Levels Of Polysaccharides Research Articles

Disorders in the CMG helicase complex increase the proliferative capacity and delay chronological aging of budding yeast

The replication of DNA requires specialized and intricate machinery. This machinery is known as a replisome and is highly evolutionarily conserved, from simple unicellular organisms such as yeast to human cells. The replisome comprises multiple protein complexes responsible for various steps in the replication process. One crucial component of the replisome is the Cdc45-MCM-GINS (CMG) helicase complex, which unwinds double-stranded DNA and coordinates the assembly and function of other replisome components, including DNA polymerases. The genes encoding the CMG helicase components are essential for initiating DNA replication. In this study, we aimed to investigate how the absence of one copy of the CMG complex genes in heterozygous Saccharomyces cerevisiae cells impacts the cells' physiology and aging. Our data revealed that these cells exhibited a significant reduction in transcript levels for the respective CMG helicase complex proteins, as well as disruptions in the cell cycle, extended doubling times, and alterations in their biochemical profile. Notably, this study provided the first demonstration that cells heterozygous for genes encoding subunits of the CMG helicase exhibited a significantly increased reproductive potential and delayed chronological aging. Additionally, we observed a noteworthy correlation between RNA and polysaccharide levels in yeast and their reproductive potential, as well as a correlation between fatty acid levels and cell doubling times. Our findings also shed new light on the potential utility of yeast in investigating potential therapeutic targets for cancer treatment.

Bioinformatic and functional characterization of cyclic-di-GMP metabolic proteins in Vibrio alginolyticus unveils key diguanylate cyclases controlling multiple biofilm-associated phenotypes.

The biofilm lifestyle is critical for bacterial survival and proliferation in the fluctuating marine environment. Cyclic diguanylate (c-di-GMP) is a key second messenger during bacterial adaptation to various environmental signals, which has been identified as a master regulator of biofilm formation. However, little is known about whether and how c-di-GMP signaling regulates biofilm formation in Vibrio alginolyticus, a globally dominant marine pathogen. Here, a large set of 63 proteins were predicted to participate in c-di-GMP metabolism (biosynthesis or degradation) in a pathogenic V. alginolyticus strain HN08155. Guided by protein homology, conserved domains and gene context information, a representative subset of 22 c-di-GMP metabolic proteins were selected to determine which ones affect biofilm-associated phenotypes. By comparing phenotypic differences between the wild-type and mutants or overexpression strains, we found that 22 c-di-GMP metabolic proteins can separately regulate different phenotypic outputs in V. alginolyticus. The results indicated that overexpression of four c-di-GMP metabolic proteins, including VA0356, VA1591 (CdgM), VA4033 (DgcB) and VA0088, strongly enhanced rugose colony morphotypes and strengthened Congo Red (CR) binding capacity, both of which are indicators of biofilm matrix overproduction. Furthermore, rugose enhanced colonies were accompanied by increased transcript levels of extracellular polysaccharide (EPS) biosynthesis genes and decreased expression of flagellar synthesis genes compared to smooth colonies (WTpBAD control), as demonstrated by overexpression strains WTp4033 and ∆VA4033p4033. Overall, the high abundance of c-di-GMP metabolic proteins in V. alginolyticus suggests that c-di-GMP signaling and regulatory system could play a key role in its response and adaptation to the ever-changing marine environment. This work provides a robust foundation for the study of the molecular mechanisms of c-di-GMP in the biofilm formation of V. alginolyticus.

Comparison and improvement of RNA extraction methods in Sargassum (Phaeophyta).

