Effectiveness and Safety of Coils Plus Glue in Slope Embankment Technology Versus Coils Plus Sclerosant in Embolization Therapy for Reflux-Type Pelvic Venous Disorders.

OBJECTIVE: This study aimed to evaluate the effectiveness and safety of coils plus glue in slope embankment technology versus coils plus sclerosant in treating reflux-type pelvic venous disorders. METHODS: The analysis included patients diagnosed with reflux-type pelvic venous disorders who were treated with coils plus glue (CPG) or coils plus sclerosant (CPS) from 2019 to 2021. The inclusion criteria were non-cyclic pain lasting more than 6 months, atypical varicose, and transvaginal Doppler ultrasonography (TVDUS) and computed tomographic venography（CTV) confirming the diagnosis and excluding compression factors and other diseases. Propensity score matching was performed at a 1:1.1 ratio based on the following covariates: age, pregnancy, BMI, pretreatment VAS, dysmenorrhea, dyspareunia, urinary urgency, tenesmus, low back pain, vulvar varicosities, vaginal varicosities and lower limb varices. The pain was relieved by embolizing the target lesions with different embolic materials. The efficacy and safety of the different embolization materials were compared by visual analog scale (VAS) and TVDUS examinations at 1, 3, 6, 12, 24, and 36 months. RESULTS: From a total of 495 patients, 88 patients were selected from the CPG group and 77 patients from the CPS group by propensity score matching. The patients were followed up for 36 months. The preoperative VAS score of the Coils Plus Glue(CPG)group was 8 (range 6-8), and the Coils Plus Sclerosant(CPS) score was 8 (range 7-8),P=0.64. The post-embolization VAS score of the CPG group was 2.05±0.37, and the CPS score was 2.14±0.35(P=0.55). A total of 28 cases (16.9%) showed complications, most of which were transient pain after embolization. No serious complications such as coil embolization to the lungs occurred. In addition, the CPG group used fewer coils than the CPS group by using the Slope Embankment Technique(SET) . The mean coil length of the CPG group was 77.18±33.82 cm, and the CPS group was 105.29±71 cm(P=0.001). The CPG group had an average operation time of 44.49±5.72 min, while the CPS group took 43.45±4.18 min on average(P=0.19). The radiation dose of the CPG group was 398.40±76.16mGy, and the radiation dose of the CPS group was 388±44.23mGy(P=0.30). The median recurrence free survival(RFS) of the CPG group was 34.23 months (95% CI 33.2-35.2), and the median RFS of the CPS group was 30.39 months (95% CI 28.2-32.6), LogRank=0.018. CONCLUSIONS: Embolization therapy for refluxing PeVD was safe and effective, and proficient use of SET with coils plus glue increased efficacy and reduced complications.

Enhancing water retention and mechanisms of citrus and soya bean dietary fibres in pre-fermented frozen dough.

In recent years, the production of prepared and frozen foods has increased with economic development. However, during freezing, moisture migration forms ice crystals that damage food structure and reduce quality. This study investigates moisture migration changes in pre-fermented dough during frozen storage and effectiveness of Citrus fibre (CF) and Soya dietary fibre (SDF) on quality improvement. Pre-fermented frozen dough properties were evaluated at different freezing storage days with CF and SDF. Results showed frozen storage reduced water retention, converting deeply bound water to weakly bound and free water. Freezable water content increased significantly from 53% (fresh) to 56.95% (60d-control), forming disruptive ice crystals in gluten protein structure. SDF had superior water flow restriction compared to CF, preventing large ice crystal accumulation, enhancing water-holding capacity, and maintaining gluten protein structure. These findings lay a theoretical foundation for improving quality and industrial applications of pre-fermented frozen dough.

Efficacy Analysis of Drug-Coated Balloons in the Treatment of Ultra-Long Segment Lesions of Femoral Popliteal Artery.

