ETV7 promotes colorectal cancer progression through upregulation of IFIT3.

Members of the E26 transformation-specific (ETS) variant transcription factor family act as either tumor suppressors or oncogenic factors in numerous types of cancer. ETS variant transcription factor 7 (ETV7) participates in the development of malignant tumors, whereas its involvement in colorectal cancer (CRC) is less clear. In this study, The Cancer Genome Atlas (TCGA) and immunochemistry staining were applied to check the clinical relevance of ETV7 and interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) in CRC patients. Overexpression and knockdown of ETV7 and IFIT3 were conducted by transfecting the cells with pCDNA3.1 plasmids and siRNAs, respectively. Western blotting was used to detect the protein expression of ETV7 in CRC cells. Cell Counting Kit-8, cell colony formation, and Transwell assays, as well as flow cytometry, were used to evaluate the proliferation, migration, cell cycle, and apoptosis of CRC cells. Furthermore, western blotting, RT-qPCR, and luciferase assay were used to explore the regulation of ETV7 on IFIT3. Rescue assay was used to investigate the significance of ETV7/IFIT3 axis on CRC progression. We found that ETV7 was upregulated in CRC tissues and cells. Overexpression of ETV7 stimulated the proliferation, migration, and cell cycle amplification, and reduced the apoptosis of CRC cells. Downregulation of ETV7 exerted the opposite effect on CRC cell progression. Moreover, we demonstrated that ETV7 stimulated the transcription activity, the mRNA and protein expression of IFIT3 in CRC cells. There was a positive correlation between ETV7 and IFIT3 in CRC patients. IFIT3 knockdown reversed the promotive effect exerted by overexpression of ETV7 on the amplification and migration of CRC cells. By contrast, overexpression of IFIT3 blocked the inhibitory effect of ETV7-targeting siRNA. In summary, ETV7 induces progression of CRC by activating the transcriptional expression of IFIT3. The EVT7/IFIT3 axis may be a novel target for CRC therapy.

Flow-through electrode system (FES): An effective approach for biofouling control of reverse osmosis membranes for municipal wastewater reclamation.

Emerging electrochemical disinfection techniques provide a promising pathway to the biofouling control of reverse osmosis (RO) process. However, the comparative effectiveness and mechanism of it under flow-through conditions with low voltage remains unclear. This study investigated the effect of a flow-through electrode system (FES) with both direct current (DC) and alternating pulse current (AC) on RO biofouling control compared with chlorine disinfection. At the initial stage of biofouling development, the normalized flux of AC-FES (67% on Day 5) was saliently higher than the control group (56% on Day 5). Subsequently, the normalized fluxes of each group tended similarity in their differences until the 20th day. After mild chemical cleaning, the RO membrane in the AC-FES group reached the highest chemical cleaning efficiency of 58%, implying its foulant was more readily removable and the biofouling was more reversible. The biofouling layer in the DC-FES group was also found to be easily cleanable. Morphological analysis suggested that the thickness and compactness of the fouling layers were the major reasons for the fouling behavior difference. The abundance of 4 fouling-related abundant genera (>1%), which were Pseudomonas, Thiobacillus, Sphingopyxis, and Mycobacterium exhibited a salient correlation with the biofouling degree. The operating cost of FES was also lower than that of chlorine disinfection. In summary, AC-FES is a promising alternative to chlorine disinfection in RO biofouling control, as it caused less and easy-cleaning biofouling layer mainly due to two advantages: a) reducing the regrowth potential after disinfection of the bacteria, leading to alleviated initial fouling, (b) reshaping the microbial community to those with weaker biofilm formation capacity.

The response of nitrifying activated sludge to chlorophenols: Insights from metabolism and redox homeostasis.

