Slr4, a newly identified S-layer protein from marine Gammaproteobacteria, is a major biofilm matrix component.

S-layers are paracrystalline proteinaceous lattices that surround prokaryotic cells, forming a critical interface between the cells and their extracellular environment. Here, we report the discovery of a novel S-layer protein present in the Gram-negative marine organism, Pseudoalteromonas tunicata D2. An uncharacterized protein (EAR28894) was identified as the most abundant protein in planktonic cultures and biofilms. Bioinformatic methods predicted a beta-helical structure for EAR28894 similar to the Caulobacter S-layer protein, RsaA, despite sharing less than 20% sequence identity. Transmission electron microscopy revealed that purified EAR28894 protein assembled into paracrystalline sheets with a unique square lattice symmetry and a unit cell spacing of ~9.1 nm. An S-layer was found surrounding the outer membrane in wild-type cells and completely removed from cells in an EAR28894 deletion mutant. S-layer material also appeared to be "shed" from wild-type cells and was highly abundant in the extracellular matrix where it is associated with outer membrane vesicles and other matrix components. EAR28894 and its homologs form a new family of S-layer proteins that are widely distributed in Gammaproteobacteria including species of Pseudoalteromonas and Vibrio, and found exclusively in marine metagenomes. We propose the name Slr4 for this novel protein family.

Anaerobic digestibility of estrogens in wastewater sludge: effect of ultrasonic pretreatment.

BACKGROUND: Estrogenic compounds have been detected in the secondary effluents and in the biosolids from conventional wastewater treatment plants, which are not designed for their removal. Furthermore, existing limited studies on anaerobic digestibility of estrogens report conflicting results. The objective of the present work was to determine the fate and anaerobic digestibility of estrogenic compounds in various types of sludge including primary sludge (PS), waste activated sludge (WAS), and anaerobically digested sludge (seed). METHODS: Estrone (E1) and 17-ß estradiol (E2) were chosen as the model estrogenic compounds. Initially batch adsorption was conducted to determine the extent of adsorption and isotherm of E1 on various sludge. Thereafter, batch anaerobic digestion of E1 and E2 was conducted in various sludge using So/X ratio of 4 gCOD/gVSS in 250 ml bottles. The effect of earlier optimized ultrasonication dosage on the anaerobic digestion of E1 and E2 was also characterized. Estrogenicity of the digested samples was determined by the YES assay. RESULTS: Most of E1 and E2 was adsorbed on the biosolids and the Freundlich isotherm fitted the experimental data well. No anaerobic digestion of E1 and E2 was found in any of the sludge tested, and the estrogenicity of the sludge measured by YES assay increased during digestion due to the formation of E2 from E1 in a reduced environment. Ultrasonication decreased the initial mass of E1 and E2 by 20% in the sonicated digester as compared to control digester, however, there was no further decrease in E1 and E2 during digestion. CONCLUSIONS: Most of the estrogenic compounds partitioned onto the solids and remained there during digestion. Ultrasonication pretreatment reduced the estrogen burden for the digester due to advance oxidation, but no further removal of the estrogens occurred in the digester.

Degradation of estrone in water and wastewater by various advanced oxidation processes.

A comprehensive study was conducted to determine the relative efficacy of various advanced oxidation processes such as O3, H2O2, UV, and combinations of UV/O3, UV/H2O2 for the removal of estrone (E1) from pure water and secondary effluent. In addition to the parent compound (E1) removal, performance of the advanced oxidation processes was characterized using removal of total organic carbon (TOC), and estrogenicity of the effluent. Although E1 removal was high for all the AOPs, intermediates formed were more difficult to degrade leading to slow TOC removal. Energy calculations and cost analysis indicated that, although UV processes have low electricity cost, ozonation is the least cost option ($ 0.34/1000 gallons) when both capital and operating costs were taken into account. Ozonation also is superior to the other tested AOPs due to higher removal of TOC and estrogenicity. The rate of E1 removal decreased linearly with the background TOC in water, however, E1 degradation in the secondary effluent from a local wastewater treatment plant was not affected significantly due to the low COD values in the effluent.

Effects of NMDA receptor antagonists with different subtype selectivities on retinal spreading depression.

BACKGROUND AND PURPOSE: Spreading depression (SD) is a local, temporary disruption of cellular ionic homeostasis that propagates slowly across the cerebral cortex and other neural tissues such as the retina. Spreading depolarization associated with SD occurs in different types of stroke, and this phenomenon correlates also with the initiation of classical migraine aura. The aim of this study was to investigate how NMDA receptor antagonists with different subtype selectivity alter SD. EXPERIMENTAL APPROACH: Immunoblotting was applied to the chick retina for NMDA receptor subunit protein analysis, and an efficient in vitro chick retinal model used with SD imaging for NMDA receptor pharmacology. KEY RESULTS: The prominent NMDA receptor subtypes GluN1, GluN2A and GluN2B were found highly expressed in the chick retina. Nanomolar concentrations of NVP-AAM077 (GluN2A-preferring receptor antagonist) markedly suppressed high K(+) -induced SD; that is, ∼30 times more effectively than MK801. At sub-micromolar concentrations, Ro 25-6981 (GluN2B-preferring receptor antagonist) produced a moderate SD inhibition, whereas CP-101,606 (also GluN2B-preferring receptor antagonist) and UBP141 (GluN2C/2D-preferring receptor antagonist) had no effect. CONCLUSIONS AND IMPLICATIONS: The expression of major NMDA receptor subtypes, GluN1, GluN2A and GluN2B in the chick retina makes them pertinent targets for pharmacological inhibition of SD. The high efficacy of NVP-AAM077 on SD inhibition suggests a critical role of GluN2A-containing receptors in SD genesis. Such high anti-SD potency suggests that NVP-AAM077, and other GluN2A-selective drug-like candidates, could be potential anti-migraine agents.