Sargassum (Sargassaceae) is widely distributed globally and plays an important role in regulating climate change, but the landscape of genomes and transcripts is less known. High-quality nucleic acids are the basis for molecular biology experiments such as high-throughput sequencing. Although extensive studies have documented methods of RNA extraction, these methods are not very applicable to Sargassum, which contains high levels of polysaccharides and polyphenols. To find a suitable method to improve the quality of RNA extracted, we compared and modified several popular RNA extraction methods and screened one practical method with three specific Sargassum spp. The results showed that three CTAB methods (denoted as Methods 1, 2, and 3) and the RNAprep Pure Plant Kit (denoted as Method 4) could, with slight modifications, effectively isolate RNA from Sargassum species, except for Method 4 used with S. fusiforme. By performing further screening, we determined Method 4 was the best choice for S. hemiphyllum and S. henslowianum, as revealed by RNA yields, RNA Integrity Number (RIN), extraction time, and unigene mapped ratio. For S. fusiforme, Methods 1, 2, and 3 showed no obvious differences among the yields, quality, or time to perform. In addition, one other method was tested, but we found the quality of the RNA extracted by TRIzol reagent methods (denoted as Method 5) performed the worst when compared with the above four methods. Therefore, our study provides four suitable methods for RNA extraction in Sargassum and is essential for future genetic exploration of Sargassum.

Effect of Combined Infrared Hot Air Drying on Yam Slices: Drying Kinetics, Energy Consumption, Microstructure, and Nutrient Composition.

Using hot air drying (HAD) and combined infrared hot air drying (IR-HAD) test devices, the drying kinetics, unit energy consumption, color difference values, rehydration rate, microstructure, and changes in polysaccharide and allantoin contents of yam slices were examined at various temperatures (50 °C, 55 °C, 60 °C, 65 °C, and 70 °C). The findings demonstrated that each of the aforementioned parameters was significantly impacted by the drying temperature. IR-HAD dries quicker and takes less time to dry than HAD. The Deff of IR-HAD is higher than that of HAD at the same temperature and increases with the increase in temperature. The activation energy required for IR-HAD (26.35 kJ/mol) is lower than that required for HAD (32.53 kJ/mol). HAD uses more energy per unit than IR-HAD by a factor of greater than 1.3. Yam slices treated with IR-HAD had higher microscopic porosity, better rehydration, lower color difference values, and higher polysaccharide and allantoin levels than HAD-treated yam slices. The IR-HAD at 60 °C had the greatest comprehensive rating after a thorough analysis of the dried yam slices using the coefficient of variation method. Three statistical indicators were used to evaluate six thin-layer drying models, and the Weibull model was most applicable to describe the variation of drying characteristics of yam slices.

Dynamic Formation of Green Tea Cream and the Identification of Key Components Using the "Knock-Out/Knock-In" Method.

The composition of green tea cream is extremely complex, and identification of key components is a prerequisite for elucidating its microstructure formation mechanism. This study examined the dynamic changes in the content of components and properties of colloid particles during the formation process of tea cream by chemical analysis and dynamic laser scattering (DLS). A "knock-out/knock-in" method was developed and used to further explore the relationship between the interaction of these components and the microstructure formation of tea cream. The results revealed that polysaccharides, proteins, epigallocatechin gallate (EGCG), and caffeine were the main components involved in tea cream formation. These components participated in the formation process in the form of polysaccharide-protein and EGCG-caffeine colloidal particles. Consequently, there were synchronized dynamic changes in the levels of polysaccharides, proteins, EGCG, and caffeine. The "knock-out/knock-in" experiment revealed that the interactions between EGCG or caffeine and macro-molecule components were not the key factors in tea cream microstructure formation. However, it was found that the complexation between EGCG and caffeine played a crucial role in the formation of tea cream. The findings suggested that decreasing the concentrations of EGCG and caffeine could be useful in controlling tea cream formation during tea beverage processing and storage.

High-quality DNA isolation protocol for detection of Khapra beetle (Dermestidae: Trogoderma granarium Everts, 1898) in standard wheat germ trap.

