The covalent NLRP3-inflammasome inhibitor Oridonin relieves myocardial infarction induced myocardial fibrosis and cardiac remodeling in mice.

BACKGROUND: Myocardial infarction (MI) triggers a strong inflammatory response that is associated with myocardial fibrosis and cardiac remodeling. Interleukin (IL)-1ß and IL-18 are key players in this response and are controlled by NLRP3-inflammatory bodies. Oridonin is a newly reported NLRP3 inhibitor with strong anti-inflammatory activity. We hypothesized that the covalent NLRP3 inhibitor Oridonin could reduce IL-1ß and IL-18 expression and ameliorate myocardial fibrosis after myocardial infarction in mice, improve poor heart remodeling, and preserve heart function. METHODS: Male C57BL/6 mice were subjected to left coronary artery ligation to induce MI and then treated with Oridonin (1, 3, or 6 mg/kg), MCC950 (10 mg/kg), CY-09 (5 mg/kg) or saline three times a week for two weeks. Four weeks after MI, cardiac function and myocardial fibrosis were assessed. In addition, myocardial expressions of inflammatory factors and fibrotic markers were analyzed by western blot, immunofluorescence, enzyme-linked immunosorbent assay, and quantitative real-time polymerase chain reaction. RESULTS: Oridonin treatment preserved left ventricular ejection fraction and fractional shortening, and markedly limited the myocardial infarct size in treated mice. The myocardial fibrosis was lower in the 1 mg/kg group (15.98 ± 1.64)%, 3 mg/kg group (17.39 ± 2.45)%, and 6 mg/kg group (16.76 ± 3.06)% compared to the control group (23.38 ± 1.65)%. Moreover, similar with the results of Oridonin, MCC950 and CY-09 also preserved cardiac function and reduced myocardial fibrosis. The expression levels of NLRP3, IL-1ß and IL-18 were decreased in the Oridonin treatment group compared to non-treated group. In addition, myocardial macrophage and neutrophil influxes were attenuated in the Oridonin treated group. CONCLUSIONS: The covalent NLRP3-inflammasome inhibitor Oridonin reduces myocardial fibrosis and preserves cardiac function in a mouse MI model, which indicates potential therapeutic effect of Oridonin on acute MI patients.

TOPSIS-based entropy measure for intuitionistic trapezoidal fuzzy sets and application to multi-attribute decision making.

As an extension of intuitionistic fuzzy numbers, intuitionistic trapezoidal fuzzy numbers (ITrFNs) are useful in expressing complex fuzzy information with an 'interval value'. This study focuses on multi-attribute decision-making (MADM) problems with unknown attribute weights under an ITrFN environment. We initially present an entropy measure for ITrFNs by using the relative closeness of technique for order preference by similarity to an ideal solution. From the view of the reliability and certainty of decision data, we present an approach to determine the attribute weights. Subsequently, a new method to solve intuitionistic trapezoidal fuzzy MADM problems with unknown attribute weight information is proposed. A numerical example is provided to verify the practicality and effectiveness of the proposed method.

Two-steps gas double-dynamic solid-state fermentation enhances growth and spore production of Conithyrium minitans.

Gas double-dynamic solid-state fermentation (SSF) is a promising strategy with the potential in transforming open-pattern fermentation into closed-pattern fermentation. This paper investigated gas double-dynamic SSF performance in cultivating Coniothyrium minitans (C. minitans), as well as its effect on physiology of C. minitans. Results showed that gas double-dynamic increased biomass content by 48.6%. High temperature impeded pycnidia formation and increased glycine production. More pycnidia formed in solid matrix at 20 °C, which was responsible for higher conidia production (1.5 (±0.03) × 1010 spores/g dry mass), indicating decisive role of high temperature in pycnidia formation of C. minitans in solid-state fermentation. Higher glycine content may be the response of high temperature stress which has close relationship with pycnidia and conidia production. Based on the findings, a two-step strategy for gas double-dynamic SSF was proposed and an satisfactory conidia production was obtained while fermentation period shortened.

Comparison of kokumi γ-[Glu](n>1)-Val and γ-[Glu](n>1)-Met synthesized through transpeptidation catalyzed by glutaminase from Bacillus amyloliquefaciens.

