FA Pretreatment Research Articles

Optimization of softwood pretreatment by microwave-assisted deep eutectic solvents at high solids loading

Microwave-assisted deep eutectic solvent (DES) has received attention as an ultrafast pretreatment method in lignocellulose fractionation. This study investigated the improvement of milled softwood mixture (MSM) fractionation with chlorine chloride-formic acid (ChCl:FA) to obtain residues with high glucan retention and purity while removing majority of the lignin and hemicelluloses. At the optimum pretreatment conditions i.e., ChCl:FA (1:4), 140 °C, 14 min, 800 W and 15 % (w/v), 96.2 % hemicellulose removal, 90.1 % delignification and 93.5 % glucan retention were achieved. About 85 % lignin was recovered with a 95 % purity when solid loading was 10–20 % (w/v). This study showed that microwave assisted ChCl:FA pretreatment was a suitable means to fractionate MSM to achieve high quality glucan and lignin at high solid loading.

Semi-continuous anaerobic digestion of secondary sludge with free ammonia pretreatment: Focusing on volatile solids destruction, dewaterability, pathogen removal and its implications

Our previous work has reported the pretreatment of secondary sludge with free ammonia (NH3, FA) enhanced the methane production in batch biochemical methane potential tests. However, the batch biochemical methane potential test could only provide conservative results compared to continuous/semi-continuous anaerobic digestion. Also, the impacts of FA pretreatment on the key anaerobic digestion parameters, including volatile solids (VS) destruction, sludge dewaterability and pathogen removal, are still unknown. This study for the first time investigated these impacts using semi-continuous anaerobic digestion systems for 130 days. Pretreatment of secondary sludge for 24 h at an FA concentration of 560 mg NH3-N/L improved VS destruction by 26.4% (from 22.0 to 27.8%), supported by a similar increase of 28.6% in methane production (from 126.7 to 162.9 ml CH4/g VSfed). Model based analysis revealed that FA pretreatment improved the sludge degradability extent, which may be the reason for the enhanced VS destruction. Equally importantly, the dewaterability of the digested sludge with FA pretreatment was also enhanced by 9.2% (from 12.0 to 13.1% in solids content of the dewatered digested sludge), which could be partly attributed to the increased zeta potential from -16.7 to -14.5 mV. Anaerobic digestion with FA pretreatment enhanced the removals of Fecal Coliform and E. Coli by 1.3 and 1.4 log MPN/g TS (MPN: Most Probable Number; TS: Total Solids), indicating FA pretreatment was effective in enhancing pathogen removal. With inorganic solids representing 21% of the sludge used, the volume of dewatered sludge to be disposed of was reduced by 14.5% via FA pretreatment. This will substantially decrease the cost as evaluated by economic analysis. In brief, this study provides a promising strategy to enhance sludge reduction in anaerobic digestion and is of great significance in promoting the application of FA pretreatment strategy in the real world.

Free ammonia pretreatment enhances the removal of antibiotic resistance genes in anaerobic sludge digestion

Sludge has been recognized as a reservoir of antibiotic resistance genes (ARGs) in the wastewater treatment plants. Our previous study has demonstrated that free ammonia (FA, i.e., NH3–N) pretreatment is an effective method for enhancing anaerobic digestion of sludge. However, the effects of FA pretreatment on the removal of ARGs in the anaerobic sludge digestion is still unknown. In this study, several ARGs representing various antibiotic classes and integrase gene (intI1) which is crucial for horizontal transfer of ARGs were chosen. This study demonstrated that combined FA pretreatment (420 mg NH3–N/L for 24 h, under which the highest anaerobic sludge biodegradability was achieved in our previous study) and anaerobic digestion could enhance the removal of aac(6′)-Ib-cr, blaTEM, sul2, tetA, tetB and tetX from sludge by 17–74% compared with anaerobic digestion without FA pretreatment, resulting in a lower ARGs abundance in the anaerobically digested sludge. This is caused by the removal of tested ARGs during FA pretreatment and the reduced abundance of potential microbial hosts of ARGs due to FA pretreatment during anaerobic digestion. The removal of IntI1 was not significantly affected by FA pretreatment and intI1 did not play a large role in the fate of the tested ARGs in this study. This study indicated that FA pretreatment for anaerobic digestion could potentially reduce the spread of ARGs from the sludge to the natural environment during sludge disposal or reuse.

Fibrinogen and Albumin Score Changes during Preoperative Treatment Can Predict Prognosis in Patients with Locally Advanced Rectal Cancer.

