Effect of Temperature on Anaerobic Fermentation of Poplar Ethanol Wastewater: Performance and Microbial Communities.

Temperature plays an important role in anaerobic digestion (AD), and different substrates have different optimum temperatures in AD. However, the effect of temperature on the performance of AD when cellulosic ethanol wastewater was used as a substrate was rarely reported. Therefore, the digestion characteristics of cellulosic ethanol wastewater at 25, 35, 45, and 55 °C were investigated, and the microbial communities of the sludge sample were analyzed after fermentation. The results showed that the cumulative methane production was the highest at 55 °C, 906.40 ± 50.67 mL/g VS, which was 81.06, 72.42, and 13.33% higher than that at 25, 35, and 45 °C, respectively. The content of methane was 68.13, 49.26, 70.46, and 85.84% at the terminal period of fermentation at temperatures of 25, 35, 45, and 55 °C, respectively. The testing of volatile fatty acids (VFAs) indicated that the accumulation of VFAs did not occur when the fermentation was carried out at 25, 35, and 45 °C; however, the VFA content at 55 °C was much larger than that in the three groups (25, 35, and 45 °C), and the ratio of propionic acid to acetic acid was larger than 1.4 at the late stage of fermentation, so it inhibited the fermentation. The diversity of the microbial community indicated that the floral structure and metabolic pathway of fermentation were alike at 25 and 35 °C. Firmicutes and Proteobacteria were the main flora covering the 25-55 °C-based phylum or below it. The relative abundance of Methanosaeta was the highest when fermentation temperatures were 25 and 35 °C; however, its relative abundance decreased sharply and the relative abundance of Methanosarcina increased substantially when the temperature increased from 35 to 45 °C, which indicated that Methanosarcina can exist in higher temperatures. At the same time, hydrogenotrophic methanogens such as Methanoculleus and Methanothermobacter were dominant when fermentation temperatures were 45 and 55 °C, which indicated that the metabolic pathway changed from acetoclastic methanogenesis to hydrogenotrophic methanogenesis.

Conservation tillage increases surface soil organic carbon stock by altering fungal communities and enzyme activity.

Fungal communities play a key role in the decomposition of crop residues and affect soil organic carbon (SOC) dynamics. Conservation tillage enhances SOC sequestration and mitigate global climate change. However, the impact of long-term tillage practices on fungal community diversity and its relation to SOC stock remains unclear. The objectives of this study were to evaluate the relationship between extracellular enzyme activities and fungal community diversity and SOC stock under different tillage practices. A field experiment was conducted with four tillage practices: (i) no-tillage with straw removal (NT0), (ii) no-tillage with straw retention (NTSR, conservation tillage), (iii) plough tillage with straw retention (PTSR), and (iv) rotary tillage with straw retention (RTSR). The results showed that the SOC stock in NTSR was higher than other treatments in the 0-10 cm soil layer. Compared to NT0, NTSR significantly increased soil ß-glucosidase, xylosidase, cellobiohydrolase, and chitinase activities at 0-10 cm soil depth (P < 0.05). However, different tillage methods with straw returning had no significant effects on enzyme activity at 0-10 cm soil depth. The observed species and Chao1 index of the fungal communities under NTSR were 22.8% and 32.1% lower than under RTSR in the 0-10 cm soil layer, respectively. The composition, structure, and co-occurrence network of fungal communities differed across tillage practices. A partial least squares path model (PLS-PM) analysis indicated that C-related enzymes were the most influential factors associated with SOC stock. Soil physicochemical properties and fungal communities affected extracellular enzyme activities. Overall, conservation tillage can promote surface SOC stock, which was associated with increased enzyme activity.

Structure and properties of nanoparticles: DES-lignin-g-PNVCL coated aspirin by self-assembly.

