-based gels. Characterization of the gels was conducted through AFM, SEM, rheological analysis, and CD spectrum. These analyses confirmed the fibrous mesh structure and impressive mechanical strength of the peptide-based gels. The cytotoxicity evaluation using MTT and hemolysis analysis showed high biocompatibility. Encapsulation efficiency analysis and fluorescence microscopy images demonstrated successful and efficient encapsulation of anthocyanins in all four peptide-based gels, with uniform distribution. Moreover, systematic investigations were conducted to assess the impact of peptide-based gels on the stability of natural anthocyanins under environmental stressors such as temperature, pH variations, and exposure to metal ions. Notably, the results revealed a significant enhancement in stability, including improved long-term storage and antioxidant activity. In conclusion, this study successfully developed four novel peptide-based gels that effectively protect natural anthocyanins from environmental stressors, highlighting their potential in various fields such as food and biology.

Metabolic profiling of the fluorinated liquid-crystal monomer 1-ethoxy-2,3-difluoro-4-(trans-4-propylcyclohexyl)benzene.

1-Ethoxy-2,3-difluoro-4-(trans-4-propylcyclohexyl)benzene (EDPrB) is a typical fluorinated liquid-crystal monomer (LCM). LCMs contaminants are becoming increasingly concerning due to their potential persistence, bioaccumulation, toxicity, and broad prevalence in environmental and human samples. However, LCM metabolism is poorly understood. Herein, by introducing selected EDPrB into the appropriate liver microsomes in vitro, we examined the metabolic pathways of LCM in humans, rats, pigs, Cyprinus carpio, crucian carp, and Channa argus. A total of 20 species-dependent metabolites were identified and structurally elucidated by gas and liquid chromatography-high resolution mass spectrometry for the first time. Dealkylation, H-abstraction, and hydroxylation reactions are the primary metabolic pathways. Half of these in vitro metabolites were found in the urine, serum, and fecal samples of Sprague-Dawley rats exposed to EDPrB. Toxicity predictions indicate that 17 metabolites can be classified as toxic. According to the Ecological Structure Activity Relationships (ECOSAR), a number of metabolites exhibit equivalent or greater aquatic toxicity to that of EDPrB. Toxicity Estimation Software Tool (T.E.S.T.) predicts that some metabolites exhibit developmental toxicity and mutagenicity in rats. These findings suggest that biotransformation should be particularly emphasized, and more toxicological and monitoring studies should be performed to assess the ecological and human safety of LCMs.

Caffeic acid phenethyl ester loaded in a targeted delivery system based on a solid-in-oil-in-water multilayer emulsion: Characterization, stability, and fate of the emulsion during in vivo digestion.

Many studies have shown that caffeic acid phenethyl ester (CAPE) has various functions, such as antioxidant, anti-inflammatory and anticancer activity, but its low bioavailability and stability limit its application. In this study, the colorectal targeted delivery system for CAPE based on a solid-in-oil-in-water (S/O/W) multilayer emulsion was prepared using CAPE-loaded nanoparticles as the solid phase, coconut oil as the oil phase, and a mixture of lecithin and sodium caseinate as the aqueous phase. The stability of the O/W interfacial layer was improved by using a sodium casein-lecithin mixture as the aqueous surface layer in the preparation. This S/O/W emulsion is a spherical droplet with an S/O/W trilayer structure with a particle size of 155.5 ± 0.72 nm and a polydispersity index (PDI) of 0.24 ± 0.01. The Fourier transform infrared (FTIR) results confirmed that CAPE was successfully loaded into the S/O/W emulsion. This S/O/W emulsion was able to maintain a stable liquid state at pH 6.00-7.4 or cholate concentration of 0-50 mg/mL but showed a gel state at pH 2.0-3.0. The storage experiments demonstrated that the S/O/W emulsion was stable for 15 days at 4 °C, but was prone to agglomeration and emulsion breakage at 25 °C. The in vivo digestion process indicated that the S/O/W emulsion was gradually digested in the digestive tract and released solid phase nanoparticles in the large intestine. Therefore, this newly developed targeted delivery system can effectively deliver CAPE to the colorectum and achieve a 12-hour delayed release, which improved the bioavailability and activity of CAPE.