A series of γ-[Glu](n=2,3,4)-Val or γ-[Glu](n=2,3,4)-Met were synthesized in the presence of donor (Gln) and corresponding acceptor (Val or Met) through transpeptidation catalyzed by the glutaminase from Bacillus amyloliquefaciens. Gln in excess significantly (p < .05) improved the yield of γ-[Glu](n>1)-Val/Met except for γ-Glu-Val/Met. The Km values for transpeptidase activity to yield γ-[Glu](n=0,1,2,3)-Val increased with an elevated n, but remained essentially the same irrespective of n value for γ-[Glu](n=0,1,2)-Met (which were 31-44% of that for γ-[Glu]3-Met). The highest Km value appearing when n = 3 (γ-[Glu]3-Val or γ-[Glu]3-Met) suggested the rising difficulty for synthesis when the number of donor increases. All the γ-[Glu]n-Val/Met substances exhibited kokumi properties and enhanced the continuity and umami taste of soy sauce as well as the thickness, mouthfulness and umaminess of model chicken broth. These results indicate the potential of the γ-[Glu]n-Val and γ-[Glu]n-Met as food flavor enhancers.

Enzymatic hydrolysis of lignocellulosic biomass from low to high solids loading.

Solid state enzymatic hydrolysis (SSEH) has many advantages, such as higher sugar concentration, lower operating costs, and less energy input. It should be a potential approach for the industrial application of lignocellulosic ethanol. The purpose of this work is to review the enzymatic hydrolysis of lignocellulosic biomass from low to high solids loading and introduce its both challenges and perspectives. The limitations of SSEH, including inhibition effects, water constraint, and rheology characteristic, are summarized firstly. Various strategies for overcoming these limitations are proposed correspondingly. Fed batch process and its feeding strategy to improve the SSEH efficiency are then discussed. Finally, several intensification methods, hydrolysis reactor, and pilot- and demonstration-scale operations of SSEH are described. In-depth analysis of main limitations and development of novel intensification methods and reactors should provide an effective way to achieve large-scale implementation of SSEH.

Two-step size reduction and post-washing of steam exploded corn stover improving simultaneous saccharification and fermentation for ethanol production.

The simultaneous saccharification and fermentation (SSF) of corn stover biomass for ethanol production was performed by integrating steam explosion (SE) pretreatment, hydrolysis and fermentation. Higher SE pretreatment severity and two-step size reduction increased the specific surface area, swollen volume and water holding capacity of steam exploded corn stover (SECS) and hence facilitated the efficiency of hydrolysis and fermentation. The ethanol production and yield in SSF increased with the decrease of particle size and post-washing of SECS prior to fermentation to remove the inhibitors. Under the SE conditions of 1.5MPa and 9min using 2.0cm particle size, glucan recovery and conversion to glucose by enzymes were 86.2% and 87.2%, respectively. The ethanol concentration and yield were 45.0g/L and 85.6%, respectively. With this two-step size reduction and post-washing strategy, the water utilization efficiency, sugar recovery and conversion, and ethanol concentration and yield by the SSF process were improved.

Mechanical property of different corn stover morphological fractions and its correlations with high solids enzymatic hydrolysis by periodic peristalsis.

Selective structure fractionation combined with periodic peristalsis was exploited to improve the conversion performance of corn stover. The increase of glucan and lignin content and the decrease of xylan content in stem pith were highest after SE, whereas they were lowest in stem node. Glucan conversion increased in this order: steam node

Enzymatic Pretreatment Coupled with the Addition of p-Hydroxyanisole Increased Levulinic Acid Production from Steam-Exploded Rice Straw Short Fiber.

Levulinic acid production, directly from lignocellulosic biomass, resulted in low yields due to the poor substrate accessibility and occurrence of side reactions. The effects of reaction conditions, enzymatic pretreatment, and inhibitor addition on the conversion of steam-exploded rice straw (SERS) short fiber to levulinic acid catalyzed by solid superacid were investigated systematically. The results indicated that the optimal reaction conditions were temperature, time, and solid superacid concentration combinations of 200 °C, 15 min, and 7.5 %. Enzymatic pretreatment improved the substrate accessibility to solid superacid catalyst, and p-hydroxyanisole inhibitor reduced the side reactions during reaction processes, which helped to increase levulinic acid yield. The levulinic acid yield reached 25.2 % under the optimal conditions, which was 61.5 % higher than that without enzymatic pretreatment and inhibitor addition. Therefore, enzymatic pretreatment coupled with the addition of p-hydroxyanisole increased levulinic acid production effectively, which contributed to the value-added utilization of lignocellulosic biomass.

Periodic peristalsis releasing constrained water in high solids enzymatic hydrolysis of steam exploded corn stover.