Khapra beetle (Dermestidae: Trogoderma granarium Everts, 1898) is an internationally significant pest of grain crops and stored grain products. Wheat germ traps, routinely used in surveillance sampling of Khapra beetle provide feed-substrates used by the pest throughout its life cycle. However, Khapra beetle larvae, eggs and other traces of the pest, such as larval frass and exuviae, in wheat germ traps are difficult to sort and taxonomically identify. Additionally, high levels of polysaccharides in wheat germ can inhibit PCR based molecular detection of this pest captured in the traps. We have developed a sensitive and low-cost protocol for extracting trace levels of Khapra beetle DNA from an entire wheat germ trap. Overnight digestion of entire trap contents in 6 mL of ATL buffer, followed by a 40min lysis step was optimal for DNA extraction. Paired with reported qPCR assays, this protocol allows the detection of a few hairs of T. granarium in a typical 2-gram wheat germ trap. This DNA extraction protocol makes it possible to perform a more rapid identification of the pest following wheat germ sample collection. The protocol has potential to improve international efforts for Khapra beetle surveillance.

Preference of Bacterial Rhamnosyltransferases for 6-Deoxysugars Reveals a Strategy To Deplete O-Antigens.

Bacteria synthesize hundreds of bacteria-specific or "rare" sugars that are absent in mammalian cells and enriched in 6-deoxy monosaccharides such as l-rhamnose (l-Rha). Across bacteria, l-Rha is incorporated into glycans by rhamnosyltransferases (RTs) that couple nucleotide sugar substrates (donors) to target biomolecules (acceptors). Since l-Rha is required for the biosynthesis of bacterial glycans involved in survival or host infection, RTs represent potential antibiotic or antivirulence targets. However, purified RTs and their unique bacterial sugar substrates have been difficult to obtain. Here, we use synthetic nucleotide rare sugar and glycolipid analogs to examine substrate recognition by three RTs that produce cell envelope components in diverse species, including a known pathogen. We find that bacterial RTs prefer pyrimidine nucleotide-linked 6-deoxysugars, not those containing a C6-hydroxyl, as donors. While glycolipid acceptors must contain a lipid, isoprenoid chain length, and stereochemistry can vary. Based on these observations, we demonstrate that a 6-deoxysugar transition state analog inhibits an RT in vitro and reduces levels of RT-dependent O-antigen polysaccharides in Gram-negative cells. As O-antigens are virulence factors, bacteria-specific sugar transferase inhibition represents a novel strategy to prevent bacterial infections.

Effects of substituting wheat by rye in diets for young fattening pigs on nutrient digestibility, performance, products of intestinal fermentation, and fecal characteristics.

Climate change and increasing demands to reduce the environmental impact of feed production are major challenges for animal nutritionists. Compared to wheat, which is commonly used in high levels in European piglet diets, rye is more efficient in using limited resources, most importantly, water and phosphorus. As a result, its cultivation has a relatively low carbon footprint. The high amounts of non-starch polysaccharides of rye might lead to an increased intestinal fermentation with potential beneficial effects on gut health. However, the high levels of non-starch polysaccharides in rye, which have a major impact on the physico-chemical conditions of the digesta, might affect digestibility and performance especially in young animals. It was therefore of interest to compare the effects of isoenergetic diets with increasing levels of rye as a replacement for wheat fed to young fattening pigs (bodyweight: 16-40 kg). The control diet contained 69% of wheat, while in the other three experimental diets, the amount of wheat was gradually replaced (by a third in each case) with rye. Thus, the experimental diets contained 23, 46, and 69% of rye. A total of 40 young pigs were housed individually in four dietary treatment groups. During a 4 week trial, effects on performance, digestibility, products of intestinal fermentation, and fecal characteristics were evaluated. There were no negative effects on feed intake and gains, even though the feed conversion ratio increased with the highest dietary rye level (69%). Digestibility rates of organic matter and crude protein did not differ significantly. Without affecting the characteristics of the feces, numerically higher amounts of intestinal fermentation products and higher colonic digesta mass were observed.

High light triggers flavonoid and polysaccharide synthesis through DoHY5-dependent signaling in Dendrobium officinale.

