Hydrated lime promoted the polysaccharide content and affected the transcriptomes of Lentinula edodes during brown film formation.

Brown film formation, a unique developmental stage in the life cycle of Lentinula edodes, is essential for the subsequent development of fruiting bodies in L. edodes cultivation. The pH of mushroom growth substrates are usually adjusted with hydrated lime, yet the effects of hydrated lime on cultivating L. edodes and the molecular mechanisms associated with the effects have not been studied systemically. We cultivated L. edodes on substrates supplemented with 0% (CK), 1% (T1), 3% (T2), and 5% (T3) hydrated lime (Ca (OH)2), and applied transcriptomics and qRT-PCR to study gene expression on the brown film formation stage. Hydrated lime increased polysaccharide contents in L. edodes, especially in T2, where the 5.3% polysaccharide content was approximately 1.5 times higher than in the CK. The addition of hydrated lime in the substrate promoted laccase, lignin peroxidase and manganese peroxidase activities, implying that hydrated lime improved the ability of L. edodes to decompose lignin and provide nutrition for its growth and development. Among the annotated 9,913 genes, compared to the control, 47 genes were up-regulated and 52 genes down-regulated in T1; 73 genes were up-regulated and 44 were down-regulated in T2; and 125 genes were up-regulated and 65 genes were down-regulated in T3. Differentially expressed genes (DEGs) were enriched in the amino acid metabolism, lipid metabolism and carbohydrate metabolism related pathways. The carbohydrate-active enzyme genes up-regulated in the hydrated lime treatments were mostly glycosyl hydrolase genes. The results will facilitate future optimization of L. edodes cultivation techniques and possibly shortening the production cycle.

Highly sensitive SERS detection in a non-volatile liquid-phase system with nanocluster-patterned optical fiber SERS probes.

The use of surface-enhanced Raman scattering (SERS) spectroscopy for the detection of substances in non-volatile systems, such as edible oil and biological cells, is an important issue in the fields of food safety and biomedicine. However, traditional dry-state SERS detection with planar SERS substrates is not suitable for highly sensitive and rapid SERS detection in non-volatile liquid-phase systems. In this paper, we take contaminant in edible oil as an example and propose an in situ SERS detection method for non-volatile complex liquid-phase systems with high-performance optical fiber SERS probes. Au-nanorod clusters are successfully prepared on optical fiber facet by a laboratory-developed laser-induced dynamic dip-coating method, and relatively high detection sensitivity (LOD of 2.4 × 10-6 mol/L for Sudan red and 3.6 × 10-7 mol/L for thiram in sunflower oil) and good reproducibility (RSD less than 10%) are achieved with a portable Raman spectrometer and short spectral integration time of 10 s even in complex edible oil systems. Additionally, the recovery rate experiment indicates the reliability and capability of this method for quantitative detection applications. This work provides a new insight for highly sensitive and rapid SERS detection in non-volatile liquid-phase systems with optical fiber SERS probes and may find important practical applications in food safety and biomedicine.

Coupled sulfur and electrode-driven autotrophic denitrification for significantly enhanced nitrate removal.

Elemental sulfur (S0)-based autotrophic denitrification (SAD) has gained intensive attention in the treatment of secondary effluent for its low cost, high efficiency, and good stability. However, in practice, the supplementary addition of limestone is necessary to balance the alkalinity consumption during SAD operation, which increases water hardness and reduces the effective reaction volume. In this study, a coupled sulfur and electrode-driven autotrophic denitrification (SEAD) process was proposed with superior nitrate removal performance, less accumulation of sulfate, and self-balance of acidity-alkalinity capacity by regulating the applied voltage. The dual-channel electron supply from S0 and electrodes made the nitrate removal rate constant k in the SEAD process 3.7-5.1 and 1.4-3.5 times higher than that of the single electrode- and sulfur-driven systems, respectively. The S° contributed to 75.3%-83.1% of nitrate removal and the sulfate yield during SEAD (5.67-6.26 mg SO42-/mg NO3--N) was decreased by 17%-25% compared with SAD. The S0 particle and electrode both as active bio-carriers constructed collaborative denitrification communities and functional genes. Pseudomonas, Ralstonia and Brevundimonas were the dominant denitrifying genera in S0 particle biofilm, while Pseudomonas, Chryseobacterium, Pantoea and Comamonas became dominant denitrifying genera in the cathode biofilm. The narG/Z/H/Y/I/V, nxrA/B, napA/B, nirS/K, norB/C and nosZ were potential functional genes for efficient nitrate reduction during the SEAD process. Metagenomic sequencing indicated that S0 as an electron donor has greater potential for complete denitrification than the electrode. These findings revealed the potential of SEAD for acting as a highly efficient post denitrification process.

