Simultaneous Pretreatment Research Articles

A novel liquid-liquid phase-change solvent with upper critical solution temperature for simultaneous pretreatment and separation of lignin, cellulose and hemicellulose from lignocellulosic biomass

The compact dimensional aggregate structure and the complex chemical components of lignocellulose go against its high value utilization. The mixture of dilute hydrochloric acid and n-butanol, which had a characteristic of homogeneous phase at high temperature and phase separation at low temperature, was proposed for lignocellulosic hydrolysis and hydrolysates separation. The results showed that lignocellulose was hydrolyzed under the synergy effect of hydrochloric acid and n-butanol in homogeneous condition, and hemicellulose and lignin hydrolysates were transferred automatically into dilute hydrochloric acid phase and n-butanol phase during the phase separation of the solvent after pretreatment, respectively. N-butanol accelerated lignin hydrolysis and protected β-O-4 linkage within lignin during the pretreatment. Meanwhile, the cellulose within solid phase product from the pretreatment was of high crystallinity, and its glucose yield in enzymatic saccharification at 48 and 72 h were 73.5% and 97.3%, respectively. This study provides a new method for lignocellulose hydrolysis and hydrolysates separation, and lays a theoretical foundation for lignocellulosic utilization of all component.

Rapid screening of toxicity to thermotolerant yeasts: inhibition of growth and fermentation by ionic liquids and zwitterions

We here established rapid screening methods for toxicity of zwitterions and investigated the toxicity to growth and fermentation.

Mercuric Chloride Induced Nephrotoxicity: Ameliorative Effect of Carica papaya Leaves Confirmed by Histopathology, Immunohistochemistry, and Gene Expression Studies.

The present study analyzes the efficacy of the ethanolic extract of C. papaya leaves (ECP) against HgCl2-induced nephrotoxicity. The effects on the biochemical and percentage of body and organ weight against HgCl2-induced nephrotoxicity in female Wistar rats were studied. Wistar rats were divided into five groups with six animals in each group: control, HgCl2 (2.5 mg/kg b.w.), N-acetylcysteine (NAC 180 mg/kg) + HgCl2, ECP (300 mg/kg b.w.) + HgCl2, and ECP (600 mg/kg) + HgCl2 groups. After 28 days of study, animals were sacrificed on the 29th day to harvest the blood and kidneys for further analysis. The effect ECP was analyzed by immunohistochemistry (NGAL) and real-time PCR (KIM-1 and NGAL mRNA) in HgCl2-induced nephrotoxicity. The results revealed that the HgCl2 group showed prominent damage in the proximal tubules and glomerulus of nephrons and enormous expression of NGAL in immunohistochemistry and KIM-1 and NGAL in real-time PCR compared to the control group. The simultaneous pretreatment with NAC (180 mg/kg) and ECP (600 and 300 mg/kg) reduced renal damage and expression of NGAL in immunohistochemistry and KIM-1 and NGAL gene in real-time PCR. This study attests to the nephroprotective effect of ECP against HgCl2-induced toxicity.

Simultaneous pretreatment with ultraviolet light and alkaline H2O2 to promote enzymatic hydrolysis of corn stover

Pretreatment processes are essential for the preparation of biofuels from lignocellulosic feedstocks. Based on alkaline H2O2 pretreatment, photocatalytic alkaline H2O2 pretreatment (U-AHP) was investigated to examine the effects of reaction time, alkali concentration, and H2O2 concentration on the enzymatic digestion of corn stover. The optimum process conditions were determined by orthogonal tests: 1% NaOH, 2% H2O2, and reaction time of 8 h. Under these conditions, the lignin removal efficiency of UH-AHP was 90.2% and the saccharification yield was 94.7%. Furthermore, FT-IR, XRD, and SEM analyses showed that U-AHP pretreatment caused structural damage to the maize straw and increased the crystallinity of the cellulose, and it was speculated that the U-AHP pretreatment reaction was a complex mechanism, which might be a multiple synergistic reaction. This study shows that U-AHP pretreatment is a simple, green and effective method to promote lignin removal.