The specialized wastewater treatment plants for the chemical industry are rapidly developed in China and many other countries. But there is a common bottleneck in that the toxic pollutants in chemical wastewater often cause shock impacts on biological nitrogen removal systems, which directly affects the stability and cost of operation. As the research on nitrification inhibition characteristics is not sufficient till now, there is a great lack of theoretical guidance on the control of the inhibition. This study investigated the response of nitrifying activated sludge to chlorophenols (CPs) inhibition in terms of metabolism disorder and oxidative stress. At the initial stage of reaction (i.e., 1 h), reactive oxygen species (ROS)-induced membrane damage which might account for declining nitrification performance. Simultaneously excessive extracellular polymeric substances (EPS) were secreted to alleviate oxidative stress injury and protected microorganisms to some extent. In particular tyrosine-like substances in LB-EPS with a Fmax increase of 242.30% were confirmed to efficiently resist phenols inhibition. Thus, as the inhibition proceeded, metabolism disorder replaced oxidative stress as the main cause of nitrification inhibition. The affected metabolic processes include weakened enzyme catalysis, restricted electron transport and lessened energy generation. At 4 h, nitrifying production of sludge amended with 5 mg/L chlorophenols was 89.27 ± 9.51%-98.15 ± 9.60% lower than blank, the inhibition could be attributed to comprehensively affected metabolism. The structural equation modeling indicated that phenols restricted nitrification enzymes and bacterial electron transport efficiency which was critical to nitrification performance. Moreover, the lessened energy generation weakens enzyme activity to further suppress nitrification. These findings enriched our knowledge of nitrifiers' responses to CPs inhibition and provided the basis for addressing nitrification inhibition.

Enhanced denitrification of the AO-MBBR system used for expressway service area sewage treatment: A new perspective on decentralized wastewater treatment.

Decentralized wastewater treatment warrants considerable development in numerous countries and regions. Owing to the unique characteristics of high ammonia nitrogen concentrations and low carbon/nitrogen ratio, nitrogen removal is a key challenge in treating expressway service area sewage. In this study, an anoxic/oxic-moving bed biofilm reactor (A/O-MBBR) and a traditional A/O bioreactor were continuously operated for 115 days and their outcomes were compared to investigate the enhancement effect of carriers on the total nitrogen removal (TN) for expressway service area sewage. Results revealed that A/O-MBBR required lower dissolved oxygen, exhibited higher tolerance toward harsh conditions, and demonstrated better shock load resistance than traditional A/O bioreactor. The TN removal load of A/O-MBBR reached 181.5 g‧N/(m3‧d), which was 15.24% higher than that of the A/O bioreactor. Furthermore, under load shock resistance, the TN removal load of A/O-MBBR still reached 327.0 g‧N/(m3‧d), with a TN removal efficiency of above 80%. Moreover, kinetics demonstrated that the denitrification rate of the A/O-MBBR was 121.9% higher than that of the A/O bioreactor, with the anoxic tank biofilm contributing 60.9% of the total denitrification rate. Community analysis results revealed that the genera OLB8, uncultured_f_Saprospiraceae and OLB12 were the dominant in biofilm loaded on carriers, and OLB8 was the key for enhanced denitrification. FAPROTAX and PICRUSt2 analyses confirmed that more bacteria associated with nitrogen metabolism were enriched by the A/O-MBBR carriers through full denitrification metabolic pathway and dissimilatory nitrate reduction pathway. This study offers a perspective into the development of cost-effective and high-efficiency treatment solutions for expressway service area sewage.

Effects of salinity on growth, physiology, biochemistry and gut microbiota of juvenile grass carp (Ctenopharyngodon idella).

Grass carp (Ctenopharyngodon idella) is among the most important freshwater fish species in China. However, it remained unclear how salinity could affect grass carp. Two experiments were performed. The first experiment was a 4-day acute salt tolerance experiment with six salinities (0, 4, 8, 12, 16, and 20 ppt). The second experiment was an 8-week chronic salt stress experiment with three salinities (0, 2 and 6 ppt). To investigate the intestinal bacterial community of grass carp from three salinities (0, 2, and 6 ppt), the 16S rDNA sequencing was performed. The results showed that grass carp exhibited great adaptability to low salinity (2 ppt), with no significant difference in growth and maintained stable physiological and immune status. However, exposed to high salinity (6 ppt) caused significant deleterious effects on grass carp, including growth inhibition as well as physiological and immune-related changes. The gut microbiota in grass carp changed with salinity. With the increase of salinity, the proportion of beneficial bacteria in the gut of grass carp gradually decreased, while some harmful bacteria gradually occupied the dominant position. Changes in gut microbial composition ultimately affected the growth of grass carp. This study helps further clarify the effects of salinity on grass carp.