Metabolic profiling of bisphenol A diglycidyl ether in vitro and in vivo.

Bisphenol A diglycidyl ethers (BADGE) is one class of human-made chemicals, and it is one of the most widely used raw materials for epoxy resins. As an active compound, BADGE undergoes biotransformation in vitro and in vivo. However, there is a limited understanding of the biotransformation of BADGE and toxicity studies on transformation products. We conducted comprehensive research on the metabolic transformation of BADGE in vitro and in vivo. The results showed that 12 metabolites and 7 metabolites were identified in vitro and in vivo, respectively. Four biotransformation products, including M1 (hydrolysis), M3 (dehydroxylation), M10 (carboxylation), and M11 (glucose conjugation), can be found in both in vitro and in vivo samples. The main metabolic pathways were hydroxylation, carboxylation, cysteine (Cys) conjugation, and glucose conjugation. Besides, our results suggested the existence of metabolic differences in BADGE between species and gender. Further, we investigated toxicities of BADGE metabolites in-silico. Importantly, some hydrolysis (M1, M2), hydroxylation (M7), and oxidation (M8) products showed similar or even higher potential toxicity than BADGE depending on the endpoint. These results enrich the biotransformation profiles of BADGE and provide useful information for understanding its biotransformation in humans and a reference for the comprehensive assessment for human health risk.

Polysaccharide selection and mechanism for prevention of protein-polyphenol haze formation in beverages.

Polysaccharides have been considered as a group of promising candidate for preventing the protein-polyphenol haze formation in beverages. In order to select effective polysaccharides to prevent the haze formation, four protein-polyphenol haze model systems were successfully established using two proteins (i.e., gelatin and bovine serum albumin) and two polyphenols (i.e., procyanidin [PC] and epigallocatechin gallate [EGCG]). Among seven common polysaccharides, 0.5 mg/mL pectin, 0.05 mg/mL xanthan gum, and 0.01 mg/mL guar gum demonstrated the maximum potential for preventing the formation of four protein-polyphenol hazes. Ultraviolet-visible spectrophotometry confirmed that polysaccharides affected protein-polyphenol interactions. Fluorescence spectrophotometry combined with microscale thermophoresis data indicated the relative affinities of polyphenol to protein and polysaccharide determined the mechanism of polysaccharide for preventing the haze formation. In bovine serum albumin (BSA)/gelatin-EGCG system, polysaccharides (pectin, xanthan gum and guar gum) competed with BSA/gelatin to bind EGCG for prevention the formation of BSA/gelatin-EGCG haze. However, in BSA/gelatin-PC system, polysaccharides (pectin, xanthan gum, and guar gum) formed a ternary complex (protein-tannin-polysaccharide) for increasing the solubility of protein-polyphenol aggregation. From apple juice results, the reduction rates of guar gum in two apple juice systems (gelatin-PC, BSA-PC) were 21% and 56% within 8 weeks, indicating guar gum might be the most effective polysaccharide in preventing the haze formation. PRACTICAL APPLICATION: This experiment data could be used for development of polysaccharide products for prevention of protein-polyphenol haze formation in beverages.

Synergistic chemopreventive effect of allyl isothiocyanate and sulforaphane on non-small cell lung carcinoma cells.

Isothiocyanates from cruciferous vegetables are known for their potential anti-carcinogenic activities. These isothiocyanates are frequently consumed together as part of a regular diet, but their combined effects on carcinogenesis have not been well studied. Herein, we tested the hypothesis that combination of two isothiocyanates, i.e. allyl isothiocyanate and sulforaphane, produced a synergy in inhibiting the growth of A549 lung cancer cells. Our results showed that the combination treatment led to a stronger growth inhibition than the singular treatment. Isobologram analysis proved that the enhanced inhibitory effect of the combination treatment was synergistic. Flow cytometry demonstrated that the combination treatment caused more extensive cell cycle arrest and apoptosis than the singular treatment with modified expression of key proteins regulating these cellular processes. The combined treatment resulted in the production of intracellular reactive oxygen species, which might contribute to the inhibitory effects on cancer cells. Moreover, a synergy between allyl isothiocyanate and sulforaphane was also observed in anti-cell migration. Collectively, our results have demonstrated the potential of different isothiocyanates used in combination to produce enhanced protective effects against carcinogenesis.