BACKGROUND: To evaluate the midterm clinical efficacy of paclitaxel drug-coated balloons (DCBs) in the treatment of femoral artery TransAtlantic Inter-Society Consensus (TASC) grades C/D lesions. METHODS: The clinical data of 73 cases with TASC grades C/D lesions of femoral artery treated with paclitaxel DCBs at the Department of Vascular Surgery, the First Hospital of Fujian Medical University from August 2016 to January 2020 were retrospectively analyzed. The primary endpoint was the primary patency rate. The secondary endpoints were freedom from reintervention, Rutherford classification, ankle-brachial index (ABI), amputation events, and all-cause death. RESULTS: A total of 73 cases of limb lesions received endoluminal treatment. The mean age of the patients including 49 males and 24 females was (72.66 ± 11.1) years, with an initial Rutherford classification of 2-5 and an ABI of 0.4 ± 0.1. The mean Rutherford classification was 3.70 ± 0.95. The mean lesion length was (25.75 ± 9.67) cm, including 61.64% chronic occlusive lesions and 27.39% stenotic lesions, the remaining 10.97% were mixed lesions, containing multiple segments of stenosis and chronic total occlusion lesions. 43.8% of the lesions were associated with severe calcification. Stent implantation rate was 8%. Overall mortality at follow-up was 4% at 1 year and 8% at 2 years, and no amputations seen. The ABI was 0.83 ± 0.07 at 1-year follow-up and 0.78 ± 0.05 at 2-year follow-up. The Kaplan-Meier survival curve predicted the 1-year phase I patency rate was 75.3% ± 5% and the 2-year patency rate was 63.3% ± 5.7%. Freedom from target lesion revascularization was 78.4 ± 4.9% at 1 year and 69.2% ± 3.6% at 2 years. Logistic regression analysis showed that diabetes mellitus, severe calcification, chronic renal insufficiency, and restenosis were the significant factors affecting the patency of target lesions. CONCLUSIONS: Paclitaxel DCBs in the treatment of femoral artery with TASC grades C/D lesions can achieve relatively satisfactory midterm clinical safety and efficacy results, provided there is an acceptable result on completion angiogram.

Antioxidant Benefits and Potential Mechanisms of Slightly Acidic Electrolyzed Water Germination in Sesame.

Slightly acidic electrolytic water (SAEW) treatment for seed germination is a promising technique for sustainable agriculture. This study investigated the antioxidant activity of germinated sesame seeds treated with SAEW for the first time. Specifically, the impact and correlation of SAEW on the activities of total phenols, total flavonoids, and antioxidant oxidase in sesame seeds were examined. The results showed that SAEW with low ACC inhibited sesame germination, SAEW with high ACC promoted sesame germination, and sesame buds treated with SAEW with 30 mg/L and 50 mg/L ACC showed lower antioxidant activity and total phenolic and flavone content compared to tap water. In contrast, SAEW with 30 mg/L ACC had no significant effect on sesame growth but positively influenced the antioxidant activity of sesame seed germination by promoting phenolic compound synthesis through increased phenylalanine ammonia-lyase (PAL) activity and enhancing antioxidant activity by boosting PAL, polyphenol oxidase (PPO), and peroxidase (POD) activities. Generally, antioxidant ability was the most prominent in SAEW with 30 mg/L ACC, and positive correlations between antioxidation and total phenols and flavonoids content were found in sesame. These findings provide valuable insights into the mechanisms underlying the enhanced antioxidant capacity observed in germinated sesame seeds under SAEW stress.

Mechanism of acid and alkali electrolyzed water on the elimination of Listeria monocytogenes biofilm based on proteomic analysis.

Acidic electrolyzed water is a relatively mature bactericide, which has a certain inhibitory effect on a variety of microorganisms, and is widely used in the field of food processing for cleaning, sterilization and disinfection. This study investigated the deactivation mechanisms of Listeria monocytogenes by Tandem Mass Tags quantitative proteomics analysis. Samples were treated through A1S4 (Alkaline electrolytic water treatment for 1 min and Acid electrolytic water treatment for 4 min), S3A1S1 (Acid electrolyzed water treatment 3 min, Alkaline electrolyzed water treatment 1 min and Acid electrolyzed water treatment 1 min), S5 (Acid electrolytic water treatment for 5 min). Proteomic analysis showed that the mechanism of acid alkaline electrolyzed water treatment to eliminate the inactivation of the biofilm of L. monocytogenes was related to protein transcription and extension, RNA processing and synthesis, gene regulation, sugar and amino acid transport and metabolism, signal transduction and ATP binding. The study on the influence mechanism and action mechanism of the combination of acidic and alkaline electrolyzed water to remove L. monocytogenes biofilm is helpful to understand the development of the process of removing biofilm by electrolyzed water, and provides theoretical support for the treatment of other microbial contamination problems in food processing by electrolyzed water.

