A New Systematic Construction of Novel Three-Dimensional Spin Crossover Coordination Polymers Based on the [AgI 2(CN)3] Building Unit.

New three-dimensional spin crossover (SCO) coordination polymers systematically constructed by the novel building unit [AgI 2(CN)3], FeII(3-Br-5-CH3pyridine)2[AgI 2(CN)3][AgI(CN)2] (1), FeII(3-Br-5-Clpyridine)2[AgI 2(CN)3][AgI(CN)2] (2), and FeII(3,5-Brpyridine)2[AgI 2(CN)3][AgI(CN)2] (3), have been synthesized and characterized. The bismonodentate binuclear [Ag2(CN)3]- and mononuclear [AgI(CN)2]- units and FeII atoms assemble to form a 3D network structure. The structures of 1-3 are crystallographically identical, which made up the triply interpenetration combined with complicated intermolecular interactions including Ag···Ag, Ag···X (pyridine substituents) and π-stacking interactions. Magnetic and differential scanning calorimetry studies were performed for 1-3. These compounds display a similar SCO behavior, while the critical temperatures (T c) are shifted by the substituent effect. Due to the identical structures of 1-3, the order of T c clearly corresponds with the Hammett constant.

Screening of protein-ligand interactions under crude conditions by native mass spectrometry.

A convenient analytical system for protein-ligand interactions under crude conditions was developed using native mass spectrometry (MS). As a model protein, Escherichia coli (E. coli) dihydrofolate reductase (DHFR) with and without a histidine tag was used for the study. First, overexpressed DHFR with a His-tag was roughly purified with a Ni-sepharose resin and subjected to native mass spectrometry with or without incubation with an inhibitor, Methotrexate (MTX). Even only with the minimum cleanup by the Ni-sepharose resin, intact ions of DHFR-nicotinamide adenine dinucleotide phosphate (NADPH) and DHFR-NADPH-ligand complexes were successfully observed. By optimizing the preparation procedures of the crude sample for native MS, e.g., avoiding sonication for cell lysis, we successfully observed intact ions of the specific DHFR-NADPH-MTX ternary complex starting with cultivation of E. coli in ≤ 25 mL medium. When the crude DHFR sample was mixed with two, four, or eight candidate compounds, only ions of the specific protein-ligand complex were observed. This indicates that the present system can be used as a rapid and convenient method for the rough determination of binding of specific ligands to the target protein without the time-consuming purification of protein samples. Moreover, it is important to rapidly determine specific interactions with target proteins under conditions similar to those in "real" biological systems. Graphical abstract.

Biological treatment of selenate-containing saline wastewater by activated sludge under oxygen-limiting conditions.

Selenium often coincides with high salinity in certain industrial wastewaters, which can be a limitation in the practical application of biological treatment. However, there are no studies on the biological treatment of selenate-containing saline wastewater. A sequencing batch reactor inoculated with activated sludge was applied to treat selenate in the presence of 3% (w/v) NaCl. Start-up of the sequencing batch reactor with a 7-day cycle duration and excessive acetate as the sole carbon source succeeded in removing above 98% and 72% soluble and solid selenium, respectively, under oxygen-limiting conditions. Further selenium removal experiments with a shorter cycle duration of 3 days and a stepwise decrease of acetate addition achieved soluble and total selenium removal efficiencies in most batches above 96% and 80%, respectively. Mass balance analysis revealed that selenate was converted into elemental selenium, most of which was accumulated in the sludge. Microscopic analyses also found that elemental selenium particles were primarily present as approximately 2 µm large rods, with some extremely large particles above 10 µm. Although the bacterial populations responsible for selenium removal, especially selenate reduction, could not be identified by microbial community analysis, this study reported for the first time that selenate could be biologically treated in the presence of considerable salinity, offering implications for the practical treatment of selenium in certain industrial wastewaters.

Depolarization-induced rapid generation of 2-arachidonoylglycerol, an endogenous cannabinoid receptor ligand, in rat brain synaptosomes.

2-arachidonoylglycerol (2-AG) is an endogenous ligand for the cannabinoid receptors with a variety of potent biological activities. In this study, we first examined the effects of potassium-induced depolarization on the level of 2-AG in rat brain synaptosomes. We found that a significant amount of 2-AG was generated in the synaptosomes following depolarization. Notably, depolarization did not affect the levels of other molecular species of monoacylglycerols. Furthermore, the level of anandamide was very low and did not change markedly following depolarization. It thus appeared that the depolarization-induced accelerated generation is a unique feature of 2-AG. We obtained evidence that phospholipase C is involved in the generation of 2-AG in depolarized synaptosomes: U73122, a phospholipase C inhibitor, markedly reduced the depolarization-induced generation of 2-AG, and the level of diacylglycerol was rapidly elevated following depolarization. A significant amount of 2-AG was released from synaptosomes upon depolarization. Interestingly, treatment of the synaptosomes with SR141716A, a CB1 receptor antagonist, augmented the release of glutamate from depolarized synaptosomes. These results strongly suggest that the endogenous ligand for the cannabinoid receptors, i.e. 2-AG, generated through increased phospholipid metabolism upon depolarization, plays an important role in attenuating glutamate release from the synaptic terminals by acting on the CB1 receptor.

2-Arachidonoyl-sn-glycero-3-phosphate, an arachidonic acid-containing lysophosphatidic acid: occurrence and rapid enzymatic conversion to 2-arachidonoyl-sn-glycerol, a cannabinoid receptor ligand, in rat brain.

A substantial amount of lysophosphatidic acid (LPA) (15.66 nmol/g tissue) was found to occur in the brain isolated from rats killed in liquid nitrogen. We found that a significant portion of brain LPA was accounted for by the arachidonic acid-containing species (5.4%). We obtained evidence that both 2-arachidonoyl species and 1-arachidonoyl species of LPA are present. The occurrence of 2-arachidonoyl LPA in the brain (0.53 nmol/g tissue) is a notable observation, because of its structural resemblance to 2-arachidonoyl-sn-glycerol (2-AG), an endogenous cannabinoid receptor ligand. We then examined the biological activity of 2-arachidonoyl LPA and compared it with that of 2-AG using neuroblastoma x glioma hybrid NG108-15 cells which express both the LPA receptor and cannabinoid CB1 receptor. We found that 2-arachidonoyl LPA interacts with the LPA receptor(s) to elicit the elevation of intracellular free Ca(2+) concentrations, whereas 2-AG interacts exclusively with the cannabinoid CB1 receptor. Next, we examined the possible metabolic relationship between 2-arachidonoyl LPA and 2-AG and obtained clear evidence that rapid enzymatic conversion of 2-arachidonoyl LPA to 2-AG took place in the brain homogenate. It is noteworthy that two types of endogenous ligands, that interact with different types of receptors, are closely related metabolically and rapidly interconvert.