"Lollipop" particle counting immunoassay based on antigen-powered CRISPR-Cas12a dual signal amplification for the sensitive detection of deoxynivalenol in the environment and food samples

Deoxynivalenol is one of the most widely distributed mycotoxins in cereals and poses tremendous threats to the agricultural environment and public health. Therefore, it is particularly important to develop sensitive and interference-resistant deoxynivalenol analysis methods. Here, we establish a “Lollipop” particle counting immunoassay (LPCI) based on antigen-powered CRISPR-Cas12a dual signal amplification. LPCI achieves high sensitivity and accuracy through antigen-powered CRISPR-Cas dual signal amplification combined with particle counting immunoassay. This strategy not only broadens the applicability of the CRISPR-Cas system in the field of non-nucleic acid target detection; it also improves the sensitivity of particle counting immunoassay. The introduction of a polystyrene "lollipop" immunoassay carrier further enables efficiently simultaneous pre-treatment of multiple samples and overcomes complex matrix interference in real samples. The linear detection range of LPCI for deoxynivalenol was 0.1–500 ng/mL with a detection limit of 0.061 ng/mL. The platform greatly broadens the scope of the CRISPR-Cas sensor for the detection of non-nucleic acid hazards in the environment and food samples.

Eco-friendly and rapid one-step fermentable sugar production from raw lignocellulosic biomass using enzyme mimicking nanomaterials: A novel cost-effective approach to biofuel production

Cerium-doped iron oxide nanoparticles (CeFe3O4-NPs) possessing exceptional laccase and cellulase/hemicellulase mimicking properties were exploited for simultaneous pretreatment (i.e. delignification) and saccharification (SPS) of raw corn cob (CC) biomass. Here, we developed an eco-friendly single step SPS strategy for conversion of raw CC biomass into C6 and C5 sugars using CeFe3O4-NPs with in-houseproduced cellulase andhemicellulaseenzymes which generated maximum glucose (20.3 ± 1.01 g/L) and xylose (22.0 ± 2.22 g/L) within 24 h of hydrolysis at 55 °C and further 21.7 g/L of bioethanol was produced by Saccharomyces cerevisiae. These results clearly elucidated the role of CeFe3O4-NPs as lignocellulose degrading laccase and cellulase/hemicellulase enzymes. Remarkably, the presence of CeFe3O4-NPs has triggered xylose utilization in S. cerevisiae which is completely a new finding. This study can provide new insights towards the probable role of CeFe3O4-NPs in the up-regulation of significant genes responsible for xylose utilization which is a noteworthy finding in the field of ‘biomass to biofuel’ conversion.

Integrated Method of Pretreatment, Saccharification and Electro-Fermentation of Saccharum Spontaneum to Produce Bioethanol Using Microbial Consortium

Production of lignocellulosic ethanol using microbial consortium can be an economical strategy. Consortium of ligninolytic and cellulolytic microbe(s) that can depolymerize lignin and hydrolyze cellulose along with fermenting yeast (CDBT-2) can replace the separate techniques used for biomass pretreatment and hydrolysis prior to fermentation. So, the aim of this study was to screen fungi capable of producing laccase (lignin depolymerizing enzyme) and cellulase (converts cellulose to fermentable sugars), and use the strains along with Saccharomyces cerevisiae (CDBT-2) to develop an integrated method of simultaneous pretreatment saccharification and electro-fermentation of Saccharum spontaneum powder to produce ethanol. 18-s rRNA sequencing and BLAST analysis of screened isolates confirmed, cellulolytic isolate (F1) resembled as Aspergillus niger and, ligninocellulolytic isolate (F2) resembled to Ganoderma sessile. These fungi were used along with CDBT2 for ethanol production. Different consortiums sets of these isolates were tested for fermentation efficiency. All the analysis were done using crude untreated as well as hot water pretreated S. spontaneum powder to compare the ethanol production efficiency. In controlled environment of 28oC, shaking at 80 rpm and 5.0 pH, culture with consortium of G. sessile and CDBT2 produced higher ethanol with yield 57 mg/g biomass powder followed by consortium of G. sessile, A. niger and CDBT2 with yield of 44±0.18 mg/g biomass. Monoculture of G. sessile produced higher amount of phenol ie 292.48±17.74 µg/mL. Similarly, G. sessile, A. niger and CDBT2 consortium produced higher furfural of 58.48±3.86 µg/mL. However, the amount of phenol and furfural generated were very low in both cases. The ethanol yield was higher in crude biomass compared to hot water pretreated S. spontaneum powder. In addition, electro-fermentation of crude S. spontaneum enhanced ethanol production by 12% with G. sessile and CDBT2 consortium while 18% with the set of G. sessile, A. niger and CDBT2 consortium. This monophasic method of integrated pretreatment saccharification and electro-fermentation can be an alternative approach for lignocellulosic bioethanol production as it does not require separate pretreatment and fermentation approach.Keywords: Bioethanol, Saccharum spontaneum, microbial consortium, simultaneous saccharification and fermentation, and electro-fermentation Figure 1

