In order to investigate the enzyme production mechanism of yak rumen-derived anaerobic fungus Orpinomyces sp. YF3 under the induction of different carbon sources, anaerobic culture tubes were used for in vitro fermentation. 8 g/L of glucose (Glu), filter paper (Flp) and avicel (Avi) were respectively added to 10 mL of basic culture medium as the sole carbon source. The activity of fiber-degrading enzyme and the concentration of volatile fatty acid in the fermentation liquid were detected, and the enzyme producing mechanism of Orpinomyces sp. YF3 was explored by transcriptomics. It was found that, in glucose-induced fermentation solution, the activities of carboxymethyl cellulase, microcrystalline cellulase, filter paper enzyme, xylanase and the proportion of acetate were significantly increased (P < 0.05), the proportion of propionate, butyrate, isobutyrate were significantly decreased (P < 0.05). The results of transcriptome analysis showed that there were 5 949 differentially expressed genes (DEGs) between the Glu group and the Flp group, 10 970 DEGs between the Glu group and the Avi group, and 6 057 DEGs between the Flp group and the Avi group. It was found that the DEGs associated with fiber degrading enzymes were significantly up-regulated in the Glu group. Gene ontology (GO) function enrichment analysis identified that DEGs were mainly associated with the xylan catabolic process, hemicellulose metabolic process, β-glucan metabolic process, cellulase activity, endo-1,4-β-xylanase activity, cell wall polysaccharide metabolic process, carbohydrate catabolic process, glucan catabolic process and carbohydrate metabolic process. Moreover, the differentially expressed pathways associated with fiber degrading enzymes enriched by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were mainly starch and sucrose metabolic pathways and other glycan degradation pathways. In conclusion, Orpinomyces sp. YF3 with glucose as carbon source substrate significantly increased the activity of cellulose degrading enzyme and the proportion of acetate, decreased the proportion of propionate, butyrate and isobutyrate. Furthermore, the degradation ability and energy utilization efficiency of fungus in the presence of glucose were improved by means of regulating the expression of cellulose degrading enzyme gene and participating in starch and sucrose metabolism pathway, and other glycan degradation pathways, which provides a theoretical basis for the application of Orpinomyces sp. YF3 in practical production and facilitates the application of Orpinomyces sp. YF3 in the future.
Animals ; Cattle ; Neocallimastigales/metabolism* ; Anaerobiosis ; Rumen/microbiology* ; Propionates/metabolism* ; Isobutyrates/metabolism* ; Cellulose/metabolism* ; Fungi ; Starch/metabolism* ; Glucose/metabolism* ; Acetates ; Sucrose/metabolism* ; Cellulases ; Cellulase

BACKGROUND: No convincing modalities have been shown to completely prevent postdural puncture headache (PDPH) after accidental dural puncture (ADP) during obstetric epidural procedures. We aimed to evaluate the role of epidural administration of hydroxyethyl starch (HES) in preventing PDPH following ADP, regarding the prophylactic efficacy and side effects. METHODS: Between January 2019 and February 2021, patients with a recognized ADP during epidural procedures for labor or cesarean delivery were retrospectively reviewed to evaluate the prophylactic strategies for the development of PDPH at a single tertiary hospital. The development of PDPH, severity and duration of headache, adverse events associated with prophylactic strategies, and hospital length of stay postpartum were reported. RESULTS: A total of 105 patients experiencing ADP received a re-sited epidural catheter. For PDPH prophylaxis, 46 patients solely received epidural analgesia, 25 patients were administered epidural HES on epidural analgesia, and 34 patients received two doses of epidural HES on and after epidural analgesia, respectively. A significant difference was observed in the incidence of PDPH across the groups (epidural analgesia alone, 31 [67.4%]; HES-Epidural analgesia, ten [40.0%]; HES-Epidural analgesia-HES, five [14.7%]; P <0.001). No neurologic deficits, including paresthesias and motor deficits related to prophylactic strategies, were reported from at least 2 months to up to more than 2 years after delivery. An overall backache rate related to HES administration was 10%. The multivariable regression analysis revealed that the HES-Epidural analgesia-HES strategy was significantly associated with reduced risk of PDPH following ADP (OR = 0.030, 95% confidence interval: 0.006-0.143; P < 0.001). CONCLUSIONS The incorporated prophylactic strategy was associated with a great decrease in the risk of PDPH following obstetric ADP. This strategy consisted of re-siting an epidural catheter with continuous epidural analgesia and two doses of epidural HES, respectively, on and after epidural analgesia. The efficacy and safety profiles of this strategy have to be investigated further.
Pregnancy ; Female ; Humans ; Post-Dural Puncture Headache/epidemiology* ; Anesthesia, Obstetrical/adverse effects* ; Retrospective Studies ; Punctures ; Starch ; Blood Patch, Epidural