Periodic peristalsis was used to release water constraint and increase high solids enzymatic hydrolysis efficiency. Glucan and xylan conversion in periodic peristalsis enzymatic hydrolysis (PPEH) at 21% solid loading increased by 5.2-6.4% and 6.8-8.8% compared with that in incubator shaker enzymatic hydrolysis (ISEH), respectively. Hydrolysis kinetics suggested that sugars conversion significantly increased within 24h in PPEH compared with ISEH. The peak height of main water pool increased by 7.7-43.1% within 24h in PPEH compared with ISEH. The increases in peak height of main water pool were consistent with the increases in glucan conversion. Submicroscopic particulates and macro granule residues contributed greatly to water constraint compared with glucose, xylose, ethanol, and Tween 80. Smaller particle size and longer residence time resulted in lower water constraint and facilitated the enzymatic hydrolysis performance. Periodic peristalsis was an effective method to reduce water constraint and increase high solids enzymatic hydrolysis efficiency.

Physical structure changes of solid medium by steam explosion sterilization.

Physical structure changes of solid medium were investigated to reveal effects of steam explosion sterilization on solid-state fermentation (SSF). Results indicated that steam explosion changed the structure of solid medium at both molecular and three-dimensional structural levels, which exposed hydrophilic groups and enlarged pores and cavities. It was interesting to find that pores where capillary water located were the active sites for SSF, due to the close relationship among capillary water relaxation time, specific surface area and fermentation performance. Therefore, steam explosion sterilization increased the effective contact area for microbial cells on solid medium, which contributed to improving SSF performance. Combined with the previous research, mechanisms of SSF improvement by steam explosion sterilization contained both chemical and physical effects.

High-throughput screening of high Monascus pigment-producing strain based on digital image processing.

This work proposed a new method which applied image processing and support vector machine (SVM) for screening of mold strains. Taking Monascus as example, morphological characteristics of Monascus colony were quantified by image processing. And the association between the characteristics and pigment production capability was determined by SVM. On this basis, a highly automated screening strategy was achieved. The accuracy of the proposed strategy is 80.6 %, which is compatible with the existing methods (81.1 % for microplate and 85.4 % for flask). Meanwhile, the screening of 500 colonies only takes 20-30 min, which is the highest rate among all published results. By applying this automated method, 13 strains with high-predicted production were obtained and the best one produced as 2.8-fold (226 U/mL) of pigment and 1.9-fold (51 mg/L) of lovastatin compared with the parent strain. The current study provides us with an effective and promising method for strain improvement.

Simultaneous saccharification and co-fermentation for improving the xylose utilization of steam exploded corn stover at high solid loading.

Simultaneous saccharification and co-fermentation (SSCF) of steam exploded corn stover (SECS) was investigated at 5-25% solid loadings compared with other conversion processes. SECS was washed with a 15-fold excess of deionized water to remove inhibitors of hydrolysis and fermentation. The concentration, yield, and productivity of ethanol was 34.3g/L, 90.0%, 2.61g/L/h in the co-fermentation of 60g/L glucose and 10g/L xylose by Saccharomyces cerevisiae IPE003. Ethanol concentration and productivity increased with increasing solid loading while ethanol yield decreased in all conversion processes of SECS. Glucan and xylan conversion was 82.0% and 82.1% in SSCF at 20% solid loading, respectively, while the concentration, yield and productivity of ethanol was 60.8g/L, 75.3% and 0.63g/L/h. The feeding strategy of SECS addition within 24h improved the SSCF performance. Therefore, SSCF increased ethanol productivity and was an effective conversion process for ethanol production at high solid loading.

Periodic-peristole agitation for process enhancement of butanol fermentation.

BACKGROUND: Mass transfer plays an important role in determining the efficiency of the biofuel conversion. However, adverse effect of shear stress from traditional agitation inhibits the cell growth and production of biofuels. How to enhance the mass transfer with less adverse effect is considered as one of the important bioengineering issues. RESULTS: In this study, a novel agitation type, named periodic-peristole was applied to butanol fermentation with Clostridium acetobutylicum ATCC 824. Meanwhile, the enhancement mechanism was studied. Initially, the fermentation performance of periodic-peristole agitation was compared with the traditional Rushton impeller and stationary cultivation. Result showed that the biomass, butanol and total solvent in periodic-peristole group (PPG) was enhanced to 1.92-, 2.06-, and 2.4-fold of those in the traditional Rushton impeller group (TIG), as well as 1.64-, 1.19- and 1.41-fold of those in the stationary group (SG). Subsequently, to get in-depth insight into enhancement mechanism, hydromechanics analysis and metabolic flux analysis (MFA) were carried out. The periodic-peristole agitation exhibits significant difference on velocity distribution, shear force, and mixing efficiency from the traditional Rushton impeller agitation. And the shear force in PPG is only 74 % of that in TIG. According to MFA result, fructose 6-phosphate, pyruvate, acetyl-CoA, oxaloacetate and α-ketoglutarate were determined the key nodes of cells in response to hydrodynamic mechanical stress. Based on such key information, rational enhancement strategies were proposed and butanol production was further improved. CONCLUSION: The agitation associated with three issues which resulted in significant changes in cell metabolic behaviors: first, a rebalanced redox status; second, the energy (ATP) acquirement and consumption; third, the tolerance mechanism of the cell for survival of solvent. Periodic-peristole agitation provides an answer to address a long-standing problem of biofuel engineering. Key information derived from current study deepens the understanding of agitation, which can guide the designment of new bioreactors and development of enhancement strategies for biofuel refinery.