Identification and characterization of a novel carboxylesterase from Phaseolus vulgaris for detection of organophosphate and carbamates pesticides.

BACKGROUND: Organophosphate and carbamate pesticide residues in food and the environment pose a great threat to human health and have made the easy and rapid detection of these pesticide residues an important task. Discovering new enzyme sources from plants can help reduce the cost of large-scale applications of rapid pesticide detection via enzyme inhibition. RESULTS: Plant esterase from kidney beans was purified. Kidney bean esterase is identified as a carboxylesterase by substrate and inhibitor specificity tests and mass spectrometry identification. The kidney bean esterase demonstrates optimal catalytic activity at 40 °C, pH 6.5 and an enzyme concentration of 0.30 µg mL-1 . The kidney bean esterase can be inhibited by organophosphate and carbamate pesticides, which can be substituted for acetylcholinesterase. The limit of detection of the purified kidney bean esterase was two- to 20-fold higher than that of the crude one. The method detection limit meets the detection requirement for the maximum residue limits (MRL) in actual samples. CONCLUSION: The findings of the present study provide a new source of enzymes for pesticides detection by enzyme inhibition. © 2018 Society of Chemical Industry.

Oxidation pretreatment by calcium hypochlorite to improve the sensitivity of enzyme inhibition-based detection of organophosphorus pesticides.

BACKGROUND: Enzyme inhibition-based detection is the most widely used method for rapid detection of organophosphorus pesticides (OPs) in food and agricultural products. However, the accuracy of the method is negatively affected by low inhibitory activities of OPs with PS moiety on acetylcholinesterase. RESULTS: We demonstrated that oxidation pretreatments with bromine, hydrogen peroxide, or calcium hypochlorite significantly enhanced the enzyme inhibitory activities of these OPs. Especially, calcium hypochlorite (0.05%) pretreatment converted the PS moiety in OPs to PO and produced the most potent and steady inhibitory effect on the enzyme. This, in turn, resulted in a dramatic increase in the sensitivity of enzyme inhibition-based detection of these OPs by as much as 2 to 7 orders of magnitude. Importantly, this enhanced detection of OPs was validated in various vegetable samples. CONCLUSION: Our findings provide a solid basis to use calcium hypochlorite pretreatment for the improved detection of OPs by the enzyme inhibition-based method. © 2017 Society of Chemical Industry.

Sirtuin 5: a review of structure, known inhibitors and clues for developing new inhibitors.

Sirtuins (SIRTs) are nicotinamide adenine dinucleotide (NAD+)-dependent protein deacetylases, which regulate important biological processes ranging from apoptosis, age-associated pathophysiologies, adipocyte and muscle differentiation, and energy expenditure to gluconeogenesis. Very recently, sirtuin 5 (SIRT5) has received considerable attention due to that it was found to have weak deacetylase activity but strong desuccinylase, demalonylase and deglutarylase activities, and it was also found to be associated with several human diseases such as cancer, Alzheimer's disease, and Parkinson's disease. In this review, we for the first time summarized the structure characteristics, known peptide and smallmolecule inhibitors of SIRT5, extracted some clues from current available information and introduced some feasible, practical in silico methods, which might be useful in further efforts to develop new SIRT5 inhibitors.

An integrated molecular docking and rescoring method for predicting the sensitivity spectrum of various serine hydrolases to organophosphorus pesticides.