Purification, characterization and inactivation kinetics of polyphenol oxidase extracted from Cistanche deserticola.

MAIN CONCLUSION: PPO was purified from Cistanche deserticola, and its enzymatic characteristics were clarified. It was found that microwave treatment was an efficient way to inactivate PPO. Polyphenol oxidase (PPO) from Cistanche deserticola was obtained and purified through an acetone precipitation and anion exchange column, the enzymatic characteristics and inactivation kinetics of PPO were studied. The specific activity of PPO was 73135.15 ± 6625.7 U/mg after purification, the purification multiple was 48.91 ± 4.43 times, and the recovery was 30.96 ± 0.27%. The molecular weight of the PPO component is about 66 kDa by SDS-PAGE analysis. The optimum substrate of PPO was catechol (Vmax = 0.048 U/mL, Km = 21.70 mM) and the optimum temperature and pH were 30 °C and 7, respectively. When the temperature is above 50 °C, pH < 3 or pH > 10, the enzyme activity can be significantly inhibited. The first-order kinetic fitting shows that microwave inactivation has lesser k values, larger D values and shorter t1/2. It was found that microwave treatment is considered as an efficient and feasible way to inactive PPO by comparing the Z values and Ea values of the two thermal treatments.

Bactericidal Effect and Associated Properties of Non-Electrolytic Hypochlorite Water on Foodborne Pathogenic Bacteria.

This study investigated the broad-spectrum bactericidal activity of non-electrolytic hypochlorite water (NEHW) and detected its hydroxyl radical content compared with that of slightly acidic electrolytic water (SAEW). Based on the results of UV scanning and storage stability, higher hypochlorite content and stronger oxidation were found to be responsible for the stronger bactericidal effect of NEHW. NEHW can achieve 99% bacterial disinfection effect by treating with 10 mg/L available chlorine concentration for more than 5 minutes. At the same time, the storage stability of NEHW was higher than that of SAEW. After 20 days of storage under sealed and dark conditions, the pH value only increased by 7.9%, and the effective chlorine concentration remained nearly 80%. The results showed that NEHW had higher germicidal efficacy and storage stability than SAEW.

Mechanically Strong and Thermally Stable Chemical Cross-Linked Polyimide Aerogels for Thermal Insulator.

High-performance thermal insulating materials are highly desirable in several fields, especially for thermal insulation of buildings to reduce energy consumption. Owing to the remarkable thermal stability, high porosity, low density, and outstanding mechanical features, polyimide (PI) aerogels have attracted great attention. In this work, chemical cross-linked PI (CCPI) aerogels were fabricated via freeze-drying and thermal imidization, which possess outstanding mechanical properties, good thermal stability, and excellent thermal insulation characteristics. The chemically cross-linked structure can effectively inhibit shrinkage, while retaining the structural integrity, resulting in the lower density and lower shrinkage of the materials. In this paper, completely imidized and highly cross-linked polyimide aerogels were synthesized by using p-phenylenediamine (PDA), 3,3',4,4'-biphenyltetracarboxylic dianhydride (BPDA), and the cross-linker 2,3,6,7,14,15-hexaaminotriptycene (HMT). The CCPI aerogels with excellent properties, such as covalently cross-linked chemical structure, low density (0.069 g/cm3), low volume shrinkage (10%), high decomposition temperature (Td5% = 587 °C), and low thermal conductivity (25 mW m-1K-1) are in high demand in the field of thermal insulation. This work furnishes a new method for the development of polymer-based thermal insulation materials for various prospective applications.

Cobalt-doping of molybdenum phosphide nanofibers for trapping-diffusion-conversion of lithium polysulfides towards high-rate and long-life lithium-sulfur batteries.

Rational design of separators is especially critical to solve the "shuttle effect" of lithium polysulfides (LiPSs) and the sluggish redox kinetics in lithium-sulfur batteries (LSBs). Here, the multi-functional nanocomposite involving Co-doped molybdenum phosphide (Co-MoP) nanofibers and porous carbon nanofibers (PCNFs) is designed and prepared through electro-blow spinning and phosphating process, which possesses multiple adsorption and catalytic sites and is acted as the functional material for LSBs separators. In this multifunctional nanocomposite, the prepared Co-MoP nanofibers can provide internal adsorption and catalytic sites for LiPSs conversion. And the interconnected nitrogen-doped PCNFs can be elaborated an efficient LiPSs mediator and accommodate the huge volume changes in the reaction process for LSBs. Benefiting from the multiple adsorptive and catalytic sites of the developed functional materials, the assembled LSBs with a Co-MoP/PCNFs modified separator display outstanding electrochemical performances, including an admirable capacity retention of 770.4 mAh g-1 after 400 cycles at 1.0 C, only 0.08 % capacity decay per cycle at 2.0 C, rate performance up to 5 C, and also decent areal capacity even under a high sulfur loading of 4.9 mg cm-2. The work provides a facile pathway towards multifunctional separators in LSBs, and it may also help deepen preparation method of MoP through the electrostatic blowing/electrospinning technology in other related energy storage fields.