α-amylase is an endonucleoside hydrolase that hydrolyzes the α-1, 4-glycosidic bonds inside polysaccharides, such as starch, to generate oligosaccharides, dextrins, maltotriose, maltose and a small amount of glucose. Due to the importance of α-amylase in food industry, human health monitoring and pharmaceuticals, detection of its activity is widely required in the breeding of α-amylase producing strains, in vitro diagnosis, development of diabetes drugs, and the control of food quality. In recent years, many new α-amylase detection methods have been developed with improved speed and sensitivity. This review summarized recent processes in the development and applications of new α-amylase detection methods. The major principle of these detection methods were introduced, and their advantages and disadvantages were compared to facilitate future development and applications of α-amylase detection methods.
Humans ; alpha-Amylases/chemistry* ; Polysaccharides ; Oligosaccharides ; Starch ; Maltose

Sweet potato is an important food crop that can also be used as an industrial raw material. Sucrose is the main form of long-distance carbohydrate transport in plants, and sucrose transporter (SUT) regulates the transmembrane transport and distribution of sucrose during plant growth and metabolism. Moreover, SUT plays a key role in phloem mediated source-to-sink sucrose transport and physiological activities, supplying sucrose for the sink tissues. In this study, the full-length cDNA sequences of IbSUT₆₂₇₈₈ and IbSUT₈₁₆₁₆ were obtained by rapid amplification of cDNA ends (RACE) cloning according to the transcripts of the two SUT coding genes which were differentially expressed in sweet potato storage roots with different starch properties. Phylogenetic analysis was performed to clarify the classification of IbSUT₆₂₇₈₈ and IbSUT₈₁₆₁₆. The subcellular localization of IbSUT₆₂₇₈₈ and IbSUT₈₁₆₁₆ was determined by transient expression in Nicotiana benthamiana. The function of IbSUT₆₂₇₈₈ and IbSUT₈₁₆₁₆ in sucrose and hexose absorption and transport was identified using yeast functional complementarity system. The expression pattern of IbSUT₆₂₇₈₈ and IbSUT₈₁₆₁₆ in sweet potato organs were analyzed by real-time fluorescence quantitative PCR (RT-qPCR). Arabidopsis plants heterologous expressing IbSUT₆₂₇₈₈ and IbSUT₈₁₆₁₆ genes were obtained using floral dip method. The differences in starch and sugar contents between transgenic and wild-type Arabidopsis were compared. The results showed IbSUT₆₂₇₈₈ and IbSUT₈₁₆₁₆ encoded SUT proteins with a length of 505 and 521 amino acids, respectively, and both proteins belonged to the SUT1 subfamily. IbSUT₆₂₇₈₈ and IbSUT₈₁₆₁₆ were located in the cell membrane and were able to transport sucrose, glucose and fructose in the yeast system. In addition, IbSUT₆₂₇₈₈ was also able to transport mannose. The expression of IbSUT₆₂₇₈₈ was higher in leaves, lateral branches and main stems, and the expression of IbSUT₈₁₆₁₆ was higher in lateral branches, stems and storage roots. After IbSUT₆₂₇₈₈ and IbSUT₈₁₆₁₆ were heterologously expressed in Arabidopsis, the plants grew normally, but the biomass increased. The heterologous expression of IbSUT₆₂₇₈₈ increased the soluble sugar content, leaf size and 1 000-seed weight of Arabidopsis plants. Heterologous expression of IbSUT₈₁₆₁₆ increased starch accumulation in leaves and root tips and 1 000-seed weight of seeds, but decreased soluble sugar content. The results obtained in this study showed that IbSUT₆₂₇₈₈ and IbSUT₈₁₆₁₆ might be important genes regulating sucrose and sugar content traits in sweet potato. They might carry out physiological functions on cell membrane, such as transmembrane transport of sucrose, sucrose into and out of sink tissue, as well as transport and unloading of sucrose into phloem. The changes in traits result from their heterologous expression in Arabidopsis indicates their potential in improving the yield of other plants or crops. The results obtained in this study provide important information for revealing the functions of IbSUT₆₂₇₈₈ and IbSUT₈₁₆₁₆ in starch and glucose metabolism and formation mechanism of important quality traits in sweet potato.
Ipomoea batatas/metabolism* ; Arabidopsis/metabolism* ; Sucrose/metabolism* ; Saccharomyces cerevisiae/metabolism* ; DNA, Complementary ; Phylogeny ; Plants, Genetically Modified/genetics* ; Membrane Transport Proteins/metabolism* ; Starch/metabolism* ; Plant Proteins/metabolism* ; Gene Expression Regulation, Plant