Background Fibrinogen (Fib) and albumin (Alb) levels are indicators of systemic inflammatory responses. Elevated Fib and decreased Alb levels are considered negative prognostic factors in different types of cancer. Here, we explored the prognostic value of changes in pre- and post- neoadjuvant chemoradiotherapy (NCRT) plasma fibrinogen and serum albumin (FA) scores in patients with locally advanced rectal cancer (LARC). Methods A total of 106 patients with LARC who underwent NCRT followed by surgical resection at Jinhua Municipal Central Hospital between 2011 and 2015 were analyzed. In addition, plasma Fib and serum Alb levels before and after NCRT were collected. FA scores were calculated based on the Fib and Alb levels dichotomized by clinical reference values. Patients were classified into two groups based on the changes in FA scores during NCRT: in group A, FA scores decreased or remained unchanged (n = 84), and in group B, FA scores increased (n = 22). Changes in FA scores were compared with patient outcomes. Results Increased FA scores were associated with worse disease-free survival (DFS) and overall survival (OS) in patients with LARC. The occurrence of systemic failure was higher in group B than in group A (40.9% vs. 19%, P = 0.032). In multivariate analysis, changes in FA scores, pretreatment carcinoembryonic antigen (CEA) levels, and pathologic differentiation were independent prognostic parameters for DFS and changes in FA scores and pretreatment CEA levels were independent prognostic parameters for OS. Conclusions Increased FA score after NCRT was an independent negative prognostic factor for DFS and OS in patients with NCRT-treated LARC.

Free ammonia pretreatment improves anaerobic methane generation from algae

Anaerobic methane generation from algae is hindered by the slow and poor algae biodegradability. A novel free ammonia (NH3 i.e. FA) pretreatment technology was proposed in this work to enhance anaerobic methane generation from algae cultivated using a real secondary effluent. The algae solubilisation was 0.05-0.06 g SCOD/g TCOD (SCOD: soluble chemical oxygen demand; TCOD: total chemical oxygen demand) following FA pretreatment of 240-530 mg NH3-N/L for 24 h, whereas the solubilisation was only 0.01 g SCOD/g TCOD for the untreated algae. This indicates that FA pretreatment at 240-530 mg NH3-N/L could substantially enhance algae solubilisation. Biochemical methane potential tests revealed that FA pretreatment on algae at 240-530 mg NH3-N/L is able to significantly enhance anaerobic methane generation. The hydrolysis rate (k) and biochemical methane potential (P0) of algae increased from 0.21 d-1 and 132 L CH4/kg TCOD to 0.33-0.50 d-1 and 140-154 L CH4/kg TCOD, respectively, after the algae was pretreated by FA at 240-530 mg NH3-N/L. Further analysis indicated that FA pretreatment improved k of both quickly and slowly biodegradable substrates, and also increased P0 of the slowly biodegradable substrate although it negatively affected P0 of the quickly biodegradable substrate. This FA technology is a closed-loop technology.

Free Ammonia Pretreatment Improves Degradation of Secondary Sludge During Aerobic Digestion

Aerobic digestion is commonly used to achieve secondary sludge reduction in the small-size wastewater treatment plants. Nevertheless, secondary sludge degradation is usually restricted by the slow hydrolysis rate and low degradable percentage of secondary sludge. Here, we present an innovative approach using pretreatment of free ammonia (FA, i.e. NH3), a renewable chemical from wastewater, to improve the degradation of secondary sludge during aerobic digestion. The secondary sludge was degraded by 36 ± 4% (volatile solids (VS) basis) within 15 days of aerobic digestion while being pretreated at 300 mg NH3–N/L (pH 9.0; total ammonia nitrogen = 800 mg N/L) for 24 h, whereas only 23 ± 3% (VS basis) of the secondary sludge without FA pretreatment was degraded over the same period. Similarly, the production of inorganic nitrogen also increased from 27 ± 2 to 38 ± 2 mg N/g VS after implementing FA pretreatment, corroborating the idea that degradation of secondary sludge was effectively improved by FA pretreatme...