Effect of carbon source conditions on response of nitrifying sludge to 3,5-dichlorophenol.

Nutritional conditions of activated sludge had a significant influence on nitrification inhibition response. This study comprehensively investigated the inhibition of 3,5-dichlorophenol (3,5-DCP) on nitrification of activated sludge with different C/N ratios and carbon source types. The corresponding extracellular polymeric substances (EPS), microbial communities and functional genes were analysed. The results indicated that the addition of carbon source would reduce the nitrification activity and nitrification sensitivity to 3,5-DCP, and the order of the EC50 was sequenced as sodium acetate > methanol > glucose. The response mechanisms of activated sludge under diverse carbon source conditions to 3,5-DCP were summarised as follows. When the 3,5-DCP content was increased from 0.4 mg/L to 0.8 mg/L, the protein content increased from 73.2 ± 2.6 mg/g SS ∼122.4 ± 4 mg/g SS to 92.2 ± 11.2 mg/g SS ∼130.8 ± 9.6 mg/g SS in the tightly bound EPS (TB-EPS). The increase of protein content was attributed to cellular self-protection mechanisms. Furthermore, fluorescence characteristic analysis revealed that tyrosine and tryptophan in loosely bound EPS (LB-EPS) might account for higher EC50 in activated sludge fed with methanol and sodium acetate. In addition, the redundancy analyses (RDA) showed activated sludge with organics enriched the resistant species, such as Proteobacteria and Patescibacteria, while activated sludge without organics enriched the sensitive species, such as Ferruginibacter. Finally, the nitrification genes were found to be consistent with nitrification activity. Thus, the findings provide new insights into nitrification inhibition mechanism under different carbon source conditions.

Performances and enhanced mechanisms of nitrogen removal in a submerged membrane bioreactor coupled sponge iron system.

Sponge iron is a potential material for nitrogen removal, but lack of a study about nitrogen removal in a membrane bioreactor (MBR) coupled with sponge iron. The performances and mechanisms of nitrogen removal of SI-MBR were investigated and compared it with that in GAC-MBR. The results showed that the average rate of organic matter removal in the SI-MBR was 92.74%, which was higher than that in the GAC-MBR (87.48%). And the average effluent NO2--N and NO3--N concentration in the SI-MBR (0.02 mg/L and 3.73 mg/L) was lower than that in the GAC-MBR (0.05 mg/L and 7.51 mg/L). Meanwhile, the highest nitrification rate and denitrification rate was respectively 3.544 ± 0.25 mg/(g VSS·h) and 6.643 ± 0.2 mg/(g VSS·h) in the SI-MBR, which was higher than that (3.094 ± 0.25 mg/(g VSS·h) and (6.376 ± 0.2 mg/(g VSS·h)) in the GAC-MBR. Additionally, the bacterial activities (e.g., DHA activity and respiratory activity) were obviously enhanced through the iron ion from sponge iron. The bacterial community in the SI-MBR system was more richness and diverse than that in the GAC-MBR. Ultimately, the mechanisms of enhanced biological nitrogen removal with sponge iron in MBR were analyzed. On the surface of sponge iron, the DIRB and FOB could use the iron ion from sponge iron as the electron transfer to improve the nitrogen and organic removal. With sponge iron, there is not only the nitrification bacteria and heterotrophic denitrifying microorganism enriched, but also the autotrophic denitrifying bacteria abounded obviously. The autotrophic denitrifying bacteria could use Fe(II) as an electron donor to achieve denitrification and enhance the nitrogen removal.

Coupling of membrane-based bubbleless micro-aeration for 2,4-dinitrophenol degradation in a hydrolysis acidification reactor.