Dendrobium officinale is edible and has medicinal and ornamental functions. Polysaccharides and flavonoids, including anthocyanins, are important components of D. officinale that largely determine the nutritional quality and consumer appeal. There is a need to study the molecular mechanisms regulating anthocyanin and polysaccharide biosynthesis to enhance D. officinale quality and its market value. Here, we report that high light (HL) induced the accumulation of polysaccharides, particularly mannose, as well as anthocyanin accumulation, resulting in red stems. Metabolome and transcriptome analyses revealed that most of the flavonoids showed large changes in abundance, and flavonoid and polysaccharide biosynthesis was significantly activated under HL treatment. Interestingly, DoHY5 expression was also highly induced. Biochemical analyses demonstrated that DoHY5 directly binds to the promoters of DoF3H1 (involved in anthocyanin biosynthesis), DoGMPP2, and DoPMT28 (involved in polysaccharide biosynthesis) to activate their expression, thereby promoting anthocyanin and polysaccharide accumulation in D. officinale stems. DoHY5 silencing decreased flavonoid- and polysaccharide-related gene expression and reduced anthocyanin and polysaccharide accumulation, whereas DoHY5 overexpression had the opposite effects. Notably, naturally occurring red-stemmed D. officinale plants similarly have high levels of anthocyanin and polysaccharide accumulation and biosynthesis gene expression. Our results reveal a previously undiscovered role of DoHY5 in co-regulating anthocyanin and polysaccharide biosynthesis under HL conditions, improving our understanding of the mechanisms regulating stem color and determining nutritional quality in D. officinale. Collectively, our results propose a robust and simple strategy for significantly increasing anthocyanin and polysaccharide levels and subsequently improving the nutritional quality of D. officinale.

Ginsenosides and Polysaccharides from Ginseng Co-Fermented with Multi-Enzyme-Coupling Probiotics Improve In Vivo Immunomodulatory Effects.

The active components of ginseng, such as ginsenosides and polysaccharides, have high therapeutic value in treating cancer, decreasing obesity, and enhancing immunity. However, simple primary ginseng treatment cannot maximize this medicinal potential. Therefore, in this study, Panax ginseng was co-fermented with multi-enzyme-coupling probiotics to obtain a fermentation broth with higher levels of ginsenosides, polysaccharides, and probiotics. When compared to other treatment methods for cyclophosphamide-induced immunosuppression in mice, the results reveal that the P. ginseng fermentation broth treated with multi-enzyme-coupling probiotics could significantly improve the immune function of immunosuppressive mice and restore intestinal flora stability. Overall, this processing method will provide a novel strategy for promoting the application of ginseng and the relief of immunosuppression.

Estimation of baseline IgG antibody levels to 23 pneumococcal vaccine-type capsular polysaccharides in healthy vaccine naïve Indian adults

Since immunological responses to pneumococcal vaccines are assessed by a fold-increase in antibody levels relative to pre-immunization levels, it is therefore critical to determine baseline antibody levels to establish putative threshold as a measure of normal response. Herein, for the first time, we measured baseline IgG antibody levels in 108 healthy unvaccinated Indian adults using WHO-recommended ELISA. Median baseline IgG concentration ranged between 0.54 µg/mL to 12.35 µg/mL. Highest levels of baseline capsule polysaccharide (cPS)-specific IgG were found against types 14, 19A, and 33F. Whereas, lowest baseline IgG levels were observed against types 3, 4, and 5. Overall, ∼79% of study population had median baseline IgG levels ≥1.3 µg/mL against 74% of cPS’s. Substantial baseline antibody levels in unvaccinated adults were observed. The study would be critical in bridging gaps in baseline immunogenicity data and may offer a valuable foundation for evaluating immune response of Indian adults to pneumococcal vaccination.

Genome sequencing and analysis of Xylaria sp. L1 provides novel insights into its utilization in IDF decomposition in okara