Ferulic acid relaxed rat aortic, small mesenteric and coronary arteries by blocking voltage-gated calcium channel and calcium desensitization via dephosphorylation of ERK1/2 and MYPT1

Ferulic acid, a natural ingredient presents in several Chinese Materia Medica such as Radix Angelicae Sinensis, has been identified as an important multifunctional and physiologically active small molecule. However, its pharmacological activity in different blood vessel types and underlying mechanisms are unclear. The present study was to investigate the vascular reactivity and the possible action mechanism of FA on aorta, small mesenteric arteries and coronary arteries isolated from Wistar rats. We found FA dose-dependently relieved the contraction of aorta, small mesenteric arteries and coronary arteries induced by different contractors, U46619, phenylephrine (Phe) and KCl. The relaxant effect of FA was not affected by L-NAME (eNOS inhibitor), ODQ (soluble guanylate cyclase inhibitor), and mechanical removal of endothelium in thoracic aortas. The contraction caused by 60mM KCl (60K) was concentration-dependently hindered by FA pretreatment in all three types of arteries. In Ca2+-free 60K solution, FA weakened Ca2+-related contraction in a concentration dependent manner. And FA relaxed both fluoride and phorbol ester which were PKC, ERK and Rho-kinase activators induced contraction in aortic rings with or without Ca2+ in krebs solution. Western blotting experiments in A7r5 cells revealed that FA inhibited calcium sensitization via dephosphorylation of ERK1/2 and MYPT1. Furthermore, the relaxation effect of FA was attenuated by verapamil (calcium channel blocker), ERK inhibitor, and fasudil (ROCK inhibitor). These results provide evidence that FA exhibits endothelium-independent vascular relaxant effect in different types of arteries. The molecular mechanism of vasorelaxation activity of FA probably involved calcium channel inhibition and calcium desensitization.

A novel free ammonia based pretreatment technology to enhance anaerobic methane production from primary sludge.

This study proposed a novel free ammonia (FA, i.e., NH3 ) pretreatment technology to enhance anaerobic methane production from primary sludge for the first time. The solubilization of primary sludge was substantially enhanced following 24 h FA pretreatment (250-680 mg NH3 -N/L), by which the release of soluble chemical oxygen demand (SCOD) (i.e., 0.4 mg SCOD/mg VS added; VS: volatile solids) was approximately 10 times as much as that without pretreatment (i.e., 0.03 mg SCOD/mg VS added). Then, biochemical methane potential (BMP) tests demonstrated that FA pretreatment of 250-680 mg NH3 -N/L was capable of enhancing anaerobic methane production while the digestion time was more than 7 days. Model based analysis indicated that the improved anaerobic methane production was due to an increased biochemical methane potential (B0 ) of 8-17% (i.e., from 331 to 357-387 L CH4 /kg VS added), with the highest B0 achieved at 420 mg NH3 -N/L pretreatment. However, FA pretreatment of 250-680 mg NH3 -N/L decreased hydrolysis rate (k) by 24-38% compared with control (i.e., from 0.29 d-1 to 0.18-0.22 d-1 ), which explained the lower methane production over the first 7 days' digestion period. Economic analysis and environmental evaluation demonstrated that FA pretreatment technology was environmentally friendly and economically favorable. Biotechnol. Bioeng. 2017;114: 2245-2252. © 2017 Wiley Periodicals, Inc.

Free ammonia pre-treatment of secondary sludge significantly increases anaerobic methane production

Energy recovery in the form of methane from sludge/wastewater is restricted by the poor and slow biodegradability of secondary sludge. An innovative pre-treatment technology using free ammonia (FA, i.e. NH3) was proposed in this study to increase anaerobic methane production. The solubilisation of secondary sludge was significantly increased after FA pre-treatment at up to 680 mg NH3-N/L for 1 day, under which the solubilisation (i.e. 0.4 mg SCOD/mg VS; SCOD: soluble chemical oxygen demand; VS: volatile solids) was >10 times higher than that without FA pre-treatment (i.e. 0.03 mg SCOD/mg VS). Biochemical methane potential assays showed that FA pre-treatment at above 250 mg NH3-N/L is effective in improving anaerobic methane production. The highest improvement in biochemical methane potential (B0) and hydrolysis rate (k) was achieved at FA concentrations of 420–680 mg NH3-N/L, and was determined as approximately 22% (from 160 to 195 L CH4/kg VS added) and 140% (from 0.22 to 0.53 d−1) compared to the secondary sludge without pre-treatment. More analysis revealed that the FA induced improvement in B0 and k could be attributed to the rapidly biodegradable substances rather than the slowly biodegradable substances. Economic and environmental analyses showed that the FA-based technology is economically favourable and environmentally friendly. Since this FA technology aims to use the wastewater treatment plants (WWTPs) waste (i.e. anaerobic digestion liquor) to enhance methane production from the WWTPs, it will set an example for the paradigm shift of the WWTPs from ‘linear economy’ to ‘circular economy’.

