Phycoremediation and valorization of hypersaline pickled mustard wastewater via Chaetoceros muelleri and indigenous bacteria.

Hypersaline pickled mustard wastewater (PMW), a typical food wastewater with high nutrient content, was successfully bioremediated via the co-treatment of Chaetoceros muelleri and indigenous bacteria in this study. Chemical oxygen demand, ammonia nitrogen, total nitrogen and total phosphorus in 10 % PMW could be effectively reduced by 82 %, 90 %, 94 % and 96 %, respectively, after 12 days treatment. Oxygen species activities, malondialdehyde content, microalgal biomass, photosynthesis and extracellular polymeric substances were characterized during the treatment to determine the responses of the consortium when exposed to different concentration of PMW. Microbial community analysis demonstrated a significant increase in the relative abundance of Halomonas and Marinobacter in the 10 % PMW after 12 days treatment, which was beneficial for nutrients recycling by the diatoms. Meanwhile, C. muelleri was effective in reducing the relative abundance of potentially pathogenic bacteria Malaciobacter. In conclusion, the work here offers a promising and environmentally friendly approach for hypersaline wastewater treatment.

Combined Treatment of Salinity Stress and Fruit Thinning Effect on Tomato.

This was an experimental investigation of the combined treatments of salinity (SAL) stress and fruit thinning (FT) on the growth, yield, fruit quality, and water use efficiency (WUE) of tomatoes with non-soil cultivation. The experiment was carried out in a plastic tunnel, Japan. Tomato (Solanum lycopersicum) cv. Momotaro seedlings were transplanted in a randomized complete block (RCB) manner with six plants/treatment, and an overall 36 plants in 18 pots (2 plants/pot). The experiment involved varying SAL treatment (no-SAL, moderate SAL, and serious SAL, with electroconductivity of 0.8, 3.0, and 4.5 dS m-1, separately) and FT treatment (NT: no thinning and 3FT: three-fruit treatment). The tomato growth, yield, and WUE were significantly suppressed with increasing SAL. In comparison, FT treatment had less effect on tomato growth and water consumption. Either SAL stress or FT treatment significantly improved fruit quality. The combined treatment proved better than single treatment of either SAL stress or FT, avoided the subsize fruit following SAL stress treatment, reduced fruit cracking found with FT treatment, and greatly improved fruit quality. The SAL thresholds of WUEs in relation to biomass, yield, and marketable yield were approximately 3.0 dS m-1 under these soilless conditions. Path analysis showed that biomass and water consumption were important indexes affecting yield. Logistic equation fitting showed that SAL stress tended to inhibit and delay plant growth; however, FT tended to advance and shorten the period of plant growth.

Resveratrol inhibits bile acid-induced gastric intestinal metaplasia via the PI3K/AKT/p-FoxO4 signalling pathway.

Gastric intestinal metaplasia (GIM) is the essential pre-malignancy of gastric cancer. Chronic inflammation and bile acid reflux are major contributing factors. As an intestinal development transcription factor, caudal-related homeobox 2 (CDX2) is key in GIM. Resveratrol has potential chemopreventive and anti-tumour effects. The aim of the study is to probe the effect of resveratrol in bile acid-induced GIM. We demonstrated that resveratrol could reduce CDX2 expression in a time- and dose-dependent manner in gastric cell lines. A Cignal Finder 45-Pathway Reporter Array and TranSignal Protein/DNA Array Kit verified that resveratrol could increase Forkhead box O4 (FoxO4) activity and that Chenodeoxycholic acid (CDCA) could reduce FoxO4 activity. Furthermore, bioinformatics analysis showed that FoxO4 could bind to the CDX2 promoter, and these conjectures were supported by chromatin-immunoprecipitation (ChIP) assays. Resveratrol can activate FoxO4 and decrease CDX2 expression by increasing phospho-FoxO4 nucleus trans-location. Resveratrol could increase FoxO4 phosphorylation through the PI3K/AKT pathway. Ectopic FoxO4 expression can up-regulate FoxO4 phosphorylation and suppress CDCA-induced GIM marker expression. Finally, we found a reverse correlation between p-FoxO4 and CDX2 in tissue arrays. This study validates that resveratrol could reduce bile acid-induced GIM through the PI3K/AKT/p-FoxO4 signalling pathway and has a potential reversing effect on GIM, especially that caused by bile acid reflux.

A high-throughput, homogeneous, fluorescence polarization assay for inhibitors of hedgehog protein autoprocessing.

Hedgehog (Hh) signaling plays an important role in embryonic patterning and adult stem cell renewal but has recently been found also to be involved in certain stem cell cancers. One of the first steps in Hh signaling is the autoprocessing of Hh protein, in which the C-terminal domain (Hh-C) catalyzes a cholesterol-dependent autocleavage reaction that leads to the production of the cholesterol ester of the N-terminal Hh domain (Hh-N), thereby yielding a signaling molecule that activates the Hh pathway by binding to the Patched receptor. This article describes an in vitro, homogeneous assay system that measures changes in fluorescence polarization that accompany the cholesterol-dependent autocleavage of Hh protein. The assay system makes use of a modified Hh protein in which Hh-N, which is not essential for autocleavage, is replaced by a 25-residue peptide containing a tetracysteine motif, complexed with a bisarsenical fluorophore. The assay is quite robust and easily adapted to high-throughput screening in 384-well plates with Z' factors above 0.8. It has been used to screen the National Institutes of Health Clinical Collection, which has led to the identification of 2 compounds that inhibit the cholesterol-dependent autocleavage of Hh protein at micromolar concentrations.

An in vitro screening system for protein splicing inhibitors based on green fluorescent protein as an indicator.

This paper describes an in vitro fluorometric assay system for protein splicing based on the RecA intein of Mycobacterium tuberculosis and a modified green fluorescent protein (GFP). The assay takes advantage of the fact that polypeptides inserted adjacent to residue 129 of GFP cause the protein to form inclusion bodies when expressed in Escherichia coli and to be incapable of fluorophore formation. However, when the inserted polypeptide is an intein, the renatured fusion protein can undergo protein splicing and chromophore formation. Comparison of chromophore formation by renatured GFP-intein fusion and renatured GFP showed that under optimal conditions (pH 6.5 and 20 degrees C) protein splicing is significantly slower than GFP chromophore formation. Taking advantage of the reversible inhibition of protein splicing by zinc ion, a fluorometric protein splicing assay was developed in which the denatured fusion protein of GFP and the RecA intein was purified on a metal ion affinity column and renatured in the presence of 2 mM ZnCl2. When diluted into appropriate buffers, protein splicing could be initiated by the addition of a molar excess of EDTA and followed fluorometrically. This assay should be valuable as a high-throughput screening system for protein splicing inhibitors as potential antimycobacterial agents and as tools for studying the mechanism of protein splicing.