Evaluation of anti-inflammatory and antioxidant for chrysanthemum stem and leaf in zebrafish inflammatory bowel disease model and identification of the bioactive compositions by UPLC-TQ/MS

Background Modern studies have shown that chrysanthemum has anti-inflammatory, regulating intestinal function and other effects, chrysanthemum stem and leaf as a nonmedicinal part of chrysanthemum, has similar chemical components with chrysanthemum, so it is speculated that chrysanthemum stem and leaf also has the effect of regulating intestinal inflammation. The purpose of this study was to evaluate the antiinflammatory and antioxidant effect of chrysanthemum stem and leaf extract through zebrafish inflammatory bowel disease model, and to detect flavonoids, phenolic acids and polysaccharides in chrysanthemum stem and leaf extract. Methods DSS induced inflammatory bowel disease model of zebrafish was used. Aliciin blue staining was used to observe the secretion of intestinal acid mucin, and H&E staining was used to detect the inflammatory cell infiltration. Superoxide dismutase activity was determined by kit, and the expression levels of key inflammatory cytokines IL-1 β , IL8 and MMP9 were detected by quantitative polymerase chain reaction. Furthermore, UPLC-TQ /MS method was used to detect the contents of flavonoids and phenolic acids in chrysanthemum stem and leaf extracts. Neutral and acidic polysaccharides were determined by the phenol-sulfuric acid method and the carbazol-sulfuric acid method. Results H&E staining showed that extracts from chrysanthemum stem and leaf inhibited inflammatory cell infiltration to varying degrees. The expressions of inflammatory cytokines IL-1 β , IL8 and MMP9 were significantly increased in DSS induced zebrafish. The extracts inhibited the expression of IL-1 β , IL8 and MMP9 in DSS induced zebrafish. The water extract 0.2mg/ mL and alcohol extract 0.04mg/ mL had the most significant inhibition. Superoxide dismutase activity in extract treatment group was also up-regulated compared with model group. The results showed that the contents of total flavonoids and phenolic acids in the alcohol extract of chrysanthemum stem and leaf were significantly higher than those in the water extract of chrysanthemum stem and leaf, but the water-soluble polysaccharides were significantly more in the water extract of chrysanthemum stem and leaf. Conclusions In conclusion, this study suggests that chrysanthemum stem and leaf extract can improve inflammatory bowel disease of zebrafish through antiinflammatory and antioxidant activities.

Whole-Genome Sequencing and Potassium-Solubilizing Mechanism of Bacillus aryabhattai SK1-7

To analyze the whole genome of Bacillus aryabhattai strain SK1-7 and explore its potassium solubilization characteristics and mechanism, thus providing a theoretical basis for analyzing the utilization and improvement of insoluble potassium resources in soil. Genome information for Bacillus aryabhattai SK1-7 was obtained by using Illumina NovaSeq second-generation sequencing and GridION Nanopore ONT third-generation sequencing technology. The contents of organic acids and polysaccharides in fermentation broth of Bacillus aryabhattai SK1-7 were determined by high-performance liquid chromatography and the anthrone sulfuric acid method, and the expression levels of the potassium solubilization-related genes ackA, epsB, gltA, mdh and ppc were compared by real-time fluorescence quantitative PCR under different potassium source culture conditions. The whole genome of the strain consisted of a complete chromosome sequence and four plasmid sequences. The sequence sizes of the chromosomes and plasmids P1, P2, P3 and P4 were 5,188,391 bp, 136,204 bp, 124,862 bp, 67,200 bp and 12,374 bp, respectively. The GC contents were 38.2, 34.4, 33.6, 32.8, and 33.7%. Strain SK1-7 mainly secreted malic, formic, acetic and citric acids under culture with an insoluble potassium source. The polysaccharide content produced with an insoluble potassium source was higher than that with a soluble potassium source. The expression levels of five potassium solubilization-related genes with the insoluble potassium source were higher than those with the soluble potassium source.

Application of the phenol-sulfuric acid method for the determination of glycogen in skeletal muscles and liver of rats

The possibility of using the phenol-sulfuric acid method for the determination of total glycogen, its acid-soluble and acid-insoluble fractions in the liver and skeletal muscles of rats was studied. It was found that the use of a precipitant in the isolation of total glycogen and its fractions increases the yield of the investigated substances. Key words: phenol-sulfate method, rats, liver, muscles, total glycogen, acid-soluble glycogen, acid-insoluble glycogen.

Quantification of total sugars and reducing sugars of dragon fruit-derived sugar-samples by UV-Vis spectrophotometric method

In the present work, the phenol-sulfuric acid method and the 3,5-dinitrosalicylic acid (DNS) method were developed with the aim to quantitatively analyze total sugars and reducing sugars, respectively. In regard with the phenol-sulfuric acid assay, 1.0 mL of sample was treated with 1.0 mL of 5% phenol, 5.0 mL of concentrated H2SO4 and measured at 485 nm, with the linearity range of 10–100 ppm for total sugars. The DNS method was performed on 2.0 mL of sample, using 1.5 mL of DNS at 80 °C for 10 minutes and measured at 510 nm, with the linearity range of 50–400 ppm for reducing sugars. The sugar contents of white dragon fruit-derived sugar-samples (extracted from species in Binh Thuan province, Vietnam) were also estimated by the above measured methods, exhibiting the total sugars of above 90% and the reducing sugars of above 5%. The methods were well-performed with the acceptable relative standard deviations of repeatability in accordance with the Association of Official Analytical Chemists (AOAC).