Fenofibrate improves high-fat diet-induced and palmitate-induced endoplasmic reticulum stress and inflammation in skeletal muscle

AimsFenofibrate (FF) is commonly used clinically as a lipid-lowering drug, but whether it participates in endoplasmic reticulum (ER) stress and decreases inflammation in skeletal muscle is still unknown. The aim of this study is to determine whether FF treatment reduces insulin resistance (IR) by alleviating ER stress and downstream inflammation in skeletal muscle tissues and cells. Main methodsFemale SD rats were randomly divided into groups receiving the standard chow diet (SCD), a high-fat diet (HFD), or HFD plus FF (HFD+FF). The rats in the latter two groups were subjected to a standard HFD for 20weeks, then the HFD+FF rats were administered FF (30mg/kg once daily via gavage) for another 8weeks. Whole-body IR, expression of peroxisome proliferator-activated receptor α (PPARα), ER stress-related genes, and inflammatory genes in the soleus muscle were assessed. The differentiated C2C12 myotubes were treated with palmitic acid or pretreated with fenofibric acid or 4-phenylbutyric acid (4-PBA), etomoxir, and the expression of ER stress, beta-oxidation-related genes, inflammatory genes, Toll-like receptor 4 (TLR4), and insulin-signaling-related molecules were determined. Key findingsEight weeks of FF treatment attenuated HFD-induced IR by decreased tribbles 3 (TRB3) expression, ER stress and inflammation in skeletal muscle. FA pretreatment markedly inverted the PA-induced expression of TLR4 and downstream inflammatory genes, activated ER stress, improved β-oxidation and insulin signaling in differentiated myotube cells. SignificanceFF treatment significantly improved HFD-induced IR in skeletal muscle and PA-induced IR in myotube cells, which may be related to reduced ER stress-induced inflammation.

LPS-stimulated RAW264.7 macrophage CAT-2–mediated l-arginine uptake and nitric oxide biosynthesis is inhibited by omega fatty acid lipid emulsion

BackgroundOmega-3 fatty acid (ω-3 FA) lipid emulsion has been reported to inhibit nitric oxide (NO) production and alter inducible nitric oxide synthase (iNOS) protein expression in lipopolysaccharide (LPS)-stimulated murine macrophages. However, the role of cellular uptake of l-arginine and iNOS transcription in ω-3 FA emulsion-induced inhibition of NO has not been explored. In addition, cationic amino acid transporter-2 (CAT-2) can regulate iNOS activity. The effect of ω-3 FA emulsion on CAT-2 expression is unknown. In the present study, we hypothesized that ω-3 FA emulsion pretreatment would decrease the production of NO in LPS-stimulated macrophages and that this effect would occur through alterations in the cellular uptake of l-arginine and CAT-2 expression, in addition to iNOS expression. MethodsConfluent immortalized murine macrophages (RAW264.7cells) were incubated with Dulbecco’s modified Eagle’s medium, ω-3 FA emulsion, or an isoenergetic ω-6 lipid emulsion for 4 h. The cells were washed and then stimulated with LPS (1 μg/mL) or media alone for 12 or 24 h before harvesting. Greiss reagent was used to assess NO production of plate well supernatants. Cellular uptake of l-arginine was assessed through [3H]-l-arginine. The expression of iNOS and CAT-2 mRNA in harvested RAW264.7 was quantified by reverse transcriptase-polymerase chain reaction. ResultsNO production of unstimulated RAW264.7 cells was similar in all groups. After LPS stimulation, ω-3 FA pretreatment at 12 and 24 h produced significantly less NO (P < 0.05) compared with ω-6 FA or media only. ω-3 FA pretreatment at 12 and 24 h resulted in less l-arginine uptake. iNOS and CAT-2 mRNA was significantly decreased with ω-3 FA pretreatment compared with ω-6 FA or media-only treatment (P < 0.05). ConclusionsThese experiments demonstrated that, in addition to other anti-inflammatory effects, ω-3 FA lipid emulsion also significantly lowers NO production and l-arginine transport through altered expression of iNOS and CAT-2 in LPS-stimulated RAW264.7 macrophage cells.

Receptor-mediated gene delivery by folic acid-modified stearic acid-grafted chitosan micelles