An absorbable hemostatic material based on polysaccharide was prepared. The concentration of blood cells and coagulation factors was increased by reducing the water content in the blood, so as to reduce the coagulation time and achieve the purpose of rapid hemostasis. The specific surface area of starch was increased by using hydrochloric acid to hydrolyze potato starch, which made it easier to combine with α-amylase and increased the degradation rate. Starch was crosslinked into microspheres by crosslinking agent, which made the particle size uniform and greatly improved the water absorption. The surface modification of crosslinked starch microspheres with carboxymethyl group can further improve the water absorption of hemostatic materials. The results showed that the water absorption rate of our hemostatic material was more than 800%, and the average hemostatic time in the animal model was 138.7s. Compared with the imported products on the market, our hemostatic material have better hemostatic performance.
Animals ; Hemostasis ; Hemostatics/pharmacology* ; Polysaccharides/pharmacology* ; Starch/pharmacology* ; Water/pharmacology*

Fluid resuscitation is an essential intervention in critically ill patients, and its ultimate goal is to restore tissue perfusion. Critical illnesses are often accompanied by glycocalyx degradation caused by inflammatory reactions, hypoperfusion, shock, and so forth, leading to disturbed microcirculatory perfusion and organ dysfunction. Therefore, maintaining or even restoring the glycocalyx integrity may be of high priority in the therapeutic strategy. Like drugs, however, different resuscitation fluids may have beneficial or harmful effects on the integrity of the glycocalyx. The purpose of this article is to review the effects of different resuscitation fluids on the glycocalyx. Many animal studies have shown that normal saline might be associated with glycocalyx degradation, but clinical studies have not confirmed this finding. Hydroxyethyl starch (HES), rather than other synthetic colloids, may restore the glycocalyx. However, the use of HES also leads to serious adverse events such as acute kidney injury and bleeding tendencies. Some studies have suggested that albumin may restore the glycocalyx, whereas others have suggested that balanced crystalloids might aggravate glycocalyx degradation. Notably, most studies did not correct the effects of the infusion rate or fluid volume; therefore, the results of using balanced crystalloids remain unclear. Moreover, mainly animal studies have suggested that plasma may protect and restore glycocalyx integrity, and this still requires confirmation by high-quality clinical studies.
Animals ; Colloids ; Crystalloid Solutions/therapeutic use* ; Fluid Therapy ; Glycocalyx ; Humans ; Hydroxyethyl Starch Derivatives ; Isotonic Solutions ; Microcirculation ; Resuscitation

Starch is composed of glucose units linked by α-1, 4-glucoside bond and α-1, 6-glucoside bond. It is the main component of foods and the primary raw material for starch processing industry. Pullulanase can effectively hydrolyze the α-1, 6-glucoside bond in starch molecules. Combined with other starch processing enzymes, it can effectively improve the starch utilization rate. Therefore, it has been widely used in the starch processing industry. This paper summarized the screening of pullulanase-producing strain and its encoding genes. In addition, the effects of expression elements and fermentation conditions on the production of pullulanase were summarized. Moreover, the progress in crystal structure elucidation and molecular modification of pullulanase was discussed. Lastly, future perspectives on pullulanase research were proposed.
Glycoside Hydrolases/genetics* ; Starch/metabolism*

