Identification of aroma active compounds in walnut oil by monolithic material adsorption extraction of RSC18 combined with gas chromatography-olfactory-mass spectrometry.

A novel extraction method of volatile aroma compounds (VACs) from walnut oil (WO) using monolithic material adsorption extraction (MMSE) was developed herein, and its efficiency was compared with the conventional extraction method based on solid-phase microextraction (SPME) combined with gas chromatography-olfactory-mass spectrometry (GC-O-MS). After protocol optimization of MMSE-RSC18, 77 volatile compounds (38 ones can be sniffed) were extracted from WO, thus having a better performance compared to SPME (56 volatile compounds, 35 ones can be sniffed). Subsequently, based on flavor dilution factors (FDs), odor activity values (OAVs), and recombination and omission experiments, key aroma-active compounds in WO were 1-octen-3-ol (OAV = 82.58), (E)-2-decenal (OAV = 4.10), linalool (OAV = 2.37), γ-dodecalactone (OAV = 2.30), 2-pentylfuran (OAV = 1.62), (E)-2-nonenal (OAV = 1.14) and pentanal (OAV = 1.04). Collectively, compared to HS-SPME, MMSE-RSC18 provided higher capture efficiency of VACs, enabling efficient extraction and analysis of sample aromas.

The combination sequence effect on nitrogen removal pathway in hybrid constructed wetlands treating raw sewage from multiple perspectives.

The combination sequence of traditional hybrid constructed wetlands (HCWs) affects the removal of nitrogen in raw sewage, but the effect of the combination sequence on nitrogen removal pathway have seldom been reported, especially the specific conditions allowing anammox to occur. Three-stage HCWs, namely vertical flow (VF), horizontal flow (HF) and surface flow (SF) constructed wetlands, were arranged in six different sequences to investigate nitrogen removal efficiencies and microbial removal pathways using metagenomic and stable isotope analyses. Results showed that the combination sequence significantly affected nitrogen removal pathways in HCWs. We found the best removal of total nitrogen (~50%) and ammonium (NH4+-N, ~99%) in HCWs with a VFCW in the 1st stage. Metagenomic results and stable isotope analyses further indicated that simultaneous nitrification and heterotrophic denitrification were the main pathways in unsaturated VFCW, which depended on the energy substance and electron donor supplied by chemical oxygen demand (CODCr) in raw sewage. Nitrifier, anammox bacteria and autotrophic denitrifies prevailed in the subsequent saturated CWs, which tend to nitrogen loss by partial nitrification and anammox in HFCW when fed with NH4+-N wastewater with low CODCr. Providing NH4+-N and oxygen in low CODCr wastewater was the essential step to facilitate anammox process in HFCW. It implied that the problem of poor nitrogen removal due to carbon limitation could be overcome by optimizing conditions in anammox's favor.

Irrigation using hybrid constructed wetland treated domestic sewage: Uptake of phthalic acid esters and antibiotics by Ipomoea aquatica forssk.

Irrigation with treated wastewater (WW) has been promoted to meet global water demands. This study investigates the occurrence and accumulation of targeted phthalic acid esters (PAEs) and antibiotics in soil and Ipomoea aquatica Forssk. irrigated with WW discharged from six hybrid constructed wetlands (HCWs), with evaluation of the associated human health risks. Results revealed that HCWs can effectively reduce the transfer of PAEs and antibiotics to soil and I. aquatica. HCW2 (VF-SF-HF) was found to be most efficient for the removal of PAEs (68.4%-95.3%) and antibiotics (28.5%-99.4%). Among the targeted PAEs, the concentration of bis (2-ethyl) hexylphthalate (DEHP) was the highest in irrigation water, soil and I. aquatica, while benzylphthalate (BBP) exhibited the highest bioconcentration factor (BCFF). Among the targeted antibiotics, the concentration of sulfapyridine (SPD) was highest in various environmental media, while norfloxacin (NFX) exhibited the highest BCFF. The properties of PAEs and antibiotics were found to be responsible for the differential uptake patterns. The estimation of the threshold of toxicological concern and hazard quotient showed that I. aquatica irrigated with HCWs treated wastewater presented a minor risk to human health. However, comprehensive safety evaluation is required for the widespread use of HCWs treated wastewater for irrigation purposes.

