Spectrophotometric determinationof trace nitrite with a novel self-coupling diazotizing reagent: J-acid

A simple and sensitive method for the spectrophotometric determination of nitrite was described and optimum reaction conditions along with other important analytical parameters were established. In the presence of potassium bromide at 25°C, nitrite reacted with J-acid in hydrochloric acid producing diazonium salt and then coupled with excess J-acid in the sodium carbonate solution yielding red colored azo compounds. At wavelength of 500 nm, Beer’s law was obeyed over the concentration range of 0,02 – 0,60 mg∙L⁻¹. The molar absorptivity was 3,92∙10⁴ L∙mol⁻¹∙cm⁻¹. This method was easily applied to the determination of trace nitrite in environmental water with recoveries of 9₈,7 – 101,2%

Binding site profiles and N-terminal minor groove interactions of the master quorum-sensing regulator LuxR enable flexible control of gene activation and repression

LuxR is a TetR family master quorum sensing (QS) regulator activating or repressing expression of hundreds of genes that control collective behaviors in Vibrios with underlying mechanism unknown. To illuminate how this regulator controls expression of various target genes, we applied ChIP-seq and DNase I-seq technologies. Vibrio alginolyticus LuxR controls expression of ∼280 genes that contain either symmetric palindrome (repDNA) or asymmetric (actDNA) binding motifs with different binding profiles. The median number of LuxR binding sites for activated genes are nearly double for that of repressed genes. Crystal structures of LuxR in complex with the respective repDNA and actDNA motifs revealed a new mode of LuxR DNA binding that involves contacts of its N-terminal extension to the minor groove. The N-terminal contacts mediated by Arginine-9 and Arginine-11 differ when LuxR binds to repDNA vs actDNA, leading to higher binding affinity at repressed targets. Moreover, modification of LuxR binding sites, binding profiles, and N-terminal extension have important consequences on QS-regulated phenotypes. These results facilitate fundamental understanding of the high flexibility of mechanisms of LuxR control of gene activation and repression in Vibrio QS, which may facilitate to design QS inhibiting chemicals that interfere with LuxR regulation to effectively control pathogens

Porcine Circovirus Type 2 Activates CaMMKβ to Initiate Autophagy in PK-15 Cells by Increasing Cytosolic Calcium

Porcine circovirus type 2 (PCV2) induces autophagy via the 5′ adenosine monophosphate-activated protein kinase (AMPK)/extracellular signal-regulated kinase (ERK)/tuberous sclerosis complex 2 (TSC2)/mammalian target of rapamycin (mTOR) pathway in pig kidney PK-15 cells. However, the underlying mechanisms of AMPK activation in autophagy induction remain unknown. With specific inhibitors and RNA interference (RNAi), we show that PCV2 infection upregulated calcium/calmodulin-dependent protein kinase kinase-beta (CaMKKβ) by increasing cytosolic Ca2+ via inositol 1,4,5-trisphosphate receptor (IP3R). Elevation of cytosolic calcium ion (Ca2+) did not seem to involve inositol 1,4,5-trisphosphate (IP3) release from phosphatidylinositol 4,5-bisphosphate (PIP2) by phosphoinositide phospholipase C-gamma (PLC-γ). CaMKKβ then activated both AMPK and calcium/calmodulin-dependent protein kinase I (CaMKI). PCV2 employed CaMKI and Trp-Asp (WD) repeat domain phosphoinositide-interacting protein 1 (WIPI1) as another pathway additional to AMPK signaling in autophagy initiation. Our findings could help better understanding of the signaling pathways of autophagy induction as part of PCV2 pathogenesis. Further research is warranted to study if PCV2 interacts directly with IP3R or indirectly with the molecules that antagonize IP3R activity responsible for increased cytosolic Ca2+ both in PK-15 cells and PCV2-targeted primary cells from pigs