Simultaneous pre-treatment of dietary Moringa oleifera leaf meal with polyethylene glycol and fibrolytic enzymes: Effect on growth performance, physiological indices, and meat quality parameters in jumbo quail

The utility of Moringa oleifera leaf meal (MOLM) as a source of biologically active substances and nutrients for Jumbo quail is limited by high concentrations of condensed tannins and fiber. Simultaneous application of polyethylene glycol (PEG), a tannin-binding compound, and exogenous fibrolytic multi-enzymes could ameliorate antinutritional effects of condensed tannins and fiber thus improving MOLM utilization in quail diets. This study investigated the effect of pre-treating dietary MOLM with PEG and fibrolytic enzymes on live performance, blood parameters, visceral organs, and carcass and meat quality characteristics in Jumbo quail. A total of 381, two-week-old quail chicks (57.5 ± 3.95 g live-weight) were randomly distributed to six dietary treatments replicated six times. The treatments were: T1 = a standard grower diet containing untreated MOLM (10%); T2 = a standard grower diet containing MOLM (10%) pre-treated with 5.4% PEG; and a standard grower diet containing MOLM (10%) pre-treated with 5.4% PEG and 1.25% (T3), 1.50% (T4), 1.75% (T5) and 2.0% (T6) fibrolytic multi-enzymes. Graded levels of enzymes did not induce linear or quadratic effects for overall feed intake, feed conversion efficiency, hematological, carcass, and meat quality parameters in response to increasing fibrolytic enzyme levels. However, weight gain in week 3 quadratically responded [R2 = 0.117, P = 0.043]. Three-week old birds reared on T3 had lower (p < 0.05) weight gains (40.9 g/bird) than those reared on the other treatment groups. Birds reared on T6 diet had longer caecum (14.1 cm) than those reared on T2, T3, T4 and T5 whose caeca lengths did not differ (P > 0.05). Birds reared on diet T5 had shorter small intestines (59.5 cm) than those reared on T1 and T2 diets. It can be concluded that simultaneous pre-treatment of dietary MOLM with PEG and fibrolytic enzymes did not improve live performance, blood parameters, and carcass and meat quality traits, but affected some visceral organ sizes in Jumbo quail.

Activation of estrogen receptor ESR1 and ESR2 induces proliferation of the human testicular embryonal carcinoma NT2/D1 cells

The aims of the present study were to investigate the expression of the classic estrogen receptors ESR1 and ESR2, the splicing variant ESR1-36 and GPER in human testicular embryonal carcinoma NT2/D1 cells, and the effects of the activation of the ESR1 and ESR2 on cell proliferation. Immunostaining of ESR1, ESR2, and GPER were predominantly found in the nuclei, and less abundant in the cytoplasm. ESR1-36 isoform was predominantly expressed in the perinuclear region and cytoplasm, and some weakly immunostained in the nuclei. In nonstimulated NT2/D1 cells (control), proteins of the cell cycle CCND1, CCND2, CCNE1 and CDKN1B are present. Activation of ESR1 and ESR2 increases, respectively, CCND2 and CCNE1 expression, but not CCND1. Activation of ESR2 also mediates upregulation of the cell cycle inhibitor CDKN1B. This protein co-immunoprecipitated with CCND2. Also, E2 induces an increase in the number and viability of the NT2/D1 cells. These effects are blocked by simultaneous pretreatment with ESR1-and ESR2-selective antagonists, confirming that both estrogen receptors regulate NT2/D1 cell proliferation. In addition, E2 increases SRC phosphorylation, and SRC mediates cell proliferation. Our study provides novel insights into the signatures and molecular mechanisms of estrogen receptor in NT2/D1 cells.