This work was carried out in order to broaden the application field of lignin and improve its additional value. The degraded deep eutectic solvent lignin-grafted poly(N-vinyl caprolactam) (DES-lignin-g-PNVCL) was synthesized by using modified DES-lignin and NVCL via activators regenerated by electron transfer-atom transfer radical polymerization (ARGET-ATRP). Aspirin was coated with DES-lignin-g-PNVCL through self-assembly by an ethanol/water anti-solvent method to obtain lignin thermosensitive polymer nanoparticle coated aspirin (aspirin@LTNP). X-ray electron spectroscopy (XPS), elemental analysis, scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), dynamic light scattering (DLS), and ultraviolet visible spectroscopy (UV) were used to characterize the composition, structure and morphology of DES-lignin-g-PNVCL and aspirin@LTNP. The releasing behavior of aspirin@LTNP at different temperatures and pH values was investigated. The safety was evaluated by cytotoxicity tests. The results indicated that aspirin@LTNP was mainly accumulated by the hydrophobic effect and π-π interaction in the process of self-assembly, and its morphology was an ellipsoid stacked layer by layer. The aspirin@LTNP hydrophilic chains were increased and had externally hydrophilic and internally hydrophobic structures. The particle size decreased slightly during the self-assembly process. The red-shift occurred at the π-π interaction wavelength of the lignin aromatic ring, which indicated a physical coating process. The coating rate of aspirin@LTNP was 88.87%. Aspirin@LTNP showed an obvious temperature response; the 96 h cumulative release rate at the LCST was 73.75 ± 1.16%, while the 96 h cumulative release rate above the LCST was 28.10 ± 0.92%. The 96 h cumulative release rate was 63.21 ± 0.57% at pH = 1.5 and 49.56 ± 0.48% at pH = 7.4. The dosage of aspirin@LTNP used in the experiment was safe. This study provided a strategy for drug coating and controlled release.

Foxhead box D1 promotes the partial epithelial-to-mesenchymal transition of laryngeal squamous cell carcinoma cells via transcriptionally activating the expression of zinc finger protein 532.

The presence of cervical lymph node metastases has been considered as the most important adverse prognostic factor for patients with laryngeal squamous cell carcinoma (LSCC). However, the underlying mechanisms remain to be fully revealed. In this study, we explored the expression profile of Foxhead box D1 (FOXD1), its association with epithelial-to-mesenchymal transition (EMT), and its downstream targets in LSCC. Bioinformatic analysis was performed based on the LSCC subset of The Cancer Genome Atlas-Head and Neck Squamous Cell Carcinoma (TCGA-HSNC) and Chromatin immunoprecipitation (ChIP)-seq data from Cistrome Data Browser. LSCC cell lines AMC-HN-8 and TU212 were used for in vitro studies. Results showed that FOXD1 upregulation was associated with poor prognosis of LSCC. FOXD1 knockdown reduced N-cadherin and Vimentin expression but increased E-cadherin expression in AMC-HN-8 cells. Its overexpression showed opposite effects in TU212 cells. FOXD1 could bind to the promoter of ZNF532 and activate its transcription. ZNF532 overexpression enhanced the invasion of both AMC-HN-8 and TU212 cells. In comparison, its knockdown significantly impaired their invasion. ZNF532 knockdown nearly abrogated the alterations of EMT markers caused by FOXD1 overexpression. Its overexpression largely rescued the phenotypes caused by FOXD1 knockdown. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that ZNF532 correlated genes are largely enriched in extracellular matrix regulations. LSCC patients with high ZNF532 expression (top 50%) had a significantly worse progression-free survival. In summary, this study confirmed that FOXD1 promotes partial-EMT of LSCC cells via transcriptionally activating the expression of ZNF532.

Preparation of LCST regulable DES-lignin-g-PNVCL thermo-responsive polymer by ARGET-ATRP.