Micro-aeration hydrolysis acidification (HA) is an effective method to enhance the removal of toxic and refractory organic matter, but the difficulty in stable dosing control of trace oxygen limits its wide application. Membrane-based bubbleless aeration has been proved as an ideal aeration method because of its higher oxygen transfer rate, more uniform mass transfer, and lower cost than HA. However, the available information on its application in HA is limited. In this study, membrane-based bubbleless micro-aeration coupled with hydrolysis acidification (MBL-MHA) was exploited to investigate the performance of 2,4-dinitrophenol (2,4-DNP) degradation via comparing it with bubble micro-aeration HA (MHA) and anaerobic HA. The results indicated that the performances in MBL-MHA and MHA were higher than those in HA during the experiment. 2,4-DNP degradation rates under redox microenvironments caused by counter-diffusion in MBL-MHA (84.43∼97.28%) were higher than those caused by co-diffusion in MHA (82.41∼94.71%) under micro-aeration of 0.5-5.0 mL air/min. The 2,4-DNP degradation pathways in MBL-MHA were nitroreduction, deamination, aromatic ring cleavage, and fermentation, while those in MHA were hydroxylation, aromatic ring cleavage, and fermentation. Reduction/oxidation-related, interspecific electron transfer-related species, and fermentative species in MBL-MHA were more abundant than that in MHA. Ultimately, more reducing/oxidizing forces formed by more redox proteins/enzymes from these rich species could enhance 2,4-DNP degradation in MBL-MHA.

Inhibitory effect of individual and mixtures of nitrophenols on anaerobic toxicity assay of anaerobic systems: Metabolism and evaluation modeling.

The single and combined inhibitory effects of different nitrophenols on the anaerobic toxicity assay (ATA) of anaerobic sludge and the variations in the content of extracellular polymeric substances (EPS) were investigated. The results indicated that 2,4-dinitrophenol (2,4-DNP) demonstrated the highest inhibitory effect, followed by 4-nitrophenol (4-NP) and 2-nitrophenol (2-NP), and the combined effects of binary and ternary nitrophenols induced additive toxicity. Furthermore, 2,4-DNP, the dominant toxic nitrophenol, at various concentrations and toxicant ratios, was the major contributor to the combined inhibitory effects of the nitrophenol mixtures. Abundant EPS could be secreted by the anaerobic sludge under the inhibitory effects of toxic 2-NP, 4-NP, and 2,4-DNP at concentrations from 0 to 200 mg/L to resist the adverse effects of the external environment. The protein contents of both loosely bound EPS (LB-EPS) and tightly bound EPS (TB-EPS) exhibited a better linear positive correlation relationship (R2 > 0.92) with the inhibitory rates of 2-NP, 4-NP, and 2,4-DNP, indicating that the proteins generated in the EPS of anaerobic sludge could be a stress response. Therefore, increasing the concentration of the toxic nitrophenols could enhance the stress response and increase protein production. Parallel factor (PARAFAC) analysis for TB-EPS and LB-EPS further confirmed that the major proteins were tyrosine, tryptophan, and aromatic proteins. Moreover, with an increase in the concentrations of 2-NP, 4-NP, and 2,4-DNP from 0 to 200 mg/L, microbial cell lysis and death in anaerobic sludge could be increasingly severe. Thus, this study provides new insights into the inhibitory effects of nitrophenol mixtures, which are frequently found in pharmaceutical and petrochemical effluents, on anaerobic sludge.

Pleckstrin 2 is a potential drug target for colorectal carcinoma with activation of APC/ß­catenin.

The tumor suppressor gene adenomatous polyposis coli (APC) is frequently inactivated or absent in colorectal carcinoma (CRC). Loss­of­function of APC promotes the expression of ß­catenin, which is critical for CRC development. Since ß­catenin acts as an important transcription factor, blockage of ß­catenin may have side effects, including impairment of tissue homeostasis and regeneration, thus limiting the application of ß­catenin inhibitors for the treatment of patients with CRC. Therefore, identifying a novel substrate of APC/ß­catenin may provide essential clues to develop effective drugs. Small interfering RNA technology and lentivirus­mediated overexpression were performed for knockdown and overexpression of pleckstrin 2 (PLEK2) in CRC cells. Cell Counting Kit­8 and colony formation assays, and cell cycle analysis and cell apoptosis detection were used to detect the capacity of cell proliferation, cell cycle distribution and apoptosis. The present study demonstrated that the APC/ß­catenin signaling cascade transcriptionally activated PLEK2 in CRC cells. PLEK2 expression was markedly increased in CRC tissues. There was an inverse correlation between APC and PLEK2 expression in patients with CRC. In vitro, overexpression of PLEK2 increased the proliferation of CRC cells. Opposite results were observed in the cells with knockdown of PLEK2. Furthermore, PLEK2 promoted cell cycle progression and suppressed apoptosis. In summary, upregulation of PLEK2 contributed to CRC proliferation and colony formation activated by the APC/ß­catenin signal pathway. Targeting PLEK2 may be important for the treatment of patients with CRC with activation of the APC/ß­catenin signaling pathway.