Numerical study on the influence of wall temperature gradient on aerodynamic characteristics of low aspect ratio flying wing configuration.

With the aim for a low-aspect-ratio flying wing configuration, this study explores the influence of wall temperature gradient on the laminar and turbulent boundary layers of aircraft surface and determines the effect on the transition Reynolds number and wall friction drag. A four-equation turbulence model with transition mode is used to numerically simulate the flow around the model. The variation of wall friction coefficient, transition Reynolds number, and turbulent boundary layer flow with wall temperature are emphatically investigated. Results show that when the wall temperature increases from 288 to 500 K, the boundary layer transition Reynolds number for the wing section increased by approximately 28% and the surface friction drags decreases by approximately 10.7%. The hot wall enhances the viscous effects of the laminar temperature boundary layer, reduces the Reynolds shear stress and turbulent kinetic energy, and increases the flow stability. However, the velocity gradient and shear stress in the bottom of the turbulent boundary layer decreases, which leads to reduced friction shear stress on the wall surface. Therefore, for the low-aspect-ratio flying wing model, the hot wall can delay the boundary layer transition and reduce the friction drag coefficient in the turbulent region.

Thermal and Mechanical Performances of the Superflexible, Hydrophobic, Silica-Based Aerogel for Thermal Insulation at Ultralow Temperature.

A superflexible hydrophobic silica-based aerogel (FHSA) was prepared via a facile sol-gel process and ambient pressure drying method. The FHSA was treated at different temperatures varying from -196 to 450 °C to evaluate its thermal and mechanical performances. The evolutions of the physical property, hydrophobicity, microstructure, pore structure, and chemical structure of the FHSA with the various treatment temperatures were investigated comprehensively. The structure of the FHSA did not show an obvious change after treatment in the liquid nitrogen. The bulk density of the FHSA increased from 0.047 to 0.077 g cm-3 when the thermal treatment temperature increased from 25 to 450 °C. The specific surface area and pore volume of the FHSA increased with the treatment temperature owing to the decomposition of the organic moieties. The Fourier transform infrared spectra showed that the methyl groups in the FHSA had excellent thermostability up to 400 °C. The water contact angles of the FHSA after treatment at -196, 25, 200, 300, 350, 400, and 450 °C were 131, 151, 162, 150, 132, 119, and 34°, respectively. The thermal conductivity of the FHSA at a low temperature of -10 °C was 0.022 W m-1 K-1. The reversible deformation rate of the FHSA was more than 80% within 100 compression cycles. After treatment in liquid nitrogen, the reversible deformation rate of the FHSA remained at 50%. The synthesis method of the FHSA is simple, the resulting FHSA showed good performance both in thermostability and flexibility, and it is promisingly applied for thermal insulation and sealing in ultralow-temperature environments.

High-level expression and enzymatic properties of a novel thermostable xylanase with high arabinoxylan degradation ability from Chaetomium sp. suitable for beer mashing.

A novel thermostable xylanase gene from Chaetomium sp. CQ31 was cloned and codon-optimized (CsXynBop). The deduced protein sequence of the gene shared the highest similarity of 75% with the glycoside hydrolase (GH) family 10 xylanase from Achaetomium sp. Xz-8. CsXynBop was over-expressed in Pichia pastoris GS115 by high-cell density fermentation, with the highest xylanase yield of 10,017 U/mL. The recombinant xylanase (CsXynBop) was purified to homogeneity and biochemically characterized. CsXynBop was optimally active at pH 6.5 and 85 °C, respectively, and stable over a broad pH range of 5.0-9.5 and up to 60 °C. The enzyme exhibited strict substrate specificity towards oat-spelt xylan (2, 489 U/mg), beechwood xylan (1522 U/mg), birchwood xylan (1067 U/mg), and showed relatively high activity towards arabinoxylan (1208 U/mg), but exhibited no activity on other tested polysaccharides. CsXynBop hydrolyzed different xylans to yield mainly xylooligosaccharides (XOSs) with degree of polymerization (DP) 2-5. The application of CsXynBop (200 U/g malt) in malt mashing substantially decreased the filtration time and viscosity of malt by 42.3% and 8.6%, respectively. These excellent characteristics of CsXynBop may make it a good candidate in beer industry.