This study investigated the genome ofXylaria sp. L1 and its application in assisting the degradation of okara, which is an agricultural waste. The fungus process showed that there was a 50.31% cellulose loss and 56.25% hemicellulose loss, respectively. In merely 7 days, compared with untreated control, the content of insoluble dietary fiber (IDF) in the substrate was significantly decreased by 62% whereas that of soluble dietary fiber (SDF) significantly increased by 413%. Importantly, the activity of cellulase showed a continuous increase trend and it reached the maximum activity at 11.37U/g on the 7th day. The pretreated biomass showed much higher levels of polysaccharides, especially for glucuronic acid (63.63%), than the untreated controls after 7 days. Additionally, Xylaria sp. L1 sporocarps had higher quantities of macronutrients, especially for Zn. Furthermore, we conducted the first annotation of Xylaria sp. L1 genome. Illumina and nanopore sequencing revealed that the genome size of Xylaria sp. L1 was 36.61 Mb. The high activity of cellulase was in accordance with the annotation the Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Eukaryotic Orthologous Groups (KOG), and carbohydrate-active enzymes (CAZy) databases, which are related to cellulose and hemicellulose degradation. GHs were the most abundant enzymes and 203 genes were annotated to GHs. Cellulose and hemicellulose decomposition was likely performed by an expressed GH43 and GH5 protein families containing their binding domains and genes A0779, A0819, A7717 played an important role in pentose and glucuronate interconversions. Therefore, the higher IDF decomposition by Xylaria sp. L1 makes it a prospective feedstock for okara degradation.

SUPPLEMENTATION OF NSPASE IN FULL-FAT SOYBEAN-BASED DIETS IN BROILER CHICKEN

The present study aimed to assess the impact of incorporating commercially available non-starch polysaccharide enzyme (NSPase) in soybean meal (SBM) and full-fat soybean-based diets on broiler bird development, nutritional digestibility, and gut health. Feed costs account for a significant portion, up to 70%, of overall production expenses in poultry businesses. Broiler diets predominantly consist of cereal grains, which contain varying levels of non-starch polysaccharides (NSPs), a fibrous component. The presence of NSPs in broiler diets hampers overall performance by reducing nutrient absorption due to increased digesta viscosity, thereby decreasing feed efficiency. It is estimated that 400-450 kcal/kg of feed remains unmetabolized in broilers fed a standard corn-soy diet. Exogenous enzymes function by making previously indigestible substances accessible for digestion by endogenous enzymes. NSPase enzyme has the potential to enhance digestion by reducing viscosity and releasing energy through the breakdown of undigested feed components.

The Mechanism of Manganese Ferrite Nanomaterials Promoting Drought Resistance in Rice.

Strategies to reduce the risk of drought damage are urgently needed as intensified climate change threatens agricultural production. One potential strategy was using nanomaterials (NMs) to enhance plant resistance by regulating various physiological and biochemical processes. In the present study, 10 mg kg-1 manganese ferrite (MnFe2O4) NMs had the optimal enhancement to elevate the levels of biomass, photosynthesis, nutrient elements, and polysaccharide in rice by 10.9-525.0%, respectively, under drought stress. The MnFe2O4 NMs were internalized by rice plants, which provided the possibility for rice to better cope with drought. Furthermore, as compared with drought control and equivalent ion control, the introduction of MnFe2O4 NMs into the roots significantly upregulated the drought-sensing gene CLE25 (29.4%) and the receptor gene NCED3 (59.9%). This activation stimulated downstream abscisic acid, proline, malondialdehyde, and wax biosynthesis by 23.3%, 38.9%, 7.2%, and 26.2%, respectively. In addition, 10 mg·kg-1 MnFe2O4 NMs significantly upregulated the relative expressions of OR1, AUX2, AUX3, PIN1a, and PIN2, and increased IAA content significantly, resulting in an enlarged root angle and a deeper and denser root to help the plant withstand drought stresses. The nutritional quality of rice grains was also improved. Our study provides crucial insight for developing nano-enabled strategies to improve crop productivity and resilience to climate change.

Characterization of Oral Candida spp. Biofilms in Children and Adults Carriers from Eastern Europe and South America.