Background: Gold nanoparticles have been studied extensively for their potential application in the detection of important analytes. Their relative ease of synthesis through numerous procedures makes possible their implementation in a variety of assays. Cysteine (cys), a thiol-containing amino acid implicated in numerous pathologies such as obstructive sleep apnea (OSA), has been routinely detected through expensive fluorometric assay kits. Objectives: As such, this study aimed to develop a carbohydrate-based gold nanoparticle colorimetric assay for the convenient and straightforward detection of cys. Methodology: Carbohydrate-based gold nanoparticles (c-AuNPs) were synthesized following a microwaveassisted procedure. The as-prepared c-AuNPs were used to detect cys by plotting the ratio of the absorbances of the aggregated and dispersed gold nanoparticles against the concentration of cys. Results: The c-AuNP solutions were able to detect cys in the micromolar range, with the glucose-based AuNPs (glc-AuNPs) showing the widest linear range (16.7 μm to 167 μm), and the fructose-based gold nanoparticles (frc-AuNPs) exhibiting the lowest detection limit (9.0 μm) for cys. Aside from being able to detect cys, the c-AuNPs were also responsive to tyr and lys. Conclusion This study demonstrates that carbohydrate-based gold nanoparticles prepared following a microwave-assisted procedure using sugars as reducing agents and capping agents can be used successfully in the detection of cysteine.
Cysteine ; Carbohydrates ; Starch

Coupling sugar is a kind of new sweetener which can substitute sucrose. It has a good application prospect in food, medicine and other fields because of its good coloration, water retention and anti caries. The purpose of this study was to find cheap and easily available donor and acceptor, and to optimize the preparation process of coupling sugar by using β-cyclodextrin glycosyltransferase from Bacilluscirculans 251. Using sucrose as acceptor, the factors of preparing coupling sugar was optimized, including enzyme dosage, starch types, temperature, pH, ratio of starch/sucrose, and cooperation of isoamylase and β-CGTase. When 105 g/L potato starch and 95 g/L sucrose was used as substrates, the yield of coupling sugar reached 88.4%, which was catalyzed by 13.5 U/g immobilized β-CGTase and 45.0 U/g isoamylase under the conditions of pH 5.5 and 40 °C for 21 h. In this study, isoamylase and β-CGTase were used to prepare coupling sugar innovatively. This method had obvious advantages in yield and cost, which laid both theoretical and experimental foundation for the industrial enzymatic preparation of coupling sugars.
Glucosyltransferases ; Hydrogen-Ion Concentration ; Isoamylase ; Starch

Production of biofuels such as ethanol from non-grain crops may contribute to alleviating the global energy crisis and reducing the potential threat to food security. Tobacco (Nicotiana tabacum) is a commercial crop with high biomass yield. Breeding of starch-rich tobacco plants may provide alternative raw materials for the production of fuel ethanol. We cloned the small subunit gene NtSSU of ADP-glucose pyrophosphorylase (NtAGPase), which controls starch biosynthesis in tobacco, and constructed a plant expression vector pCAMBIA1303-NtSSU. The NtSSU gene was overexpressed in tobacco upon Agrobacterium-mediated leaf disc transformation. Phenotypic analysis showed that overexpression of NtSSU gene promoted the accumulation of starch in tobacco leaves, and the content of starch in tobacco leaves increased from 17.5% to 41.7%. The growth rate and biomass yield of the transgenic tobacco with NtSSU gene were also significantly increased. The results revealed that overexpression of NtSSU gene could effectively redirect more photosynthesis carbon flux into starch biosynthesis pathway, which led to an increased biomass yield but did not generate negative effects on other agronomic traits. Therefore, NtSSU gene can be used as an excellent target gene in plant breeding to enrich starch accumulation in vegetative organs to develop new germplasm dedicated to fuel ethanol production.
Biomass ; Gene Expression Regulation, Plant ; Plant Breeding ; Plant Leaves/genetics* ; Plants, Genetically Modified/metabolism* ; Starch ; Tobacco/metabolism*