NPPB prevents postoperative peritoneal adhesion formation by blocking volume-activated Cl- current.

5-Nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB) is a non-specific chloride channel blocker. Peritoneal adhesion is an inevitable complication of abdominal surgery and remains an important clinical problem, leading to chronic pain, intestinal obstruction, and female infertility. The aim of this study is to observe the effects of NPPB on peritoneal adhesions and uncover the underlying mechanism. The formation of postoperative peritoneal adhesions was induced by mechanical injury to the peritoneum of rats. MTT assay and wound-healing assay were used to evaluate proliferation and migration of primary cultured adhesion fibroblasts (AFB) respectively. Whole-cell chloride currents were measured using a fully automated patch-clamp workstation. Cell volume changes were monitored by light microscopy and video imaging. Our results demonstrated that NPPB could significantly prevent the formation of peritoneal adhesion in rats and inhibit the proliferation of AFB in a concentration-dependent manner. NPPB also reduced the migration of AFB cells with an IC50 of 53.09 µM. A 47% hypotonic solution successfully activated the ICl,vol in AFB cells. The current could be blocked by extracellular treatment with NPPB. Moreover, 100 µM NPPB almost completely eliminated the capacity of regulatory volume decrease (RVD) in these cells. These data indicate that NPPB could prevent the formation of postoperative peritoneal adhesions. The possible mechanism may be through the inhibition of the proliferation and migration of AFB cells by modulating ICl,vol and cell volume. These results suggest a potential clinical use of NPPB for preventing the formation of peritoneal adhesions.

Curcumin Promoted miR-34a Expression and Suppressed Proliferation of Gastric Cancer Cells.

Background: To investigate the effects of curcumin on miR-34a and proliferation of gastric cancer cells. Materials and Methods: Human gastric cancer cell line SGC-7901 was divided into control, curcumin, miR-34a agomir (miR-34a), miR-34a agomir negative control, and curcumin combined miR-34a antagomir (combine) groups. 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide assay, scratch damage test, and transwell assay were used to detect cell proliferation, migration, and invasion. The cell apoptosis and cell cycle were detected by flow cytometry. Western blot was used to detect the expression of B-cell lymphoma-2 (Bcl-2), cyclin-dependent kinase 4 (CDK4), and cyclin D1. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was used to detect apoptosis of tumors and Western blot was used to detect the expression of Bcl-2, CDK4, and cyclin D1 in tumors. Results: The results showed that curcumin markedly increased the content of miR-34a microRNA (mRNA) in SGC-7901 cells, inhibited proliferation, migration, and invasion of SGC-7901 cells, when compared to control group (p < 0.05). Compared with control group, curcumin significantly inhibited cell cycle progression in G0/G1-S phase, increased the cell number of G0/G1 phase, and downregulated the Bcl-2, CDK4, and cyclin D1 protein expression in cells and tissues (p < 0.05). After transfection of miR-34a agomir or antagomir into cells it was found that miR-34a agomir and curcumin had similar effects on resisting malignant biological behavior. Curcumin combined with miR-34a antagomir could weaken or reverse the above results. Conclusions: Curcumin could inhibit the proliferation and induce apoptosis of SGC-7901 cells. Its mechanism might be related to the miR-34a expression in cells, thus affecting the expression of Bcl-2, CDK4, and cyclin D1.

Biochemical characterization of a novel exo-oligoxylanase from Paenibacillus barengoltzii suitable for monosaccharification from corncobs.