Candida albicans and non-Candida albicans Candida species (NCACs) are known to colonize and invade various tissues, including the oral mucosa. In this work, we aimed to characterize mature biofilms of several Candida spp. clinical isolates (n = 33) obtained from the oral mucosa of children, adults, and elders of Eastern Europe and South America. Each strain was evaluated for its capacity to form biofilms in terms of total biomass using the crystal violet assay and for matrix components production (proteins and carbohydrates) using the BCA and phenol-sulfuric tests, respectively. The effect of different antifungals on biofilm formation was studied. in the children's group, a predominance of C. krusei (81%) was observed, while, among adults, the main species was C. albicans (59%). Most strains showed a reduced response to antimicrobial drugs when in biofilm form (p < 0.01). Moreover, it was observed that strains isolated from children produced more matrix, with higher levels of protein and polysaccharides. children were more likely to be infected by NCACs than adults. More importantly, these NCACs were able to form biofilms richer in matrix components. This finding is of clinical importance, particularly in pediatric care, since stronger biofilms are highly associated with antimicrobial resistance, recurrent infections, and higher therapeutic failure.

Long-term exposure of polytetrafluoroethylene-nanoplastics on the nitrogen removal and extracellular polymeric substances in sequencing batch reactor

The impact of polytetrafluoroethylene-nanoplastics (PTFE-NPs) on biological sewage disposal was delved, containing nitrogen remotion, microbiological activity and composition of extracellular polymer (EPS). The addition of PTFE-NPs reduced the removal efficiencies of chemical oxygen demand (COD) and ammonia nitrogen (NH4+-N) by 3.43 % and 2.35 %, respectively. In comparison with no PTFE-NPs, the specific oxygen uptake rate (SOUR), specific ammonia oxidation rate (SAOR), specific nitrite oxidation rate (SNOR) and specific nitrate reduction rate (SNRR) decreased by 65.26 %, 65.24 %, 41.77 % and 54.56 %, respectively. The PTFE-NPs inhibited the activities of nitrobacteria and denitrobacteria. It was worth noting that, nitrite oxidized bacterium was more resistant to adverse environments than ammonia oxidizing bacterium. Compared with no PTFE-NPs, the reactive oxygen species (ROS) content and lactate dehydrogenase (LDH) grew by 130 % and 50 % under PTFE-NPs pressure. The appearance of PTFE-NPs affected the normal function of microorganisms by inducing endocellular oxidative stress and destroying the completeness of the cytomembrane. The protein (PN) and polysaccharide (PS) levels in loosely bound EPS (LB-EPS) and tightly bound EPS (TB -EPS) increased by 4.96, 0.70, 3.07 and 0.71 mg g−1 VSS, under PTFE-NPs. Meanwhile, the PN/PS ratios of LB-EPS and TB -EPS increased from 6.18 and 6.41–11.04 and 9.29, respectively. The LB-EPS might provide sufficient binding sites for PTFE-NPs adsorption due to its loose and porous structure. The defense mechanism of bacteria against PTFE-NPs was mainly the PN in loosely bound EPS. Moreover, the functional groups referred to the complexation of EPS with PTFE-NPs were mainly related to N-H, CO, and C-N in proteins and O-H in polysaccharides.

Non-Starch Polysaccharide-Degrading Enzymes Increase the Performance of Broiler Chickens Fed Wheat of Low Apparent Metabolizable Energy

The effect of a commercial glycanase product (Avizyme TX) on the performance of 4-wk-old broiler chickens fed wheats with low and normal apparent metabolizable energy values was studied. Controls were fed a corn-based diet. Supplementation with the enzyme product significantly (P < 0.01) increased the apparent metabolizable energy of the low metabolizable energy wheat from 12.02 to 14.94 MJ/kg dry matter. The apparent metabolizable energy value of the normal wheat was increased from 14.52 to 14.83 MJ/kg dry matter; this was, however, not significant. Birds fed the low metabolizable energy wheat diet had significantly (P < 0.01) higher digesta viscosity and lower small intestinal starch and protein digestibilities than birds fed the normal wheat diet. Chickens fed the low metabolizable energy wheat tended to grow less than those fed the normal wheat diet. When the low metabolizable energy wheat + enzyme diet was fed, digesta viscosity was significantly (P < 0.01) lower (20.28 vs. 10.36 mPa·s), and small intestinal digestibility coefficient of starch was significantly (P < 0.01) greater (0.584 vs. 0.861) relative to values in birds fed the low metabolizable energy wheat diet alone. Although the protein digestibility coefficient also increased from 0.689 to 0.745, the difference was not significant. Weight gain and feed efficiency of birds fed the low metabolizable energy wheat + enzyme equaled those of controls. The enzyme product significantly (P < 0.01) increased the solubilization of non-starch polysaccharides within the gastrointestinal tract of birds fed both types of wheat diets. This experiment demonstrated that non-starch polysaccharide-degrading enzymes markedly increase the nutritive value of low metabolizable energy wheat broiler chicken diets, suggesting that increased level of viscous non-starch polysaccharides may cause the low apparent metabolizable energy contents of some wheats.

