Urea Excretion and Arginase Activity as New Biomarkers for Nitrite Stress in Freshwater Aquatic Animals

Background: In recent years, the concern has been growing on increasing aquatic nitrite levels due to anthropogenic activities. Crustaceans and fish easily uptake nitrite via the chloride uptake system of gills. High nitrite body levels may interfere with nitric oxide (NO) production by nitric oxide synthase (NOS). The arginase, which catalyzes arginine conversion to ornithine and urea, is central to NO homeostasis. In vivo, changes in the arginase activity alter urea body levels and urea excretion and modulate NOS by altering arginine availability for NO synthesis. Excess arginase activity may uncouple NOS and induce oxidative stress. Methods: We tested muscle arginase activity and urea excretion in two fish species, zebrafish and convict cichlid, and the crustacean Yamato shrimp, under sub-lethal nitrite stress. Results: Exposure to nitrite (2 mM in the fish, 1 mM in the shrimp) significantly increased blood nitrite concentration in all species. Concomitantly, nitrite stress significantly increased arginase activity, urea excretion, and urea levels in the blood. In Yamato shrimp, urea levels also increased in muscle. Conclusion: Our results agree with the hypothesis that nitrite stress affects NO homeostasis by arginase stimulation and urea excretion. These parameters might function as markers of sub-lethal nitrite stress in freshwater fish and crustaceans.

Durability Characterisation of Portland Cement–Carbon Nanotube Nanocomposites

Multiwalled carbon nanotubes have outstanding mechanical properties that, when combined with Portland cement, can provide cementitious composites that could lead to the innovative construction of stronger, lighter, and thinner built infrastructure. This paper addresses a knowledge gap that relates to the durability of CNT–cement composites. The durability to corrosive chloride, uptake of water by sorption, and flow of the permeability of water acting under high water pressure are addressed. Flow simulations were undertaken through segmented 3D pore networks, based on X-ray computed microtomography measurements, the creation of a virtual microstructure, and fluid simulations that were compared with larger-scale samples. The investigation showed decreased water sorptivity of CNT–cement mixtures, indicating improved durability for the cover zone of concrete that is prone to the uptake of water and water-borne corrosives. Chloride diffusion of CNT–cement composites provided up to 63% improvement compared with control samples. The favourable durability bodes well for the construction of long-life CNT-reinforced concrete infrastructure.

Experimental Investigation of Chloride Uptake Performances of Hydrocalumite-Like Ca-Al LDHs with Different Microstructures

In this study, hydrocalumite-like Ca2Al-NO3− layered double hydroxides (Ca-Al LDHs) with different microstructures were synthesized. The crystalline properties, structure composition, morphology and particle size distribution of the Ca-Al LDH (CAL) samples were illustrated. To obtain the chloride uptake performances of CAL, the influences of contact time, initial concentration of Cl−, pH of reaction solution and coexistence anions on the chloride uptake were examined systematically. Compared to the CAL samples obtained at a higher aging temperature, CAL synthesized at 60 °C demonstrated the minimum average particle size (6.148 μm) and the best Cl− adsorption capacity (211.324 mg/g). Based on the test results, the main adsorption mechanism of chloride ion on CAL was recognized as an interlayer anion exchanging reaction other than the dissolution-precipitate mode. With the increase in the pH value of reaction solution from 7 to 13, it was found that the amount of chloride ion adsorbed by CAL increased slightly, and the solution could remain at relatively high pH value even after the adsorption. The presence of CO32− and SO42− reduced the adsorption capacity of CAL dramatically as compared with OH− due to the destruction of layered structure and the formation of precipitates (CaCO3 or CaSO4). The interference sequence of the investigated anions on the chloride uptake of CAL was SO42−, CO32− and OH−, and the order of interlayer anionic affinity was Cl− > OH− > NO3−. The results illustrated that the synthesized CAL could be used as a promising chloride ion adsorbent for the corrosion inhibition of reinforcement embedded cement-based materials.

Ser262 determines the chloride-dependent colour tuning of a new halorhodopsin from Haloquadratum walsbyi

Light is an important environmental signal for all organisms on earth because it is essential for physiological signalling and the regulation of most biological systems. Halophiles found in salt-saturated ponds encode various archaeal rhodopsins and thereby harvest various wavelengths of light either for ion transportation or as sensory mediators. HR (halorhodopsin), one of the microbial rhodopsins, senses yellow light and transports chloride or other halides into the cytoplasm to maintain the osmotic balance during cell growth, and it exists almost ubiquitously in all known halobacteria. To date, only two HRs, isolated from HsHR (Halobacterium salinarum HR) and NpHR (Natronomonas pharaonis HR), have been characterized. In the present study, two new HRs, HmHR (Haloarcula marismortui HR) and HwHR (Haloquadratum walsbyi HR), were functionally overexpressed in Escherichia coli, and the maximum absorbance (λmax) of the purified proteins, the light-driven chloride uptake and the chloride-binding affinity were measured. The results showed them to have similar properties to two HRs reported previously. However, the λmax of HwHR is extremely consistent in a wide range of salt/chloride concentrations, which had not been observed previously. A structural-based sequence alignment identified a single serine residue at 262 in HwHR, which is typically a conserved alanine in all other known HRs. A Ser262 to alanine replacement in HwHR eliminated the chloride-independent colour tuning, whereas an Ala246 to serine mutagenesis in HsHR transformed it to have chloride-independent colour tuning similar to that of HwHR. Thus Ser262 is a key residue for the mechanism of chloride-dependent colour tuning in HwHR.