BACKGROUND: The enzymatic chemistry method is currently the most widely used method for the rapid detection of organophosphorus (OP) pesticides, but the enzymes used, such as cholinesterases, lack sufficient sensitivity to detect low concentrations of OP pesticides present in given samples. Serine hydrolase is considered an ideal enzyme source in seeking high-sensitivity enzymes used for OP pesticide detection. However, it is difficult to systematically evaluate sensitivities of various serine hydrolases to OP pesticides by in vitro experiments. This study aimed to establish an in silico method to predict the sensitivity spectrum of various serine hydrolases to OP pesticides. RESULTS: A serine hydrolase database containing 219 representative serine hydrolases was constructed. Based on this database, an integrated molecular docking and rescoring method was established, in which the AutoDock Vina program was used to produce the binding poses of OP pesticides to various serine hydrolases and the ID-Score method developed recently by us was adopted as a rescoring method to predict their binding affinities. In retrospective case studies, this method showed good performance in predicting the sensitivities of known serine hydrolases to two OP pesticides: paraoxon and diisopropyl fluorophosphate. The sensitivity spectrum of the 219 collected serine hydrolases to 37 commonly used OP pesticides was finally obtained using this method. CONCLUSION: Overall, this study presented a promising in silico tool to predict the sensitivity spectrum of various serine hydrolases to OP pesticides, which will help in finding high-sensitivity serine hydrolases for OP pesticide detection.

Actinopolyspora lacussalsi sp. nov., an extremely halophilic actinomycete isolated from a salt lake.

A novel halophilic, filamentous actinobacterium, designated strain TRM 40139(T), was isolated from a hypersaline habitat in Xinjiang Province, north-west China. Its taxonomic status was determined using a polyphasic approach. Phylogenetic analysis based on the almost-complete 16S rRNA gene sequence of the strain showed that it formed a well-separated sub-branch within the radiation of the genus Actinopolyspora and the organism was related most closely to the type strains of Actinopolyspora alba (97.6 % similarity), Actinopolyspora xinjiangensis (97.6 %) and Actinopolyspora erythraea (97.1 %). However, it had relatively lower mean DNA-DNA relatedness values with the above strains (36.4, 31.3 and 26.1 %, respectively). Optimal growth occurred at 35 °C, at pH 7.0 and in the presence of 12 % (w/v) NaCl. The whole-cell sugar pattern consisted of xylose, glucose, ribose and arabinose. The major fatty acids were iso-C16 : 0 (28.0 %) and anteiso-C17 : 0 (27.6 %). The diagnostic phospholipids detected were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, phosphatidylinositol and two unknown phospholipids. The predominant menaquinones were MK-9(H4) (49.8 %) and MK-10(H4) (24.2 %). The G+C content of the genomic DNA was 66.4 mol%. Strain TRM 40139(T) therefore represents a novel species of the genus Actinopolyspora, for which the name Actinopolyspora lacussalsi sp. nov. is proposed. The type strain is TRM 40139(T) (= KCTC 19657(T) = CCTCC AA 2012020(T)).

Myceligenerans salitolerans sp. nov., a halotolerant actinomycete isolated from a salt lake in Xinjiang, China.

A halotolerant actinomycete strain, designated XHU 5031(T), was isolated from a salt lake in Xinjiang Province, northwest China. Its taxonomic status was determined using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the organism was most closely related to Myceligenerans xiligouense DSM 15700(T) (98.4 %), Myceligenerans halotolerans XJEEM 11063(T) (98.0 %) and Myceligenerans crystallogenes DSM 17134(T) (97.5 %). However, it had relatively low values for DNA-DNA relatedness with the above strains (46.2, 39.4 and 36.5 %, respectively). The peptidoglycan type was A4α. This organism contained glucose, mannose and galactose as the major whole cell sugars. The predominant menaquinone was MK-9(H(4)). The major fatty acids were iso-C(15:0,) anteiso-C(15:0) and iso-C(16:0). The polar lipids detected were diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylinositol (PI), one unknown phospholipid (PL) and two unknown glycolipids (GL). The G+C content of genomic DNA was 71.2 mol %. Phenotypic data clearly distinguished the isolate from its closest relatives. The combined genotypic and phenotypic data indicated that the isolate XHU 5031(T) represented a novel species of the genus Myceligenerans. The proposed name for this organism is Myceligenerans salitolerans sp. nov., with type strain XHU 5031(T) (=KCTC 29128(T) = CCTCC AB 2012908(T)).