Porcine teschovirus 2 induces an incomplete autophagic responsein PK?15 cells

AbstractAutophagy is a homeostatic process that has been shown to be vital in the innate immune defense against pathogens. However,little is known about the regulatory role of autophagy in porcine teschovirus 2 (PTV-2) replication. In this study, wefound that PTV-2 infection induces a strong increase in GFP-LC3 punctae and endogenous LC3 lipidation. However, PTV-2infection did not enhance autophagic protein degradation. When cellular autophagy was pharmacologically inhibited bywortmannin or 3-methyladenine, PTV-2 replication increased. The increase in virus yield via autophagy inhibition wasfurther confirmed by silencing atg5, which is required for autophagy. Furthermore, PTV-2 replication was suppressed whenautophagy was activated by rapamycin. Together, the results suggest that PTV-2 infection activates incomplete autophagyand that autophagy then inhibits further PTV-2 replication

Hardware in the loop simulation test platform of fuel cell backup system

Based on an analysis of voltage mechanistic model, a real-time simulation model of the proton exchange membrane (PEM) fuel cell backup system is developed, and verified by the measurable experiment data. The method of online parameters identification for the model is also improved. Based on the software LabVIEW/VeriStand real-time environment and the PXI Express hardware system, the PEM fuel cell system controller hardware in the loop (HIL) simulation plat-form is established. Controller simulation test results showed the accuracy of HIL simulation platform

Hardware in the loop simulation test platform of fuel cell backup system

Based on an analysis of voltage mechanistic model, a real-time simulation model of the proton exchange membrane (PEM) fuel cell backup system is developed, and verified by the measurable experiment data. The method of online parameters identification for the model is also improved. Based on the software LabVIEW/VeriStand real-time environment and the PXI Express hardware system, the PEM fuel cell system controller hardware in the loop (HIL) simulation plat-form is established. Controller simulation test results showed the accuracy of HIL simulation platform

Preparation of Sludge-Derived Activated Carbon by Fenton Activation and the Adsorption of Eriochrome Black T

Sludge-derived activated carbon (SAC) was prepared by Fenton activation and calcination, and used as adsorbent to eliminate Eriochrome Black T (EBT) dye from aqueous media. The characterization results indicated that the produced SAC had a porous structure, high specific surface area, and abundant functional groups on its surface. The adsorption process was affected by pH, adsorbent dosage, time, and temperature. The adsorption capacity increased with temperature, and the highest adsorption capacity reached 178.2 mg·g−1 in 48 h at 318 K and pH 6. The results of the adsorption isotherm, kinetic, and thermodynamic analyses revealed that the adsorption of EBT onto SAC was naturally endothermic and spontaneous, involved both physical and chemical processes, and belonged mostly to the multilayer type of adsorption

Porcine Circovirus 2 Deploys PERK Pathway and GRP78 for Its Enhanced Replication in PK-15 Cells

Porcine circovirus type 2 (PCV2) infection induces autophagy and apoptosis. These cellular responses could be connected with endoplasmic reticulum (ER) stress. It remains unknown if PCV2 induces ER stress and if autophagy or apoptosis is primary to PCV2 infection or secondary responses following ER stress. Here, we demonstrate that PCV2 triggered unfolded protein response (UPR) in PK-15 cells by activating the PERK/eIF2α pathway without concomitant activation of IRE1 or ATF6. Since ATF4 and CHOP were induced later than PERK/eIF2α, it is clear that persistent PCV2 infection could lead to selective activation of PERK via the PERK-eIF2α-ATF4-CHOP axis. Therefore, PERK activation could be part of the pro-apoptotic signaling via induced expression of CHOP by PCV2. Since PERK inhibition by GSK2606414 or RNA silencing or suppression of eIF2α dephosphorylation by salubrinal limited viral replication, we suppose that PCV2 deploys UPR to enhance its replication. Over-expression of GRP78 or treatment with tauroursodeoxycholic acid could enhance viral capsid expression and/or viral titers, indicating that these chaperones, endogenous or exogenous, could help correct folding of viral proteins. Our findings provide the first evidence that ER stress plays a role in the pathogenesis of PCV2 infection probably as part of autophagic and apoptotic responses