Electrical selection for planktonic sludge microbial community function and assembly.

Electrostimulated hydrolysis acidification (eHA) has been used as an efficient biological pre-treatment of refractory industrial wastewater. However, the effects of electrostimulation on the function and assembly of planktonic anaerobic sludge microbial communities are poorly understood. Using 16S rRNA gene and metagenomic sequencing, we investigated planktonic sludge microbial community structure, composition, function, assembly, and microbial interactions in response to electrostimulation. Compared with a conventional hydrolysis acidification (HA) reactor, the planktonic sludge microbial communities selected by electrostimulation promoted biotransformation of the azo dye Alizarin Yellow R. The taxonomic and functional structure and composition were significantly shifted upon electrostimulation with azo dyes degraders (e.g. Acinetobacter and Dechloromonas) and electroactive bacteria (e.g. Pseudomonas) being enriched. More microbial interactions between fermenters and decolorizing and electroactive bacteria, as well as fewer interactions between different fermenters evolved in the eHA microbial communities. Moreover, the decolorizing bacteria were linked to the higher abundance of genes encoding for azo- and nitro-reductases and redox mediator (e.g. ubiquinone) biosynthesis involved in the transformation of azo dye. Microbial community assembly was more driven by deterministic processes upon electrostimulation. This study offers new insights into the effects of electrostimulation on planktonic sludge microbial community function and assembly, and provides a promising strategy for the manipulation of anaerobic sludge microbiomes in HA engineering systems.

Yin Yang 1 Is Essential for Transcriptional Activation of the Bovine Sirt2 Gene in Preadipocytes.

Sirtuin 2 (Sirt2) belongs to the NAD+-dependent deacetylase family, is more highly expressed than other family members in adipocytes, and plays crucial roles in a wide range of biological processes. However, the mechanisms underlying Sirt2 expression during adipogenesis are poorly studied. In this study, the transcriptional start site (TSS) of Sirt2 was identified and two alternative transcript variants were spliced from Sirt2. The 5'-regulatory region of Sirt2 was also characterized; no TATA-box or CCAAT-box was presented in the 5'-flanking region. Two cytosine-phosphate diester-guanine (CpG) islands were also identified between nucleotides -563 and +4. A dual-luciferase reporter assay revealed that a 178 base pair sequence upstream from the TSS (+1) was the core promoter of Sirt2. Results from a site-directed mutagenesis experiment, electrophoretic mobility shift assay, and chromatin immunoprecipitation assay indicated Yin Yang 1 (YY1) to be a positive regulator of bovine Sirt2 in preadipocytes. YY1 is likely to suppress adipogenesis in two different ways by regulating peroxisome proliferator-activated receptor gamma expression. Our results expand the information on the regulatory network of adipogenesis, which is an important basis for improving beef quality, treating obesity, and other related diseases.

Bio-Optimization of Chemical Parameters and Earthworm Biomass for Efficient Vermicomposting of Different Palm Oil Mill Waste Mixtures.

The present study reports mathematical modelling of palm oil mill effluent and palm-pressed fiber mixtures (0% to 100%) during vermicomposting process. The effects of different mixtures with respect to pH, C:N ratio and earthworms have been optimized using the modelling parameters. The results of analysis of variance have established effect of different mixtures of palm oil mill effluent plus palm press fiber and time, under selected physicochemical responses (pH, C:N ratio and earthworm numbers). Among all mixtures, 60% mixture was achieved optimal growth at pH 7.1 using 16.29 C:N ratio in 15 days of vermicomposting. The relationship between responses, time and different palm oil mill waste mixtures have been summarized in terms of regression models. The obtained results of mathematical modeling suggest that these findings have potential to serve a platform for further studies in terms of kinetic behavior and degradation of the biowastes via vermicomposting.