To expand the field of high-value utilization of lignin. The degraded deep eutectic solvent lignin-grafted poly (N-Vinyl caprolactam) (DES-lignin-g-PNVCL) was synthesized by modified DES-lignin and NVCL via the combination of activators regenerated by electron transfer-atom transfer radical polymerization (ARGET-ATRP). Fourier transform infrared spectroscopy (FT-IR), 1H NMR, X-ray electron spectroscopy (XPS), dynamic light scattering (DLS), differential scanning calorimeter (DSC) were used to characterize the structure and performance of DES-lignin-g-PNVCL. The results indicated that the PNVCL and DES-lignin-g-PNVCL were successfully prepared by ARGET-ATRP. The lowest critical solution temperature (LCST) of PNVCL was 35.75 °C. Due to different strength of hydrogen bond, different energies were required, so the LCST of the polymer can be regulated. When the molar ratio of phenolic hydroxyl group in degraded DES-lignin to 2-bromoisobutyryl bromide (BiBB) was increased from 1:1 to 1:7, the grafting rate of DES-lignin-Br was increased from 32.87% to 60.84%, and the LCST of DES-lignin-g-PNVCL was decreased from 47.98 °C to 27.88 °C. The LCST of DES-lignin-g-PNVCL was increased from 30.98 °C to 44.64 °C when the addition amount of DES-lignin-Br was increased from 20 mg to 200 mg. The LCST of DES-lignin-g-PNVCL was increased from 27.20 °C to 39.86 °C when the ratio of DMF/water was increased from 1:4 to 4:1. The LCST of DES-lignin-g-PNVCL was decreased from 52.10 °C to 31.02 °C when the concentration of DES-lignin-g-PNVCL was increased from 0.5 mg/mL to 2.5 mg/mL. The equation represented the relationship between LCST and influencing factors was obtained, the good predictability provided a tactics for preparing desired LCST thermo-responsible polymer.

Correction: An electroporation-free method based on Red recombineering for markerless deletion and genomic replacement in the Escherichia coli DH1 genome.

[This corrects the article DOI: 10.1371/journal.pone.0186891.].

LMX1B mRNA expression and its gene body CpG methylation are valuable prognostic biomarkers for laryngeal squamous cell carcinoma.

This study aimed to explore the prognostic value of LMX1B mRNA expression and the methylation of its CpG sites in patients with laryngeal squamous cell carcinoma (LSCC). An in-silicon analysis was performed using data from the cancer genome atlas (TCGA)-Head and Neck Squamous Carcinoma (HNSC). After screening, 112 LSCC and 10 adjacent normal tissues were identified as eligible samples for analysis. Results showed that LMX1B expression was significantly upregulated in the cancer tissues (p < 0.01) and was an independent prognostic indicator in terms of OS (HR: 1.233, 95%CI: 1.082-1.405, p = 0.002) and RFS (HR: 1.200, 95%CI: 1.002-1.438, p = 0.048). By examining the methylation profile of 55 CpG sites in LMX1B locus, we found that the promoter methylation status was irrelevant to LMX1B expression. In comparison, LMX1B expression was generally positively correlated with gene body methylation. Among the gene body CpG sites, cg13600622 methylation showed a better predictive value than LMX1B expression in terms of OS (HR: 12.363, 95%CI: 1.076-142.033, p = 0.043), while cg14204784 methylation was a better marker of shorter RFS (HR: 12.363, 95%CI: 1.076-142.033, p = 0.043). Among the known downstream genes of LMX1B, only NR4A2 expression showed a moderately negative correlation (Pearson's r = -0.54) with it in LSCC tissues. However, this correlation was inconsistent with previous publications those reported a positive correlation between them. Based on these findings, we infer that upregulated LMX1B mRNA expression had an independent prognostic value in LSCC patients. Increased gene body methylation might be an important mechanism of its upregulation. Among the gene body CpG sites, cg13600622 and cg14204784 methylation level might be better prognostic markers than LMX1B mRNA expression in terms of OS and RFS respectively.

An electroporation-free method based on Red recombineering for markerless deletion and genomic replacement in the Escherichia coli DH1 genome.

The λ-Red recombination system is a popular method for gene editing. However, its applications are limited due to restricted electroporation of DNA fragments. Here, we present an electroporation-free λ-Red recombination method in which target DNA fragments are excised by I-CreI endonuclease in vivo from the landing pad plasmid. Subsequently, the I-SceI endonuclease-cutting chromosome and DNA double-strand break repair were required. Markerless deletion and genomic replacement were successfully accomplished by this novel approach. Eight nonessential regions of 2.4-104.4 kb in the Escherichia coli DH1 genome were deleted separately with selection efficiencies of 5.3-100%. Additionally, the recombination efficiencies were 2.5-45%, representing an order of magnitude improvement over the electroporation method. For example, for genomic replacement, lycopene expression flux (3.5 kb) was efficiently and precisely integrated into the chromosome, accompanied by replacement of nonessential regions separately into four differently oriented loci. The lycopene production level varied approximately by 5- and 10-fold, corresponding to the integrated position and expression direction, respectively, in the E. coli chromosome.