Developing a Procedure of Extracting Total Polysaccharide from Dendrobium Nobile Lindl. in Vietnam

Dendrobium nobile Lind., also known as Hoang thao dui ga, is a species of plant belonging to the genus Dendrobium, a large genus of the Orchidaceae. This is a medical plant used in many remedies in China and Vietnam. Polysaccharide from this species has been shown many valuable biological activities such as hypoglycemia antioxidant and anti-cancer. In this paper, a procedure for extracting total polysaccharide by aqueous solvent from D.nobile based on the survey of factors: temperature, time, material/solvent ratio affecting this extraction process was proposed. The total polysaccharide content was also determined according to phenol-sulfuric acid method.

Recovery of Lithium from Lepidolite by Sulfuric Acid and Separation of Al/Li by Nanofiltration

The recovery and leaching kinetics of lithium from lepidolite by sulfuric acid method were investigated in this study, and a new method of nanofiltration to separate Al/Li from lepidolite leaching solution was coupled. The results indicated the optimal conditions about leaching lithium from lepidolite: leaching at 433 K for 4 h with the agitation rate of 120 r min−1, sulfuric acid concentration of 60 wt%, liquid-solid mass ratio of 2.5:1, under which the Li yield could reach at 97%. The kinetics observations revealed that the leaching process was controlled by the hybrid control of solid product layer diffusion and the chemical reaction, and dominated by chemical reaction step, which improved the conclusion of single-step control in the previous literature. A successful attempt was made to couple nanofiltration separation with sulfuric acid extraction of lithium, and DK membrane was used to separate Al/Li from lepidolite leaching solution. DK membrane has shown excellent retention of Al3+ and Ca2+ and also can effectively permeate Li+, which may bring a new inspiration for lithium extraction from lepidolite in the future.

Purification of acrylamide from polymerization inhibitors in the manufacture of high quality flocculants based on polyacrylamide

Polyacrylamide and its copolymers are widely used as flocculating agents for the separation of industrial suspensions. The formation of high molecular weight polymers depends on the content of various impurities present in the monomer. The article presents the scientific and practical information on the production of acrylamide by sulfuric acid method of hydration of nitrile acrylic acid in the form of an aqueous solution of different concentrations and a more modern heterogeneously catalytic method of hydration of acrylonitrile using as catalysts with variable valence. Ways to get different impurities in the stages of production of acrylamide with the purpose of applying appropriate methods for its purification. Laboratory studies of the purification of an aqueous solution of acrylamide from iron ions were carried out as an element of inhibition of the premature polymerization process.

A brief exploration of EPS composition in biofilms of Staphylococcus spp ATCC reference strains

<p>Antibiotic resistance is expected to cause 10 million deaths per year worldwide by 2050. One of the mechanisms for the resilient nature of bacteria toward antibiotics is through the formation of biofilm. Bacterial biofilms are sessile communities of microorganisms, which exist in a matrix of proteins, carbohydrates, eDNA and other various components – collectively known as extracellular polymeric substances. Biofilms slow the penetration of drugs, and also contribute to the development of a resistant phenotype known as persisters. Thus, understanding biofilm composition might contribute to the development of anti-biofilm strategies. The aim of this study was to explore biofilm formed by five <em>Staphylococcus</em> spp ATCC strains, commonly used in research as references: <em>S. aureus</em> 25923, <em>S. aureus</em> 29213, <em>S. aureus</em> 43300 (methicillin-resistant), <em>S. aureus</em> 6538 and <em>S. epidermidis</em> 12228. Biofilm mass and its components were analysed after 24h and 72h of biofilm growth. Bacterial biofilm was prepared in 96-well microtiter plates, in Trypticase Soy Broth supplemented with 1% glucose. After incubation at 37°C, absorbance measurements and crystal violet staining were performed and the specific biofilm formation determined for each strain. Extracellular polymeric substances were extracted using a combination of physical and chemical methods; including centrifugation, vortexing and the use of 1.5M NaCl. In these assays, biofilms were grown in polystyrene tubes containing 10 ml of same media mentioned above. The concentration of protein, carbohydrate and eDNA was determined using the Bicinchoninic acid assay, phenol-sulfuric acid method and DNeasy<sup>®</sup> Blood and Tissue Kit, respectively, followed by spectroscopy. Our data demonstrated heterogeneity between the biofilm-forming capabilities and EPS components within staphylococcal strains and species. Strains 25923 and 6538 had the highest value for biofilm formation at both time points. Interestingly, strain 43300 was the only one to show a significant increase in biofilm after 72h. Contradictory to previous findings, <em>S. epidermidis</em> 12228 was found to be a good biofilm producer. At both time points studied, strains demonstrated considerably higher concentrations of protein (varying from 172 µg/mL – 345 µg/mL) and carbohydrate (56 µg/mL - 372µg/mL) in EPS compared to eDNA (2.74 µg/mL – 8.12 µg/mL). On average, strains 43300 and 12228 had the highest concentration of protein, and the latter also had the highest carbohydrate and eDNa amounts at 72h. Strains 25923 and 6538 had a significant decrease in eDNA concentration over time. Based on this brief study, the relative quantities of EPS components investigated is similar to that of other studies with protein being the most plentiful component followed by carbohydrate and then considerably lower amounts of eDNA. Differences in specific biofilm formation did not directly reflect variations observed in abundance of a particular constituent in the matrix of EPS. This study also showed that <em>S. epidermidis</em> 12228, usually classified as a weak or non-biofilm former, was able to grow a relatively substantial biofilm under the conditions tested here.</p>