Erratum to: A novel solid state fermentation coupled with gas stripping enhancing the sweet sorghum stalk conversion performance for bioethanol.

[This corrects the article DOI: 10.1186/1754-6834-7-53.].

Comparative study on occurrence characteristics of matrix water in static and gas double-dynamic solid-state fermentations using low-field NMR and MRI.

The water in a solid substrate is generally divided into three forms: hygroscopic, capillary, and free. However, there are few methods available for detecting the contents of different states of water in substrates. In this paper, low-field NMR and MRI were used to analyze the water occurrence characteristics of steam-exploded corn straw in solid-state fermentation (SSF). A significant linear relationship was found between the total NMR peak areas and the total water contents with a correlation coefficient of 0.993. It was further proved to be successful in comparing the contents and distributions of different states of water in static SSF and gas double-dynamic SSF (GDD-SSF). The results showed that among the three states of water, capillary water was the main form of water present and lost in substrates during fermentation. Total water and capillary water contents did not significantly differ as a result of different sample treatments, but hygroscopic water and free water contents in static SSF were respectively 0.38 and 2.98 times that in GDD-SSF with a packing height of 3 cm after fermentation. A relatively uniform water distribution and deep-depth region for microbial growth were found in GDD-SSF, suggesting that GDD-SSF was more suitable for industrialization. This technology has great potential for achieving efficient on-line water supply through water loss detection in SSF.

Xylose production from corn stover biomass by steam explosion combined with enzymatic digestibility.

A novel conversion process using steam explosion combined with enzymatic digestibility was exploited to increase sugar yield. Results showed that glucan and xylan recovery decreased with the increase of holding temperature and residence time in SE, respectively, while glucan and xylan conversion exhibited an opposite trend. The optimal conditions of steam explosion were 160 °C and 48 min, under which glucan and xylan recovery was 93.4% and 71.6%, respectively. Glucan and xylan conversion at 18% solid loading by periodic peristalsis increased by 3.4-5.8% and 4.5-6.2%, respectively, compared with that by water baths shaker. In the whole process, glucose, xylose and total sugar yield reached to 77.3%, 62.8% and 72.3%, respectively. The yield of hydroxymethyl furfural, furfural and lignin-derived products was 6.3 × 10(-2), 7.5 × 10(-2) and less than 3.7 × 10(-2) g/100 g feedstock, respectively. This novel conversion process increased sugar recovery, reduced degradation products formation, improved digestibility efficiency, and hence increased sugar yield.

A novel steam explosion sterilization improving solid-state fermentation performance.

Traditional sterilization of solid medium (SM) requires lengthy time, degrades nutrients, and even sterilizes inadequately compared with that of liquid medium due to its low thermal conductivity. A novel sterilization strategy, high-temperature and short-time steam explosion (SE), was exploited for SM sterilization in this study. Results showed that SE conditions for complete sterilization were 172 °C for 2 min and 128 °C for 5 min. Glucose and xylose contents in medium after SE sterilization increased by 157% and 93% respectively compared with those after conventional sterilization (121 °C, 20 min) while fermentation inhibitors were not detected. FTIR spectra revealed that the mild SE conditions helped to release monosaccharides from the polysaccharides. Bacillus subtilis fermentation productivity on medium after SE sterilization was 3.83 times of that after conventional sterilization. Therefore, SE shortened sterilization time and improved SM nutrition, which facilitated fermentability of SM and should promote economy of solid-state fermentation.

Steam explosion and its combinatorial pretreatment refining technology of plant biomass to bio-based products.