BACKGROUND: Xylan is the major component of hemicelluloses, which are the second most abundant polysaccharides in nature, accounting for approximately one-third of all renewable organic carbon resources on earth. Efficient degradation of xylan is the prerequisite for biofuel production. Enzymatic degradation has been demonstrated to be more attractive due to low energy consumption and environmental friendliness, when compared with chemical degradation. Exo-xylanases, as a rate-limiting factor, play an important role in the xylose production. It is of great value to identify novel exo-xylanases for efficient bioconversion of xylan in biorefinery industry. RESULTS: A novel glycoside hydrolase (GH) family 8 reducing-end xylose-releasing exo-oligoxylanase (Rex)-encoding gene (PbRex8) was cloned from Paenibacillus barengoltzii and heterogeneously expressed in Escherichia coli. The deduced amino acid sequence of PbRex8 shared the highest identity of 74% with a Rex from Bacillus halodurans. The recombinant enzyme (PbRex8) was purified and biochemically characterized. The optimal pH and temperature of PbRex8 were 5.5 and 55 °C, respectively. PbRex8 showed prominent activity on xylooligosaccharides (XOSs), and trace activity on xylan. It also exhibited ß-1,3-1,4-glucanase and xylobiase activities. The enzyme efficiently converted corncob xylan to xylose coupled with a GH family 10 endo-xylanase, with a xylose yield of 83%. The crystal structure of PbRex8 was resolved at 1.88 Å. Structural comparison suggests that Arg67 can hydrogen-bond to xylose moieties in the -1 subsite, and Asn122 and Arg253 are close to xylose moieties in the -3 subsite, the hypotheses of which were further verified by mutation analysis. In addition, Trp205, Trp132, Tyr372, Tyr277 and Tyr369 in the grove of PbRex8 were found to involve in glucooligosaccharides interactions. This is the first report on a GH family 8 Rex from P. barengoltzii. CONCLUSIONS: A novel reducing-end xylose-releasing exo-oligoxylanase suitable for xylose production from corncobs was identified, biochemically characterized and structurally elucidated. The properties of PbRex8 may make it an excellent candidate in biorefinery industries.

Chloride channel-3 mediates multidrug resistance of cancer by upregulating P-glycoprotein expression.

Chloride channel-3 (ClC-3), a member of the ClC family of voltage-gated Cl- channels, is involved in the resistance of tumor cells to chemotherapeutic drugs. Here, we report a new mechanism for ClC-3 in mediating multidrug resistance (MDR). ClC-3 was highly expressed in the P-glycoprotein (P-gp)-dependent human lung adenocarcinoma cell line (A549)/paclitaxel (PTX) and the human breast carcinoma cell line (MCF-7)/doxorubicin (DOX) resistant cells. Changes in the ClC-3 expression resulted in the development of drug resistance in formerly drug-sensitive A549 or MCF-7 cells, and drug sensitivity in formerly drug-resistant A549/Taxol and MCF-7/DOX cells. Double transgenic MMTV-PyMT/CLCN3 mice with spontaneous mammary cancer and ClC-3 overexpression demonstrated drug resistance to PTX and DOX. ClC-3 expression upregulated the expression of MDR1 messenger RNA and P-gp by activating the nuclear factor-κB (NF-κB)-signaling pathway. These data suggest that ClC-3 expression in cancer cells induces MDR by upregulating NF-κB-signaling-dependent P-gp expression involving another new mechanism for ClC-3 in the development of drug resistance of cancers.

Biochemical characterization of a novel xylanase from Paenibacillus barengoltzii and its application in xylooligosaccharides production from corncobs.

A novel xylanase gene (PbXyn10A) from Paenibacillus barengoltzii was cloned and expressed in Escherichia coli. PbXyn10A had an open reading frame of 3,063 bp, and its deduced amino acid sequence shared the highest identity of 72% with a xylanase from Paenibacillus curdlanolyticus. The recombinant xylanase (PbXyn10A) was purified and biochemically characterized. PbXyn10A was most active at pH 6.5 and 60 °C, respectively. It exhibited strict substrate specificity towards birchwood xylan, beechwood xylan and oat-spelt xylan, with Km values of 2.19, 2.04 and 2.51 mg/mL, respectively. The enzyme hydrolyzed xylan to yield mainly xylooligosaccharides (XOS) with degree of polymerization 2-4. A new strategy for XOS production from corncobs pretreated by steam explosion using acidic electrolyzed water, followed by enzymatic hydrolysis was developed. The highest XOS yield of 75% (based on xylan in raw corncobs) was achieved. This is the first report on a xylanase from Paenibacillus barengoltzii.

High level expression of ß-mannanase (RmMan5A) in Pichia pastoris for partially hydrolyzed guar gum production.