Single and joint inhibitory effect of nitrophenols on activated sludge.

In this study, single and joint inhibitory effects of nitrophenols on activated sludge and variations on the content of extracellular polymeric substances (EPS) were investigated. Results indicate that the nitrophenols adversely affected the organic and NH3-N removal of activated sludge and the adverse effect of nitrophenols on autotrophic bacteria was higher than that on heterotrophic bacteria. Further, 2,4-dinitrophenol (2,4-DNP) demonstrated the highest inhibitory effect, followed by 4-nitrophenol (4-NP) and 2-nitrophenol (2-NP), and the combined effects of binary and ternary nitrophenols induced additive toxicity. At various concentrations and toxicant ratios, 2,4-DNP, as the dominant toxic nitrophenol, was the major contributor to the joint inhibition effects of the mixed nitrophenols. At lower concentrations of 2-NP (below 100 mg/L), 4-NP (below 50 mg/L), and 2,4-DNP (below 10 mg/L), large amounts of both tightly bound EPS (TB-EPS) and loosely bound EPS (LB-EPS) were secreted for the normal physiological activities of the microbiological cells. After further stimulation with higher concentrations of 2-NP (above 100 mg/L), 4-NP (above 50 mg/L), and 2,4-DNP (above 10 mg/L), the inhibitory effect of nitrophenols on bacterial metabolism evidently increased. However, the EPS production sharply reduced, particularly with respect to protein production. Parallel factor analysis for TB-EPS and LB-EPS further confirmed that the major proteins were tyrosine, tryptophan, and aromatic proteins. Thus, this study provides new insights into the inhibitory effects of mixed nitrophenols, which are frequently found in pharmaceutical and petrochemical effluents.

Effects of 25(OH)D3 supplementation during late gestation on the serum biochemistry and reproductive performance of aged sows and newborn piglets.

The purpose of this study was to investigate the effects of diet type (normal or low Ca and P diets) and 25(OH)D3 supplementation (with or with not 2000 IU/kg 25(OH)D3 ) during late gestation on the serum biochemistry and reproductive performance of aged sows and newborn piglets. A total of 40 sows, which are at their 7th parity, were divided into four groups: control group (standard diet), low Ca group, 25(OH)D3 group and low Ca plus 25(OH)D3 group respectively (10 in each group). The blood of sows on day 100 and 114 of gestation and newborn piglets was collected for serum biochemical analyses. Results showed that the reproductive performance of sows was not influenced by diet type or 25(OH)D3 supplementation (p > 0.05). And the addition of 25(OH)D3 to diet low Ca group caused that the content of serum TG in sows on day 100 of gestation was not different from that of the control group (p > 0.05). The addition of 25(OH)D3 significantly decreases the content of serum TG in sows on day 114 of gestation (p < 0.05). The addition of 25(OH)D3 significantly increased the content of serum UREA and CREA in newborn piglets (p < 0.05). Overall, feeding 2000 IU/kg 25(OH)D3 to aged sows at late gestation had no effects on reproductive performance, but partly contributed to keeping serum TG balance in sows and may indicate increased pressure on kidneys in newborn piglets.

Copper induces oxidative stress and apoptosis through mitochondria-mediated pathway in chicken hepatocytes.