Pretreatment is a key unit operation affecting the refinery efficiency of plant biomass. However, the poor efficiency of pretreatment and the lack of basic theory are the main challenges to the industrial implementation of the plant biomass refinery. The purpose of this work is to review steam explosion and its combinatorial pretreatment as a means of overcoming the intrinsic characteristics of plant biomass, including recalcitrance, heterogeneity, multi-composition, and diversity. The main advantages of the selective use of steam explosion and other combinatorial pretreatments across the diversity of raw materials are introduced. Combinatorial pretreatment integrated with other unit operations is proposed as a means to exploit the high-efficiency production of bio-based products from plant biomass. Finally, several pilot- and demonstration-scale operations of the plant biomass refinery are described. Based on the principle of selective function and structure fractionation, and multi-level and directional composition conversion, an integrated process with the combinatorial pretreatments of steam explosion and other pretreatments as the core should be feasible and conform to the plant biomass refinery concept. Combinatorial pretreatments of steam explosion and other pretreatments should be further exploited based on the type and intrinsic characteristics of the plant biomass used, the bio-based products to be made, and the complementarity of the processes.

Multilevel composition fractionation process for high-value utilization of wheat straw cellulose.

BACKGROUND: Biomass refining into multiple products has gained considerable momentum due to its potential benefits for economic and environmental sustainability. However, the recalcitrance of biomass is a major challenge in bio-based product production. Multilevel composition fractionation processes should be beneficial in overcoming biomass recalcitrance and achieving effective conversion of multiple compositions of biomass. The present study concerns the fractionation of wheat straw using steam explosion, coupled with ethanol extraction, and that this facilitates the establishment of sugars and lignin platform and enables the production of regenerated cellulose films. RESULTS: The results showed that the hemicellulose fractionation yield was 73% under steam explosion at 1.6 MPa for 5.2 minutes, while the lignin fractionation yield was 90% by ethanol extraction at 160°C for 2 hours and with 60% ethanol (v/v). The cellulose yield reached up to 93% after steam explosion coupled with ethanol extraction. Therefore, cellulose sugar, hemicellulose sugar, and lignin platform were established effectively in the present study. Long fibers (retained by a 40-mesh screening) accounted for 90% of the total cellulose fibers, and the glucan conversion of short fibers was 90% at 9.0 hours with a cellulase loading of 25 filter paper units/g cellulose in enzymatic hydrolysis. Regenerated cellulose film was prepared from long fibers using [bmim]Cl, and the tensile strength and breaking elongation was 120 MPa and 4.8%, respectively. The cross-section of regenerated cellulose film prepared by [bmim]Cl displayed homogeneous structure, which indicated a dense architecture and a better mechanical performance. CONCLUSIONS: Multilevel composition fractionation process using steam explosion followed by ethanol extraction was shown to be an effective process by which wheat straw could be fractionated into different polymeric fractions with high yields. High-value utilization of wheat straw cellulose was achieved by preparing regenerated cellulose film using [bmim]Cl.

A novel solid state fermentation coupled with gas stripping enhancing the sweet sorghum stalk conversion performance for bioethanol.

BACKGROUND: Bioethanol production from biomass is becoming a hot topic internationally. Traditional static solid state fermentation (TS-SSF) for bioethanol production is similar to the traditional method of intermittent operation. The main problems of its large-scale intensive production are the low efficiency of mass and heat transfer and the high ethanol inhibition effect. In order to achieve continuous production and high conversion efficiency, gas stripping solid state fermentation (GS-SSF) for bioethanol production from sweet sorghum stalk (SSS) was systematically investigated in the present study. RESULTS: TS-SSF and GS-SSF were conducted and evaluated based on different SSS particle thicknesses under identical conditions. The ethanol yield reached 22.7 g/100 g dry SSS during GS-SSF, which was obviously higher than that during TS-SSF. The optimal initial gas stripping time, gas stripping temperature, fermentation time, and particle thickness of GS-SSF were 10 h, 35°C, 28 h, and 0.15 cm, respectively, and the corresponding ethanol stripping efficiency was 77.5%. The ethanol yield apparently increased by 30% with the particle thickness decreasing from 0.4 cm to 0.05 cm during GS-SSF. Meanwhile, the ethanol yield increased by 6% to 10% during GS-SSF compared with that during TS-SSF under the same particle thickness. The results revealed that gas stripping removed the ethanol inhibition effect and improved the mass and heat transfer efficiency, and hence strongly enhanced the solid state fermentation (SSF) performance of SSS. GS-SSF also eliminated the need for separate reactors and further simplified the bioethanol production process from SSS. As a result, a continuous conversion process of SSS and online separation of bioethanol were achieved by GS-SSF. CONCLUSIONS: SSF coupled with gas stripping meet the requirements of high yield and efficient industrial bioethanol production. It should be a novel bioconversion process for bioethanol production from SSS biomass.