Comprehensive evaluation and anti-inflammatory activity of “Zhi” polygonatum cyrtonema produced by the classical steaming approach

IntroductionPolygonati Rhizoma, known as “Huangjing”, was first documented in the well-known book Mingyi Bielu, written in the Chinese Han Dynasty. A "Nine-Steam-Nine-Bask" method is the best way to increase their tonic function by steaming and drying the raw rhizomes. This study was conducted with the primary objective of assessing "Zhi" P. cyrtonema qualitatively and quantitatively. Additionally, in vitro studies of the anti-inflammatory activity of "Zhi" P. cyrtonema revealed its pharmacodynamic effects and possible mechanisms of action. MethodsP. cyrtonema was steamed and dried nine times and subsequently subjected to hot water extraction and ethanol precipitation to obtain the desired P. cyrtonema polysaccharides (PCPs). Several approaches, including HPGPC-RID, NMR, and GC-MS, were used to evaluate the molecular weight, composition changes, and monosaccharide composition of PCPs during steaming. Multivariate data analysis, including PCA and HCA, was applied for the divergence analysis of "Zhi" P. cyrtonema. The anti-inflammatory effect of 4-Z PCPs on MH7A cells was evaluated using a cell viability assay. ELISA assays, DAPI staining, cell cycle assay, and scratching experiments revealed the potential mechanism. ResultsWith increasing steaming time, the color of the herb gradually changes from yellow to black and finally to dark black. In processed roots, the levels of polysaccharides decreased, while monosaccharides, such as galactose and glucose, increased. The high-molecular-weight polysaccharide was partially hydrolyzed, forming smaller polysaccharides, monosaccharides, and trace oligosaccharides. In the HPGPC profile, peak 1, with a retention time of 5.20 min and a molecular mass of 1140.51 kDa, can transform into peak 2 (6.10 min, 374.00 kDa) and peak 3 (9.71 min, 9.73 kDa). Using NMR analysis, it has been shown that 5-hydroxymethylfurfural (5-HMF), a toxic substance formed by the thermal degradation of saccharides, is present from the start of the fifth processed herb but is rare before 5-Z P. cyrtonema. The GC-MS analysis also revealed the presence of various monosaccharides, including mannose, galactose, glucose, fructose, rhamnose, arabinose, and galacturonic acid in "Zhi" P. cyrtonema. Using saccharides as variables, we were able to classify "Zhi" P. cyrtonema into four groups: fresh and 1-Z PCPs in groups A, 2-Z, 3-Z, 4-Z, 5-Z, 6-Z, 7-Z and 8-Z PCPs in group C, and 9-Z PCPs in group D. Results of the MH7A cell experiment indicate that 4-Z PCPs are more efficient than 0-Z and 9-Z PCP. After the fourth processing cycle, the differences between metabolic profiles became smaller. ConclusionsA combination of UV–Vis spectra, HPGPC-RID analysis, and NMR techniques were used in this study to achieve qualitative and quantitative analyses of "Zhi" PCPs, and we were able to make a comprehensive evaluation of "Zhi" P. cyrtonema. Meanwhile, using saccharides as variables by the HCA and PCA method, we were able to divide "Zhi" P. cyrtonema into four groups, namely fresh and 1-Z PCPs as group A, and 2-Z, 3-Z and 4-Z PCPs as group B, as well as 5-Z, 6-Z, 7-Z, 8-Z as group C, and 9-Z PCPs as group D. The anti-inflammatory activity studies indicated that the 4-Z PCPs better inhibited the growth of MH7A cells and induced apoptosis of synovial fibroblasts, suggesting that the 4-Z PCPs could be used as an anti-inflammatory reagent.