Apigenin inhibits cell migration through MAPK pathways in human bladder smooth muscle cells.

Apigenin, a nonmutagenic flavonoid, has been shown to possess free radical scavenging activities, anticarcinogenic properties, antioxidant and anti-inflammatory effects. Recently, apigenin was reported to cause gastric relaxation in murine. To assess possible effects of apigenin on migration of bladder smooth muscle (SM) cell, we isolated SM cells from peri-cancer tissue of human bladder and established a cell model that was capable to overexpress transiently MEKK1 (MEK kinase 1). Results showed that overexpression of active human MEKK1 by adenoviruses infection induced migration of human bladder smooth muscle (hBSM) cells and phosphorylation of MAPKs, ERK, JNK and p38, which are the downstream molecules of MEKK1. Then, hBSM cell overexpressing MEKK1 were exposed to apigenin (50 microM). Our data indicated that apigenin inhibited significantly activation/phosphorylation of MAPKs and migration of hBSM cells induced by MEKK1 overexpression. Besides, apigenin inhibited actin polymerization, which underlines muscle contraction and cell migration. The results suggest that apigenin inhibits activation of MAPKs and thereby the cell migration. The mechanism might be that apigenin blocks signal transmission from MEKK1 to MAPKs.

[Effects of exogenerous nucleotides on the apoptosis of intestinal epithelial cells IEC-6].

OBJECTIVE: To investigate the effects of exogenous nucleotides on apoptosis of a normal rat small intestinal epithelial cell line, IEC-6. METHODS: Cultured IEC-6 cells were treated by four kinds of monophosphate nucleotides and their mixture prepared according to their composition in human milk, then the cell apoptosis was determined by flow cytometry measurement, morphologic characterization, and electron-microscope observation. RESULTS: IEC-6 cells treated with AMP or GMP showed a apotosis peak in flow cytometry measurement, but only AMP produce typical apoptosis characteristics in electron-microscope observation. Pyrimidine nucleotides (UMP and CMP)and nucleotides mixture could not induce apoptosis. However, UMP could significantly eliminate the apoptosis-inducing effects of AMP or GMP. CONCLUSION: Purine nucleotides induce apoptosis of IEC-6, inducing effects of purine nucleotides. pyrimidine nucleotides UMP could abolish the apoptosis-inducing effects of purine nucleotides.

[Effects of exogenous nucleotides on the proliferation and migration of intestinal epithelial cells].

The effect of exogenous nucleotides on proliferation and migration of a normal rat small intestinal epithelial cell line IEC-6 is studied. The concentration-effects as well as interaction of exogenous nucleotides on proliferation of IEC-6 are measured by MTT. Migration of IEC-6 after wounded is determined by an in vitro model of intestinal epithelial restitution of IEC-6 monolayer. Expression of TGF beta is detected by immunohistochemistry. The results show AMP and GMP remarkably inhibit proliferation of IEC-6 in concentration-dependent manner respectively at 30 mumol/L or 150 mumol/L and more. CMP, UMP and nucleotides mixture can not enhance or inhibit the growth with the exception of inhibition of CMP on proliferation at very high concentration (1440 mumol/L). In contrast, CMP, especially UMP, can remarkably abolish the proliferation-inhibiting effects of AMP or GMP on the cell, when AMP or GMP is supplemented. Nucleotides mixture significantly facilitate migration of IEC-6 after wounded but fail to promote the expression of TGF beta. It is concluded that purine nucleotides inhibits proliferation of IEC-6. Pyrimidine nucleotides can abolish the inhibitive effects of purine nucleotides, and Nucleotides mixture promotes migration of IEC-6 after wounded by a TGF beta-independented way.