Effect of Near-Infrared Blood Photobiomodulation on Red Blood Cell Damage from the Extracorporeal Circuit during Hemodialysis In Vitro

The contact of blood with the bioincompatible membranes of the dialyzer, which is part of the extracorporeal circuit during hemodialysis (HD), causes upregulation of various cellular and non-cellular processes, including massive generation and release of reactive oxygen species (ROS), (which is one of the primary causes of anemia in chronic renal failure). We hypothesize that near-infrared (NIR) radiation possesses antioxidant properties and is considered to protect the red blood cell (RBC) membrane by enhancing its resilience to negative pressures. Our experimental setup consisted of an HD machine equipped with a dialyzer with a polyamide membrane; whole bovine blood was examined in vitro in blood-treated circulation. Blood samples were taken at 0, 5, 15, and 30 min during the HD therapy. We also assessed osmotic fragility, hematocrit, hemolysis, and oxidative stress as a concentration of reactive thiobarbituric acid substances (TBARS). Our results have shown that RBC membrane peroxidation increased significantly after 30 min of circulation, whereas the TBARS level in NIR-treated blood remained relatively steady throughout the experiment. The osmotic fragility of NIR-irradiated samples during dialysis was decreased compared to control samples. Our studies confirm that in vitro, blood photobiomodulation using NIR light diminishes oxidative damage during HD and can be considered a simultaneous pretreatment strategy for HD.

On-site sample pretreatment: Natural deep eutectic solvent-based multiple air-assisted liquid–liquid microextraction

Chemical protection is one of the most important methods to control forest pests. Since pesticides used in chemical protection might have significant adverse effects on aquatic organisms, it is essential to monitor and detect trace pesticides in environmental water. In this study, an on-site natural deep eutectic solvent-based multiple air-assisted liquid–liquid microextraction coupled with gas chromatography-electron capture detection was developed for determining triadimefon, bifenthrin, bromopropylate, and permethrin in water samples in the forest. A series of menthol-based deep eutectic solvents were synthesized for the first time to find green and efficient extractants. Moreover, a deep eutectic solvent with a 1:2 molar ratio of DL-menthol to citric acid was selected as the extractant due to its high stability and extraction performance. A portable modular array device based on air-assisted liquid–liquid microextraction was designed and manufactured to enable the simultaneous pretreatment of several samples. Although no electric equipment is required throughout the process, up to six samples were pretreated on-site within 20 min. Moreover, the average sample pretreatment time was less than 3.5 min, and the process was substantially simplified and enhanced. In addition, the parameters affecting the extraction performance were optimized. The extraction recoveries were in the range of 92–95% by using the optimized conditions, and the relative standard deviations of the intra-device, inter-device, intra-day, and inter-day factors were all less than 4.9%. The detection limits of the method ranged from 0.34 to 3.3 µg L−1. Good linearities (R2>0.999) were shown in the linear range of 1.13 to 2500 µg L−1 for the studied pesticides. Moreover, the proposed method was proved to be highly environmentally friendly by the Analytical Eco-Scale and Green Analytical Procedure Index. In summary, the proposed method is green, efficient, and flexible for on-site water sample pretreatment. The fact that tedious operations can be converted into standard operations provides a new perspective for on-site sample processing.

Ethanol production from lignocellulosic biomass by co-fermentation with Pecoramyces sp. F1 and Zymomonas mobilis ATCC 31821 in an integrated process

A novel, integrated process for the production of ethanol from lignocellulosic biomass is reported which involves simultaneous pretreatment, saccharification and fermentation using environmentally friendly technologies. The lignocellulosic biomass (8.33 g/L rice straw) was pretreated and saccharified with the anaerobic fungus Pecoramyces sp. F1, while the facultative anaerobic bacterium Zymomonas mobilis ATCC 31821 was employed to simultaneously ferment to produce ethanol. Without the need for additional pretreatment, the conversion from lignocellulosic biomass to ethanol was completed in four days with a production yield of 0.32 g ethanol/g glucose and a volumetric productivity of 0.010 g/L/h (0.98 g/L). Meanwhile, other metabolites were produced, including 0.024 g/L/h (2.28 g/L) acetate, 0.002 g/L/h (0.22 g/L) lactate and 0.002 g/L/h (0.15 g/L) formate. Results showed that the integrated process has the potential for ethanol production from lignocellulosic biomass.