Long-term performance study applying a tandem AnSBR-ASBR to treat wastewater containing N, N-dimethylformamide: Sludge physicochemical properties, microecology, and functional genes

The effective treatment of industrial pollutant N, N-dimethylformamide (DMF) containing wastewater is a matter of worldwide concern. This study investigated the performance and microecology of anaerobic granular sludge (AGS) and aerobic sludge (AS) in treating DMF wastewater. It was also explored the degradation pathways of DMF in an anaerobic sequencing batch reactor series aerobic sequencing batch reactor (AnSBR-ASBR). Results showed that 5 mg/L DMF facilitated the degradation of chemical oxygen demand (COD) by AGS with AS, while 100 mg/L DMF reduced the efficiency of AS. The physicochemical properties of the sludge showed that the polysaccharide and protein levels in extracellular polymeric substances and key enzyme activity (acetate kinase, coenzyme F420) deteriorated when DMF increased from 25 to 100 mg/L. As the DMF dose increased (0–100 mg/L), the Euryarchaeota abundance in the AGS decreased from 28.41% to 11.06%, and the predominant methanogenic pathway shifted from a combination of aceticlastic methanogenesis (AM) and hydrogenotrophic methanogenesis (HM) to HM. The metabolic pathway to effectively remove DMF was as follows: hydrolysis of DMF to dimethylamine, methylamine, and finally mineralisation to CO2. The abundance of dmd-tmd, a functional gene encoding dimethylamine/trimethylamine dehydrogenase, increased as the DMF increased in the AS. This study systematically examined the operating efficiency and stability of the AnSBR-ASBR, and summarized the AGS and AS stress response, microbial community evolution, key metabolic pathways to DMF, also provided useful information for the treatment of industrial wastewater containing DMF.

Oral Wound Healing Potential of Polygoni Cuspidati Rhizoma et Radix Decoction-In Vitro Study.

Polygoni Cuspidati Rhizoma et Radix (syn. rhizomes of Reynoutria japonica Houtt.) is a pharmacopoeial raw material in Europe and China. In traditional medicine, one of the applications for Reynoutria japonica rhizomes is wound healing. In a recent in vitro study, we demonstrated that ethanol and acetone extracts from this herbal drug have the potential to heal oral gum wounds. However, considering that a majority of herbal medicines have been traditionally administered as water decoctions, in the present study, a decoction of Reynoutria japonica rhizomes was prepared and detailed tests to determine its in vitro gingival wound healing activity were conducted. We used the primary human gingival fibroblasts (HGF) incubated with a decoction to determine cell viability (MTT assay), cell proliferation (the confocal laser scanning microscope-CLSM), and cell migration (wound healing assay). Moreover, the collagen type III expression was examined using immunocytochemical staining. The studied decoction was qualitatively and quantitatively characterized using the validated HPLC/DAD/ESI-HR-QTOF-MS method. The Folin-Ciocalteu assay was used to determine the total phenols and tannins content. Additionally, HPLC-RI analysis of decoction and the previously obtained ethanol and acetone extracts was used to determine the composition of saccharides. Low concentration (from 50 to 1000 µg/mL) of decoction after 24 h incubation caused a significant increase in HGF cell viability. No cytotoxic effect was observed at any tested concentration (up to 2000 µg/mL). The lowest active concentration of decoction (50 µg/mL) was selected for further experiments. It significantly stimulated human gingival fibroblasts to proliferate, migrate, and increase the synthesis of collagen III. Phytochemical analysis showed significantly fewer polyphenols in the decoction than in the ethanol and acetone extracts tested earlier. In contrast, high levels of polysaccharides were observed. In our opinion, they may have a significant effect on the oral wound healing parameters analyzed in vitro. The results obtained encourage the use of this raw material in its traditional, safe form-decoction.