Enhanced performance of pilot-scale hybrid constructed wetlands with A/O reactor in raw domestic sewage treatment.

The aim of this study was to improve the nutrient removal efficiency by optimizing aeration time, hydraulic loading rate (HLR) and combination type in hybrid constructed wetlands (HCWs) with anoxic/oxic (A/O) reactor. The results showed that, the highest removal percentages of TN, NH4-N, TP, PO4-P and COD were 87.9%, 98.1%, 86.1%, 85.3% and 95.2%, respectively, in horizontal subsurface flow (HF) - surface flow - vertical subsurface flow CW with A/O reactor at 0.1 m/d HLR and 2 h aeration. HLR, aeration time and combination type had a significant impact on the removal of nutrients, though plant diversity did not have any significant influence. Presence of the A/O reactor improved the removal of TN in the HCWs by reducing influent loading and changing influent characteristics, thereby affecting the role of nitrification, anammox, and denitrification processes. The anammox process was the dominant pathway in the 1st HF CWs during the aeration period, where the highest removal of phosphorus was observed.

[Distribution and Pollution Assessment of Nutrient and Heavy Metals in Surface Sediments from Lake Gehu in Southern Jiangsu Province, China].

This study investigated the horizontal distribution characteristics of nutrients and heavy metals (Zn, As, Cr, Cu, Ni, Pb, Cd and Hg) in January, 2014, and assessed the potential ecological risk of Lake Gehu. It was found that the average contents of TN and TP were 2,207.94 and 708.62 mg · kg⁻¹ respectively. TN and TP contents of the sediments at the centre were significantly highei than those in the north, while the TN content in the south was also significantly higher than that in the north of Lake Gehu. The average contents of Zn, As, Cr, Cu, Ni, Pb, Cd, Hg were 766.59, 350.66, 307.98, 59.54, 122.67, 168.97, 2.34, 0.41 mg · kg⁻¹, respectively. The content of Cu at the centre was significantly higher than that in the north, and the Zn content at the centre was significantly greater than that in the south of Lake Gehu, however the difference in the content of other heavy metals at these three areas was not significant. Furthermore, the obvious correlation between elements and granularity was only found in the aspect of TP, Cu and Hg. The comprehensive pollution index (PI) indicated that the Lake Gehu was heavily polluted, especially the centre and south areas. The potential ecological risk index (RI) showed that Cd, As and Hg had caused serious pollution in Lake Gehu while the other heavy metals only induced slight or medium pollution. According to the contribution of Cd, As and Hg to RI, it was concluded that the sediments in Lake Gehu were at a serious potential ecological risk.

Development of a time-resolved fluoroimmunoassay of CFP-10 for rapid diagnosis of tuberculous pleural effusion.

Tuberculous pleural effusion is the second most common form of extrapulmonary tuberculosis, which is very difficult to rapidly distinguish from malignant pleural effusion in the clinical setting. A time-resolved fluoroimmunoassay (TRF) of CFP-10, a low molecular weight protein secreted by pathogenic Mycobacterium tuberculosis, was developed to differentiate tuberculous pleural effusion from malignant one. The measuring range was 0.3-187.5 ng/ml with the dose-response coefficient of 0.9998 and detection limit of 0.036 ng/ml. The intra-assay and inter-assay coefficients of variation were 3.6-9.2% and 10.0-12.4%, respectively. The concentration of CFP-10 in malignant pleural effusion was less than 0.8 ng/ml. The negative predictive value was 93.1% in malignant pleural effusion (n = 247) while the positive predictive value was 83.0% in tuberculous pleural effusion (n = 235). Moreover, there was a statistically significant difference in the CFP-10 concentration of pleural effusion between the groups before and after clinical therapy of tuberculosis (P < 0.001, n = 81). In addition, the stability of the diagnostic reagents lasted at least 1 year at 4 °C. Therefore, the TRF of CFP-10 may be used for the rapid diagnosis of tuberculous pleural effusion and further monitoring the clinical therapeutic efficacy of tuberculosis.

The polymorphism in the let-7 targeted region of the Lin28 gene is associated with increased risk of type 2 diabetes mellitus.