An integrated biorefinery approach for bioethanol production from sugarcane tops

Bioethanol from lignocellulosic biomass is a promising alternative to petroleum-based fuels to alleviate greenhouse gas emissions and reduce the dependency on fossil fuels. The lignocellulosic biomass is exploited for ethanol production due to its sustainability and abundance. Sugarcane tops, an agricultural residue, was employed in the present investigation to assess its potential as a feedstock for bioethanol production by adopting different fermentation approaches, namely separate fermentation, simultaneous saccharification and fermentation (SSF) and partially consolidated bioprocessing (PCBP). The present study demonstrated the potential of mono and co-fermentation for ethanol production. Comparison between separate fermentation and SSF using S.cerevisiae showed the latter to be more efficient with ethanol production of 5.69% (v/v) in 30.67 h of fermentation time than separate fermentation with 3.76% (v/v) ethanol in 48 h. An integrated fermentation strategy stated as partially consolidated bioprocessing (PCBP) was investigated to improve fermentation efficiency. This process integrates simultaneous pretreatment and saccharification (SPS), conducted by enzyme blends of laccase and cellulase followed by co-fermentation using S. cerevisiae and xylose-fermenting yeast AKBR 212. This approach resulted in a 7.57% (v/v) maximum ethanol concentration in 24.30 h. The different fermentation strategies involved enzymes and yeasts only, thus offering a green biotechnology approach towards converting sugarcane tops into bioethanol.

Determination of 12 anti-obesity drugs in human plasma by a 96-well protein precipitation plate using HPLC-MS.

An analytical method was developed and validated for the simultaneous determination of 12 anti-obesity drugs (methylephedrine (MER), amphetamine (AMP), fenfluramine (FEN), bupropion (BUP), fluoxetine (FLU), sibutramine (SIBU), bisacodyl (BISA), bumetanide (BUM), lovastatin (LOVA), simvastatin (SIM), rimonabant (RIMO), and fenofibrate (FENO)) in human plasma by a 96-well protein precipitation plate combined with high-performance liquid chromatography-mass spectrometry (HPLC-MS/MS). The 96-well protein precipitation plate was chosen for simultaneous pretreatment of large sample volumes, making the whole process more efficient and faster. Drugs were separated on an Agilent Poroshell 120 EC-C18 column, and detected by MS/MS under multiple reaction monitoring (MRM) mode. The developed method was validated in terms of linearity, matrix effect, accuracy and precision. A good linearity was obtained in the range of 0.1–20.0 ng mL−1 for fenfluramine, bupropion, fluoxetine, sibutramine, bisacodyl, and rimonabant; and 0.5–20.0 ng mL−1 for methylephedrine, amphetamine, bumetanide, lovastatin, simvastatin, and fenofibrate with a correlation coefficient above 0.995. The method was fully validated with an acceptable accuracy of 75.63–108.21%, matrix effect of 80.41–117.71% except for fenofibrate (76.07% at low concentration levels), and precision of 0.32–13.12%. Owing to the advantages of simple operation, high accuracy and sensitivity, this method is suitable for the rapid and simultaneous detection of 12 anti-obesity drugs in human plasma, providing support for clinically monitoring the development of adverse reactions and guiding the rational and appropriate use of weight-loss drugs for obese people.

Simultaneous Pretreatment of Aspirin and Omega-3 Fatty Acid Attenuates Nuclear Factor-κB Activation in a Murine Model with Ventilator-Induced Lung Injury.

Ventilator-induced lung injury (VILI) is an important critical care complication. Nuclear factor-κB (NF-κB) activation, a critical signaling event in the inflammatory response, has been implicated in the tracking of the lung injury. The present study aimed to determine the effect of simultaneous pretreatment with enteral aspirin and omega-3 fatty acid on lung injury in a murine VILI model. We compared the lung inflammation after the sequential administration of lipopolysaccharides and mechanical ventilation between the pretreated simultaneous enteral aspirin and omega-3 fatty acid group and the non-pretreatment group, by quantifying NF-κB activation using an in vivo imaging system to detect bioluminescence signals. The pretreated group with enteral aspirin and omega-3 fatty acid exhibited a smaller elevation of bioluminescence signals than the non-pretreated group (p = 0.039). Compared to the non-pretreated group, the pretreatment group with simultaneous enteral aspirin and omega-3 fatty acid showed reduced expression of the pro-inflammatory cytokine, tumor necrosis factor-α, in bronchoalveolar lavage fluid (p = 0.038). Histopathological lung injury scores were also lower in the pretreatment groups compared to the only injury group. Simultaneous pretreatment with enteral administration of aspirin and omega-3 fatty acid could be a prevention method for VILI in patients with impending mechanical ventilation therapy.