BackgroundCationic polymers have been accepted as effective nonviral vectors for gene delivery with low immunogenicity unlike viral vectors. However, the lack of organ or cell specificity sometimes hampers their application and the modification of polymeric vectors has also shown successful improvements in achieving cell-specific targeting delivery and in promoting intracellular gene transfer efficiency.MethodsA folic acid-conjugated stearic acid-grafted chitosan (FA-CS-SA) micelle, synthesized by a 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide-coupling reaction, was designed for specific receptor-mediated gene delivery.ResultsDue to the cationic properties of chitosan, the micelles could compact the plasmid DNA (pDNA) to form micelle/pDNA complexes nanoparticles. The particle size and zeta potential of the FA-CS-SA/pDNA complexes with different N/P ratios were 100–200 nm and −20 to −10 mV, respectively. The DNase I protection assay indicated that the complexes can efficiently protect condensed DNA from enzymatic degradation by DNase I. A cytotoxicity study indicated that the micelles exhibited less toxicity in comparison with LipofectamineTM 2000. Using SKOV3 and A549 as model tumor cells, the cellular uptake of micelles was investigated.ConclusionIt was found that cellular uptake of FA-CS-SA in SKOV3 cells with higher folate receptor expression was faster than that in A549 cells with a short incubation time. Luciferase assay and green fluorescent protein detection were used to confirm that FA-CS-SA could be an effective gene vector. Transfection efficiency of the FA-CS-SA/pDNA complexes in SKOV3 cells was enhanced up to 2.3-fold compared with that of the CS-SA/pDNA complexes. However, there was no significant difference between the transfection efficiencies of the two complexes in A549 cells. Importantly, the transfection efficiency of FA-CS-SA/pDNA decreased with free FA pretreatment in SKOV3 cells. It was concluded that the increase in transfection efficiency of the FA-CS-SA/pDNA complexes was attributed to folate receptor-mediated endocytosis.

Metabolic switching of human myotubes is improved by n-3 fatty acids

The aim of the present study was to examine whether pretreatment with different fatty acids, as well as the liver X receptor (LXR) agonist T0901317, could modify metabolic switching of human myotubes. The n-3 FA eicosapentaenoic acid (EPA) increased suppressibility, the ability of glucose to suppress FA oxidation. Substrate-regulated flexibility, the ability to increase FA oxidation when changing from a high glucose, low fatty acid condition ("fed") to a high fatty acid, low glucose ("fasted") condition, was increased by EPA and other n-3 FAs. Adaptability, the capacity to increase FA oxidation with increasing FA availability, was enhanced after pretreatment with EPA, linoleic acid (LA), and palmitic acid (PA). T0901317 counteracted the effect of EPA on suppressibility and adaptability, but it did not affect these parameters alone. EPA per se accumulated less, however, EPA, LA, oleic acid, and T0901317 treatment increased the number of lipid droplets (LD) in myotubes. LD volume and intensity, as well as mitochondrial mass, were independent of FA pretreatment. Microarray analysis showed that EPA regulated more genes than the other FAs and that specific pathways involved in carbohydrate metabolism were induced only by EPA. The present study suggests a favorable effect of n-3 FAs on skeletal muscle metabolic switching and glucose utilization.

Lipopolysaccharide-Stimulated RAW 264.7 Macrophage Inducible Nitric Oxide Synthase and Nitric Oxide Production Is Decreased by an Omega-3 Fatty Acid Lipid Emulsion

Omega-3 fatty acids (omega-3 FA) have been demonstrated to have anti-inflammatory properties, postulated to occur through several principal mechanisms, including (1) displacement of arachidonic acid from the cellular membrane; (2) shifting of prostaglandin E(2) and leukotriene B(4) production; and (3) molecular level alterations including decreased activation of nuclear factor kappa B and activator protein-1. An additional regulator that is likely associated is the production of nitric oxide (NO) by nitric oxide synthetase. NO is a short-lived free radical involved in many biological functions. However, excessive NO production can lead to complications, suggesting that decreased NO production is a potential target for some inflammatory diseases. We hypothesized that pretreating with an omega-3 FA lipid emulsion would decrease the production of NO in macrophages and that this effect would occur through alterations in inducible nitric oxide synthetase (iNOS). Greiss reagent was used to assess NO production in RAW 264.7 macrophages following omega-3 or omega-6 FA treatment alone or in combination with lipopolysaccharide (LPS) stimulation for 12 h/24 h. iNOS levels were determined by Western blot. Tumor necrosis factor-alpha levels were determined by enzyme-linked immunosorbent assay. Following LPS-stimulation, omega-3 FA pretreatment at 12 and 24 h produced significantly less NO (P < 0.05) compared to omega-6 FA or media-only conditions. omega-3 FA pretreatment at 12 and 24 h also had less iNOS protein expression compared to omega-6 FA or media-only conditions. Tumor necrosis factor-alpha production was significantly decreased with omega-3 FA treatment compared to omega-6 FA treatment (P < 0.05) after 24 h LPS stimulation. These experiments demonstrate that, in addition to other anti-inflammatory effects, omega-3 FA lipid emulsions also significantly lower NO production in LPS-stimulated macrophages through altered iNOS protein expression.