Genetic polymorphisms in the miRNAs pathway of the pathogenesis of disease might contribute to the risk of disease. However, it is unclear whether these polymorphisms about miRNAs are associated with the risk of type 2 diabetes mellitus (T2DM). We performed a case-control study to investigate two polymorphisms in the let-7/Lin28 pathway based on 588 T2DM patients and 588 age and sex matched controls. The results showed that the rs3811463 polymorphism was associated with increased risk of T2DM (odds ratio (OR)=1.47, 95% confidence inference (95%CI)=1.13-1.93, P=0.005), while the rs3811464 not (OR=1.04, 95%CI=0.79-1.36, P=0.78). For the rs3811463 polymorphism, the variant genotypes were associated with increased risk of disease in females; statistically differences were observed in the clinical features of age at diagnosis, hypertension and peripheral neuropathy for the variant and wild genotype of the rs3811463 in T2DM. In summary, the results indicated that the rs3811463 polymorphism in the let-7/Lin28 pathway could significantly increase the risk of T2DM.

[Comparison of two types of cell cultures for preparation of sTNFRII-gAD fusion protein].

In this study we used two types of cell cultures, i.e., anchorage-dependent basket and full suspension batch cultures of sTNFRII-gAD-expressing CHO cells in the CelliGen 310 bioreactor (7.5 L) to compare their yields in order to optimize the culturing conditions for efficient expression of sTNFRII-gAD fusion protein consisting of soluble tumor necrosis factor receptor II and globular domain of adiponectin. The anchorage-dependent basket culture was performed in 4L 10% serum-containing medium with the final inoculating concentration of 3 x 10(5) to 4 x 10(5) cells/mL of sTNFRII-gAD-expressing CHO cells for 3 days, and then switched to 4 L serum-free LK021 medium to continue the culture for 4 days. The full suspension batch culture was carried out in the 4 L serum-free LK021 medium with the final inoculating concentration of 3 x 10(5) to 4 x 10(5) cells/mL of sTNFRII-gAD-expressing CHO cells for 7 days. The culturing conditions were monitored in real-time to maintain pH and dissolved oxygen stability through the whole process. The supernatants were collected by centrifuge, and the protein was concentrated through Pellicon flow ultrafiltration system and then purified by DEAE anion exchange. The results showed that the yields of sTNFRII-gAD fusion protein were 8.0 mg/L with 95% purity and 7.5 mg/L with 98% purity in the anchorage-dependent basket and the full suspension batch cultures, respectively. The study provided the framework for the pilot production of sTNFRII-gAD fusion protein.

Protective effects of a novel trimerized sTNFRII on acute liver injury.

TNF α plays a central role in the pathogenesis of inflammatory diseases such as rheumatoid arthritis and murine acute liver injury induced by injection of D-galactosamine and subsequent LPS. Recombinant Fc-fused soluble TNF receptor II (sTNFRII-Fc) has been used in the treatment of rheumatoid arthritis for a decade. We have recently constructed a novel fusion protein sTNFRII-gAD, which is composed of a soluble TNF receptor II and a globular domain of adiponectin. Utilizing the inclination of gAD to form homologous trimer naturally, we sought to explore TNFα antagonism of the novel trimerized sTNFRII-gAD and meantime compare TNFα-neutralizing effects in vitro and in vivo between sTNFRII-Fc and sTNFRII-gAD. Here, we evaluated the TNFα-antagonizing activity of sTNFRII-gAD with TNFα-induced L929 cytotoxicity assay. Furthermore, sTNFRII-Fc or sTNFRII-gAD was administered simultaneously with d-galactosamine 1h prior to LPS injection in the murine model of acute liver injury. Serum TNFα and TNFα-sTNFRII-gAD complex were measured by ELISA and the liver injury was assessed through alanine transaminase measurement and liver histological analysis. sTNFRII-gAD was shown to have higher TNFα-neutralizing activity than sTNFRII-Fc (p<0.05) in the L929 cytotoxicity assay. With a significant attenuation of murine lethality (p<0.05), sTNFRII-gAD showed more protective effects than sTNFRII-Fc in the murine model of acute liver injury. These results demonstrated that sTNFRII-gAD was more efficacious than sTNFRII-Fc as a TNFα antagonist, highlighting the potential of sTNFRII-gAD for the treatment of diseases associated with excessive TNFα.