Simultaneous Pretreatment Process on The Isolation of Cellulose Mycrocrystalline from Oil Palm empty Fruit Bunches

This research was aimed to evaluate the isolation of cellulose derived from empty fruit bunches (EFB), especially to increase the purity of cellulose content by subsequent pretreatment process involving delignification using NaOH followed by bleaching process using hydrogen peroxide (H2O2). The result showed that the cellulose content of the raw EFB content at 37.6% ± 0.3 w/w increased to 89.2% ± 0.4 w/w after the simultaneous pretreatment process. Moreover, after the simultaneous of bleaching process using H2O2, the lignin content can be reduced up to 3.1% ± 0.1 w/w which similar to cellulose standard (2.7% ± w/w). The utilization of simultaneous pretreatment process can also reduce the hemicellulose content from 23.9% ± 0.3 w/w (raw EFB) to 7.7% ± 0.2 w/w even lower than the cellulose standard (13.1% w/w). FTIR spectrum showed that the peak absorption of cellulose increased significantly upon the simultaneous process of delignification-bleaching. Morphological changes of the raw EFB compared to pre-treated EFB as a note by SEM analysis showed the significant transformation from the larger size and tightly bundles of micro-fibril cellulose into smaller size and individually separated micro-fibrils. Further through the simultaneous process exhibited yield at 80-85%, high crystallinity (70-80%), and good thermal stability in terms of Tonset at about 230-290 °C. From these results, it can be concluded that the subsequent pretreatment process involving NaOH delignification and H2O2 bleaching processes more likely offer a high selectivity of cellulose microcrystalline from lignocellulosic biomass material by simultaneous reduction of the hemicellulose and lignin content without disrupting the structure of cellulose.Keywords: bleaching; cellulose; delignified; microcrystalline cellulose; oil palm empty fruit bunches

Simultaneous pretreatment and catalytic conversion of polyolefins into hydrocarbon fuels over acidic zeolite catalysts

The effect of pre-degradation treatment in catalytic pyrolysis of polymer was assessed using thermogravimetric analysis (TGA) and pyrolysis reactor experiments of different polymers on various catalysts. Intimate contact between the polymer and catalyst was achieved using physical, mechanical and thermal treatments. The results from the TGA analysis show that pre-degradation treatment has greatly improved the performance of the catalyst attaining maximum degradation at lower temperature. Pre-degradation treatment had increased the liquid yield and lowered the coke yield at various extents depending on the catalyst structure and acidity. The ZSM-5 catalyst with high Si/Al ratio showed maximum amount of (C5–C9) and ZSM-5 with lower Si/Al ratio has maximum percentage of (C14–C20). Based on the performance of the pre-degradation treatment methods, normal mixing produced the maximum amount of lighter fractions and therefore pre-degradation treatment can serve to enhance the gasoline fraction while the diesel fraction can be optimised using the co-pressing method, where polymer and catalyst particles were thoroughly mixed and co-pressed together into mixed particles. Coke characterisation showed coke formed by linear low density polyethylene contained more volatile coke components while polypropylene coke had higher percentage of hard coke. ZSM-5 had lower retention of coke components with volatile coke precursors. The volatility of the coke increases while coke concentration and decreases as the catalyst amount decreases. Pre-degradation treatment facilitated the formation of soft coke components that are easy to remove in inert atmosphere.

Effects of M-1, a Major Metabolite of Sarpogrelate, on 5-HT-Induced Constriction of Isolated Human Internal Thoracic Artery.