Partially hydrolyzed guar gum (PHGG), an important supplemental dietary fiber, has been used as food ingredient in many industries. In this study, a novel ß-mannanase gene (RmMan5A) from Rhizomucor miehei was successfully expressed in Pichia pastoris and subjected for PHGG production. Enzyme activity of fermentation supernatant reached 85,200UmL-1 after 168h high cell density fermentation. The purified RmMan5A exhibited the highest enzyme activity at pH 7.0 and 65°C. RmMan5A was then employed for guar gum hydrolysis and PHGG obtained demonstrated a weight-average molecular weight (Mw) of 2.5×104Da. Total dietary fiber accounted 90.6% of PHGG and 24.9% (w/w) of PHGG were identified as manno-oligosaccharides with degree of polymerization<7. PHGG was further fractionated (F1-F4) by gradual ethanol precipitation. PHGG F1 with an Mw value of 3.6×104Da and a mannose/galactose (M/G) ratio of 1.47 was precipitated initially, followed by PHGG F2 and F3 which showed lower Mw and higher M/G ratio. According to the structure analysis, the distribution of α-d-galactose of PHGG F1 was compact and regular, and that of other fractions was more random. A suitable ß-mannanase for PHGG production and some useful information of PHGG are provided in this paper.

Directed evolution of a ß-mannanase from Rhizomucor miehei to improve catalytic activity in acidic and thermophilic conditions.

BACKGROUND: ß-Mannanase randomly cleaves the ß-1,4-linked mannan backbone of hemicellulose, which plays the most important role in the enzymatic degradation of mannan. Although the industrial applications of ß-mannanase have tremendously expanded in recent years, the wild-type ß-mannanases are still defective for some industries. The glycoside hydrolase (GH) family 5 ß-mannanase (RmMan5A) from Rhizomucor miehei shows many outstanding properties, such as high specific activity and hydrolysis property. However, owing to the low catalytic activity in acidic and thermophilic conditions, the application of RmMan5A to the biorefinery of mannan biomasses is severely limited. RESULTS: To overcome the limitation, RmMan5A was successfully engineered by directed evolution. Through two rounds of screening, a mutated ß-mannanase (mRmMan5A) with high catalytic activity in acidic and thermophilic conditions was obtained, and then characterized. The mutant displayed maximal activity at pH 4.5 and 65 °C, corresponding to acidic shift of 2.5 units in optimal pH and increase by 10 °C in optimal temperature. The catalytic efficiencies (kcat/Km) of mRmMan5A towards many mannan substrates were enhanced more than threefold in acidic and thermophilic conditions. Meanwhile, the high specific activity and excellent hydrolysis property of RmMan5A were inherited by the mutant mRmMan5A after directed evolution. According to the result of sequence analysis, three amino acid residues were substituted in mRmMan5A, namely Tyr233His, Lys264Met, and Asn343Ser. To identify the function of each substitution, four site-directed mutations (Tyr233His, Lys264Met, Asn343Ser, and Tyr233His/Lys264Met) were subsequently generated, and the substitutions at Tyr233 and Lys264 were found to be the main reason for the changes of mRmMan5A. CONCLUSIONS: Through directed evolution of RmMan5A, two key amino acid residues that controlled its catalytic efficiency under acidic and thermophilic conditions were identified. Information about the structure-function relationship of GH family 5 ß-mannanase was acquired, which could be used for modifying ß-mannanases to enhance the feasibility in industrial application, especially in biorefinery process. This is the first report on a ß-mannanase from zygomycete engineered by directed evolution.

Phenytoin silver: a new nanocompound for promoting dermal wound healing via comprehensive pharmacological action.

Phenytoin, an antiepileptic drug, has been widely used for wound healing. Inspired by previous studies, phenytoin silver (PnAg), a sparingly soluble silver nanocompound, was synthesized which exhibited good therapeutic efficacy in tissue repair with low toxicity (LD50 >5 g/kg). In vivo studies showed that PnAg could accelerate dermal wound healing and strong inflammation control in Sprague-Dawley rats (SD rat) and Bama minipigs. Due to its low solubility, PnAg led to low toxicity and blood enrichment in animals. Furthermore, PnAg could upregulate the promoter activity of Jak, Stat3, and Stat3 downstream proteins. Therefore, PnAg may serve as an effective therapeutic compound for wound healing through regulating the gp130/Jak/Stat3 signaling pathway.