Total flavonoid aglycones extract in Radix Scutellariae induces cross-regulation between autophagy and apoptosis in pancreatic cancer cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Radix Scutellariae (RS), the dried root of Scutellariae baicalensis Georgi, known as a herbal medicine in several Asian countries including China, has been widely used to treat inflammation, hypertension, cardiovascular disease as well as cancer. The total flavonoid aglycone extracted (TFAE) was extracted by ethyl acetate and this extraction methodology was optimized and obtained the protection of Chinese patents. AIM OF THE STUDY: To investigate the underlying mechanism of the chemotherapeutic effects of TFAE in inducing autophagy and apoptosis in pancreatic cancer cells in vitro and in vivo. MATERIALS AND METHODS: We performed CCK8 assays, AnnexinV-FITC/PI staining, flow cytometry assays, transmission electron microscopy, immunofluorescence analysis and Western blot to study the molecular mechanism of TFAE in inducing autophagy and apoptosis in pancreatic cancer cells in vitro and in vivo. RESULTS: In vitro, TFAE exhibits significant anti-tumor activity against pancreatic cancer cell lines, especially for BxPC3 (IC50 = 6.5 µg mL-1). Moreover, TFAE induces apoptosis and autophagy as evidenced by the increased apoptosis or autophagy-related protein level, the increased the fraction of apoptotic cells and the punctuate patterns of LC3 II. Furthermore, TFAE induce autophagy through PI3K/Akt/mTOR inhibition. Interestingly, pharmacological block autophagy by 3-MA enhanced TFAE-induced apoptosis, indicating that TFAE induced autophagy functions as a cytoprotective process against apoptosis. In vivo, 150 mg/kg TFAE inhibited the BxPC3 tumor growth in immune deficient mice with the inhibitory rate of 66.87% and induced both apoptosis and autophagy. CONCLUSION: TFAE have anti-tumor activity against pancreatic cancer and can induce apoptosis and autophagy through PI3K/Akt/mTOR signal pathway. TFAE might be a potential anticancer drug to be further developed for human pancreatic cancer therapy.

Impact of orientation and flexibility of peptide linkers on T. maritima lipase Tm1350 displayed on Bacillus subtilis spores surface using CotB as fusion partner.

Fusion protein construction often requires peptide linkers for prolonged conformation, extended stability and enzyme activity. In this study a series of fusion between Thermotoga maritima lipase Tm1350 and Bacillus subtillis coat protein CotB, comprising of several peptide linkers, with different length, flexibility and orientations were constructed. Effects of temperature, pH and chemicals were examined, on the activity of displayed enzyme. The fusion protein with longer flexible linkers L9 [(GGGGS)4] and L7 (GGGGS-GGGGS-EAAAK-EAAAK-GGGGS-GGGGS) possess 1.29 and 1.16-fold higher activity than the original, under optimum temperature and pH respectively. Moreover, spore surface displaying Tm1350 with L3 (EAAAK-GGGGS) and L9 ((GGGGS)4) showed extended thermostably, maintaining 1.40 and 1.35-fold higher activity than the original respectively, at 80 °C after 5 h of incubation. The enzyme activity of linkers with different orientation, including L5, L6 and L7 was determined, where L7 maintained 1.05 and 1.27-fold higher activity than L5 and L6. Effect of 0.1% proteinase K, bromelain, 20% ethanol and 30% methanol was investigated. Linkers with appropriate Glycine residues (flexible) showed higher activity than Alanine residues (rigid). The activity of the displayed enzyme can be improved by maintaining orientation and flexibility of peptide linkers, to evaluate high activity and stability in industrial processes.

Citric acid and ethylene diamine tetra-acetic acid as effective washing agents to treat sewage sludge for agricultural reuse.

This paper presents the effects of different concentrations of citric acid (CA) and ethylene diamine tetra-acetic acid (EDTA) when used as additive reagents for the treatment of sewage sludge for agricultural use. Herein, both the retention of nutrients and removal of metals from the sewage sludge are examined. The average removal rate for the metals after treatment by CA decreased in the order Cu>Pb>Cd>Cr>Zn, while the rates after treatment by EDTA decreased in the order of Pb>Cu>Cr>Cd>Zn. After treatment with CA and EDTA, total nitrogen and total phosphorus concentrations in the sludge decreased, while the content of available nitrogen and Olsen-P increased. In addition, a multi-criteria analysis model-fuzzy analytic network process method (with 3 main factors and 12 assessment sub-factors) was adopted to evaluate the effectiveness of different treatment methods. The results showed that the optimal CA and EDTA concentrations for sewage sludge treatment were 0.60 and 0.125 mol/L, respectively.

[Chemical constituents from the fruits of Areca catechu].

OBJECTIVE: To investigate the chemical constituents of the fruits of Areca catechu. METHODS: The chemical constituents were isolated by silica gel, RP-18 and Sephadex LH-20 column chromatographies. Their structures were determined by chemical and spectroscopic methods. RESULTS: Thirteen compounds were isolated from the 95% ethanol extract of the fruits of Areca catechu. Their structures were identified as isorhamnetin (1), quercetin (2), liquiritigenin (3), 5,7,4'-trihydroxy-3',5'-dimethoxyflavanone (4), (+)-catechin (5), resveratrol (6), ferulic acid (7), vanillic acid (8), 5,8-epidioxiergosta-6,22-dien-3beta-ol (9), stigmasta-4-en-3-one (10), beta-sitosterol (11), cycloartenol (12), and de-O-methyllasiodiplodin (13), respectively. CONCLUSION: Compounds 2-4,6,7,9,10, 12,13 are isolated from this plant for the first time.