Sarpogrelate, a selective 5-hydroxytryptamine (5-HT)2A receptor antagonist, inhibits 5-HT-induced platelet aggregation and vasoconstriction. It improves ischemic symptoms in patients with arteriosclerosis obliterans. M-1 is a major metabolite of sarpogrelate, and has been reported to show a higher affinity for the 5-HT2A receptor on platelets than sarpogrelate. However, the effects of M-1 on 5-HT-induced constrictive response in human blood vessels have not been investigated. The internal thoracic artery (ITA) is the key conduit for coronary artery bypass grafting (CABG). 5-HT has been implicated as playing an important role in the pathogenesis of vasospasm. Thus, in the present study, the effects of M-1 on 5-HT-induced vasoconstriction were examined in isolated human endothelium denuded ITA. M-1 inhibited 5-HT-induced vasoconstriction in a concentration-dependent manner. At the highest concentration, M-1 almost completely inhibited the 5-HT-induced vasoconstriction. Expression of 5-HT2A and 5-HT1B receptor proteins in the membrane fraction of ITA smooth muscle cells was confirmed by Western blot analysis. Individually, supramaximal concentrations of sarpogrelate and SB224289, a selective 5-HT1B receptor antagonist, only partially inhibited the 5-HT-induced vasoconstriction. However, simultaneous pretreatment with both these antagonists almost completely inhibited the 5-HT-induced vasoconstriction. The inhibitory effect of M-1 pretreatment mimicked the inhibitory effect of simultaneous pretreatment with sarpogrelate and SB224289. These results suggest that M-1 has antagonistic effects not only on the 5-HT2A receptor but also on the 5-HT1B receptor in human ITA smooth muscle cells. M-1 may be useful as a lead compound for the development of drugs for the treatment of 5-HT-induced vasospasms in CABG.

Simultaneous pretreatment and saccharification process for fermentable sugars production from Casuarina equisetifolia biomass using transgenic Trichoderma atroviride

ABSTRACT With the increase in the cognizance toward the wide and abundant lignocellulosic biomass, a great interest has been garnered toward the production of value-added products from the biomass. Hence, by capitalizing the Casuarina equisetifolia biomass, the current work developed a simultaneous pre-treatment and saccharification (SPS) process using transgenic Trichoderma atroviride. The ability of T. atroviride to produce lignolytic and cellulolytic enzymes was enhanced by optimizing key process parameters. Under the optimized conditions, a maximum of 1245.6 U/kg of cellulase and 1203.36 U/kg of xylanase, 183.2 U/kg of laccase along with 392.36 g/kg of fermentable sugars were obtained. On comparing with acid and alkaline pre-treatment methods, the T. atroviride -mediated SPS process exhibited trace formation of fermentative inhibitors, which resulted in a minimal inhibition of Escherichia coli. Overall, the current work implements the biorefinery concept on Casuarina equisetifolia biomass by advocating circular economy. Implications: Valorization of lignocellulosic waste biomass into value added compound and as biofuel is considered as a promising alternative resource, owing to its availability and low production cost. However, the presence of chemically resistant lignin demands an intensive treatment process, which sometimes leads to the formation of fermentative inhibitors. Casuarina equisetifolia is a deciduous commercial plant, and an average of 125 tonnes/hector of waste is generated annually in India. By considering the demerit of delignification and the wide availability of Casuarina equisetifolia biomass (CB), the current work aimed at the development of a single-pot simultaneous pre-treatment and saccharification (SPS) of CB by transgenic Trichoderma atroviride.

Simultaneous pretreatment and saccharification of oil palm empty fruit bunch using laccase-cellulase cocktail

Abstract Oil palm empty fruit bunch (OPEFB) can be categorized as one of the toughest lignocellulosic biomass materials to be degraded naturally due to its complexity in structure. A new biological pretreatment approach of utilizing laccase-cellulase cocktail to simultaneously pretreat and saccharify OPEFB for reducing sugars production was presented in this study. Combination of laccase and cellulase has shown to exhibit minimum inhibition at 5% and 8%, respectively. This simultaneous pretreatment and saccharification of OPEFB using laccase-cellulase cocktail comprised of laccase (25–105 U/g) and Acremonium cellulase (10–30 FPU/g) was carried out at 50 °C for 72 h. As a result, the laccase-cellulase cocktail at a ratio of 45 U/g:25 FPU/g successfully produced 10.90 g/L of reducing sugars and 32.62% of hydrolysis yield, without the need of substrate washing step after pretreatment, in which it shortens the time from a total of 144 h–72 h and reduces the utilization of extra vessels.