[Analysis of allergic sieving detection results and clinical significance in children with allergic disease].

OBJECTIVE: To explore the clinical use of sieving detection among the childhood with allergic disease. METHOD: The sieving detection about allergen inhalant allergens, Fx5 in the CAP anaphylactogen detection system, and serum specific IgE were detected in three hundred and thirty-one cases of children (aged from 1 year to 14 years old) with allergic disease. Patients were divided into group 1, group 2 and group 3 according to the age from 0 to 3, 3 to 6, and 6 to 14 years old. All datas were statistical analysed among different age groups. RESULT: Among the 331 patients, the positive rate of allergic sieving detection was 67.98%, the elevation rate of IgE was 53.78%. Inhalant allergen positive rate was 60.42%, while the food allergen positive rate was 28.10%. Inhalant allergen positive rate of the group 3 (aged from 6 to 14 years old) was significant higher than the other two age groups (68.45%). And the food allergen positive rate of the age group 1 (aged from 0 to 3 years old) was significant higher than the other two age groups (62.50%). Positive rate for simply inhalant allergen was 39.88%, while positive rate for simply food allergen was 7.55% and mixed allergen was 20.54%. CONCLUSION: Inhalant allergen was the main allergen of the children with allergic disease aged over 3 years old, while food allergen was the main allergen of the children with allergic disease aged below 3 years old. It was safe, sensible and effective to use Uni CAP anaphylactogen detection system for rapid assay of specific allergens.

[Analyzing of the inhaled allergens profiles of allergic rhinitis patients in district of Jingmen].

OBJECTIVE: To investigate the profiles of inhaled allergens in allergic rhinitis patients in district of Jingmen, and to provide basic epidemiologic information for the prevention and treatment of allergic rhinitis in this area. METHOD: Retrospectively analyze 10 allergens skin prick test results of 1800 allergic rhinitis patients recruited. RESULT: The most common inhaled allergens in allergic rhinitis patients in our area were house dust mite (76.83%), dermatophagoides farinae (75.50%), mugwort (10.50%), ragweed (7. 39%), fungus II (6.28%), fungus I (5.83%), spring-pollen I (4.44%), spring pollen II (3.72%), poly-animal hair (3.56%), poly-feather (1.89%). Moreover, the positivity frequencies of house dust mite and dermatophagoides farinae were decreased with age increasing. CONCLUSION: House dust mite and dermatophagoides farinae were the most common allergens in allergic rhinitis patients in Jingmen area.

[Investigation and analysis of the common inhaled allergens in allergic diseases in Jingmen area of Hubei Province].

OBJECTIVE: In order provide important evidences for epidemiologic survey, clinical diagnosis, treatment and preservation of allergic diseases in this area, we investigate the distribution of the common allergens in allergic rhinitis and allergic asthma in Jingmen area and the sensitization condition of the major allergen house dust mite. METHOD: Four hundred and twenty-five patients with suspected allergic rhinitis and asthma in our area accepted skin prick tests of 10 standard allergen agents and specificity IgE detection. The different positivity ratios of allergens in different age groups were compared and the correlation between the two different technique methods testing the dust mite allergen was analyzed. RESULT: Among 425 patients with suspected allergic rhinitis and asthma in our area, 298 patients were positive and 89 patients suffered a combined allergic asthma. House dust mite (292 cases) and dermatophagoides farinae (289 cases) were the commonest allergens. The other common allergens were mugwort (45), ragweed (31), fungus I (19), fungus II (15), spring-pollen II (14), poly-animal hair (14), spring-pollen I (9), poly-feather (3). There was a correlation between skin test and sIgE test for house dust mite, dermatophagoides farinae in children group, and the positivity ratios were significantly higher than the adults group. CONCLUSION: Dust mite is the most important allergen in allergic rhinitis and asthma in Jingmen.