The aim of this study was to investigate the effects of excessive copper (Cu)-induced cytotoxicity on oxidative stress and mitochondrial apoptosis in chicken hepatocytes. Chicken hepatocytes were cultured in medium in the absence and presence of copper sulfate (CuSO4) (10, 50, 100 µM), in N-acetyl-L-cysteine (NAC) (1 mM), and the combination of CuSO4 and NAC for 24 h. Morphologic observation and function, reactive oxygen species (ROS) level, antioxidant indices, nitric oxide (NO) content, mitochondrial membrane potential (MMP), and apoptosis-related mRNA and protein levels were determined. These results indicated that excessive Cu could induce release of intracellular lactate dehydrogenase (LDH), aspartate aminotransferase (AST), and alanine aminotransferase (ALT); increase levels of ROS, superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT), lipid peroxidation (LPO), and NO; decrease glutathione (GSH) content and MMP; upregulated Bak1, Bax, CytC, and Caspase3 mRNA and protein expression, inhibited Bcl2 mRNA and protein expression, and induced cell apoptosis in a dose effect. The Cu-caused changes of all above factors were alleviated by treatment with NAC. These results suggested that excessive Cu could induce oxidative stress and apoptosis via mitochondrial pathway in chicken hepatocytes.

Design and synthesis of natural product derivatives with selective and improved cytotoxicity based on a sesquiterpene scaffold.

Brasilamide E (1) is a bisabolane sesquiterpenoid isolated from the solid-substrate fermentation cultures of a plant endophytic fungus Paraconiothyrium brasiliense. The compound specifically inhibited proliferation of the MCF-7 cells, but did not show cytotoxicity towards the negative controls HaCaT and NIH3T3 cells (IC50>50 µM). To improve its potency while maintain selectivity, a total of 27 derivatives of 1 were designed, synthesized, and evaluated for in vitro cytotoxicity against six tumor cell lines and the negative control NIH3T3 cells. Among these compounds, compound 12b showed significantly improved potency against the MCF-7, HeLa, and HO8910 cells with IC50 values of 0.13-0.25 µM compared to 1 (IC50 8.47-18.00 µM), and remained nontoxic to the NIH3T3 cells.

Copper-Induced Spermatozoa Head Malformation Is Related to Oxidative Damage to Testes in CD-1 Mice.

The molecular mechanism for copper toxicity on spermatozoa quality in mice is not well understood. In a 4-week experiment, we challenged 24, 6-week-old male CD-1 mice with twice-a-week intraperitoneal copper chloride injections and evaluated spermatozoa quality, copper levels in the testes, serum testosterone, the expression of key antioxidant glutathione peroxidase 5 (GPx5), and the regulated androgen receptor (AR) in the mice testes. We compared these outcomes for four groups of six mice given doses of 0, 1.25, 2.5, 5.0 mg/kg weight copper chloride twice a week for 4 weeks. The mice demonstrated a copper increase spermatozoa head malformation in a dose-response manner. However, we observed no changes in spermatozoa viability and acrosome integrity in the ratio of mouse body weight to testes weight or in the histomorphology of the testes as the average copper level increased. Results of our RT-PCR assays, immunohistochemical tests, ELISA, and histochemistry analyses indicated that testis GPx5 expression was increased, AR expression in the testes was decreased, serum testosterone was decreased, and the activity of 3ß-hydroxysteroid dehydrogenase was decreased as the copper dose increased. In conclusion, these data show that sublethal exposure to copper induces spermatozoa head malformation and influences both mRNA and protein levels of GPx5 and AR which is related to copper resides in the testes.

Two new ramulosin derivatives from the entomogenous fungus Truncatella angustata.

Two new ramulosin derivatives, 7α-hydroxy-8-dihydroramulosin (1) and 7-ketoramulosin (2), along with three known metabolites, (+)-ramulosin (3), 6-hydroxyramulosin (4), and 8-dihydroramulosin (5), were isolated from the crude extract of Truncatella angustata, an entomogenous fungus isolated from the Septobasidium-infected insect Aspidiotus sp. The structures of 1 and 2 were elucidated by nuclear magnetic resonance experiments, and 1 was further confirmed by X-ray crystallography. The absolute configuration of 1 was assigned by single-crystal X-ray diffraction analysis using Cu Kα radiation, whereas that of 2 was determined by electronic circular dichroism (ECD) calculations. Compounds 1-5 were tested for cytotoxicity against four human carcinoma cell lines, HeLa, A549, MCF-7, and T24. Compound 4 showed weak cytotoxic effects against A549 and T24.